Sample records for bewo cell permeability

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

    DEFF Research Database (Denmark)

    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...

  2. Iron metabolism in BeWo chorion carcinoma cells. Transferrin-mediated uptake and release of iron

    NARCIS (Netherlands)

    van der Ende, A.; du Maine, A.; Simmons, C. F.; Schwartz, A. L.; Strous, G. J.


    Growing human choriocarcinoma BeWo b24 cells contain 1.5 X 10(6) functional cell surface transferrin binding sites and 2.0 X 10(6) intracellular binding sites. These cells rapidly accumulate iron at a rate of 360,000 iron atoms/min/cell. During iron uptake the transferrin and its receptor recycle at

  3. Plasmodium falciparum parasites expressing pregnancy-specific variant surface antigens adhere strongly to the choriocarcinoma cell line BeWo

    DEFF Research Database (Denmark)

    Haase, Rikke N; Megnekou, Rosette; Lundquist, Maja


    Placenta-sequestering Plasmodium falciparum parasites causing pregnancy-associated malaria express pregnancy-specific variant surface antigens (VSA(PAM)). We report here that VSA(PAM)-expressing patient isolates adhere strongly to the choriocarcinoma cell line BeWo and that the BeWo line can...... be used to efficiently select for VSA(PAM) expression in vitro....

  4. Relevance of Wnt10b and activation of β-catenin/GCMa/syncytin-1 pathway in BeWo cell fusion. (United States)

    Malhotra, Sudha Saryu; Banerjee, Priyanka; Chaudhary, Piyush; Pal, Rahul; Gupta, Satish Kumar


    To study the involvement of specific Wnt(s) ligand during trophoblastic BeWo cell differentiation. BeWo cells on treatment with forskolin/human chorionic gonadotropin (hCG) were studied for cell fusion by desmoplakin I+II staining and/or hCG secretion by ELISA. Levels of Wnt10b/β-catenin/glial cell missing a (GCMa)/syncytin-1 were studied by qPCR/Western blotting in forskolin-/hCG-treated control siRNA and Wnt10b silenced BeWo cells. BeWo cells on treatment with hCG (5 IU/mL) led to a 94-fold increase in Wnt10b transcript. Wnt10b silencing showed significant decrease in forskolin-/hCG-mediated BeWo cell fusion and/or hCG secretion. It led to down-regulation of β-catenin (nuclear and cytoplasmic), GCMa and syncytin-1 expression. Treatment of BeWo cells with H89, protein kinase A (PKA) signaling inhibitor, significantly reduced forskolin-/hCG-induced Wnt10b, β-catenin, and syncytin-1 expression, which also resulted in reduced cell fusion. Wnt10b is involved in forskolin/hCG-mediated BeWo cell fusion via β-catenin/GCMa/syncytin pathway, which may also involve activation of PKA. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  5. Relevance of the NR4A sub-family of nuclear orphan receptors in trophoblastic BeWo cell differentiation. (United States)

    Malhotra, Sudha Saryu; Gupta, Satish Kumar


    Nur-77, a member of the NR4A sub-family of nuclear orphan receptors, is downregulated in the placentae of pre-eclamptic women. Here, we investigate the relevance of Nor-1, Nurr-1 and Nur-77 in trophoblastic cell differentiation. Their transcript levels were found to be significantly upregulated in BeWo cells treated with forskolin. The maximum increase was observed after 2 h, with a second peak in the expression levels after 48 h. The expression of NR4A sub-family members was also found to be upregulated in BeWo cells after treatment with hCG and GnRH. A similar significant increase was observed at the respective protein levels after 2 and 48 h of treatment with forskolin, hCG or GnRH. Silencing Nor-1, Nurr-1 or Nur-77 individually did not show any effect on forskolin-, hCG- and/or GnRH-mediated BeWo cell fusion and/or hCG secretion. After silencing any one member of the NR4A sub-family, an increase in the transcript levels of the other sub-family members was observed, indicating a compensatory effect due to their functional redundancy. Simultaneously silencing all three NR4A sub-family members significantly downregulated forskolin- and hCG-mediated BeWo cell fusion and/or hCG secretion. However, a considerable amount of cell death occurred after forskolin or hCG treatment as compared to the control siRNA-transfected cells. These results suggest that the NR4A sub-family of nuclear orphan receptors has a role in trophoblastic cell differentiation.

  6. Isolation of basal membrane proteins from BeWo cells and their expression in placentas from fetal growth-restricted pregnancies. (United States)

    Oh, Soo-Young; Hwang, Jae Ryoung; Lee, Yoonna; Choi, Suk-Joo; Kim, Jung-Sun; Kim, Jong-Hwa; Sadovsky, Yoel; Roh, Cheong-Rae


    The syncytiotrophoblast, a key barrier between the mother and fetus, is a polarized epithelium composed of a microvillus and basal membrane (BM). We sought to characterize BM proteins of BeWo cells in relation to hypoxia and to investigate their expression in placentas from pregnancies complicated by fetal growth restriction (FGR). We isolated the BM fraction of BeWo cells by the cationic colloidal silica method and identified proteins enriched in this fraction by mass spectrometry. We evaluated the effect of hypoxia on the expression and intracellular localization of identified proteins and compared their expression in BM fractions of FGR placentas to those from normal pregnancies. We identified BM proteins from BeWo cells. Among BM proteins, we further characterized heme oxygenase-1 (HO-1), voltage-dependent anion channel-1 (VDAC1), and ribophorin II (RPN2), based on their relevance to placental biology. Hypoxia enhanced the localization of these proteins to the BM of BeWo cells. HO-1, VDAC1, and RPN2 were selectively expressed in the human placental BM fraction. C-terminally truncated HO-1 was identified in placental BM fractions, and its BM expression was significantly reduced in FGR placentas than in normal placentas. Interestingly, a truncated HO-1 construct was predominantly localized in the BM in response to hypoxia and co-localized with VDAC1 in BeWo cells. Hypoxia increased the BM localization of HO-1, VDAC1, and RPN2 proteins. FGR significantly reduced the expression of truncated HO-1, which was surmised to co-localize with VDAC1 in hypoxic BeWo cells. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Enrofloxacin and Toltrazuril Are Able to Reduce Toxoplasma gondii Growth in Human BeWo Trophoblastic Cells and Villous Explants from Human Third Trimester Pregnancy

    Directory of Open Access Journals (Sweden)

    Rafaela J. da Silva


    Full Text Available Classical treatment for congenital toxoplasmosis is based on combination of sulfadiazine and pyrimethamine plus folinic acid. Due to teratogenic effects and bone marrow suppression caused by pyrimethamine, the establishment of new therapeutic strategies is indispensable to minimize the side effects and improve the control of infection. Previous studies demonstrated that enrofloxacin and toltrazuril reduced the incidence of Neospora caninum and Toxoplasma gondii infection. The aim of the present study was to evaluate the efficacy of enrofloxacin and toltrazuril in the control of T. gondii infection in human trophoblast cells (BeWo line and in human villous explants from the third trimester. BeWo cells and villous were treated with several concentrations of enrofloxacin, toltrazuril, sulfadiazine, pyrimethamine, or combination of sulfadiazine+pyrimethamine, and the cellular or tissue viability was verified. Next, BeWo cells were infected by T. gondii (2F1 clone or the ME49 strain, whereas villous samples were only infected by the 2F1 clone. Then, infected cells and villous were treated with all antibiotics and the T. gondii intracellular proliferation as well as the cytokine production were analyzed. Finally, we evaluated the direct effect of enrofloxacin and toltrazuril in tachyzoites to verify possible changes in parasite structure. Enrofloxacin and toltrazuril did not decrease the viability of cells and villous in lower concentrations. Both drugs were able to significantly reduce the parasite intracellular proliferation in BeWo cells and villous explants when compared to untreated conditions. Regardless of the T. gondii strain, BeWo cells infected and treated with enrofloxacin or toltrazuril induced high levels of IL-6 and MIF. In villous explants, enrofloxacin induced high MIF production. Finally, the drugs increased the number of unviable parasites and triggered damage to tachyzoite structure. Taken together, it can be concluded that

  8. Enrofloxacin and Toltrazuril Are Able to Reduce Toxoplasma gondii Growth in Human BeWo Trophoblastic Cells and Villous Explants from Human Third Trimester Pregnancy. (United States)

    da Silva, Rafaela J; Gomes, Angelica O; Franco, Priscila S; Pereira, Ariane S; Milian, Iliana C B; Ribeiro, Mayara; Fiorenzani, Paolo; Dos Santos, Maria C; Mineo, José R; da Silva, Neide M; Ferro, Eloisa A V; de Freitas Barbosa, Bellisa


    Classical treatment for congenital toxoplasmosis is based on combination of sulfadiazine and pyrimethamine plus folinic acid. Due to teratogenic effects and bone marrow suppression caused by pyrimethamine, the establishment of new therapeutic strategies is indispensable to minimize the side effects and improve the control of infection. Previous studies demonstrated that enrofloxacin and toltrazuril reduced the incidence of Neospora caninum and Toxoplasma gondii infection. The aim of the present study was to evaluate the efficacy of enrofloxacin and toltrazuril in the control of T. gondii infection in human trophoblast cells (BeWo line) and in human villous explants from the third trimester. BeWo cells and villous were treated with several concentrations of enrofloxacin, toltrazuril, sulfadiazine, pyrimethamine, or combination of sulfadiazine+pyrimethamine, and the cellular or tissue viability was verified. Next, BeWo cells were infected by T. gondii (2F1 clone or the ME49 strain), whereas villous samples were only infected by the 2F1 clone. Then, infected cells and villous were treated with all antibiotics and the T. gondii intracellular proliferation as well as the cytokine production were analyzed. Finally, we evaluated the direct effect of enrofloxacin and toltrazuril in tachyzoites to verify possible changes in parasite structure. Enrofloxacin and toltrazuril did not decrease the viability of cells and villous in lower concentrations. Both drugs were able to significantly reduce the parasite intracellular proliferation in BeWo cells and villous explants when compared to untreated conditions. Regardless of the T. gondii strain, BeWo cells infected and treated with enrofloxacin or toltrazuril induced high levels of IL-6 and MIF. In villous explants, enrofloxacin induced high MIF production. Finally, the drugs increased the number of unviable parasites and triggered damage to tachyzoite structure. Taken together, it can be concluded that enrofloxacin and

  9. Development of a model for functional studies of ABCG2 (breast cancer resistance protein) efflux employing a standard BeWo clone (B24). (United States)

    Crowe, Andrew; Keelan, Jeffrey A


    Human choriocarcinoma-derived BeWo cells express high levels of breast cancer resistance protein (BCRP/ABCG2) with no functional P-glycoprotein (P-gp) (ABCB1) activity, making them a potential model to study bidirectional ABCG2-mediated drug transport. However, the original BeWo clone (B24) available to researchers does not form confluent monolayers with tight junctions required by the model. Our aim was to adapt culture conditions to attempt to generate confluent BeWo monolayers for drug transport studies using the standard B24 clone. BeWo cells (B24; American Type Culture collection [ATCC]) were cultured in six-well plates or polycarbonate millicell inserts in a number of media formulations, growth supplements, and basement membrane substitutes. Cells were examined for confluence by microscopy, and transepithelial electrical resistance (TEER) was measured daily; monolayer permeability was assessed when TEER had stabilized. Optimal growth rates were achieved in culture conditions consisting of Medium 199 (M199) supplemented with epidermal growth factor (EGF; 20 ng/mL), vitamin supplements, and 10% fetal calf serum (FCS) with collagen coating. A TEER of 170 Ω in 0.6 cm(2) inserts was achieved 2 weeks after seeding under optimal conditions. The cell-impermeable diffusion marker 5(6) carboxy-2,7dichlorodihydrofluorescein (C-DCDHF) had a permeability coefficient of 3.5×10(-6) cm/s, indicative of minimal paracellular permeability. ABCG2 expression, as determined by immunoblotting, remained unaffected by confluency. In conclusion, we describe culture conditions for the B24 BeWo clone that facilitate the formation of monolayers with tighter junctions and reduced paracellular transport compared to previously published models. These growth conditions provide a good model of ABCG2-mediated drug transport in a human placental cell line.

  10. Film Permeability Determination Using Static Permeability Cells (United States)

    The permeability of tarps to soil fumigant pesticides varies depending on the active ingredient chemical: dimethyl disulfide (DMDS), methyl bromide, chloropicrin, or other. The diffusion rate can be represented by the mass transfer coefficient (MTC).

  11. BeWo cells stimulate smooth muscle cell apoptosis and elastin breakdown in a model of spiral artery transformation


    Harris, L. K.; Keogh, R. J.; Wareing, M.; Baker, P. N.; Cartwright, J. E.; Whitley, G. S.; Aplin, J. D.


    BACKGROUND: During pregnancy, extravillous trophoblast invades the uterine wall and enters the spiral arteries. Remodelling ensues, with loss of vascular smooth muscle cells (SMCs) to create high flow, low resistance vessels. Pregnancies complicated by pre-eclampsia are characterized by incomplete arterial remodelling. Endovascular trophoblast is not easily accessible for studies to establish the pathogenesis of pre-eclampsia, so we have developed a model appropriate to carry out mechanistic ...

  12. Regulation of pregnancy-associated plasma protein A2 (PAPPA2 in a human placental trophoblast cell line (BeWo

    Directory of Open Access Journals (Sweden)

    Christians Julian K


    Full Text Available Abstract Background Pregnancy-associated plasma protein A2 (PAPPA2 is an insulin-like growth factor-binding protein (IGFBP protease expressed at high levels in the placenta and upregulated in pregnancies complicated by preeclampsia and HELLP (Hemolytic anemia, Elevated Liver enzymes, and Low Platelet count syndrome. However, it is unclear whether elevated PAPPA2 expression causes abnormal placental development, or whether upregulation compensates for placental pathology. In the present study, we investigate whether PAPPA2 expression is affected by hypoxia, oxidative stress, syncytialization factors or substances known to affect the expression of PAPPA2's paralogue, PAPPA. Methods BeWo cells, a model of placental trophoblasts, were treated with one of the following: hypoxia (2% O2, oxidative stress (20 microM hydrogen peroxide, forskolin (10 microM and 100 microM, TGF-beta (10 and 50 ng/mL, TNF-alpha (100 ng/mL, IL-1beta (100 ng/mL or PGE2 (1 microM. We used quantitative RT-PCR (qRT-PCR to quantify the mRNA levels of PAPPA2, as well as those of PAPPA and ADAM12 since these proteases have similar substrates and are also highly expressed in the placenta. Where we observed significant effects on PAPPA2 mRNA levels, we tested for effects at the protein level using an in-cell Western assay. Results Hypoxia, but not oxidative stress, caused a 47-fold increase in PAPPA2 mRNA expression, while TNF-alpha resulted in a 6-fold increase, and both of these effects were confirmed at the protein level. PGE2 resulted in a 14-fold upregulation of PAPPA2 mRNA but this was not reflected at the protein level. Forskolin, TGF-beta and IL-1beta had no significant effect on PAPPA2 mRNA expression. We observed no effects of any treatment on PAPPA or ADAM12 expression. Conclusion Our study demonstrates that factors previously known to be highly expressed in preeclamptic placentae (PGE2 and TNF-alpha, contribute to the upregulation of PAPPA2. Hypoxia, known to occur in

  13. Cell permeability beyond the rule of 5. (United States)

    Matsson, Pär; Doak, Bradley C; Over, Björn; Kihlberg, Jan


    Drug discovery for difficult targets that have large and flat binding sites is often better suited to compounds beyond the "rule of 5" (bRo5). However, such compounds carry higher pharmacokinetic risks, such as low solubility and permeability, and increased efflux and metabolism. Interestingly, recent drug approvals and studies suggest that cell permeable and orally bioavailable drugs can be discovered far into bRo5 space. Tactics such as reduction or shielding of polarity by N-methylation, bulky side chains and intramolecular hydrogen bonds may be used to increase cell permeability in this space, but often results in decreased solubility. Conformationally flexible compounds can, however, combine high permeability and solubility, properties that are keys for cell permeability and intestinal absorption. Recent developments in computational conformational analysis will aid design of such compounds and hence prediction of cell permeability. Transporter mediated efflux occurs for most investigated drugs in bRo5 space, however it is commonly overcome by high local intestinal concentrations on oral administration. In contrast, there is little data to support significant impact of transporter-mediated intestinal absorption in bRo5 space. Current knowledge of compound properties that govern transporter effects of bRo5 drugs is limited and requires further fundamental and comprehensive studies. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Endothelial cell permeability to water and antipyrine

    International Nuclear Information System (INIS)

    Garrick, R.A.


    The endothelium provides a structural barrier between plasma constituents and the tissues. The permeability characteristics of the the endothelial cells regulate the transcellular movement of materials across this barrier while other movement is paracellular. In this study the permeability of the endothelial cells to tritiated water ( 3 HHO) and 14 C-labeled antipyrine (AP) was investigated. The cells were isolated non-enzymatically from calf pulmonary artery and were maintained in culture and used between the seventh and fifteenth passage. The cells were removed from the T-flasks with a rubber policeman, titurated with a 22g needle and centrifuged. The cells were mixed with an extracellular marker, drawn into polyethylene tubing and packed by centrifugation for use in the linear diffusion technique. All measurements were made at 37 C. The diffusion coefficients for 3 HHO through the packed cells (D), the intracellular material (D 2 ), and the extracellular material (D 1 ) were 0.682, 0.932 and 2.45 x 10 -5 cm 2 s -1 and for AP were 0.273, 0.355 and 1.13 x 10 -5 cm 2 s -1 respectively. The permeability coefficient calculated by the series-parallel pathway model for 3 HHO was higher than that for AP and for both 3 HHO and AP were lower than those calculated for isolated lung cells and erythrocytes

  15. Engineered Trehalose Permeable to Mammalian Cells.

    Directory of Open Access Journals (Sweden)

    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.

  16. A comparison of the embryonic stem cell test and whole embryo culture assay combined with the BeWo placental passage model for predicting the embryotoxicity of azoles.

    NARCIS (Netherlands)

    Dimopoulou, Myrto; Verhoef, Aart; Gomes, Caroline A; van Dongen, Catharina W; Rietjens, Ivonne M C M; Piersma, Aldert H; van Ravenzwaay, Bennard


    In the present study, we show the value of combining toxico-dynamic and -kinetic in vitro approaches for embryotoxicity testing of azoles. Both the whole embryo culture (WEC) and the embryonic stem cells test (EST) predicted the in vivo potency ranking of twelve tested azoles with moderate accuracy.

  17. Measurement of relative permeability of fuel cell diffusion media

    KAUST Repository

    Hussaini, I.S.; Wang, C.Y.


    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

  18. Measurement of relative permeability of fuel cell diffusion media

    KAUST Repository

    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.

  19. Human parvovirus B19 antibodies induce altered membrane protein expression and apoptosis of BeWo trophoblasts. (United States)

    Tzang, Bor-Show; Chiang, Szu-Yi; Chan, Hsu-Chin; Liu, Chung-Hsien; Hsu, Tsai-Ching


    Human parvovirus B19 (B19) is harmful during pregnancy since it can be vertically transmitted to the developing fetus. In addition, the anti‑B19 antibodies induced by B19 infection are believed to have a cytopathic role in B19 transmission; however, knowledge regarding the effects of anti‑B19 antibodies during pregnancy is limited. To investigate the possible roles of anti‑B19 antibodies during pregnancy, the present study examined the effects of anti‑B19‑VP1 unique region (VP1u), anti‑B19‑VP2 and anti‑B19‑nonstructural protein 1 (NS1) immunoglobulin G (IgG) antibodies on BeWo trophoblasts. Briefly, BeWo trophoblasts were incubated with purified IgG against B19‑VP1u, B19‑VP2 and B19‑NS1. Subsequently, the expression of surface proteins and apoptotic molecules were assessed in BeWo trophoblasts using flow cytometry, ELISA and western blotting. The expression levels of human leukocyte antigen (HLA)‑G were significantly increased on BeWo trophoblasts treated with rabbit anti‑B19‑VP1u IgG, and were unchanged in those treated with rabbit anti‑B19‑NS1 and anti‑B19‑VP2 IgG, as compared with the control group. Furthermore, the expression levels of globoside (P blood group antigen) and cluster of differentiation (CD)29 (β1 integrin) were significantly increased in BeWo trophoblasts treated with rabbit anti‑B19‑NS1 and anti‑B19‑VP2 IgG, whereas only CD29 was also significantly increased in cells treated with anti‑B19‑VP1u IgG. In addition, the number of cells at sub‑G1 phase; caspase‑3 activity; and the expression of intrinsic and extrinsic apoptotic molecules, including Fas‑associated death domain protein, activated caspase‑8, activated caspase‑3, B‑cell lymphoma 2‑associated X protein, cytochrome c, apoptotic peptidase activating factor 1 and activated caspase‑9, were significantly increased in BeWo trophoblasts treated with anti‑B19‑VP1u and anti‑B19‑NS1 IgG. In conclusion, the present

  20. Acamprosate permeability across Caco-2 cell monolayer is predominantly paracellular

    DEFF Research Database (Denmark)

    Antonescu, Irina-Elena; Steffansen, Bente

    support area, thickness, and porosity). Results. The mean (± SD) Papp, exp of acamprosate and [14C]-mannitol across Caco-2 cell monolayers was measured as 0.19 ± 0.07 x 10-6 cm/s (n = 2, N = 3) and 0.35 ± 0.17 x 10-6 cm/s (n = 3, N = 4), respectively. Acamprosate PUBL and Pf were estimated as 200 - 3150 x...... role in acamprosate permeability, as only a very low fraction of acamprosate is in the neutral form at pH 7.4. The estimated acamprosate Ppara accounts for nearly 100% of the mathematically determined acamprosate Papp, calc (0.20 ± 0.10 x 10-6 cm/s), which matches well with the experimentally...... to the overall acamprosate apparent permeability. Methods. Acamprosate apparent permeability (Papp, exp) was determined across Caco-2 monolayers in the apical-to-basolateral transport direction using a buffer pH of 7.4 and several cell passages (N). Acamprosate concentrations were quantified by LC...

  1. The effects of heavy metal ions on the chlorophyll content and cell membrane permeability of charophytes

    International Nuclear Information System (INIS)

    Fu Hualong; Chen Hao; Dong Bin; Qing Renwei


    The authors studied the effects of several heavy metal ions in different concentrations (Cd 2+ , Hg 2+ , Pb 2+ , Cr 6+ ) on the chlorophyll content and cell membrane permeability of Chara vulgaris L. It was discovered that the effects of heavy metal ions on the chlorophyll content and cell membrane permeability of Chara vulgaris L. changed with their different concentration. The trend was that the chlorophyll content and cell membrane permeability were decreased with the increase of the heavy metal ions. The degree of chlorophyll content affected was Cr 6+ , Cd 2+ , Hg 2+ , Pb 2+ , and that of cell membrane permeability affected was Cd 2+ , Cr 6+ , Hg 2+ , Pb 2+

  2. Cell-permeable nanobodies for targeted immunolabelling and antigen manipulation in living cells. (United States)

    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.

  3. Cell-permeable nanobodies for targeted immunolabelling and antigen manipulation in living cells (United States)

    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.

  4. Permeability of surface modified polyamidoamine (PAMAM) dendrimers across Caco-2 cell monolayers


    Yellepeddi, Venkata K.; Pisal, Dipak S.; Kumar, Ajay; Kaushik, Radhey S.; Hildreth, Michael B.; Guan, Xiangming; Palakurthi, Srinath


    Aim of this study was to prepare polyamine-conjugated PAMAM dendrimers and study their permeability across Caco-2 cell monolayers. Polyamines, namely, arginine and ornithine were conjugated to the amine terminals of the G4 PAMAM dendrimers by Fmoc synthesis. The apical-to-basolateral (AB) and basolateral-to-apical (BA) apparent permeability coefficients (Papp) for the PAMAM dendrimers increased by conjugating the dendrimers with both of the polyamines. The enhancement in permeability was depe...

  5. Drug-permeability and transporter assays in Caco-2 and MDCK cell lines. (United States)

    Volpe, Donna A


    The human colon adenocarcinoma Caco-2 and Madin-Darby canine kidney epithelial cell lines provide in vitro tools to assess a drug's permeability and transporter interactions during discovery and development. The cells, when cultured on semiporous filters, form confluent monolayers that model the intestinal epithelial barrier for permeability, transporter and drug-interaction assays. The applications of these assays in pharmaceutical research include qualitative prediction and ranking of absorption, determining mechanism(s) of permeability, formulation effects on drug permeability, and the potential for transporter-mediated drug-drug interactions. This review focuses on recent examples of Caco-2 and Madin-Darby canine kidney cells assays for drug permeability including transfected and knock-down cells, miniaturization and automation, and assay combinations to better understand and predict intestinal drug absorption.

  6. Inflammasome Inhibition Suppresses Alveolar Cell Permeability Through Retention of Neuregulin-1 (NRG-1

    Directory of Open Access Journals (Sweden)

    Rajanbabu Venugopal


    Full Text Available Background: Neuregulin (NRG-1-human epidermal receptor (HER-2 signaling pathway is a key regulator of IL-1β-mediated pulmonary inflammation and epithelial permeability. The inflammasome is a newly discovered molecular platform required for caspase-1 activation and maturation of IL-1β. However, the role of the inflammasome in NRG-1-HER2 signaling-mediated alveolar cell permeability is unknown. Methods: The inflammasome was activated or inhibited in THP-1 cells; supernatants from these cells were added to A549 cells and human small airway epithelial cells (HSAEC. The protein expression of NRG-1 and phospho-HER2 (pHER2 were measured by Western blot analysis and epithelial permeability was measured using Lucifer yellow dye. Results: Results reveal that alveolar permeability in A549 cells and HSAEC is increased when treated with supernatants of inflammasome-activated THP-1 cells. Alveolar permeability is significantly suppressed when treated with supernatant of inflammasome-inhibited THP-1 cells. Inflammasome-mediated permeability is decreased when A549 cells and HSAEC are pretreated with IL-1β receptor antagonist (IL-1βRA. In addition, HER2 kinase inhibitor AG825 or NRG-1 inhibitor TAPI inhibits inflammasome-mediated permeability in A549 cells and HSAEC demonstrating critical roles of IL-1β, NRG-1, and HER2 in inflammasome-mediated alveolar permeability. Conclusion: These findings suggest that inflammasome-induced alveolar cell permeability is mediated by NRG-1/HER2 signaling through IL-1β regulation.

  7. Stereochemistry Balances Cell Permeability and Solubility in the Naturally Derived Phepropeptin Cyclic Peptides. (United States)

    Schwochert, Joshua; Lao, Yongtong; Pye, Cameron R; Naylor, Matthew R; Desai, Prashant V; Gonzalez Valcarcel, Isabel C; Barrett, Jaclyn A; Sawada, Geri; Blanco, Maria-Jesus; Lokey, R Scott


    Cyclic peptide (CP) natural products provide useful model systems for mapping "beyond-Rule-of-5" (bRo5) space. We identified the phepropeptins as natural product CPs with potential cell permeability. Synthesis of the phepropeptins and epimeric analogues revealed much more rapid cellular permeability for the natural stereochemical pattern. Despite being more cell permeable, the natural compounds exhibited similar aqueous solubility as the corresponding epimers, a phenomenon explained by solvent-dependent conformational flexibility among the natural compounds. When analyzing the polarity of the solution structures we found that neither the number of hydrogen bonds nor the total polar surface area accurately represents the solvation energies of the high and low dielectric conformations. This work adds to a growing number of natural CPs whose solvent-dependent conformational behavior allows for a balance between aqueous solubility and cell permeability, highlighting structural flexibility as an important consideration in the design of molecules in bRo5 chemical space.

  8. Radiation Effects on the Cytoskeleton of Endothelial Cells and Endothelial Monolayer Permeability

    International Nuclear Information System (INIS)

    Gabrys, Dorota; Greco, Olga; Patel, Gaurang; Prise, Kevin M.; Tozer, Gillian M.; Kanthou, Chryso


    Purpose: To investigate the effects of radiation on the endothelial cytoskeleton and endothelial monolayer permeability and to evaluate associated signaling pathways, which could reveal potential mechanisms of known vascular effects of radiation. Methods and Materials: Cultured endothelial cells were X-ray irradiated, and actin filaments, microtubules, intermediate filaments, and vascular endothelial (VE)-cadherin junctions were examined by immunofluorescence. Permeability was determined by the passage of fluorescent dextran through cell monolayers. Signal transduction pathways were analyzed using RhoA, Rho kinase, and stress-activated protein kinase-p38 (SAPK2/p38) inhibitors by guanosine triphosphate-RhoA activation assay and transfection with RhoAT19N. The levels of junction protein expression and phosphorylation of myosin light chain and SAPK2/p38 were assessed by Western blotting. The radiation effects on cell death were verified by clonogenic assays. Results: Radiation induced rapid and persistent actin stress fiber formation and redistribution of VE-cadherin junctions in microvascular, but not umbilical vein endothelial cells, and microtubules and intermediate filaments remained unaffected. Radiation also caused a rapid and persistent increase in microvascular permeability. RhoA-guanosine triphosphatase and Rho kinase were activated by radiation and caused phosphorylation of downstream myosin light chain and the observed cytoskeletal and permeability changes. SAPK2/p38 was activated by radiation but did not influence either the cytoskeleton or permeability. Conclusion: This study is the first to show rapid activation of the RhoA/Rho kinase by radiation in endothelial cells and has demonstrated a link between this pathway and cytoskeletal remodeling and permeability. The results also suggest that the RhoA pathway might be a useful target for modulating the permeability and other effects of radiation for therapeutic gain

  9. Ascorbic acid attenuates endothelial permeability triggered by cell-free hemoglobin. (United States)

    Kuck, Jamie L; Bastarache, Julie A; Shaver, Ciara M; Fessel, Joshua P; Dikalov, Sergey I; May, James M; Ware, Lorraine B


    Increased endothelial permeability is central to shock and organ dysfunction in sepsis but therapeutics targeted to known mediators of increased endothelial permeability have been unsuccessful in patient studies. We previously reported that cell-free hemoglobin (CFH) is elevated in the majority of patients with sepsis and is associated with organ dysfunction, poor clinical outcomes and elevated markers of oxidant injury. Others have shown that Vitamin C (ascorbate) may have endothelial protective effects in sepsis. In this study, we tested the hypothesis that high levels of CFH, as seen in the circulation of patients with sepsis, disrupt endothelial barrier integrity. Human umbilical vein endothelial cells (HUVEC) were grown to confluence and treated with CFH with or without ascorbate. Monolayer permeability was measured by Electric Cell-substrate Impedance Sensing (ECIS) or transfer of 14 C-inulin. Viability was measured by trypan blue exclusion. Intracellular ascorbate was measured by HPLC. CFH increased permeability in a dose- and time-dependent manner with 1 mg/ml of CFH increasing inulin transfer by 50% without affecting cell viability. CFH (1 mg/ml) also caused a dramatic reduction in intracellular ascorbate in the same time frame (1.4 mM without CFH, 0.23 mM 18 h after 1 mg/ml CFH, p < 0.05). Pre-treatment of HUVECs with ascorbate attenuated CFH induced permeability. CFH increases endothelial permeability in part through depletion of intracellular ascorbate. Supplementation of ascorbate can attenuate increases in permeability mediated by CFH suggesting a possible therapeutic approach in sepsis. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Oxcarbazepine-loaded polymeric nanoparticles: development and permeability studies across in vitro models of the blood-brain barrier and human placental trophoblast. (United States)

    Lopalco, Antonio; Ali, Hazem; Denora, Nunzio; Rytting, Erik


    Encapsulation of antiepileptic drugs (AEDs) into nanoparticles may offer promise for treating pregnant women with epilepsy by improving brain delivery and limiting the transplacental permeability of AEDs to avoid fetal exposure and its consequent undesirable adverse effects. Oxcarbazepine-loaded nanoparticles were prepared by a modified solvent displacement method from biocompatible polymers (poly(lactic-co-glycolic acid) [PLGA] with or without surfactant and PEGylated PLGA [Resomer(®) RGPd5055]). The physical properties of the developed nanoparticles were determined with subsequent evaluation of their permeability across in vitro models of the blood-brain barrier (hCMEC/D3 cells) and human placental trophoblast cells (BeWo b30 cells). Oxcarbazepine-loaded nanoparticles with encapsulation efficiency above 69% were prepared with sizes ranging from 140-170 nm, polydispersity indices below 0.3, and zeta potential values below -34 mV. Differential scanning calorimetry and X-ray diffraction studies confirmed the amorphous state of the nanoencapsulated drug. The apparent permeability (Pe ) values of the free and nanoencapsulated oxcarbazepine were comparable across both cell types, likely due to rapid drug release kinetics. Transport studies using fluorescently-labeled nanoparticles (loaded with coumarin-6) demonstrated increased permeability of surfactant-coated nanoparticles. Future developments in enzyme-prodrug therapy and targeted delivery are expected to provide improved options for pregnant patients with epilepsy.

  11. Oxcarbazepine-loaded polymeric nanoparticles: development and permeability studies across in vitro models of the blood–brain barrier and human placental trophoblast (United States)

    Lopalco, Antonio; Ali, Hazem; Denora, Nunzio; Rytting, Erik


    Encapsulation of antiepileptic drugs (AEDs) into nanoparticles may offer promise for treating pregnant women with epilepsy by improving brain delivery and limiting the transplacental permeability of AEDs to avoid fetal exposure and its consequent undesirable adverse effects. Oxcarbazepine-loaded nanoparticles were prepared by a modified solvent displacement method from biocompatible polymers (poly(lactic-co-glycolic acid) [PLGA] with or without surfactant and PEGylated PLGA [Resomer® RGPd5055]). The physical properties of the developed nanoparticles were determined with subsequent evaluation of their permeability across in vitro models of the blood–brain barrier (hCMEC/D3 cells) and human placental trophoblast cells (BeWo b30 cells). Oxcarbazepine-loaded nanoparticles with encapsulation efficiency above 69% were prepared with sizes ranging from 140–170 nm, polydispersity indices below 0.3, and zeta potential values below -34 mV. Differential scanning calorimetry and X-ray diffraction studies confirmed the amorphous state of the nanoencapsulated drug. The apparent permeability (Pe) values of the free and nanoencapsulated oxcarbazepine were comparable across both cell types, likely due to rapid drug release kinetics. Transport studies using fluorescently-labeled nanoparticles (loaded with coumarin-6) demonstrated increased permeability of surfactant-coated nanoparticles. Future developments in enzyme-prodrug therapy and targeted delivery are expected to provide improved options for pregnant patients with epilepsy. PMID:25792832

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

    Directory of Open Access Journals (Sweden)

    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

  13. Automation of cell-based drug absorption assays in 96-well format using permeable support systems. (United States)

    Larson, Brad; Banks, Peter; Sherman, Hilary; Rothenberg, Mark


    Cell-based drug absorption assays, such as Caco-2 and MDCK-MDR1, are an essential component of lead compound ADME/Tox testing. The permeability and transport data they provide can determine whether a compound continues in the drug discovery process. Current methods typically incorporate 24-well microplates and are performed manually. Yet the need to generate absorption data earlier in the drug discovery process, on an increasing number of compounds, is driving the use of higher density plates. A simple, more efficient process that incorporates 96-well permeable supports and proper instrumentation in an automated process provides more reproducible data compared to manual methods. Here we demonstrate the ability to perform drug permeability and transport assays using Caco-2 or MDCKII-MDR1 cells. The assay procedure was automated in a 96-well format, including cell seeding, media and buffer exchanges, compound dispense, and sample removal using simple robotic instrumentation. Cell monolayer integrity was confirmed via transepithelial electrical resistance and Lucifer yellow measurements. Proper cell function was validated by analyzing apical-to-basolateral and basolateral-to-apical movement of rhodamine 123, a known P-glycoprotein substrate. Apparent permeability and efflux data demonstrate how the automated procedure provides a less variable method than manual processing, and delivers a more accurate assessment of a compound's absorption characteristics.

  14. Determination of lamivudine and zidovudine permeability using a different ex vivo method in Franz cells. (United States)

    Dezani, André Bersani; Pereira, Thaisa Marinho; Caffaro, Arthur Massabki; Reis, Juliana Mazza; Serra, Cristina Helena Dos Reis


    The major processes that control the absorption of orally administered drugs are dissolution and gastrointestinal permeation. These processes depend on two main properties: solubility and permeability. Based on these characteristics, the Biopharmaceutical Classification System (BCS) was proposed as a tool to assist in biowaiver and bioavailability prediction of drugs. The purpose of the present study was to evaluate the permeability of lamivudine (3TC) and zidovudine (AZT) using a different ex vivo method in Franz cells. A segment of jejunum was inserted in a Franz cells apparatus, in order to assess drug permeability in the apical-basolateral (A-B) and basolateral-apical (B-A) directions. Each drug was added to the donor chamber, collected from the acceptor chamber and analyzed by HPLC. Fluorescein (FLU) and metoprolol (METO) were used as low and high permeability markers, respectively. The apparent permeability (Papp) results for the A-B direction were: Papp FLU A-B=0.54×10(-4)cm·s(-1), Papp METO A-B=7.99×10(-4)cm·s(-1), Papp 3TC A-B=4.58×10(-4)cm·s(-1) and Papp AZT A-B=5.34×10(-4)cm·s(-1). For the B-A direction, the Papp results were: Papp FLU B-A=0.56×10(-4)cm·s(-1), Papp METO B-A=0.25×10(-4)cm·s(-1), Papp 3TC B-A=0.24×10(-4)cm·s(-1) and Papp AZT B-A=0.19×10(-4)cm·s(-1). For the A-B direction, the Papp results of fluorescein and metoprolol show low and high permeability, respectively, indicating that the membranes were appropriate for permeability studies. For the A-B direction, the Papp results of 3TC and AZT suggest that these antiretroviral drugs have permeability values close to metoprolol. Nevertheless, for the B-A direction the Papp results do not suggest efflux mechanism for any of the drugs. Thereby, the different ex vivo methods using Franz cells can be successfully applied in drug permeability studies, in particular for drug biopharmaceutical classification. Copyright © 2013 Elsevier Inc. All rights reserved.

  15. Regulation of endothelial cell shape and monolayer permeability by atrial natriuretic peptide

    International Nuclear Information System (INIS)

    Lofton-Day, C.E.


    Atrial natriuretic peptide (ANP), considered to be an important regulator of intravascular fluid volume, binds specifically to receptors on endothelial cells. In this study, the role of ANP-specific binding was investigated by examining the effect of ANP on the morphology and macromolecular permeability of monolayer cultures of bovine aortic endothelial cells. ANP alone had no observable effect on the monolayers. However, incubation of monolayers with ANP antagonized thrombin- or glucose oxidase-induced cell shape changes and intercellular gap formation. ANP pretreatment also opposed the effect of thrombin and glucose oxidase on actin filament distribution as observed by rhodamine-phalloidin staining and digital image analysis of F0actin staining. In addition, ANP reversed cell shape changes and cytoskeletal alterations induced by thrombin treatment but did not reverse alternations induced by glucose oxidase treatment. ANP significantly reduced increases in monolayer permeability to albumin resulting from thrombin or glucose oxidases treatment. Thrombin caused a 2-fold increase in monolayer permeability to 125 I-labeled albumin, which was abolished by 10 -8 -10 -6 M ANP pretreatment. Glucose oxidase caused similar increases in permeability and was inhibited by ANP at slightly shorter time periods

  16. The novel oral imatinib microemulsions: physical properties, cytotoxicity activities and improved Caco-2 cell permeability. (United States)

    Gundogdu, Evren; Karasulu, Hatice Yesim; Koksal, Cinel; Karasulu, Ercüment


    The objective of this study was to formulate imatinib (IM) loaded to water-in-oil (w/o) microemulsions as an alternative formulation for cancer therapy and to evaluate the cytotoxic effect of microemulsions Caco-2 and MCF-7. Moreover, permeability studies were also performed with Caco-2 cells. W/o microemulsion systems were developed by using pseudo-ternary phase diagram. According to cytotoxicity studies, all formulations did not exert a cytotoxic effect on Caco-2 cells. Furthermore, all formulations had a significant cytotoxic effect on MCF-7 cells and the cytotoxic effect of M3IM was significantly more than that of other microemulsions and IM solution (p < 0.05). The permeability studies of IM across Caco-2 cells showed that permeability value from apical to basolateral was higher than permeability value of other formulations. In conclusion, the microemulsion formulations as a drug carrier, especially M3IM formulation, may be used as an effective alternative breast cancer therapy for oral delivery of IM.

  17. Towards a Biohybrid Lung: Endothelial Cells Promote Oxygen Transfer through Gas Permeable Membranes. (United States)

    Menzel, Sarah; Finocchiaro, Nicole; Donay, Christine; Thiebes, Anja Lena; Hesselmann, Felix; Arens, Jutta; Djeljadini, Suzana; Wessling, Matthias; Schmitz-Rode, Thomas; Jockenhoevel, Stefan; Cornelissen, Christian Gabriel


    In patients with respiratory failure, extracorporeal lung support can ensure the vital gas exchange via gas permeable membranes but its application is restricted by limited long-term stability and hemocompatibility of the gas permeable membranes, which are in contact with the blood. Endothelial cells lining these membranes promise physiological hemocompatibility and should enable prolonged application. However, the endothelial cells increase the diffusion barrier of the blood-gas interface and thus affect gas transfer. In this study, we evaluated how the endothelial cells affect the gas exchange to optimize performance while maintaining an integral cell layer. Human umbilical vein endothelial cells were seeded on gas permeable cell culture membranes and cultivated in a custom-made bioreactor. Oxygen transfer rates of blank and endothelialized membranes in endothelial culture medium were determined. Cell morphology was assessed by microscopy and immunohistochemistry. Both setups provided oxygenation of the test fluid featuring small standard deviations of the measurements. Throughout the measuring range, the endothelial cells seem to promote gas transfer to a certain extent exceeding the blank membranes gas transfer performance by up to 120%. Although the underlying principles hereof still need to be clarified, the results represent a significant step towards the development of a biohybrid lung.

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

    KAUST Repository

    Hussaini, Irfan; Wang, Chao-Yang


    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.

  19. Water permeability of acinar cell membranes in the isolated perfused rabbit mandibular salivary gland. (United States)

    Steward, M C; Seo, Y; Rawlings, J M; Case, R M


    1. The diffusive water permeability of epithelial cell membranes in the perfused rabbit mandibular salivary gland was measured at 37 degrees C by a 1H nuclear magnetic resonance relaxation method using an extracellular relaxation reagent, gadolinium diethylenetriaminepentaacetic acid (Gd(DTPA)). 2. In glands perfused with a HEPES-buffered solution containing 10 mmol l-1 Gd(DTPA), the spin-lattice (T1) relaxation of the water protons showed two exponential components. The water compartment responsible for the slower component corresponded in magnitude to 71 +/- 5% of the wet weight of the gland, and was attributed to the exchangeable intracellular water of the acinar cells. 3. The rate constant for water efflux from the cells was estimated to be 4.1 +/- 0.1 s-1 which would be consistent with a diffusive membrane permeability (Pd) of approximately 3 x 10(-3) cm s-1. Stimulation with acetylcholine (10(-6) mol l-1) did not cause any detectable change in membrane water permeability. 4. Since the basolateral membrane probably provides the main pathway for water efflux, the osmotic water permeability of this barrier (expressed per gland) was estimated to be less than 6.2 cm3 s-1. This would be insufficient to account for the generation of a near-isosmotic fluid at the flow rates observed during secretion, and suggests that a substantial fraction of the flow of water occurs via a paracellular route. PMID:1966053

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

    DEFF Research Database (Denmark)

    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......: AQP2-S256D > AQP2 wild-type > AQP2-S256A. We propose that the method can be applied to study AQP function and more generally to study cell volume changes in adherent cell lines. Furthermore, it should be adaptable for AQP inhibitor screening in chemical compound libraries....

  1. Effects of gamma- and UV-radiation on DNA synthesis in permeable cells of Bacillus stearothermophilus

    International Nuclear Information System (INIS)

    Trofimenko, A.F.; Vorob'eva, A.M.; Gaziev, A.I.


    It was shown that the most of the DNA synthesis is repaired in permeable cells of Bacillus stearothermophilus not affected by injurious agents. γ-irradiation stimulates the reparative synthesis and degradation of DNA whereas UV-radiation decreases the activity of these processes. The reason for such an unusual response of thermophiles to irradiation lies perhaps in high temperatures at which the cells exist

  2. Cell-Permeable Parkin Proteins Suppress Parkinson Disease-Associated Phenotypes in Cultured Cells and Animals (United States)

    Duong, Tam; Kim, Jaetaek; Ruley, H. Earl; Jo, Daewoong


    Parkinson’s disease (PD) is a neurodegenerative disorder of complex etiology characterized by the selective loss of dopaminergic neurons, particularly in the substantia nigra. Parkin, a tightly regulated E3 ubiquitin ligase, promotes the survival of dopaminergic neurons in both PD and Parkinsonian syndromes induced by acute exposures to neurotoxic agents. The present study assessed the potential of cell-permeable parkin (CP-Parkin) as a neuroprotective agent. Cellular uptake and tissue penetration of recombinant, enzymatically active parkin was markedly enhanced by the addition of a hydrophobic macromolecule transduction domain (MTD). The resulting CP-Parkin proteins (HPM13 and PM10) suppressed dopaminergic neuronal toxicity in cells and mice exposed to 6-hydroxydopamine (6-OHDH) and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). These included enhanced survival and dopamine expression in cultured CATH.a and SH-SY5Y neuronal cells; and protection against MPTP-induced damage in mice, notably preservation of tyrosine hydroxylase-positive cells with enhanced dopamine expression in the striatum and midbrain, and preservation of gross motor function. These results demonstrate that CP-Parkin proteins can compensate for intrinsic limitations in the parkin response and provide a therapeutic strategy to augment parkin activity in vivo. PMID:25019626

  3. An alpha-adrenergic receptor mechanism controlling potassium permeability in the rat lacrimal gland acinar cell

    International Nuclear Information System (INIS)

    Parod, R.J.; Putney, J.W. Jr.


    Rat lacrimal gland slices, incubated in a balanced, buffered salt solution, were found to be physiologically stable for up to 2 hr with respect to 0 2 consumption, extracellular space, and water and ion content. The release of 86 Rb serves as a good substitute for 42 K in monitoring the movement of K through the cell membrane. Adrenaline appears to increase membrane permeability to K as evidenced by an increase in the rate of 86 Rb efflux. This response to adrenaline was blocked by phentolamine but not by propranolol and was mimicked by phenylephrine but not by isoprenaline. The magnitude of the 86 Rb release indicates that it is being released, at least in part, from the lacrimal gland acinar cell. It is concluded that the lacrimal gland acinar cell has an α-adrenergic receptor, activation of which leads to an increase in membrane permeability to K. (author)

  4. Simulating kinetic parameters in transporter mediated permeability across Caco-2 cells. A case study on estrange-3-sulphate

    DEFF Research Database (Denmark)

    Rolsted, Kamilla; Rapin, Nicolas; Steffansen, Bente


    Substances that compete for the same saturable intestinal transporters may when dosed together lead to altered permeability and hence influence bioavailability. The aim was to simulate kinetic parameters, i.e. K(m) and J(max), for transporter mediated E(1)S permeability across Caco-2 cells...

  5. Oxcarbazepine-loaded polymeric nanoparticles: development and permeability studies across in vitro models of the blood–brain barrier and human placental trophoblast

    Directory of Open Access Journals (Sweden)

    Lopalco A


    Full Text Available Antonio Lopalco,1–3,* Hazem Ali,1,* Nunzio Denora,3 Erik Rytting1,4,5 1Department of Obstretrics and Gynecology, University of Texas Medical Branch, Galveston, TX, USA; 2Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS, USA; 3Department of Pharmacy – Drug Sciences, University of Bari Aldo Moro, Bari, Italy; 4Center for Biomedical Engineering, University of Texas Medical Branch, Galveston, TX, USA; 5Department of Pharmacology and Toxicology, University of Texas Medical Branch, Galveston, TX, USA *These authors contributed equally to this work Abstract: Encapsulation of antiepileptic drugs (AEDs into nanoparticles may offer promise for treating pregnant women with epilepsy by improving brain delivery and limiting the transplacental permeability of AEDs to avoid fetal exposure and its consequent undesirable adverse effects. Oxcarbazepine-loaded nanoparticles were prepared by a modified solvent displacement method from biocompatible polymers (poly(lactic-co-glycolic acid [PLGA] with or without surfactant and PEGylated PLGA [Resomer® RGPd5055]. The physical properties of the developed nanoparticles were determined with subsequent evaluation of their permeability across in vitro models of the blood–brain barrier (hCMEC/D3 cells and human placental trophoblast cells (BeWo b30 cells. Oxcarbazepine-loaded nanoparticles with encapsulation efficiency above 69% were prepared with sizes ranging from 140–170 nm, polydispersity indices below 0.3, and zeta potential values below −34 mV. Differential scanning calorimetry and X-ray diffraction studies confirmed the amorphous state of the nanoencapsulated drug. The apparent permeability (Pe values of the free and nanoencapsulated oxcarbazepine were comparable across both cell types, likely due to rapid drug release kinetics. Transport studies using fluorescently-labeled nanoparticles (loaded with coumarin-6 demonstrated increased permeability of surfactant-coated nanoparticles

  6. Tumor necrosis factor alpha increases epithelial barrier permeability by disrupting tight junctions in Caco-2 cells

    Directory of Open Access Journals (Sweden)

    W. Cui


    Full Text Available The objectives of this study were to determine the effect of tumor necrosis factor alpha (TNF-α on intestinal epithelial cell permeability and the expression of tight junction proteins. Caco-2 cells were plated onto Transwell® microporous filters and treated with TNF-α (10 or 100 ng/mL for 0, 4, 8, 16, or 24 h. The transepithelial electrical resistance and the mucosal-to-serosal flux rates of the established paracellular marker Lucifer yellow were measured in filter-grown monolayers of Caco-2 intestinal cells. The localization and expression of the tight junction protein occludin were detected by immunofluorescence and Western blot analysis, respectively. SYBR-Green-based real-time PCR was used to measure the expression of occludin mRNA. TNF-α treatment produced concentration- and time-dependent decreases in Caco-2 transepithelial resistance and increases in transepithelial permeability to the paracellular marker Lucifer yellow. Western blot results indicated that TNF-α decreased the expression of phosphorylated occludin in detergent-insoluble fractions but did not affect the expression of non-phosphorylated occludin protein. Real-time RT-PCR data showed that TNF-α did not affect the expression of occludin mRNA. Taken together, our data demonstrate that TNF-α increases Caco-2 monolayer permeability, decreases occludin protein expression and disturbs intercellular junctions.

  7. Tc-99m Radiolabeled Alendronate Sodium Microemulsion: Characterization and Permeability Studies Across Caco-2 Cells. (United States)

    Elitez, Yetkin; Ekinci, Meliha; Ilem-Ozdemir, Derya; Gundogdu, Evren; Asikoglu, Makbule


    Alendronate sodium (ALD) is used orally but it is poorly absorbed from the gastrointestinal (GI) tract. For this reason, microemulsion system was chosen to evaluate ALD from the GI tract after oral delivery. This study was aimed to prepare water-in-oil (w/o) microemulsion formulation of ALD and evaluate the permeability of ALD microemulsion from Caco-2 cell lines with radioactive and nonradioactive studies. The ALD microemulsion was developed by using pseudo-ternary phase diagram and composed of Soybean oil, Colliphor EL, Tween 80, Transcutol and distilled water. The prepared ALD microemulsion was characterized by physical appearance, droplet size, viscosity, pH, electrical conductivity and refractive index. The stability of the formulation was investigated for 6 months at 25±2°C/60±5% of relative humidity (RH) as well as at 40±2°C/75±5% RH. After that 1 mg of ALD was radiolabeled with 99mTc and added to microemulsion. The permeability studies were performed with both 99mTc-ALD microemulsion and ALD microemulsion. The experimental results suggested that ALD microemulsion presented adequate stability with droplet size varying from 37.8±0.9 to 39.9±1.2 nm during incubation time. In addition, ALD microemulsion was radiolabeled with high labeling efficiency (>95%). In a non-radioactive study, ALD permeability was found to be 45 µg.mL-1 and microemulsion has high permeability percentage when compared to another study. The novel w/o microemulsion formulation has been developed for oral delivery of ALD. Based on the results, permeability of ALD could be significantly improved by the microemulsion formulation. In addition, 99mTc-ALD microemulsion in capsule can be used for bone disease treatment and diagnosis. Copyright© Bentham Science Publishers; For any queries, please email at

  8. Structural and permeability characterization of biosynthetic PVA hydrogels designed for cell-based therapy. (United States)

    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.

  9. Permeability, transport, and metabolism of solutes in Caco-2 cell monolayers: a theoretical study. (United States)

    Sun, Huadong; Pang, K Sandy


    We explored the properties of a catenary model that includes the basolateral (B), apical (A), and cellular compartments via simulations under linear and nonlinear conditions to understand the asymmetric observations arising from transporters, enzymes, and permeability in Caco-2 cells. The efflux ratio (EfR; P(app,B-->A)/P(app,A-->B)), obtained from the effective permeability from the A-->B and B-->A direction under linear conditions, was unity for passively permeable drugs whose transport does not involve transporters; the value was unaffected by cellular binding or metabolism, but increased with apical efflux. Metabolism was asymmetric, showing lesser metabolite accrual for the B-->A than A-->B direction because of inherent differences in the volumes for A and B. Moreover, the net flux (total - passive permeation) due to saturable apical efflux, absorption, or metabolism showed nonconformity to simple Michaelis-Menten kinetics against C(D,0), the loading donor concentration. EfR values differed with saturable apical efflux and metabolism (>1), as well as apical absorption (EfRs transport and metabolic data in Caco-2 cells.

  10. Connexin 26-mediated gap junctional intercellular communication suppresses paracellular permeability of human intestinal epithelial cell monolayers

    International Nuclear Information System (INIS)

    Morita, Hidekazu; Katsuno, Tatsuro; Hoshimoto, Aihiro; Hirano, Noriaki; Saito, Yasushi; Suzuki, Yasuo


    In some cell types, gap junctional intercellular communication (GJIC) is associated with tight junctions. The present study was performed to determine the roles of GJIC in regulation of the barrier function of tight junctions. Caco-2 human colonic cells were used as a monolayer model, and barrier function was monitored by measuring mannitol permeability and transepithelial electrical resistance (TER). The monolayers were chemically disrupted by treatment with oleic acid and taurocholic acid. Western blotting analyses were performed to evaluate the protein levels of connexins, which are components of gap junctional intercellular channels. Cx26 expression was detected in preconfluent Caco-2 cells, and its level increased gradually after the monolayer reached confluency. These results prompted us to examine whether overexpression of Cx26 affects barrier function. Monolayers of Caco-2 cells stably expressing Cx26 showed significantly lower mannitol permeability and higher TER than mock transfectants when the monolayers were chemically disrupted. The levels of claudin-4, an important component of tight junctions, were significantly increased in the stable Cx26 transfectant. These results suggest that Cx26-mediated GJIC may play a crucial role in enhancing the barrier function of Caco-2 cell monolayers

  11. Factors Determining the Oxygen Permeability of Biological Membranes: Oxygen Transport Across Eye Lens Fiber-Cell Plasma Membranes. (United States)

    Subczynski, Witold Karol; Widomska, Justyna; Mainali, Laxman


    Electron paramagnetic resonance (EPR) spin-label oximetry allows the oxygen permeability coefficient to be evaluated across homogeneous lipid bilayer membranes and, in some cases, across coexisting membrane domains without their physical separation. The most pronounced effect on oxygen permeability is observed for cholesterol, which additionally induces the formation of membrane domains. In intact biological membranes, integral proteins induce the formation of boundary and trapped lipid domains with a low oxygen permeability. The effective oxygen permeability coefficient across the intact biological membrane is affected not only by the oxygen permeability coefficients evaluated for each lipid domain but also by the surface area occupied by these domains in the membrane. All these factors observed in fiber cell plasma membranes of clear human eye lenses are reviewed here.

  12. Studies on the relationship between epidermal cell turnover kinetics and permeability of hairless mouse skin

    International Nuclear Information System (INIS)

    Han, S.R.


    The primary aim of this study was to develop non-invasive, physical means to quantitatively assess the epidermal turnover kinetics and barrier properties of the skin and relate these to the cutaneous irritation which results from ultraviolet light irradiation and mold thermal burns. After systematically injecting radiolabeled glycine, the appearance of radioactivity at the skin's surface indicated the transit time of radiolabeled cells through the skin. By plotting the data as the cumulative specific activity against time and then fitting them with a third order polynomial equation, it is possible to estimate the turnover time of the stratum corneum. The skin turnover was coordinated with non-invasive transepidermal water loss (TEWL) studies determined with an evaporimeter. In vitro diffusion studies of the permeability of hydrocortisone through UVB irradiated and thermally burned skin were also performed. The studies indicated that irritated skin offers a relatively low diffusional resistance to hydrocortisone. Depending on the severity of the trauma, the increases in hydrocortisone's permeability coefficient through irritated skin ranged from a low of about 2 times normal to a high of about 210 times normal. Trauma-induced changes in hydrocortisone permeability parallel changes in TEWL, proving that the barrier deficient state resulting from rapid epidermal turnover is a general phenomenon

  13. Curcumin modulates endothelial permeability and monocyte transendothelial migration by affecting endothelial cell dynamics. (United States)

    Monfoulet, Laurent-Emmanuel; Mercier, Sylvie; Bayle, Dominique; Tamaian, Radu; Barber-Chamoux, Nicolas; Morand, Christine; Milenkovic, Dragan


    Curcumin is a phenolic compound that exhibits beneficial properties for cardiometabolic health. We previously showed that curcumin reduced the infiltration of immune cells into the vascular wall and prevented atherosclerosis development in mice. This study aimed to investigate the effect of curcumin on monocyte adhesion and transendothelial migration (TEM) and to decipher the underlying mechanisms of these actions. Human umbilical vein endothelial cells (HUVECs) were exposed to curcumin (0.5-1μM) for 3h prior to their activation by Tumor Necrosis Factor alpha (TNF-α). Endothelial permeability, monocyte adhesion and transendothelial migration assays were conducted under static condition and shear stress that mimics blood flow. We further investigated the impact of curcumin on signaling pathways and on the expression of genes using macroarrays. Pre-exposure of endothelial cells to curcumin reduced monocyte adhesion and their transendothelial migration in both static and shear stress conditions. Curcumin also prevented changes in both endothelial permeability and the area of HUVECs when induced by TNF-α. We showed that curcumin modulated the expression of 15 genes involved in the control of cytoskeleton and endothelial junction dynamic. Finally, we showed that curcumin inhibited NF-κB signaling likely through an antagonist interplay with several kinases as suggested by molecular docking analysis. Our findings demonstrate the ability of curcumin to reduce monocyte TEM through a multimodal regulation of the endothelial cell dynamics with a potential benefit on the vascular endothelial function barrier. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. Hypoxia/reoxygenation increases the permeability of endothelial cell monolayers: Role of oxygen radicals

    International Nuclear Information System (INIS)

    Inauen, W.; Payne, D.K.; Kvietys, P.R.; Granger, D.N.


    We assessed the effect of hypoxia/reoxygenation on 14C-albumin flux across endothelial monolayers. Cultured bovine pulmonary artery endothelial cells were grown to confluence on nitrocellulose filters (pore size 12 microns). The endothelialized filters were mounted in Ussing-type chambers which were filled with cell culture medium (M 199). Equimolar amounts (33 nM) of 14C-labeled and unlabeled albumin were added to the hot and cold chambers, respectively. The monolayers were then exposed to successive periods (90 min) of normoxia (pO2 145 mmHg), hypoxia (pO2 20 mmHg), and reoxygenation (pO2 145 mmHg). A gas bubbling system was used to control media pO2 and to ensure adequate mixing. Four aliquots of culture media were taken during each period in order to calculate the 14C-albumin permeability across the endothelialized filter. In some experiments, either the xanthine oxidase inhibitor, oxypurinol (10 microM), or superoxide dismutase (600 U/mL), was added to the media immediately prior to the experiments. As compared to the normoxic control period, albumin permeability was 1.5 times higher during hypoxia (p less than 0.01) and 2.3 times higher during reoxygenation (p less than 0.01). The reoxygenation-induced increase in albumin permeability was prevented by either oxypurinol or superoxide dismutase. These data indicate that xanthine oxidase-derived oxygen radicals contribute to the hypoxia/reoxygenation-induced endothelial cell dysfunction. The altered endothelial barrier function induced by hypoxia/reoxygenation is consistent with the microvascular dysfunction observed following reperfusion of ischemic tissues

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

    International Nuclear Information System (INIS)

    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. (author)

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

    DEFF Research Database (Denmark)

    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...

  17. Oxidative stress, mitochondrial permeability transition, and cell death in Cu-exposed trout hepatocytes

    International Nuclear Information System (INIS)

    Krumschnabel, Gerhard; Manzl, Claudia; Berger, Christian; Hofer, Bettina


    We have previously shown that, in trout hepatocytes, exposure to a high dose of copper (Cu) leads to disruption of Ca 2+ homeostasis and elevated formation of reactive oxygen species (ROS), with the latter ultimately causing cell death. In the present study, we aimed at identifying, using a lower Cu concentration, the role of mitochondria in this scenario, the potential involvement of the mitochondrial permeability transition (MPT), and the mode of cell death induced by the metal. Incubation with 10 μM Cu resulted in a strong stimulation of ROS formation, and after 2 h of exposure a significant increase of both apoptotic and necrotic cells was seen. Co-incubation of Cu-treated hepatocytes with the iron-chelator deferoxamine significantly inhibited ROS production and completely prevented cell death. The origin of the radicals generated was at least partly mitochondrial, as visualized by confocal laser scanning microscopy. Furthermore, ROS production was diminished by inhibition of mitochondrial respiration, but since this also aggravated the elevation of intracellular Ca 2+ induced by Cu, it did not preserve cell viability. In a sub-population of cells, Cu induced a decrease of mitochondrial membrane potential and occurrence of the MPT. Cyclosporin A, which did not inhibit ROS formation, prevented the onset of the MPT and inhibited apoptotic, but not necrotic, cell death. Cu-induced apoptosis therefore appears to be dependent on induction of the MPT, but the prominent contribution of mitochondria to ROS generation also suggests an important role of mitochondria in necrotic cell death

  18. Effects of Streptococcus sanguinis Bacteriocin on Cell Surface Hydrophobicity, Membrane Permeability, and Ultrastructure of Candida Thallus

    Directory of Open Access Journals (Sweden)

    Shengli Ma


    Full Text Available Candida albicans (C.a and Candida tropicalis (C.t were treated with Streptococcus sanguinis bacteriocin (S.s bacteriocin, respectively; the bacteriostatic dynamics of S.s bacteriocin, their effects on cell surface hydrophobicity, leakage of inorganic phosphorus and macromolecular substance, cytosolic calcium concentration, and ultrastructure changes of Candida thallus were detected and analyzed. The results showed that inhibitory effect of S.s bacteriocin on C.a and C.t reached peak level at 24 h, the cell-surface hydrophobicity decreased significantly (P < 0.05 after S.s bacteriocin treatment, and there was leakage of cytoplasmic inorganic phosphorus and macromolecular substance from C.a and C.t; cytosolic calcium concentration decreased greatly. After 24 h treatment by S.s bacteriocin, depressive deformity and defect could be found in the cell surface of C.a and C.t; the thallus displayed irregular forms: C.a was shrunken, there was unclear margins abutting upon cell wall and cell membrane, nucleus disappeared, and cytoplasm was inhomogeneous; likewise, C.t was first plasmolysis, and then the cytoplasm was shrunk, the ultrastructure of cell wall and cell membrane was continuously damaged, and the nucleus was karyolysis. It was illustrated that S.s bacteriocin had similar antifungal effect on C.a and C.t; their cell surface hydrophobicity, membrane permeability, and ultrastructure were changed significantly on exposure to S.s bacteriocin.

  19. Acamprosate has no impact on the permeability of paracellular markers across Caco-2 cells

    DEFF Research Database (Denmark)

    Antonescu, Irina; Steffansen, Bente; Neuhoff, Sibylle

    of the paracellular markers, mannitol and Lucifer Yellow (LY), was investigated. Methods: Ppara of LY and [14C]-mannitol was investigated across filter grown human epithelial colorectal adenocarcinoma (Caco-2) cell monolayers. Changes in the transepithelial electrical resistance (TEER) across the monolayers were...... the [14C]-mannitol, Papp values of 0.71±0.2x10-6 and 0.51±0.17x10-6 cm/s were obtained. TEER values at the end of all experiments were in the range of 426-444 ohm*cm2. Summary/Conclusion: Acamprosate has no impact on the paracellular pathway across Caco-2 cell monolayers of LY and mannitol, or on the TEER......Backgrounds: The oral bioavailability of poorly permeable and non-metabolised acamprosate (BCS III) is 11%. It is controversial whether the intestinal effective permeability of the fully an-ionized acamprosate (pKa 1.83; MW 181.2 g/mol) is predominantly paracellular (Ppara) or transcellular...

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

    Energy Technology Data Exchange (ETDEWEB)

    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}.

  1. A fractal analytical model for the permeabilities of fibrous gas diffusion layer in proton exchange membrane fuel cells

    International Nuclear Information System (INIS)

    Xiao, Boqi; Fan, Jintu; Ding, Feng


    The study of water and gas transport through fibrous gas diffusion layer (GDL) is important to the optimization of proton exchange membrane fuel cells (PEMFCs). In this work, analytical models of dimensionless permeability, and water and gas relative permeabilities of fibrous GDL in PEMFCs are derived using fractal theory. In our models, the structure of fibrous GDL is characterized in terms of porosity, tortuosity fractal dimension (D T ), pore area fractal dimensions (d f ), water phase (d f,w ) and gas phase (d f,g ) fractal dimensions. The predicted dimensionless permeability, water and gas relative permeabilities based on the proposed models are in good agreement with experimental data and predictions of numerical simulations reported in the literature. The model reveals that, although water phase and gas phase fractal dimensions strongly depend on porosity, the water and gas relative permeabilities are independent of porosity and are a function of water saturation only. It is also shown that the dimensionless permeability decreases significantly with the increase of tortuosity fractal dimension. On the other hand, there is only a small decrease in the water and gas relative permeabilities when tortuosity fractal dimension increases. One advantage of the proposed analytical model is that it contains no empirical constant, which is normally required in past models

  2. Effect of heterogeneity on the characterization of cell membrane compartments: I. Uniform size and permeability. (United States)

    Hall, Damien


    Observations of the motion of individual molecules in the membrane of a number of different cell types have led to the suggestion that the outer membrane of many eukaryotic cells may be effectively partitioned into microdomains. A major cause of this suggested partitioning is believed to be due to the direct/indirect association of the cytosolic face of the cell membrane with the cortical cytoskeleton. Such intimate association is thought to introduce effective hydrodynamic barriers into the membrane that are capable of frustrating molecular Brownian motion over distance scales greater than the average size of the compartment. To date, the standard analytical method for deducing compartment characteristics has relied on observing the random walk behavior of a labeled lipid or protein at various temporal frequencies and different total lengths of time. Simple theoretical arguments suggest that the presence of restrictive barriers imparts a characteristic turnover to a plot of mean squared displacement versus sampling period that can be interpreted to yield the average dimensions of the compartment expressed as the respective side lengths of a rectangle. In the following series of articles, we used computer simulation methods to investigate how well the conventional analytical strategy coped with heterogeneity in size, shape, and barrier permeability of the cell membrane compartments. We also explored questions relating to the necessary extent of sampling required (with regard to both the recorded time of a single trajectory and the number of trajectories included in the measurement bin) for faithful representation of the actual distribution of compartment sizes found using the SPT technique. In the current investigation, we turned our attention to the analytical characterization of diffusion through cell membrane compartments having both a uniform size and permeability. For this ideal case, we found that (i) an optimum sampling time interval existed for the analysis

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

    Directory of Open Access Journals (Sweden)

    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.

  4. Isoflavones in food supplements: chemical profile, label accordance and permeability study in Caco-2 cells. (United States)

    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.

  5. Reduction of /sup 51/Cr-permeability of tissue culture cells by infection with herpes simplex virus type 1

    Energy Technology Data Exchange (ETDEWEB)

    Schlehofer, J.R.; Habermehl, K.O.; Diefenthal, W.; Hampl, H.


    Infection of different strains of tissue culture cells with herpes simplex virus type 1(HSV-1) resulted in a reduced /sup 51/Cr-permeability. A stability of the cellular membrane to Triton X-100, toxic sera and HSV-specific complement-mediated immune-cytolysis could be observed simultaneously. The results differed with respect to the cell strain used in the experiments.

  6. Chitosan-modified porous silicon microparticles for enhanced permeability of insulin across intestinal cell monolayers. (United States)

    Shrestha, Neha; Shahbazi, Mohammad-Ali; Araújo, Francisca; Zhang, Hongbo; Mäkilä, Ermei M; Kauppila, Jussi; Sarmento, Bruno; Salonen, Jarno J; Hirvonen, Jouni T; Santos, Hélder A


    Porous silicon (PSi) based particulate systems are emerging as an important drug delivery system due to its advantageous properties such as biocompatibility, biodegradability and ability to tailor the particles' physicochemical properties. Here, annealed thermally hydrocarbonized PSi (AnnTHCPSi) and undecylenic acid modified AnnTHCPSi (AnnUnTHCPSi) microparticles were developed as a PSi-based platform for oral delivery of insulin. Chitosan (CS) was used to modify the AnnUnTHCPSi microparticles to enhance the intestinal permeation of insulin. Surface modification with CS led to significant increase in the interaction of PSi microparticles with Caco-2/HT-29 cell co-culture monolayers. Compared to pure insulin, the CS-conjugated microparticles significantly improved the permeation of insulin across the Caco-2/HT-29 cell monolayers, with ca. 20-fold increase in the amount of insulin permeated and ca. 7-fold increase in the apparent permeability (P(app)) value. Moreover, among all the investigated particles, the CS-conjugated microparticles also showed the highest amount of insulin associated with the mucus layer and the intestinal Caco-2 cells and mucus secreting HT-29 cells. Our results demonstrate that CS-conjugated AnnUnTHCPSi microparticles can efficiently enhance the insulin absorption across intestinal cells, and thus, they are promising microsystems for the oral delivery of proteins and peptides across the intestinal cell membrane. Copyright © 2014 Elsevier Ltd. All rights reserved.

  7. Gliadin, zonulin and gut permeability: Effects on celiac and non-celiac intestinal mucosa and intestinal cell lines. (United States)

    Drago, Sandro; El Asmar, Ramzi; Di Pierro, Mariarosaria; Grazia Clemente, Maria; Tripathi, Amit; Sapone, Anna; Thakar, Manjusha; Iacono, Giuseppe; Carroccio, Antonio; D'Agate, Cinzia; Not, Tarcisio; Zampini, Lucia; Catassi, Carlo; Fasano, Alessio


    Little is known about the interaction of gliadin with intestinal epithelial cells and the mechanism(s) through which gliadin crosses the intestinal epithelial barrier. We investigated whether gliadin has any immediate effect on zonulin release and signaling. Both ex vivo human small intestines and intestinal cell monolayers were exposed to gliadin, and zonulin release and changes in paracellular permeability were monitored in the presence and absence of zonulin antagonism. Zonulin binding, cytoskeletal rearrangement, and zonula occludens-1 (ZO-1) redistribution were evaluated by immunofluorescence microscopy. Tight junction occludin and ZO-1 gene expression was evaluated by real-time polymerase chain reaction (PCR). When exposed to gliadin, zonulin receptor-positive IEC6 and Caco2 cells released zonulin in the cell medium with subsequent zonulin binding to the cell surface, rearrangement of the cell cytoskeleton, loss of occludin-ZO1 protein-protein interaction, and increased monolayer permeability. Pretreatment with the zonulin antagonist FZI/0 blocked these changes without affecting zonulin release. When exposed to luminal gliadin, intestinal biopsies from celiac patients in remission expressed a sustained luminal zonulin release and increase in intestinal permeability that was blocked by FZI/0 pretreatment. Conversely, biopsies from non-celiac patients demonstrated a limited, transient zonulin release which was paralleled by an increase in intestinal permeability that never reached the level of permeability seen in celiac disease (CD) tissues. Chronic gliadin exposure caused down-regulation of both ZO-1 and occludin gene expression. Based on our results, we concluded that gliadin activates zonulin signaling irrespective of the genetic expression of autoimmunity, leading to increased intestinal permeability to macromolecules.

  8. Mitochondrial permeability transition and cell death: the role of cyclophilin D

    Directory of Open Access Journals (Sweden)

    Sabzali eJavadov


    Full Text Available Mitochondria serve as a powerhouse which provides near 90% of ATP necessary for cell life. However, recent studies provide strong evidence that mitochondria also play a central role in cell death. Irreversible mitochondrial permeability transition (mPT at high conductance in response to oxidative or other cellular stresses is accompanied by formation of pathological and non-specific mPT pores (mPTP in the inner membrane of mitochondria. Mitochondrial PTP can serve as a target to prevent cell death under pathological conditions such as cardiac and brain ischemia/reperfusion injury and diabetes. On the other hand, mPTP can be used as an executioner to specifically induce cell death thus blocking tumorigenesis in cancer diseases. Despite many studies, the molecular identity of the mPTP remains unclear. At present, cyclophilin D (CyP-D represents the only mPTP protein which plays an essential role in pore formation. This review will discuss direct and indirect mechanisms underlying CyP-D interaction with a target protein of the mPTP complex. Understanding of the mechanisms of mPTP formation will be helpful to further develop new pharmacological agents targeting mitochondria-mediated cell death.

  9. Enhancing gene delivery of adeno-associated viruses by cell-permeable peptides

    Directory of Open Access Journals (Sweden)

    Yarong Liu


    Full Text Available Adeno-associated virus type 2 (AAV2 is considered a promising gene delivery vector and has been extensively applied in several disease models; however, inefficient transduction in various cells and tissues has limited its widespread application in many areas of gene therapy. In this study, we have developed a general, but efficient, strategy to enhance viral transduction, both in vitro and in vivo, by incubating viral particles with cell-permeable peptides (CPPs. We show that CPPs increase internalization of viral particles into cells by facilitating both energy-independent and energy-dependent endocytosis. Moreover, CPPs can significantly enhance the endosomal escape process of viral particles, thus enhancing viral transduction to those cells that have exhibited very low permissiveness to AAV2 infection as a result of impaired intracellular viral processing. We also demonstrated that this approach could be applicable to other AAV serotypes. Thus, the membrane-penetrating ability of CPPs enables us to generate an efficient method for enhanced gene delivery of AAV vectors, potentially facilitating its applicability to human gene therapy.

  10. Normal versus sickle red blood cells: hemodynamic and permeability characteristics in reperfusion lung injury. (United States)

    Haynes, J; Seibert, A; Shah, A; Taylor, A


    Decreased deformability and increased internal viscosity of the sickle red blood cell (SRBC) contribute to abnormal flow in the microcirculation. Since the lungs are commonly affected in sickle cell disease, we compared the hemodynamics of the normal human red blood cell (NRBC) with the SRBC in the pulmonary circulation. The SRBC has decreased antioxidant enzyme activities compared with the NRBC. Thus, using the capillary filtration coefficient (Kfc), we determined the ability of the NRBC and the SRBC to attenuate the increased permeability and resulting edema seen in the oxidant stress of reperfusion lung injury (RLI). We found that lungs perfused with a 5% SRBC perfusate had higher pulmonary arterial pressures (Ppa) and resistances than lungs perfused with a 5% NRBC perfusate. Lungs made ischemic and reperfused with a physiologic cell-free perfusate resulted in a significant increase (P less than .05) in Kfc compared with the preischemic Kfc (.45 +/- .06 to 1.4 +/- 22 H2O.100 g-1). In lungs reperfused with 5% RBC-containing perfusates, the Kfc did not change from preischemic Kfc with NRBCs and decreased from the preischemic Kfc with SRBCs. These findings suggest that the SRBC causes physiologically significant increases in Ppa and resistances and the SRBC, like the NRBC, offers apparent protection in RLI.

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

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

    DEFF Research Database (Denmark)

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


    The purpose of the present study was to evaluate different toxicity assays for use on proliferating buccal TR146 cells and on stratified TR146 epithelium and to compare these results to the permeability enhancing effect of glycocholate (GC). Both the proliferating cells and the epithelium were...... across the epithelium concurrent with a decrease in the transepithelial electrical resistance (TEER) was also determined. The robustness of the epithelium was significantly higher than that of the proliferating cells (P...

  13. Role of mitochondrial permeability transition in human renal tubular epithelial cell death induced by aristolochic acid

    International Nuclear Information System (INIS)

    Qi Xinming; Cai Yan; Gong Likun; Liu Linlin; Chen Fangping; Xiao Ying; Wu Xiongfei; Li Yan; Xue Xiang; Ren Jin


    Aristolochic acid (AA), a natural nephrotoxin and carcinogen, can induce a progressive tubulointerstitial nephropathy. However, the mechanism by which AA causes renal injury remains largely unknown. Here we reported that the mitochondrial permeability transition (MPT) plays an important role in the renal injury induced by aristolochic acid I (AAI). We found that in the presence of Ca 2+ , AAI caused mitochondrial swelling, leakage of Ca 2+ , membrane depolarization, and release of cytochrome c in isolated kidney mitochondria. These alterations were suppressed by cyclosporin A (CsA), an agent known to inhibit MPT. Culture of HK-2 cell, a human renal tubular epithelial cell line for 24 h with AAI caused a decrease in cellular ATP, mitochondrial membrane depolarization, cytochrome c release, and increase of caspase 3 activity. These toxic effects of AAI were attenuated by CsA and bongkrekic acid (BA), another specific MPT inhibitor. Furthermore, AAI greatly inhibited the activity of mitochondrial adenine nucleotide translocator (ANT) in isolated mitochondria. We suggested that ANT may mediate, at least in part, the AAI-induced MPT. Taken together, these results suggested that MPT plays a critical role in the pathogenesis of HK-2 cell injury induced by AAI and implied that MPT might contribute to human nephrotoxicity of aristolochic acid

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

    Directory of Open Access Journals (Sweden)

    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.

  15. Tritiated 2-deoxy-D-glucose as a probe for cell membrane permeability studies

    International Nuclear Information System (INIS)

    Walum, E.; Peterson, A.


    Tritiated 2-deoxy-D-glucose was taken up and phosphorylated by cultured cells of neuronal (NIE 115), glial (138 MG), muscle (L 6) and liver (BRL 123) origin. Upon perfusion the cells slowly released 2-deoxy-D-glucose 6-phosphate. The following values for rate constants, half-lives, and activation energies for the efflux were obtained: NIE 115: 0.0048 min -1 , 143 min, and 72 kJ mol -1 ; 138 MG: 0.0013 min -1 , 547 min, and 85 kJ mol -1 ; L 6: 0.0022 min -1 , 311 min, and 60 kJ mol -1 ; and BRL 123: 0.0013 min -1 , 528 min and 63 kJ mol -1 . When the cultures were perfused with buffer containing Triton X-100 a time- and concentration-dependent increase in the rate of efflux of 2-deoxy-D-glucose 6-phosphate was obtained. It is suggested that 2-deoxy-D-[ 3 H]glucose can be used as a probe in studies of general cell membrane permeability changes

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

    Directory of Open Access Journals (Sweden)

    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.

  17. The effect of pore-scale geometry and wettability on two-phase relative permeabilities within elementary cells (United States)

    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.


    NARCIS (Netherlands)



    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

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

    Directory of Open Access Journals (Sweden)

    Almudena Pérez-Sánchez

    Full Text Available 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.

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

    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

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  2. Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial (Caco-2) cells

    International Nuclear Information System (INIS)

    Artursson, P.; Karlsson, J.


    Monolayers of a well differentiated human intestinal epithelial cell line, Caco-2, were used as a model to study passive drug absorption across the intestinal epithelium. Absorption rate constants (expressed as apparent permeability coefficients) were determined for 20 drugs and peptides with different structural properties. The permeability coefficients ranged from approximately 5 x 10 - 8 to 5 x 10 - 5 cm/s. A good correlation was obtained between data on oral absorption in humans and the results in the Caco-2 model. Drugs that are completely absorbed in humans had permeability coefficients greater than 1 x 10 - 6 cm/s. Drugs that are absorbed to greater than 1% but less than 100% had permeability coefficients of 0.1-1.0 x 10 - 6 cm/s while drugs and peptides that are absorbed to less than 1% had permeability coefficients of less than or equal to 1 x 10 - 7 cm/s. The results indicate that Caco-2 monolayers can be used as a model for studies on intestinal drug absorption

  3. The effect of xanthine oxidase and hypoxanthine on the permeability of red cells from patients with sickle cell anemia. (United States)

    Al Balushi, Halima W M; Rees, David C; Brewin, John N; Hannemann, Anke; Gibson, John S


    Red cells from patients with sickle cell anemia (SCA) are under greater oxidative challenge than those from normal individuals. We postulated that oxidants generated by xanthine oxidase (XO) and hypoxanthine (HO) contribute to the pathogenesis of SCA through altering solute permeability. Sickling, activities of the main red cell dehydration pathways (P sickle , Gardos channel, and KCl cotransporter [KCC]), and cell volume were measured at 100, 30, and 0 mmHg O 2 , together with deoxygenation-induced nonelectrolyte hemolysis. Unexpectedly, XO/HO mixtures had mainly inhibitory effects on sickling, P sickle , and Gardos channel activities, while KCC activity and nonelectrolyte hemolysis were increased. Gardos channel activity was significantly elevated in red cells pharmacologically loaded with Ca 2+ using the ionophore A23187, consistent with an effect on the transport system per se as well as via Ca 2+ entry likely via the P sickle pathway. KCC activity is controlled by several pairs of conjugate protein kinases and phosphatases. Its activity, however, was also stimulated by XO/HO mixtures in red cells pretreated with N-ethylmaleimide (NEM), which is thought to prevent regulation via changes in protein phosphorylation, suggesting that the oxidants formed could also have direct effects on this transporter. In the presence of XO/HO, red cell volume was better maintained in deoxygenated red cells. Overall, the most notable effect of XO/HO mixtures was an increase in red cell fragility. These findings increase our understanding of the effects of oxidative challenge in SCA patients and are relevant to the behavior of red cells in vivo. © 2018 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.

  4. An HPLC-UV method for the measurement of permeability of marker drugs in the Caco-2 cell assay

    Directory of Open Access Journals (Sweden)

    J.M. Kratz


    Full Text Available The Caco-2 cell line has been used as a model to predict the in vitro permeability of the human intestinal barrier. The predictive potential of the assay relies on an appropriate in-house validation of the method. The objective of the present study was to develop a single HPLC-UV method for the identification and quantitation of marker drugs and to determine the suitability of the Caco-2 cell permeability assay. A simple chromatographic method was developed for the simultaneous determination of both passively (propranolol, carbamazepine, acyclovir, and hydrochlorothiazide and actively transported drugs (vinblastine and verapamil. Separation was achieved on a C18 column with step-gradient elution (acetonitrile and aqueous solution of ammonium acetate, pH 3.0 at a flow rate of 1.0 mL/min and UV detection at 275 nm during the total run time of 35 min. The method was validated and found to be specific, linear, precise, and accurate. This chromatographic system can be readily used on a routine basis and its utilization can be extended to other permeability models. The results obtained in the Caco-2 bi-directional transport experiments confirmed the validity of the assay, given that high and low permeability profiles were identified, and P-glycoprotein functionality was established.

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

    DEFF Research Database (Denmark)

    Brahm, Jesper


    This study extends permeability (P) data on chloride, urea and water in red blood cells (RBC), and concludes that the urea transporter (UT-B) does not transport water. P of chick, duck, Amphiuma means, dog and human RBC to (36)Cl(-), (14)C-urea and (3)H2O was determined under self....... Temperature dependence, as measured by apparent activation energy, EA, of PCl is 117.8 (duck), 74.9 (Amphiuma) and 89.6 kJ mol(-1) (dog). The EA of Purea is 69.6 (duck) and 53.3 kJ mol(-1) (Amphiuma), and that of Pd is 34.9 (duck) and 32.1 kJ mol(-1) (Amphiuma). The present and previous RBC studies indicate......-exchange conditions. At 25°C and pH 7.2-7.5, PCl is 0.94×10(-4)-2.15×10(-4) cm s(-1) for all RBC species at [Cl]=127-150 mmol l(-1). In chick and duck RBC, Purea is 0.84×10(-6) and 1.65×10(-6) cm s(-1), respectively, at [urea]=1-500 mmol l(-1). In Amphiuma, dog and human RBC, Purea is concentration dependent (1...

  6. Cell Type Preference of a Novel Human Derived Cell-Permeable Peptide dNP2 and TAT in Murine Splenic Immune Cells.

    Directory of Open Access Journals (Sweden)

    Sangho Lim

    Full Text Available Cell-permeable peptides (CPPs have been widely studied as an attractive drug delivery system to deliver therapeutic macromolecules such as DNA, RNA, and protein into cells. However, its clinical application is still limited and controversial due to the lack of a complete understanding of delivery efficiency in target cells. Previously we identified and characterized the novel and superior CPP, named dNP2, and here we comparatively analyzed intracellular delivery efficiency of dNP2 and TAT in various immune cells of mouse spleen to demonstrate their cell type preference. dNP2- or TAT-conjugated fluorescent proteins were most efficiently taken up by phagocytic cells such as dendritic cells and macrophages while little protein uptake was seen by lymphocytes including T cells, B cells, and NK cells. Interestingly CD8+ lymphoid dendritic cells and CD62LloCD44hi memory like T cell subsets showed significantly better uptake efficiency in vitro and in vivo relative to other dendritic cells or T cells, respectively. In addition, activated macrophages, T cells, and B cells took up the proteins more efficiently relative to when in the resting state. Importantly, only dNP2, not TAT, shows significant intracellular protein delivery efficiency in vivo. Collectively, this study provides important information regarding heterogeneous intracellular delivery efficiency of CPPs such as dNP2 and TAT with cell type preference in the spleen needed for its application in phagocytic cells or activated immune cells.

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

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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

  10. Crustal permeability (United States)

    Gleeson, Tom; Ingebritsen, Steven E.


    Permeability is the primary control on fluid flow in the Earth’s crust and is key to a surprisingly wide range of geological processes, because it controls the advection of heat and solutes and the generation of anomalous pore pressures.  The practical importance of permeability – and the potential for large, dynamic changes in permeability – is highlighted by ongoing issues associated with hydraulic fracturing for hydrocarbon production (“fracking”), enhanced geothermal systems, and geologic carbon sequestration.  Although there are thousands of research papers on crustal permeability, this is the first book-length treatment.  This book bridges the historical dichotomy between the hydrogeologic perspective of permeability as a static material property and the perspective of other Earth scientists who have long recognized permeability as a dynamic parameter that changes in response to tectonism, fluid production, and geochemical reactions. 

  11. Response of exfoliated human buccal epithelium cells to combined gamma radiation, microwaves, and magnetic field exposure estimated by changes in chromatin condensation and cell membrane permeability

    Directory of Open Access Journals (Sweden)

    K. А. Kuznetsov


    Full Text Available Modulation of the biological effects produced by ionizing radiation (IR using microwave and magnetic fields has important theoretical and practical applications. Response of human buccal epithelium cells to different physical agents (single and combined exposure to 0.5–5 Gy γ-radiation (60Co; microwaves with the frequency of 36.64 GHz and power densities of 0.1 and 1 W/m2, and static magnetic field with the intensity of 25 mT has been investigated. The stress response of the cells was evaluated by counting heterochromatin granules quantity (HGQ in the cell nuclei stained with orcein. Membrane permeability was assessed by the percentage of cells stained with indigocarmine (cells with damaged membrane. The increase of heterochromatin granules quantity (HGQ, i.e. chromatin condensation was detected at the doses of 2 Gy and higher. Changes in the cell membrane permeability to indigocarmine expressed the threshold effect. Membrane permeability reached the threshold at the doses of 2–3 Gy for the cells of different donors and did not change with the increase of the dose of γ-radiation. Cells obtained from different donors revealed some individual peculiarities in their reaction to γ-radiation. The static magnetic field and microwaves applied before or after γ-radiation decreased its impact, as revealed by means of HGQ assessment.

  12. Silver ions increase plasma membrane permeability through modulation of intracellular calcium levels in tobacco BY-2 cells. (United States)

    Klíma, Petr; Laňková, Martina; Vandenbussche, Filip; Van Der Straeten, Dominique; Petrášek, Jan


    Silver ions increase plasma membrane permeability for water and small organic compounds through their stimulatory effect on plasma membrane calcium channels, with subsequent modulation of intracellular calcium levels and ion homeostasis. The action of silver ions at the plant plasma membrane is largely connected with the inhibition of ethylene signalling thanks to the ability of silver ion to replace the copper cofactor in the ethylene receptor. A link coupling the action of silver ions and cellular auxin efflux has been suggested earlier by their possible direct interaction with auxin efflux carriers or by influencing plasma membrane permeability. Using tobacco BY-2 cells, we demonstrate here that besides a dramatic increase of efflux of synthetic auxins 2,4-dichlorophenoxyacetic acid (2,4-D) and 1-naphthalene acetic acid (NAA), treatment with AgNO 3 resulted in enhanced efflux of the cytokinin trans-zeatin (tZ) as well as the auxin structural analogues tryptophan (Trp) and benzoic acid (BA). The application of AgNO 3 was accompanied by gradual water loss and plasmolysis. The observed effects were dependent on the availability of extracellular calcium ions (Ca 2+ ) as shown by comparison of transport assays in Ca 2+ -rich and Ca 2+ -free buffers and upon treatment with inhibitors of plasma membrane Ca 2+ -permeable channels Al 3+ and ruthenium red, both abolishing the effect of AgNO 3 . Confocal microscopy of Ca 2+ -sensitive fluorescence indicator Fluo-4FF, acetoxymethyl (AM) ester suggested that the extracellular Ca 2+ availability is necessary to trigger the response to silver ions and that the intracellular Ca 2+ pool alone is not sufficient for this effect. Altogether, our data suggest that in plant cells the effects of silver ions originate from the primal modification of the internal calcium levels, possibly by their interaction with Ca 2+ -permeable channels at the plasma membrane.

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

    Directory of Open Access Journals (Sweden)

    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.

  14. Notional Permeability

    NARCIS (Netherlands)

    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

  15. Perfluorooctane sulfonate (PFOS) induces reactive oxygen species (ROS) production in human microvascular endothelial cells: role in endothelial permeability. (United States)

    Qian, Yong; Ducatman, Alan; Ward, Rebecca; Leonard, Steve; Bukowski, Valerie; Lan Guo, Nancy; Shi, Xianglin; Vallyathan, Val; Castranova, Vincent


    Perfluorooctane sulfonate (PFOS) is a member of the perfluoroalkyl acids (PFAA) containing an eight-carbon backbone. PFOS is a man-made chemical with carbon-fluorine bonds that are among the strongest in organic chemistry, and PFOS is widely used in industry. Human occupational and environmental exposure to PFOS occurs globally. PFOS is non-biodegradable and is persistent in the human body and environment. In this study, data demonstrated that exposure of human microvascular endothelial cells (HMVEC) to PFOS induced the production of reactive oxygen species (ROS) at both high and low concentrations. Morphologically, it was found that exposure to PFOS induced actin filament remodeling and endothelial permeability changes in HMVEC. Furthermore, data demonstrated that the production of ROS plays a regulatory role in PFOS-induced actin filament remodeling and the increase in endothelial permeability. Our results indicate that the generation of ROS may play a role in PFOS-induced aberrations of the endothelial permeability barrier. The results generated from this study may provide a new insight into the potential adverse effects of PFOS exposure on humans at the cellular level.

  16. Influence of local porosity and local permeability on the performances of a polymer electrolyte membrane fuel cell

    Energy Technology Data Exchange (ETDEWEB)

    Akiki, Tilda [Universite Saint Esprit Kaslik (Lebanon); Universite de Technologie de Belfort-Montbeliard, FCLAB Institute for Research on Fuel Cell Systems, 90010 Belfort (France); Charon, Willy; Iltchev, Marie-Christine; Kouta, Raed [Universite de Technologie de Belfort-Montbeliard, FCLAB Institute for Research on Fuel Cell Systems, 90010 Belfort (France); Accary, Gilbert [Universite Saint Esprit Kaslik (Lebanon)


    In the literature, many models and studies focused on the steady-state aspect of fuel cell systems while their dynamic transient behavior is still a wide area of research. In the present paper, we study the effects of mechanical solicitations on the performance of a proton exchange membrane fuel cell as well as the coupling between the physico-chemical phenomena and the mechanical behavior. We first develop a finite element method to analyze the local porosity distribution and the local permeability distribution inside the gas diffusion layer induced by different pressures applied on deformable graphite or steel bipolar plates. Then, a multi-physical approach is carried out, taking into account the chemical phenomena and the effects of the mechanical compression of the fuel cell, more precisely the deformation of the gas diffusion layer, the changes in the physical properties and the mass transfer in the gas diffusion layer. The effects of this varying porosity and permeability fields on the polarization and on the power density curves are reported, and the local current density is also investigated. Unlike other studies, our model accounts for a porosity field that varies locally in order to correctly simulate the effect of an inhomogeneous compression in the cell. (author)

  17. Reversible alterations in cultured pulmonary artery endothelial cell monolayer morphology and albumin permeability induced by ionizing radiation

    International Nuclear Information System (INIS)

    Friedman, M.; Ryan, U.S.; Davenport, W.C.; Chaney, E.L.; Strickland, D.L.; Kwock, L.


    The effects of ionizing irradiation (0, 600, 1500, or 3000 rads) on the permeability of pulmonary endothelial monolayers to albumin were studied. Pulmonary endothelial cells were grown to confluence on gelatin-coated polycarbonate filters, placed in serum-free medium, and exposed to a 60 Co source. The monolayers were placed in modified flux chambers 24 hours after irradiation; 125 I-albumin was added to the upper well, and both the upper and lower wells were serially sampled over 4 hours. The amount of albumin transferred from the upper well/hour over the period of steady-state clearance (90-240 min after addition of 125 I-albumin) was 2.8 +/- 0.2% in control monolayers and was increased in monolayers exposed to 1500 or 3000 rads (increase of 63 +/- 10% and 61 +/- 10%, respectively, P less than 0.01). No increase was found in monolayers exposed to 600 rads. The increases in endothelial albumin transfer rates were associated with morphologic evidence of monolayer disruption and endothelial injury which paralleled the changes in albumin permeability. Dose-dependent alterations in endothelial actin filament organization were also found. Incubation of the monolayers exposed to 3000 rads with medium supplemented with 10% fetal calf serum for 24 hours resulted in normalization of albumin permeability, improvement in morphologic appearance of the monolayers, and reorganization of the actin filament structure. These studies demonstrate that ionizing radiation is an active principle in the reversible disorganization of cultured pulmonary endothelial cell monolayers without the need of other cell types or serum components

  18. Retention behavior of flavonoids on immobilized artificial membrane chromatography and correlation with cell-based permeability. (United States)

    Tsopelas, Fotios; Tsagkrasouli, Maria; Poursanidis, Pavlos; Pitsaki, Maria; Vasios, George; Danias, Panagiotis; Panderi, Irene; Tsantili-Kakoulidou, Anna; Giaginis, Constantinos


    The aim of the study was to investigate the immobilized artificial membrane (IAM) retention mechanism for a set of flavonoids and to evaluate the potential of IAM chromatography to model Caco-2 permeability. For this purpose, the retention behavior of 41 flavonoid analogs on two IAM stationary phases, IAM.PC.MG and IAM.PC.DD2, was investigated. Correlations between retention factors, logk w(IAM) and octanol-water partitioning (logP) were established and the role of hydroxyl groups of flavonoids to the underlying retention mechanism was explored. IAM retention and logP values were used to establish sound linear models with Caco-2 permeability (logP app ) taken from the literature. Both stepwise regression and multivariate analysis confirmed the contribution of hydrogen bond descriptors, as additional parameters in the either logk w(IAM) or logP models. Retention factors on both IAM stationary phases showed comparable performance with n-octanol-water partitioning towards Caco-2 permeability. Copyright © 2017 John Wiley & Sons, Ltd.

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

    Directory of Open Access Journals (Sweden)

    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.

  20. Subzero water permeability parameters and optimal freezing rates for sperm cells of the southern platyfish, Xiphophorus maculatus. (United States)

    Pinisetty, D; Huang, C; Dong, Q; Tiersch, T R; Devireddy, R V


    This study reports the subzero water transport characteristics (and empirically determined optimal rates for freezing) of sperm cells of live-bearing fishes of the genus Xiphophorus, specifically those of the southern platyfish Xiphophorus maculatus. These fishes are valuable models for biomedical research and are commercially raised as ornamental fish for use in aquariums. Water transport during freezing of X. maculatus sperm cell suspensions was obtained using a shape-independent differential scanning calorimeter technique in the presence of extracellular ice at a cooling rate of 20 degrees C/min in three different media: (1) Hanks' balanced salt solution (HBSS) without cryoprotective agents (CPAs); (2) HBSS with 14% (v/v) glycerol, and (3) HBSS with 10% (v/v) dimethyl sulfoxide (DMSO). The sperm cell was modeled as a cylinder with a length of 52.35 microm and a diameter of 0.66 microm with an osmotically inactive cell volume (Vb) of 0.6 V0, where V0 is the isotonic or initial cell volume. This translates to a surface area, SA to initial water volume, WV ratio of 15.15 microm(-1). By fitting a model of water transport to the experimentally determined volumetric shrinkage data, the best fit membrane permeability parameters (reference membrane permeability to water at 0 degrees C, Lpg or Lpg [cpa] and the activation energy, E(Lp) or E(Lp) [cpa]) were found to range from: Lpg or Lpg [cpa] = 0.0053-0.0093 microm/minatm; E(Lp) or E(Lp) [cpa] = 9.79-29.00 kcal/mol. By incorporating these membrane permeability parameters in a recently developed generic optimal cooling rate equation (optimal cooling rate, [Formula: see text] where the units of B(opt) are degrees C/min, E(Lp) or E(Lp) [cpa] are kcal/mol, L(pg) or L(pg) [cpa] are microm/minatm and SA/WV are microm(-1)), we determined the optimal rates of freezing X. maculatus sperm cells to be 28 degrees C/min (in HBSS), 47 degrees C/min (in HBSS+14% glycerol) and 36 degrees C/min (in HBSS+10% DMSO). Preliminary empirical

  1. c-Kit mutation reduce intestinal epithelial cell proliferation and migration, but not influence intestinal permeability stimulated by lipopolysaccharide. (United States)

    Xue, Hong; Wang, Feng Yun; Kang, Qian; Tang, Xu Dong


    The proto-oncogene c-kit, as a marker of interstitial cells of Cajal (ICCs) in the gastrointestinal tract, plays an important role in the ICCs. Although limited evidences showed c-kit is present in the colonic epithelium but its roles remain unclear. In the present study, we aimed to investigate the expression, location and function of c-kit in the intestinal epithelium. Immunofluorescence, western blotting, and RT-PCR were performed to detect the expression and location of c-kit in the intestinal mucosa of WT mice. We investigated intestinal epithelial proliferation and migration in vivo by performing 5-Bromodeoxyuridine (BrdU) incorporation and Ki-67 staining in WT and Wads m/m mice. An Ussing chamber with fluorescein-isothiocyanate dextran 4000 was used to detect the transepithelial electric resistance (TER), short circuit current (ISC) and permeability across ex vivo colon segments under control and endotoxaemia conditions. We demonstrated that c-kit was located and expressed in the gut crypt compartment in WT mice, which was demonstrated in the c-kit mutant mice (Wads m/m ). In addition, both the number of proliferating cells and the percentage of the distance migrated were lower in the Wads m/m mice than those in the WT mice. Moreover, the intestinal permeability, TER and tight junction were unaltered in the Wads m/m mice under endotoxic conditions compared with those in both the control condition and the WT mice. Altogether, these observations imply that the expression of c-kit in the colonic epithelium is involved in the proliferation and permeability of the colonic epithelium. Copyright © 2018. Published by Elsevier GmbH.

  2. Inhibition of PCAF Histone Acetyltransferase, Cytotoxicity and Cell Permeability of 2-Acylamino-1-(3- or 4-Carboxy-phenylbenzamides

    Directory of Open Access Journals (Sweden)

    Eunsook Ma


    Full Text Available Small molecule HAT inhibitors are useful tools to unravel the role of histone acetyltransferases (HATs in the cell and they also have relevance in oncology. We synthesized a series of 2-acylamino-1-(3- or 4-carboxyphenylbenzamides 8–19 bearing C6, C8, C10, C12, C14, and C16 acyl chains at the 2-amino position of 2-aminobenzoic acid. Enzyme inhibition of these compounds was investigated using in vitro PCAF HAT assays. The inhibitory activities of compounds 8–10, 16, and 19 were similar to that of anacardic acid, and 17 was found to be more active than anacardic acid at 100 μM. Compounds 11–15 showed the low inhibitory activity on PCAF HAT. The cytotoxicity of the synthesized compounds was evaluated by SRB (sulforhodamine B assay against seven human cancer cell lines: HT-29 (colon, HCT-116 (colon, MDA-231 (breast, A549 (lung, Hep3B (hepatoma, HeLa (cervical and Caki (kidney and one normal cell line (HSF. Compound 17 was more active than anacardic acid against human colon cancer (HCT 116, IC50: 29.17 μM, human lung cancer (A549, IC50: 32.09 μM cell lines. 18 was more active than anacardic acid against human colon cancer (HT-29, IC50: 35.49 μM and HCT 116, IC50: 27.56 μM, human lung cancer (A549, IC50: 30.69 μM, and human cervical cancer (HeLa, IC50: 34.41 μM cell lines. The apparent permeability coefficient (Papp, cm/s values of two compounds (16 and 17 were evaluated as 68.21 and 71.48 × 10−6 cm/s by Caco-2 cell permeability assay.

  3. Inhibition of free radical scavenging enzymes affects mitochondrial membrane permeability transition during growth and aging of yeast cells. (United States)

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


    In this study, we investigated the change in the antioxidant enzymes activity, cell respiration, reactive oxygen species (ROS), and impairment of membrane mitochondria permeability in the Endomyces magnusii yeasts during culture growth and aging. We showed that the transition into stationary phase is the key tool to understanding interaction of these processes. This growth stage is distinguished by two-fold increase in ROS production and respiration rate as compared to those in the logarithmic phase. It results in induction of alternative oxidase (AO) in the stationary phase, decline of the main antioxidant enzymes activities, ROS-production, and mitochondria membrane permeability. Significant increase in the share of mitochondrial isoform of superoxide dismutase (SOD2) occurred in the stationary phase from 51.8% (24 h of cultivation) to 68.6% (48 h of cultivation). Upon blocking the essential ROS-scavenging enzymes, SODs and catalases (CATs) some heterogeneity of cell population was observed: 80-90% of cells displayed evident signs of early apoptosis (such as disorientation of mitochondria cristae, mitochondrial fragmentation and deformation of nuclear chromatine). However, 10-20% of the population were definitely healthy. It allowed to draw the conclusion that a complete system of cell antioxidant protection underlies normal mitochondria functioning while the E. magnusii yeasts grow and age. Moreover, this system provides unimpaired cell physiology under oxidative stress during culture aging in the stationary phase. Failures in mitochondria functions due to inhibition of ROS-scavenging enzymes of CATs and SODs could lead to damage of the cells and some signs of early apoptosis.

  4. Glycerol-3-phosphate acyltransferase 2 expression modulates cell roughness and membrane permeability: An atomic force microscopy study.

    Directory of Open Access Journals (Sweden)

    Elizabeth R Cattaneo

    Full Text Available In mammalian cells, de novo glycerolipid synthesis begins with the acylation of glycerol-3-phosphate, catalyzed by glycerol-3-phosphate acyltransferases (GPAT. GPAT2 is a mitochondrial isoform primarily expressed in testis under physiological conditions, and overexpressed in several types of cancers and cancer-derived human cell lines where its expression contributes to the tumor phenotype. Using gene silencing and atomic force microscopy, we studied the correlation between GPAT2 expression and cell surface topography, roughness and membrane permeability in MDA-MB-231 cells. In addition, we analyzed the glycerolipid composition by gas-liquid chromatography. GPAT2 expression altered the arachidonic acid content in glycerolipids, and the lack of GPAT2 seems to be partially compensated by the overexpression of another arachidonic-acid-metabolizing enzyme, AGPAT11. GPAT2 expressing cells exhibited a rougher topography and less membrane damage than GPAT2 silenced cells. Pore-like structures were present only in GPAT2 subexpressing cells, correlating with higher membrane damage evidenced by lactate dehydrogenase release. These GPAT2-induced changes are consistent with its proposed function as a tumor-promoting gene, and might be used as a phenotypic differentiation marker. AFM provides the basis for the identification and quantification of those changes, and demonstrates the utility of this technique in the study of cancer cell biology.

  5. Radioactive rare gas separation using a separation cell with two kinds of membrane differing in gas permeability tendency

    International Nuclear Information System (INIS)

    Ohno, Masayoshi; Ozaki, Osamu; Sato, Hajime; Kimura, Shoji; Miyauchi, Terukatsu.


    A separation cell embodying two kinds of membrane-porous and nonporous, i.e. differing in gas permeability - has a separation factor higher than possible with a conventional separation cell with a single kind of membrane. The performance of such separation cells and of cascades constituted thereof are analyzed theoretically and measured experimentally for different conditions of operation, to determine the applicability of the concept to the separation of rare gases from gaseous waste out of nuclear plants. Theoretical considerations indicate that, in a cascade composed of symmetric separation cells, the separation performance can be improved by recycling part of the effluent from a cell back through the same cell (recycling cascade). It is shown that its performance is better than with the arrangement of diverting another effluent several stages upstream. With the recycling cascade, the symmetric separation recycling rate is determined by the depletion separation and enrichment separation factors relevant to the respective membranes. The separation performance of a 9-stage recycling cascade composed of separation cells with silicone rubber tubular membranes and cellulose acetate tubular membranes is derived for a case of Kr separation from N 2 -Kr mixture. The experimental data coincide well with the analytical results. From both the experimental and the analytical results, it is found that the attainable separation coefficient per stage of the cascade comes to average approximately 0.97. (auth.)

  6. Knockout of the alanine racemase gene in Aeromonas hydrophila HBNUAh01 results in cell wall damage and enhanced membrane permeability. (United States)

    Liu, Dong; Zhang, Lu; Xue, Wen; Wang, Yaping; Ju, Jiansong; Zhao, Baohua


    This study focused on the alanine racemase gene (alr-2), which is involved in the synthesis of d-alanine that forms the backbone of the cell wall. A stable alr-2 knockout mutant of Aeromonas hydrophila HBNUAh01 was constructed. When the mutant was supplemented with d-alanine, growth was unaffected; deprivation of d-alanine caused the growth arrest of the starved mutant cells, but not cell lysis. No alanine racemase activity was detected in the culture of the mutant. Additionally, a membrane permeability assay showed increasing damage to the cell wall during d-alanine starvation. No such damage was observed in the wild type during culture. Scanning and transmission electron microscopy analyses revealed deficiencies of the cell envelope and perforation of the cell wall. Leakage of UV-absorbing substances from the mutants was also observed. Thus, the partial viability of the mutants and their independence of d-alanine for growth indicated that inactivation of alr-2 does not impose an auxotrophic requirement for d-alanine. © FEMS 2015. All rights reserved. For permissions, please e-mail:

  7. Enhanced Intestinal Permeability of Bufalin by a Novel Bufalin-Peptide-Dendrimer Inclusion through Caco-2 Cell Monolayer

    Directory of Open Access Journals (Sweden)

    Chi-on Chan


    Full Text Available Bufalin (BFL has excellent physiological activities such as defending tumors, improving cardiac function, and so on. However, due to its poor water-solubility and bioavailability, the clinical application of BFL remains limited. In order to improve bioavailability of BFL, in our previous research, a novel peptide-dendrimer (PD was synthesized and applied to encapsulate BFL. In the present study, we investigate the absorption property and mechanism of BFL in free form and BFL-peptide-dendrimer inclusion (BPDI delivery system by using the Caco-2 cell monolayer model in vitro. The apparent permeability coefficient (Papp values of BFL in free or BPDI form were over 1.0 × 10−6 cm/s. Meanwhile, their almost equal bi-directional transport and linear transport percentage with time and concentration course indicated that BFL in both forms was absorbed mainly through passive diffusion. The most important result is that the Papp values of BFL increased about three-fold more BPDI than those of its free form, which indicated the intestinal permeability of BFL could be improved while BFL was encapsulated in BPDI form. Therefore, PD encapsulation may be a potential delivery system to increase the bioavailability of BFL.

  8. Isthmin is a novel vascular permeability inducer that functions through cell-surface GRP78-mediated Src activation. (United States)

    Venugopal, Shruthi; Chen, Mo; Liao, Wupeng; Er, Shi Yin; Wong, Wai-Shiu Fred; Ge, Ruowen


    Isthmin (ISM) is a recently identified 60 kDa secreted angiogenesis inhibitor. Two cell-surface receptors for ISM have been defined, the high-affinity glucose-regulated protein 78 kDa (GRP78) and the low-affinity αvβ5 integrin. As αvβ5 integrin plays an important role in pulmonary vascular permeability (VP) and ISM is highly expressed in mouse lung, we sought to clarify the role of ISM in VP. Recombinant ISM (rISM) dose-dependently enhances endothelial monolayer permeability in vitro and local dermal VP when administered intradermally in mice. Systemic rISM administration through intravenous injection leads to profound lung vascular hyperpermeability but not in other organs. Mechanistic investigations using molecular, biochemical approaches and specific chemical inhibitors revealed that ISM-GRP78 interaction triggers a direct interaction between GRP78 and Src, leading to Src activation and subsequent phosphorylation of adherens junction proteins and loss of junctional proteins from inter-endothelial junctions, resulting in enhanced VP. Dynamic studies of Src activation, VP and apoptosis revealed that ISM induces VP directly via Src activation while apoptosis contributes indirectly only after prolonged treatment. Furthermore, ISM is significantly up-regulated in lipopolysaccharide (LPS)-treated mouse lung. Blocking cell-surface GRP78 by systemic infusion of anti-GRP78 antibody significantly attenuates pulmonary vascular hyperpermeability in LPS-induced acute lung injury (ALI) in mice. ISM is a novel VP inducer that functions through cell-surface GRP78-mediated Src activation as well as induction of apoptosis. It induces a direct GRP78-Src interaction, leading to cytoplasmic Src activation. ISM contributes to pulmonary vascular hyperpermeability of LPS-induced ALI in mice. Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2015. For permissions please email:

  9. Site-specific modification of genome with cell-permeable Cre fusion protein in preimplantation mouse embryo

    International Nuclear Information System (INIS)

    Kim, Kyoungmi; Kim, Hwain; Lee, Daekee


    Site-specific recombination (SSR) by Cre recombinase and its target sequence, loxP, is a valuable tool in genetic analysis of gene function. Recently, several studies reported successful application of Cre fusion protein containing protein transduction peptide for inducing gene modification in various mammalian cells including ES cell as well as in the whole animal. In this study, we show that a short incubation of preimplantation mouse embryos with purified cell-permeable Cre fusion protein results in efficient SSR. X-Gal staining of preimplantation embryos, heterozygous for Gtrosa26 tm1Sor , revealed that treatment of 1-cell or 2-cell embryos with 3 μM of Cre fusion protein for 2 h leads to Cre-mediated excision in 70-85% of embryos. We have examined the effect of the concentration of the Cre fusion protein and the duration of the treatment on embryonic development, established a condition for full term development and survival to adulthood, and demonstrated the germ line transmission of excised Gtrosa26 allele. Potential applications and advantages of the highly efficient technique described here are discussed.

  10. Protective effects of l-carnitine and piracetam against mitochondrial permeability transition and PC3 cell necrosis induced by simvastatin. (United States)

    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.

  11. Increase of radiation damage to potassium-ion permeability in E. coli cells with decrease in membrane fluidity

    International Nuclear Information System (INIS)

    Suzuki, S.


    Membrane lipids of an auxotroph of E. coli requiring unsaturated fatty acid were manipulated by supplementing the growth medium with unsaturated fatty acids of different chain lengths and/or configurations, and the radiation damage to K + -permeability of the resulting modified cells was investigated in relation with factors influencing membrane fluidity, such as temperature and procaine. Radiation had greater effects on membranes supplemented with unsaturated fatty acids of the trans configuration with a longer chain than on those of the cis configuration with a shorter chain. Radiation damage also increased with decrease in temperature. Furthermore, procaine-treated membranes showed increased resistance to radiation. All these results indicate that the damage was affected by the physical character of membrane lipids and that it was greater in membranes with decreased fluidity. (author)

  12. Arabidopsis annexin1 mediates the radical-activated plasma membrane Ca²+- and K+-permeable conductance in root cells. (United States)

    Laohavisit, Anuphon; Shang, Zhonglin; Rubio, Lourdes; Cuin, Tracey A; Véry, Anne-Aliénor; Wang, Aihua; Mortimer, Jennifer C; Macpherson, Neil; Coxon, Katy M; Battey, Nicholas H; Brownlee, Colin; Park, Ohkmae K; Sentenac, Hervé; Shabala, Sergey; Webb, Alex A R; Davies, Julia M


    Plant cell growth and stress signaling require Ca²⁺ influx through plasma membrane transport proteins that are regulated by reactive oxygen species. In root cell growth, adaptation to salinity stress, and stomatal closure, such proteins operate downstream of the plasma membrane NADPH oxidases that produce extracellular superoxide anion, a reactive oxygen species that is readily converted to extracellular hydrogen peroxide and hydroxyl radicals, OH•. In root cells, extracellular OH• activates a plasma membrane Ca²⁺-permeable conductance that permits Ca²⁺ influx. In Arabidopsis thaliana, distribution of this conductance resembles that of annexin1 (ANN1). Annexins are membrane binding proteins that can form Ca²⁺-permeable conductances in vitro. Here, the Arabidopsis loss-of-function mutant for annexin1 (Atann1) was found to lack the root hair and epidermal OH•-activated Ca²⁺- and K⁺-permeable conductance. This manifests in both impaired root cell growth and ability to elevate root cell cytosolic free Ca²⁺ in response to OH•. An OH•-activated Ca²⁺ conductance is reconstituted by recombinant ANN1 in planar lipid bilayers. ANN1 therefore presents as a novel Ca²⁺-permeable transporter providing a molecular link between reactive oxygen species and cytosolic Ca²⁺ in plants.

  13. Multiphoton-generated localized electron plasma for membrane permeability modification in single cells (United States)

    Merritt, T.; Leblanc, M.; McMillan, J.; Westwood, J.; Khodaparast, G. A.


    Successful incorporation of a specific macromolecule into a single cell would be ideal for characterizing trafficking dynamics through plasmodesmata or for studying intracellular localizations. Here, we demonstrate NIR femtosecond laser-mediated infiltration of a membrane impermeable dextran-conjugated dye into living cells of Arabidopsis thaliana seedling stems. Based on the reactions of fluorescing vacuoles of transgenic cells and artificial cell walls comprised of nanocellulose, laser intensity and exposure time were adjusted to avoid deleterious effects. Using these plant-tailored laser parameters, cells were injected with the fluorophores and long-term dye retention was observed, all while preserving vital cell functions. This method is ideal for studies concerning cell-to-cell interactions and potentially paves the way for introducing transgenes to specific cells. This work was supported by NSF award IOS-0843372 to JHW, with additional support from and U.S. Department of Agriculture Hatch Project no. 135997, and by the Institute of Critical Technology and Applied Sciences (ICTAS) at Virginia Tech.

  14. The Molecular Basis for Altered Cation Permeability in Hereditary Stomatocytic Human Red Blood Cells

    Directory of Open Access Journals (Sweden)

    Joanna F. Flatt


    Full Text Available Normal human RBCs have a very low basal permeability (leak to cations, which is continuously corrected by the Na,K-ATPase. The leak is temperature-dependent, and this temperature dependence has been evaluated in the presence of inhibitors to exclude the activity of the Na,K-ATPase and NaK2Cl transporter. The severity of the RBC cation leak is altered in various conditions, most notably the hereditary stomatocytosis group of conditions. Pedigrees within this group have been classified into distinct phenotypes according to various factors, including the severity and temperature-dependence of the cation leak. As recent breakthroughs have provided more information regarding the molecular basis of hereditary stomatocytosis, it has become clear that these phenotypes elegantly segregate with distinct genetic backgrounds. The cryohydrocytosis phenotype, including South-east Asian Ovalocytosis, results from mutations in SLC4A1, and the very rare condition, stomatin-deficient cryohydrocytosis, is caused by mutations in SLC2A1. Mutations in RHAG cause the very leaky condition over-hydrated stomatocytosis, and mutations in ABCB6 result in familial pseudohyperkalemia. All of the above are large multi-spanning membrane proteins and the mutations may either modify the structure of these proteins, resulting in formation of a cation pore, or otherwise disrupt the membrane to allow unregulated cation movement across the membrane. More recently mutations have been found in two RBC cation channels, PIEZO1 and KCNN4, which result in dehydrated stomatocytosis. These mutations alter the activation and deactivation kinetics of these channels, leading to increased opening and allowing greater cation fluxes than in wild type.

  15. Effect of laser light on the fragility and permeability of the red blood cell membrane

    International Nuclear Information System (INIS)

    El-Yassin, H. D.


    The resistance of red blood cells to hypotonic hemolysis is often characterized in terms of their osmotic fragility. The percentage of cells that hemolize when plotted as a function of different concentrations of NaCl forms fragility curve, which has a sigmoidal shape. In this study we show that the exposure of red blood cells to laser light converts the sigmoidal shape of the fragility curve to a hyperbolic one, which means that the old population of the red blood cells are the ones more affected by the light which cause their destruction. At the same time it seems that transport across the cell membrane is affected also. The biochemical and physiological implications of this finding are discussed. (author)

  16. Epigalloccatechin-3-gallate Inhibits Ocular Neovascularization and Vascular Permeability in Human Retinal Pigment Epithelial and Human Retinal Microvascular Endothelial Cells via Suppression of MMP-9 and VEGF Activation

    Directory of Open Access Journals (Sweden)

    Hak Sung Lee


    Full Text Available Epigalloccatechin-3-gallate (EGCG is the main polyphenol component of green tea (leaves of Camellia sinensis. EGCG is known for its antioxidant, anti-inflammatory, antiviral, and anti-carcinogenic properties. Here, we identify EGCG as a new inhibitor of ocular angiogenesis and its vascular permeability. Matrix metalloproteinases (MMPs and vascular endothelial growth factor (VEGF play a key role in the processes of extracellular matrix (ECM remodeling and microvascular permeability during angiogenesis. We investigated the inhibitory effects of EGCG on ocular neovascularization and vascular permeability using the retina oriented cells and animal models induced by VEGF and alkaline burn. EGCG treatment significantly decreased mRNA and protein expression levels of MMP-9 in the presence of 12-O-tetradecanoylphorbol-13-acetate (TPA and tumor necrosis factor alpha (TNF-α in human retinal pigment epithelial cells (HRPECs. EGCG also effectively protected ARPE-19 cells from cell death and attenuated mRNA expressions of key angiogenic factors (MMP-9, VEGF, VEGF Receptor-2 by inhibiting generation of reactive oxygen species (ROS. EGCG significantly inhibited proliferation, vascular permeability, and tube formation in VEGF-induced human retinal microvascular endothelial cells (HRMECs. Furthermore, EGCG significantly reduced vascular leakage and permeability by blood-retinal barrier breakdown in VEGF-induced animal models. In addition, EGCG effectively limited upregulation of MMP-9 and platelet endothelial cell adhesion molecule (PECAM/CD31 on corneal neovascularization (CNV induced by alkaline burn. Our data suggest that MMP-9 and VEGF are key therapeutic targets of EGCG for treatment and prevention of ocular angiogenic diseases such as age-related macular degeneration, diabetic retinopathy, and corneal neovascularization.

  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

    DEFF Research Database (Denmark)

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

    DEFF Research Database (Denmark)

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


    of this interaction, and here, this template is exploited for the development of blood plasma-stable and cell-permeable inhibitors. Initially, we explored both the amino acid sequence of the tetrapeptide and the nature of the N-alkyl groups, which consolidated N-cyclohexylethyl-ETAV (1) as the most potent...

  19. Acrolein Disrupts Tight Junction Proteins and Causes Endoplasmic Reticulum Stress-Mediated Epithelial Cell Death Leading to Intestinal Barrier Dysfunction and Permeability. (United States)

    Chen, Wei-Yang; Wang, Min; Zhang, Jingwen; Barve, Shirish S; McClain, Craig J; Joshi-Barve, Swati


    Increasing evidence suggests that environmental and dietary factors can affect intestinal epithelial integrity leading to gut permeability and bacterial translocation. Intestinal barrier dysfunction is a pathogenic process associated with many chronic disorders. Acrolein is an environmental and dietary pollutant and a lipid-derived endogenous metabolite. The impact of acrolein on the intestine has not been investigated before and is evaluated in this study, both in vitro and in vivo. Our data demonstrate that oral acrolein exposure in mice caused damage to the intestinal epithelial barrier, resulting in increased permeability and subsequently translocation of bacterial endotoxin-lipopolysaccharide into the blood. Similar results were seen in vitro using established Caco-2 cell monolayers wherein acrolein decreased barrier function and increased permeability. Acrolein also caused the down-regulation and/or redistribution of three representative tight junction proteins (ie, zonula occludens-1, Occludin, Claudin-1) that critically regulate epithelial paracellular permeability. In addition, acrolein induced endoplasmic reticulum stress-mediated death of epithelial cells, which is an important mechanism contributing to intestinal barrier damage/dysfunction, and gut permeability. Overall, we demonstrate that exposure to acrolein affects the intestinal epithelium by decrease/redistribution of tight junction proteins and endoplasmic reticulum stress-mediated epithelial cell death, thereby resulting in loss of barrier integrity and function. Our findings highlight the adverse consequences of environmental and dietary pollutants on intestinal barrier integrity/function with relevance to gut permeability and the development of disease. Copyright © 2017 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

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


    Tahereh Pourmirjafari Firoozabadi; Zeinab Shankayi; Azam Izadi; Seyed Mohammad Pourmirjafari Firoozabadi


    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...

  1. Development and validation of a HPLC method for the assay of dapivirine in cell-based and tissue permeability experiments. (United States)

    das Neves, José; Sarmento, Bruno; Amiji, Mansoor; Bahia, Maria Fernanda


    Dapivirine, a non-nucleoside reverse transcriptase inhibitor, is being currently used for the development of potential anti-HIV microbicide formulations and delivery systems. A new high-performance liquid chromatography (HPLC) method with UV detection was developed for the assay of this drug in different biological matrices, namely cell lysates, receptor media from permeability experiments and homogenates of mucosal tissues. The method used a reversed-phase C18 column with a mobile phase composed of trifluoroacetic acid solution (0.1%, v/v) and acetonitrile in a gradient mode. Injection volume was 50μL and the flow rate 1mL/min. The total run time was 12min and UV detection was performed at 290nm for dapivirine and the internal standard (IS) diphenylamine. A Box-Behnken experimental design was used to study different experimental variables of the method, namely the ratio of the mobile phase components and the gradient time, and their influence in responses such as the retention factor, tailing factor, and theoretical plates for dapivirine and the IS, as well as the peak resolution between both compounds. The optimized method was further validated and its usefulness assessed for in vitro and ex vivo experiments using dapivirine or dapivirine-loaded nanoparticles. The method showed to be selective, linear, accurate and precise in the range of 0.02-1.5μg/mL. Other chromatographic parameters, namely carry-over, lower limit of quantification (0.02μg/mL), limit of detection (0.006μg/mL), recovery (equal or higher than 90.7%), and sample stability at different storage conditions, were also determined and found adequate for the intended purposes. The method was successfully used for cell uptake assays and permeability studies across cell monolayers and pig genital mucosal tissues. Overall, the proposed method provides a simple, versatile and reliable way for studying the behavior of dapivirine in different biological matrices and assessing its potential as an anti

  2. Identification of two immortalized cell lines, ECV304 and bEnd3, for in vitro permeability studies of blood-brain barrier.

    Directory of Open Access Journals (Sweden)

    Shu Yang

    Full Text Available To identify suitable cell lines for a mimetic system of in vivo blood-brain barrier (BBB for drug permeability assessment, we characterized two immortalized cell lines, ECV304 and bEnd3 in the respect of the tightness, tight junction proteins, P-glycoprotein (P-gp function and discriminative brain penetration. The ECV304 monoculture achieved higher transendothelial electrical resistance (TEER and lower permeability to Lucifer yellow than bEnd3. However, co-culture with rat glioma C6 cells impaired the integrity of ECV304 and bEnd3 cell layers perhaps due to the heterogeneity among C6 cells in inducing BBB characteristics. The immunostaining of ZO-1 delivered distinct bands along cell borders on both cell lines while those of occludin and claudin-5 were diffused and weak. P-gp functionality was only proved in bEnd3 by Rhodamine 123 (R123 uptake assay. A permeability test of reference compounds displayed a similar rank order (digoxin < R123 < quinidine, verapamil < propranolol in ECV304 and bEnd3 cells. In comparison with bEnd3, ECV304 developed tighter barrier for the passage of reference compounds and higher discrimination between transcellular and paracellular transport. However, the monoculture models of ECV304 and bEnd3 fail to achieve the sufficient tightness of in vitro BBB permeability models with high TEER and evident immunostaining of tight junction proteins. Further strategies to enhance the paracellular tightness of both cell lines to mimic in vivo BBB tight barrier deserve to be conducted.

  3. Nifedipine-activated Ca(2+) permeability in newborn rat cortical collecting duct cells in primary culture. (United States)

    Valencia, L; Bidet, M; Martial, S; Sanchez, E; Melendez, E; Tauc, M; Poujeol, C; Martin, D; Namorado, M D; Reyes, J L; Poujeol, P


    To characterize Ca(2+) transport in newborn rat cortical collecting duct (CCD) cells, we used nifedipine, which in adult rat distal tubules inhibits the intracellular Ca(2+) concentration ([Ca(2+)](i)) increase in response to hormonal activation. We found that the dihydropyridine (DHP) nifedipine (20 microM) produced an increase in [Ca(2+)](i) from 87.6 +/- 3.3 nM to 389.9 +/- 29.0 nM in 65% of the cells. Similar effects of other DHP (BAY K 8644, isradipine) were also observed. Conversely, DHPs did not induce any increase in [Ca(2+)](i) in cells obtained from proximal convoluted tubule. In CCD cells, neither verapamil nor diltiazem induced any rise in [Ca(2+)](i). Experiments in the presence of EGTA showed that external Ca(2+) was required for the nifedipine effect, while lanthanum (20 microM), gadolinium (100 microM), and diltiazem (20 microM) inhibited the effect. Experiments done in the presence of valinomycin resulted in the same nifedipine effect, showing that K(+) channels were not involved in the nifedipine-induced [Ca(2+)](i) rise. H(2)O(2) also triggered [Ca(2+)](i) rise. However, nifedipine-induced [Ca(2+)](i) increase was not affected by protamine. In conclusion, the present results indicate that 1) primary cultures of cells from terminal nephron of newborn rats are a useful tool for investigating Ca(2+) transport mechanisms during growth, and 2) newborn rat CCD cells in primary culture exhibit a new apical nifedipine-activated Ca(2+) channel of capacitive type (either transient receptor potential or leak channel).

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

    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.

  5. The Blood-Brain Barrier Permeability of Six Indole Alkaloids from Uncariae Ramulus Cum Uncis in the MDCK-pHaMDR Cell Monolayer Model

    Directory of Open Access Journals (Sweden)

    Yi-Nan Zhang


    Full Text Available Uncariae Ramulus Cum Uncis (URCU is a widely used traditional Chinese medicine, and is reported to have various central nervous system effects. Alkaloids have been demonstrated to be the predominant pharmacological active components of URCU. In order to evaluate the blood-brain barrier (BBB permeability and transport mechanism of six typical indole alkaloids from URCU, the MDCK-pHaMDR cell monolayer model was used as an in vitro surrogate model for BBB. The samples were analyzed by high-performance liquid chromatography, and the apparent permeability coefficients (Papp were calculated. Among the six alkaloids, isorhynchophylline (2, isocorynoxeine (4, hirsutine (5 and hirsuteine (6 showed high permeability, with Papp values at 10−5 cm/s level in bidirectional transport. For rhynchophylline (1 and corynoxeine (3, they showed moderate permeability, with Papp values from the apical (AP side to the basolateral (BL side at 10−6 cm/s level and efflux ratio (Papp BL→AP/Papp AP→BL above 2. The time- and concentration-dependency experiments indicated that the main mechanism for 2, 4, 5 and 6 through BBB was passive diffusion. The efflux mechanism involved in the transports of compounds 1 and 3 could be reduced significantly by verapamil, and molecular docking screening also showed that 1 and 3 had strong bindings to P-glycoprotein. This study provides useful information for predicting the BBB permeability for 1–6, as well as better understanding of their central nervous system pharmacological activities.

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

    DEFF Research Database (Denmark)

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


    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...... with promising mucoadhesive properties for drug delivery to the oral cavity. The following polymers were tested: chitosan, low-methoxylated pectin (LM-pectin), high-methoxylated pectin (HM-pectin), amidated pectin (AM-pectin), Eudragit, poly(N-isopropylacrylamide-co-methacrylic acid) (p...... 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....

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

    NARCIS (Netherlands)

    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.

  8. Water permeability of Na+-K+-2Cl- cotransporters in mammalian epithelial cells

    DEFF Research Database (Denmark)

    Hamann, Steffen; Herrera-Perez, José Jaime; 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...

  9. Permeability, strength and electrochemical studies on ceramic multilayers for solid-state electrochemical cells

    DEFF Research Database (Denmark)

    Andersen, Kjeld Bøhm; Charlas, Benoit; Stamate, Eugen


    An electrochemical reactor can be used to purify flue gasses. Such a reactor can be a multilayer structure consisting of alternating layers of porous electrodes and electrolytes (a porous cell stack). In this work optimization of such a unit has been done by changing the pore former composition...

  10. Excision of HIV-1 proviral DNA by recombinant cell permeable tre-recombinase.

    Directory of Open Access Journals (Sweden)

    Lakshmikanth Mariyanna

    Full Text Available Over the previous years, comprehensive studies on antiretroviral drugs resulted in the successful introduction of highly active antiretroviral therapy (HAART into clinical practice for treatment of HIV/AIDS. However, there is still need for new therapeutic approaches, since HAART cannot eradicate HIV-1 from the infected organism and, unfortunately, can be associated with long-term toxicity and the development of drug resistance. In contrast, novel gene therapy strategies may have the potential to reverse the infection by eradicating HIV-1. For example, expression of long terminal repeat (LTR-specific recombinase (Tre-recombinase has been shown to result in chromosomal excision of proviral DNA and, in consequence, in the eradication of HIV-1 from infected cell cultures. However, the delivery of Tre-recombinase currently depends on the genetic manipulation of target cells, a process that is complicating such therapeutic approaches and, thus, might be undesirable in a clinical setting. In this report we demonstrate that E.coli expressed Tre-recombinases, tagged either with the protein transduction domain (PTD from the HIV-1 Tat trans-activator or the translocation motif (TLM of the Hepatitis B virus PreS2 protein, were able to translocate efficiently into cells and showed significant recombination activity on HIV-1 LTR sequences. Tre activity was observed using episomal and stable integrated reporter constructs in transfected HeLa cells. Furthermore, the TLM-tagged enzyme was able to excise the full-length proviral DNA from chromosomal integration sites of HIV-1-infected HeLa and CEM-SS cells. The presented data confirm Tre-recombinase activity on integrated HIV-1 and provide the basis for the non-genetic transient application of engineered recombinases, which may be a valuable component of future HIV eradication strategies.

  11. Improving enzymatic activities and thermostability of a tri-functional enzyme with SOD, catalase and cell-permeable activities. (United States)

    Luangwattananun, Piriya; Eiamphungporn, Warawan; Songtawee, Napat; Bülow, Leif; Isarankura Na Ayudhya, Chartchalerm; Prachayasittikul, Virapong; Yainoy, Sakda


    Synergistic action of major antioxidant enzymes, e.g., superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) is known to be more effective than the action of any single enzyme. Recently, we have engineered a tri-functional enzyme, 6His-MnSOD-TAT/CAT-MnSOD (M-TAT/CM), with SOD, CAT and cell-permeable activities. The protein actively internalized into the cells and showed superior protection against oxidative stress-induced cell death over native enzymes fused with TAT. To improve its molecular size, enzymatic activity and stability, in this study, MnSOD portions of the engineered protein were replaced by CuZnSOD, which is the smallest and the most heat resistant SOD isoform. The newly engineered protein, CAT-CuZnSOD/6His-CuZnSOD-TAT (CS/S-TAT), had a 42% reduction in molecular size and an increase in SOD and CAT activities by 22% and 99%, respectively. After incubation at 70°C for 10min, the CS/S-TAT retained residual SOD activity up to 54% while SOD activity of the M-TAT/CM was completely abolished. Moreover, the protein exhibited a 5-fold improvement in half-life at 70°C. Thus, this work provides insights into the design and synthesis of a smaller but much more stable multifunctional antioxidant enzyme with ability to enter mammalian cells for further application as protective/therapeutic agent against oxidative stress-related conditions. Copyright © 2017 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    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.

  13. The presence of pMRC01 promotes greater cell permeability and autolysis in lactococcal starter cultures. (United States)

    Fallico, Vincenzo; McAuliffe, Olivia; Fitzgerald, Gerald F; Hill, Colin; Ross, R Paul


    Conjugative transfer of plasmid-associated properties is routinely used to generate food-grade derivatives of lactococcal starter strains with improved technological traits. However, the introduction of one or more plasmids in a single strain is likely to impose a burden on regular cell metabolism and may affect the growth characteristics of the transconjugant culture. The aim of this study was to evaluate the impact of the 60.2-kb plasmid pMRC01 (encoding for an abortive infection bacteriophage resistance system and production of the anti-microbial, lacticin 3147) on starter performance. Five lactococcal strains (L. lactis HP, 255A, SK1, 712 and IL1403) and their pMRC01-containing derivatives were compared in terms of technological properties, including analysis of growth, acidification and autolysis rates. The transconjugants exhibited lower specific growth rates and higher generation times compared to the parental strains when grown at 30 degrees C in glucose-M17, but the presence of pMRC01 did not significantly affect the acidification capacity of strains in 11% reconstituted skimmed milk and synthetic media. Levels of lactate dehydrogenase were two-fold higher in supernatants of transconjugants than in those of parental strains, after 24 and 72 h of growth at 30 degrees C in glucose-M17, suggesting that the presence of pMRC01 somehow accelerates and promotes cellular autolysis. Analysis by flow cytometry following live/dead staining confirmed this result by showing larger populations of injured and dead cells in pMRC01-carrying cultures compared to the parental strains. The results of this study reveal that the plasmid pMRC01 places a burden on lactococcal host metabolism, which is associated with an increased cell permeability and autolysis, without significantly affecting the acidification capacity of the starter. While the magnitude of these effects appears to be strain dependent, the production of the bacteriocin lacticin 3147 may not be involved.

  14. In situ enzymology of DNA replication and ultraviolet-induced DNA repair synthesis in permeable human cells

    International Nuclear Information System (INIS)

    Dresler, S.; Frattini, M.G.; Robinson-Hill, R.M.


    Using permeable diploid human fibroblasts, the authors have studied the deoxyribonucleoside triphosphate concentration dependences of ultraviolet- (UV-) induced DNA repair synthesis and semiconservative DNA replication. In both cell types (AG1518 and IMR-90) examined, the apparent K m values for dCTP, dGTP, and dTTP for DNA replication were between 1.2 and 2.9 μM. For UV-induced DNA repair synthesis, the apparent K m values were substantially lower, ranging from 0.11 to 0.44 μM for AG1518 cells and from 0.06 to 0.24 μM for IMR-90 cells. Recent data implicate DNA polymerase δ in UV-induced repair synthesis and suggest that DNA polymerases α and δ are both involved in semiconservative replication. They measured K m values for dGTP and dTTP for polymerases α and δ, for comparison with the values for replication and repair synthesis. The deoxyribonucleotide K m values for DNA polymerase δ are much greater than the K m values for UV-induced repair synthesis, suggesting that when polymerase δ functions in DNA repair, its characteristics are altered substantially either by association with accessory proteins or by direct posttranslational modification. In contrast, the deoxyribonucleotide binding characteristics of the DNA replication machinery differ little from those of the isolated DNA polymerases. The K m values for UV-induced repair synthesis are 5-80-fold lower than deoxyribonucleotide concentrations that have been reported for intact cultured diploid human fibroblasts. For replication, however, the K m for dGTP is only slightly lower than the average cellular dGTP concentration that has been reported for exponentially growing human fibroblasts. This finding is consistent with the concept that nucleotide compartmentation is required for the attainment of high rates of DNA replication in vivo

  15. Effect of porosity heterogeneity on the permeability and tortuosity of gas diffusion layers in polymer electrolyte membrane fuel cells (United States)

    Nabovati, Aydin; Hinebaugh, James; Bazylak, Aimy; Amon, Cristina H.


    In this paper, we study the effect of porosity heterogeneity on the bulk hydrodynamic properties (permeability and tortuosity) of simulated gas diffusion layers (GDLs). The porosity distributions of the heterogeneous reconstructed samples are similar to those previously reported in the literature for Toray TGP-H 120™ GDLs. We use the lattice Boltzmann method to perform pore-level flow simulations in the reconstructed GDL samples. Using the results of pore-level simulations, the effect of porosity distribution is characterized on the predicted in- and cross-plane permeability and tortuosity. It was found that porosity heterogeneity causes a higher in-plane permeability and lower in-plane tortuosity, while the effect is opposite in the cross-plane direction, that is a lower cross-plane permeability and a higher cross-plane tortuosity. We further investigate the effect of adding poly-tetra-fluoro-ethylene (PTFE) & binder material to the reconstructed GDL samples. Three fiber volume percentages of 50, 75, and 100% are considered. Overall, increasing the fiber volume percentage reduces the predicted in- and cross-plane permeability and tortuosity values. A previously reported relationship for permeability of fibrous materials is fitted to the predicted permeability values, and the magnitude of the fitting parameter is reported as a function of fiber volume percentage.

  16. Knockout of MIMP protein in lactobacillus plantarum lost its regulation of intestinal permeability on NCM460 epithelial cells through the zonulin pathway. (United States)

    Liu, Zhihua; Kang, Liang; Li, Chao; Tong, Chao; Huang, Meijin; Zhang, Xingwei; Huang, Nanqi; Moyer, Mary Pat; Qin, Huanlong; Wang, Jianping


    Previous studies indicated that the micro integral membrane protein located within the media place of the integral membrane protein of Lactobacillus plantarum CGMCC 1258 had protective effects against the intestinal epithelial injury. In our study, we mean to establish micro integral membrane protein -knockout Lactobacillus plantarum (LPKM) to investigate the change of its protective effects and verify the role of micro integral membrane protein on protection of normal intestinal barrier function. Binding assay and intestinal permeability were performed to verify the protective effects of micro integral membrane protein on intestinal permeability in vitro and in vivo. Molecular mechanism was also determined as the zonulin pathway. Clinical data were also collected for further verification of relationship between zonulin level and postoperative septicemia. LPKM got decreased inhibition of EPEC adhesion to NCM460 cells. LPKM had lower ability to alleviate the decrease of intestinal permeability induced by enteropathogenic-e.coli, and prevent enteropathogenic-e.coli -induced increase of zonulin expression. Overexpression of zonulin lowered the intestinal permeability regulated by Lactobacillus plantarum. There was a positive correlation between zonulin level and postoperative septicemia. Therefore, micro integral membrane protein could be necessary for the protective effects of Lactobacillus plantarum on intestinal barrier. MIMP might be a positive factor for Lactobacillus plantarum to protect the intestinal epithelial cells from injury, which could be related to the zonulin pathway.

  17. Sodium butyrate attenuates soybean oil-based lipid emulsion-induced increase in intestinal permeability of lipopolysaccharide by modulation of P-glycoprotein in Caco-2 cells

    International Nuclear Information System (INIS)

    Yan, Jun-Kai; Gong, Zi-Zhen; Zhang, Tian; Cai, Wei


    Down-regulation of intestinal P-glycoprotein (P-gp) by soybean oil-based lipid emulsion (SOLE) may cause elevated intestinal permeability of lipopolysaccharide (LPS) in patients with total parenteral nutrition, but the appropriate preventative treatment is currently limited. Recently, sodium butyrate (NaBut) has been demonstrated to regulate the expression of P-gp. Therefore, this study aimed to address whether treatment with NaBut could attenuate SOLE-induced increase in intestinal permeability of LPS by modulation of P-gp in vitro. Caco-2 cells were exposed to SOLE with or without NaBut. SOLE-induced down-regulation of P-gp was significantly attenuated by co-incubation with NaBut. Nuclear recruitment of FOXO 3a in response to NaBut was involved in P-gp regulation. Transport studies revealed that SOLE-induced increase in permeability of LPS was significantly attenuated by co-incubation with NaBut. Collectively, our results suggested that NaBut may be a potentially useful medication to prevent SOLE-induced increase in intestinal permeability of LPS. - Highlights: • Caco-2 cells were used as models for studying parenteral nutrition in vitro. • NaBut restored SOLE-induced down-regulation of P-gp in Caco-2 cells. • Regulation of P-gp by NaBut was mediated via nuclear recruitment of FOXO 3a. • NaBut modulated the permeability of LPS by P-gp function, not barrier function.

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

    International Nuclear Information System (INIS)

    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

  19. The permeability characteristics and interaction of main components from Si-Ni-San in a MDCK epithelial cell monolayer model. (United States)

    Chen, Ruonan; Shen, Chenlin; Xu, Qingqing; Liu, Yaru; Li, Bo; Huang, Cheng; Ma, Taotao; Meng, Xiaoming; Wu, Maomao; Li, Jun


    1. Si-Ni-San (SNS) possesses extensive therapeutic effects, however, the extent to which main components are absorbed and the mechanisms involved are controversial. 2. In this study, MDCK cell model was used to determine the permeability characteristics and interaction between the major components of Si-Ni-San, including saikosaponin a, paeoniflorin, naringin and glycyrrhizic acid. 3. The transport of the major components was concentration-dependent in both directions. Moreover, the transport of paeoniflorin, naringin and glycyrrhizic acid was significantly reduced at 4°C or in the presence of NaN3. Additionally, the efflux of paeoniflorin and naringin were apparently reduced in the presence of P-gp inhibitor verapamil. The transport of glycyrrhizic acid was clearly inhibited by the inhibitors of MRP2, indicating that MRP2 may be involved in the transport of glycyrrhizic acid. However, the results indicated that saikosaponin a was absorbed mainly by passive diffusion. Furthermore, the combined incubation of four major components had a powerful sorbefacient effect than a single drug used alone which may be regulated by tight junctions. 4. Taken together, our study provides useful information for pharmacological applications of Si-Ni-San and offers new insights into this ancient decoction for further researches, especially in drug synergism.

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

    Directory of Open Access Journals (Sweden)

    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.

  1. Influence of 2,3-diphosphoglycerate metabolism on sodium-potassium permeability in human red blood cells: studies with bisulfite and other redox agents (United States)

    Parker, John C.


    It is known that bisulfite ions can selectively deplete red blood cells of 2,3-diphosphoglycerate (2,3-DPG). Studies of the effects of bisulfite on sodium-potassium permeability and metabolism were undertaken to clarify the physiologic role of the abundant quantities of 2,3-DPG in human erythrocytes. Treatment of cells with bisulfite results in a reversible increase in the passive permeability to Na and K ions. Metabolism of glucose to lactate is increased, with a rise in the intracellular ratio of fructose diphosphate to hexose monophosphate. Cell 2,3-DPG is quantitatively converted to pyruvate and inorganic phosphate. The permeability effects of bisulfite are countered by ethacrynic acid and by such oxidizing agents as pyruvate and methylene blue. Taken together, the results suggest that the effects on Na-K flux of bisulfite are related more to the reducing potential of this anion than to its capacity to deplete cells of 2,3-DPG. PMID:5765015

  2. Permeability testing of biomaterial membranes

    Energy Technology Data Exchange (ETDEWEB)

    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.

  3. Permeability testing of biomaterial membranes

    International Nuclear Information System (INIS)

    Dreesmann, L; Hajosch, R; Nuernberger, J Vaz; Schlosshauer, B; Ahlers, M


    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. Blood-brain barrier permeability and neuroprotective effects of three main alkaloids from the fruits of Euodia rutaecarpa with MDCK-pHaMDR cell monolayer and PC12 cell line. (United States)

    Zhang, Yi-Nan; Yang, Yan-Fang; Yang, Xiu-Wei


    The fruits of Euodia rutaecarpa (Euodiae Fructus, EF), the widely used traditional Chinese medicine, have various central nervous system effects. Alkaloids following as evodiamine (EDM), rutaecarpine (RCP) and dehydroevodiamine (DEDM) are the major substances in EF. The MDCK-pHaMDR cell monolayer model was utilized as a blood-brain barrier (BBB) surrogate model to study their BBB permeability. The transport samples were analyzed by high performance liquid chromatography and the apparent permeability coefficients (P app ) were calculated. EDM and RCP showed high permeability through BBB by passive diffusion, while DEDM showed moderate permeability with efflux mechanism related to P-glycoprotein (P-gp). EDM and RCP could also reduce the efflux of DEDM probably by inhibiting P-gp. The neuroprotective effects of the three alkaloids were then studied on the PC12 cell line injured by 1-methyl-4-phenylpyridinium ion (MPP + ) or hydrogen peroxide (H 2 O 2 ). EDM could significantly reduce MPP + or H 2 O 2 -induced cell injury dose-dependently. RCP could increase the cell viability in MPP + treated group while DEDM showed a protective effect against H 2 O 2 injury. This study predicted the permeability of EDM, RCP and DEDM through BBB and discovered the neuroprotective substance basis of EF as a potential encephalopathy drug. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  5. Syntheses of prodrug-type phosphotriester oligonucleotides responsive to intracellular reducing environment for improvement of cell membrane permeability and nuclease resistance. (United States)

    Hayashi, Junsuke; Samezawa, Yusuke; Ochi, Yosuke; Wada, Shun-Ichi; Urata, Hidehito


    We synthesized prodrug-type phosphotriester (PTE) oligonucleotides containing the six-membered cyclic disulfide moiety by using phosphoramidite chemistry. Prodrug-type oligonucleotides named "Reducing-Environment-Dependent Uncatalyzed Chemical Transforming (REDUCT) PTE oligonucleotides" were converted into natural oligonucleotides under cytosol-mimetic reductive condition. Furthermore, the REDUCT PTE oligonucleotides were robust to nuclease digestion and exhibited good cell membrane permeability. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. The oxygen effect and adaptive response of cells. Report 3. Simulation of respiratory oxygenation and oxygen permeability of cells

    International Nuclear Information System (INIS)

    Ehpshtejn, I.M.


    Variations in the oxygen concentration in extracellural [O 2 ] 0 and intracellular [Osub(2)]sub(i) media of cells small in size (d = 2 ] 0 - t-curves). It is shown that the Value of [Osub(2)]sub(i) may be expressed by four variants of its functional dependence: (a) on enzymic reaction of oxygen consumption, (b) on the order of reaction with respect to oxygen, (c) on physiological parameters of cells, and (d) on characteristic oxygen concentrations in the system. Items (c) and (d) are based on the postulated diffusion-kinetic model of oxygen consumption by an idealized cell of small size that consists of a drop of homogenous solution of the respiratory enzyme which is characterized by an equivalent Michaelis constant. The drop is enveloped in a uniform membrane that possesses a definite diffuse resistance to oxygen

  7. 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

    Directory of Open Access Journals (Sweden)

    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

  8. Proteasome-independent degradation of HIV-1 in naturally non-permissive human placental trophoblast cells

    Directory of Open Access Journals (Sweden)

    Barré-Sinoussi Françoise


    Full Text Available Abstract Background The human placenta-derived cell line BeWo has been demonstrated to be restrictive to cell-free HIV-1 infection. BeWo cells are however permissive to infection by VSV-G pseudotyped HIV-1, which enters cells by a receptor-independent mechanism, and to infection by HIV-1 via a cell-to-cell route. Results Here we analysed viral entry in wild type BeWo (CCR5+, CXCR4+ and BeWo-CD4+ (CD4+, CCR5+, CXCR4+ cells. We report that HIV-1 internalisation is not restricted in either cell line. Levels of internalised p24 antigen between VSV-G HIV-1 pseudotypes and R5 or X4 virions were comparable. We next analysed the fate of internalised virions; X4 and R5 HIV-1 virions were less stable over time in BeWo cells than VSV-G HIV-1 pseudotypes. We then investigated the role of the proteasome in restricting cell-free HIV-1 infection in BeWo cells using proteasome inhibitors. We observed an increase in the levels of VSV-G pseudotyped HIV-1 infection in proteasome-inhibitor treated cells, but the infection by R5-Env or X4-Env pseudotyped virions remains restricted. Conclusion Collectively these results suggest that cell-free HIV-1 infection encounters a surface block leading to a non-productive entry route, which either actively targets incoming virions for non-proteasomal degradation, and impedes their release into the cytoplasm, or causes the inactivation of mechanisms essential for viral replication.

  9. Permeability of cork to gases. (United States)

    Faria, David P; Fonseca, Ana L; Pereira, Helen; Teodoro, Orlando M N D


    The permeability of gases through uncompressed cork was investigated. More than 100 samples were assessed from different plank qualities to provide a picture of the permeability distribution. A novel technique based on a mass spectrometer leak detector was used to directly measure the helium flow through the central area of small disks 10 mm in diameter and 2 mm thick. The permeability for nitrogen, oxygen, and other gases was measured by the pressure rise technique. Boiled and nonboiled cork samples from different sections were evaluated. An asymmetric frequency distribution ranging 3 orders of magnitude (roughly from 1 to 1000 μmol/(cm·atm·day)) for selected samples without macroscopic defects was found, having a peak below 100 μmol/(cm·atm·day). Correlation was found between density and permeability: higher density samples tend to show lower permeability. However, boiled cork showed a mean lower permeability despite having a lower density. The transport mechanism of gases through cork was also examined. Calculations suggest that gases permeate uncompressed cork mainly through small channels between cells under a molecular flow regime. The diameter of such channels was estimated to be in the range of 100 nm, in agreement with the plasmodesmata size in the cork cell walls.

  10. Permeability and Diffusion Coefficients of Single Methyl Lactate Enantiomers in Nafion® and Cellophane Membranes Measured in Diffusion Cell.

    Czech Academy of Sciences Publication Activity Database

    Hovorka, Š.; Randová, A.; Borbášová, T.; Sysel, P.; Vychodilová, Hana; Červenková Šťastná, Lucie; Brožová, Libuše; Žitka, Jan; Storch, Jan; Kačírková, Marie; Drašar, P.; Izák, Pavel


    Roč. 158, JAN 28 (2016), s. 322-332 ISSN 1383-5866 R&D Projects: GA ČR(CZ) GAP106/12/0569 Institutional support: RVO:67985858 ; RVO:61389013 Keywords : diffusion coefficient measurement * permeability * nafion * cellophane * chirality of polymer membrane Subject RIV: CI - Industrial Chemistry, Chemical Engineering Impact factor: 3.359, year: 2016

  11. Mechanism of mucosal permeability enhancement of CriticalSorb® (Solutol® HS15) investigated in vitro in cell cultures. (United States)

    Shubber, Saif; Vllasaliu, Driton; Rauch, Cyril; Jordan, Faron; Illum, Lisbeth; Stolnik, Snjezana


    CriticalSorb™, with the principal component Solutol® HS15, is a novel mucosal drug delivery system demonstrated to improve the bioavailability of selected biotherapeutics. The intention of this study is to elucidate mechanism(s) responsible for the enhancement of trans-mucosal absorption of biological drugs by Solutol® HS15. Micelle size and CMC of Solutol® HS15 were determined in biologically relevant media. Polarised airway Calu-3 cell layers were used to measure the permeability of a panel of biological drugs, and to assess changes in TEER, tight junction and F-actin morphology. The rate of cell endocytosis was measured in vitro in the presence of Solutol® HS15 using a membrane probe, FM 2-10. This work initially confirms surfactant-like behaviour of Solutol® HS15 in aqueous media, while subsequent experiments demonstrate that the effect of Solutol® HS15 on epithelial tight junctions is different from a 'classical' tight junction opening agent and illustrate the effect of Solutol® HS15 on the cell membrane (endocytosis rate) and F-actin cytoskeleton. Solutol® HS15 is the principle component of CriticalSorb™ that has shown an enhancement in permeability of medium sized biological drugs across epithelia. This study suggests that its mechanism of action arises primarily from effects on the cell membrane and consequent impacts on the cell cytoskeleton in terms of actin organisation and tight junction opening.

  12. Comparison of the permeability of metoprolol and labetalol in rat, mouse, and Caco-2 cells: use as a reference standard for BCS classification. (United States)

    Incecayir, Tuba; Tsume, Yasuhiro; Amidon, Gordon L


    The purpose of this study was to investigate labetalol as a potential high permeability reference standard for the application of Biopharmaceutics Classification Systems (BCS). Permeabilities of labetalol and metoprolol were investigated in animal intestinal perfusion models and Caco-2 cell monolayers. After isolating specific intestinal segments, in situ single-pass intestinal perfusions (SPIP) were performed in rats and mice. The effective permeabilities (Peff) of labetalol and metoprolol, an FDA standard for the low/high Peff class boundary, were investigated in two different segments of rat intestine (proximal jejunum and distal ileum) and in the proximal jejunum of mouse. No significant difference was found between Peff of metoprolol and labetalol in the jejunum and ileum of rat (0.33 ± 0.11 × 10(-4) vs 0.38 ± 0.06 × 10(-4) and 0.57 ± 0.17 × 10(-4) vs 0.64 ± 0.30 × 10(-4) cm/s, respectively) and in the jejunum of mouse (0.55 ± 0.05 × 10(-4) vs 0.59 ± 0.13 × 10(-4) cm/s). However, Peff of metoprolol and labetalol were 1.7 and 1.6 times higher in the jejunum of mouse, compared to the jejunum of rat, respectively. Metoprolol and labetalol showed segmental-dependent permeability through the rat intestine, with increased Peff in the distal ileum in comparison to the proximal jejunum. Most significantly, Peff of labetalol was found to be concentration-dependent. Decreasing concentrations of labetalol in the perfusate resulted in decreased Peff compared to Peff of metoprolol. The intestinal epithelial permeability of labetalol was lower than that of metoprolol in Caco-2 cells at both apical pH 6.5 and 7.5 (5.96 ± 1.96 × 10(-6) vs 9.44 ± 3.44 × 10(-6) and 15.9 ± 2.2 × 10(-6) vs 23.2 ± 7.1 × 10(-6) cm/s, respectively). Labetalol exhibited higher permeability in basolateral to apical (BL-AP) compared to AP-BL direction in Caco-2 cells at 0.1 times the highest dose strength (HDS) (46.7 ± 6.5 × 10(-6) vs 14.2 ± 1.5 × 10(-6) cm/s). The P

  13. Regulation of immune responsiveness in vivo by disrupting an early T-cell signaling event using a cell-permeable peptide.

    Directory of Open Access Journals (Sweden)

    David M Guimond

    Full Text Available The inducible T cell kinase (ITK regulates type 2 (Th2 cytokines that provide defense against certain parasitic and bacterial infections and are involved in the pathogenesis of lung inflammation such as allergic asthma. Activation of ITK requires the interaction of its SH3 domain with the poly-proline region of its signaling partner, the SH2 domain containing leukocyte phosphoprotein of 76 kilodaltons (SLP-76. The specific disruption of the ITK-SH3/SLP-76 poly-proline interaction in vitro by a cell-permeable competitive inhibitor peptide (R9-QQP interferes with the activation of ITK and the transduction of its cellular functions in T lymphocytes. In the present investigation, we assessed the effects of R9-QQP treatment on the induction of an in vivo immune response as represented by lung inflammation in a murine model of allergic asthma. We found that mice treated with R9-QQP and sensitized and challenged with the surrogate allergen ovalbumin (OVA display significant inhibition of lung inflammation in a peptide-specific manner. Thus, parameters of the allergic response, such as airway hyper-responsiveness, suppression of inflammatory cell infiltration, reduction of bronchial mucus accumulation, and production of relevant cytokines from draining lymph nodes were significantly suppressed. These findings represent the first demonstration of the biological significance of the interaction between ITK and SLP-76 in the induction of an immune response in a whole animal model and specifically underscore the significance of the ITK-SH3 domain interaction with the poly-proline region of SLP-76 in the development of an inflammatory response. Furthermore, the experimental approach of intracellular peptide-mediated inhibition might be applicable to the study of other important intracellular interactions thus providing a paradigm for dissecting signal transduction pathways.

  14. Benzo(a)pyrene induced cell cycle arrest and apoptosis in human choriocarcinoma cancer cells through reactive oxygen species-induced endoplasmic reticulum-stress pathway. (United States)

    Kim, Soo-Min; Lee, Hae-Miru; Hwang, Kyung-A; Choi, Kyung-Chul


    Cigarette smoke (CS) contains over 60 well established carcinogens. In this study, we examined the effects of benzo(a)pyrene (B(a)P), a main CS component, on the viability and apoptosis of JEG-3 and BeWo human choriocarcinoma cancer cell lines. An MTT assay confirmed that B(a)P decreased the cell viability of JEG-3 and BeWo cells in a dose-dependent manner. Additionally, Western blot (WB) assay revealed that protein expression of cyclin D and cyclin E decreased, while protein expression of p21 and p27 was increased in response to B(a)P treatment for 48 h. The changes in reactive oxygen species (ROS) levels in JEG-3 and BeWo cells exposed to B(a)P were also measured by a dichlorofluorescein diacetate (DCF-DA) assay, which revealed that ROS levels increased in response to B(a)P treatment for 48 h. WB assay also confirmed that each B(a)P treatment of JEG-3 and BeWo cells for 4 h promoted the expression of phosphorylated eukaryotic initiation factor 2 alpha protein (p-eIF2α) and C/EBP homologous protein (CHOP), which are known to be involved in ROS-mediated endoplasmic reticulum stress (ER-stress) related apoptosis. Overall, the protein expression of Bax (a pro-apoptosis marker) increased, while the expression of Bcl-xl (an anti-apoptotic marker) decreased and the number of apoptotic cells increased in response to B(a)P treatment for 48 h. Taken together, these results suggest that B(a)P has the potential to induce apoptosis of JEG-3 and BeWo human choriocarcinoma cancer cells by increasing the ROS level and simultaneously activating ER-stress. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Developments in permeable and low permeability barriers

    International Nuclear Information System (INIS)

    Jefferis, S.A.; Norris, G.H.; Thomas, A.O.


    The concept of the reactive treatment zone whereby pollutants are attenuated as they move along a pathway in the ground has enabled a re-thinking of many of the concepts of containment. In particular it offers the potential for the control of the flux from a contaminated area by controlling the contaminant concentration in the pathway(s) as well as or instead of using a low permeability barrier. The paper outlines the basic concepts of the reactive treatment zone and the use of permeable and low permeability reactive systems. The paper then gives a case history of the installation of a permeable barrier using an in-situ reaction chamber

  16. The impact of cofactors and inhibitors on DNA repair synthesis after γ-irradiation in semi-permeable Escherichia coli cells

    International Nuclear Information System (INIS)

    Gaertner, C.


    The DNA-repair synthesis in tuluol-permeable E. coli cells after γ-irradiation has been investigated in dependence on the co-facotrs. ATB and NAD by means of enzyme kinetics. A partly repair-deficient mutants were taken into consideration which are well characterized in view of molecular biology; they showed which enzyme functions participate in the γ-induced DNA repair synthesis. The inhibition of the DNA-repair synthesis by the intercalary substances Adriamycin and Proflavin has been described and compared with the survival rates after irradiation and after combined treatment by irradiation and intercalary agents. (orig./AJ) [de

  17. Toxicity of diuron in human cancer cells. (United States)

    Huovinen, Marjo; Loikkanen, Jarkko; Naarala, Jonne; Vähäkangas, Kirsi


    Diuron is a substituted phenylurea used as a herbicide to control broadleaf and grass weeds and as a biocidal antifouling agent. Diuron is carcinogenic in rat urinary bladder and toxic to the reproductive system of oysters, sea urchins and lizards. The few studies carried out in human cells do not include the genotoxicity of diuron. We have investigated the toxicity of diuron in human breast adenocarcinoma (MCF-7) and human placental choriocarcinoma (BeWo) cells. The production of reactive oxygen species (ROS) was statistically significantly increased in both cell lines but only at the highest 200 μM concentration. Diuron clearly reduced the viability of BeWo, but not MCF-7 cells. The relative cell number was decreased in both cell lines indicative of inhibition of cell proliferation. In the Comet assay, diuron increased DNA fragmentation in MCF-7 but not in BeWo cells. The expressions of p53 protein, a marker for cell stress, and p21 protein, a transcriptional target of p53, were increased, but only in MCF-7 cells. In conclusion, our results suggest that diuron is cytotoxic and potentially genotoxic in a tissue-specific manner and that ROS play a role in its toxicity. Thus, exposure to diuron may exert harmful effects on fetal development and damage human health. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Effects of 5-hydroxymethyl-2-furfural on the volume and membrane permeability of red blood cells from patients with sickle cell disease (United States)

    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 Ca2+-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 Ca2+ 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. PMID:25015917

  19. Effects of 5-hydroxymethyl-2-furfural on the volume and membrane permeability of red blood cells from patients with sickle cell disease. (United States)

    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. © 2014 The Authors. The Journal of Physiology © 2014 The Physiological Society.

  20. A PKA survival pathway inhibited by DPT-PKI, a new specific cell permeable PKA inhibitor, is induced by T. annulata in parasitized B-lymphocytes. (United States)

    Guergnon, Julien; Dessauge, Frederic; Traincard, François; Cayla, Xavier; Rebollo, Angelita; Bost, Pierre Etienne; Langsley, Gordon; Garcia, Alphonse


    T. annulata, an intracellular pathogenic parasite of the Aplicomplexa protozoan family infects bovine B-lymphocytes and macrophages. Parasitized cells that become transformed survive and proliferate independently of exogenous growth factors. In the present study, we used the isogenic non parasitized BL3 and parasitized TBL3 B cell lines, as a model to evaluate the contribution of two-major PI3-K- and PKA-dependent anti-apoptotic pathways in the survival of T. annulata parasitized B lymphocytes. We found that T. annulata increases PKA activity, induces over-expression of the catalytic subunit and down-regulates the pro-survival phosphorylation state of Akt/PKB. Consistent with a role of PKA activation in survival, two pharmacological inhibitors H89 and KT5720 ablate PKA-dependent survival of parasitized cells. To specifically inhibit PKA pro-survival pathways we linked the DPTsh1 peptide shuttle sequence to PKI(5-24) and we generated DPT-PKI, a cell permeable PKI. DPT-PKI specifically inhibited PKA activity in bovine cell extracts and, as expected, also inhibited the PKA-dependent survival of T. annulata parasitized TBL3 cells. Thus, parasite-dependent constitutive activation of PKA in TBL3 cells generates an anti-apoptotic pathway that can protect T. annulata-infected B cells from apoptosis. These results also indicate that DPT-PKI could be a powerful tool to inhibit PKA pathways in other cell types.

  1. The Phosphatase PTP-PEST/PTPN12 Regulates Endothelial Cell Migration and Adhesion, but Not Permeability, and Controls Vascular Development and Embryonic Viability* (United States)

    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. PMID:23105101

  2. Inhibition of PCAF histone acetyltransferase, cytotoxicity and cell permeability of 2-acylamino-1-(3- or 4-carboxy-phenyl)benzamides. (United States)

    Park, Woong Jae; Ma, Eunsook


    Small molecule HAT inhibitors are useful tools to unravel the role of histone acetyltransferases (HATs) in the cell and they also have relevance in oncology. We synthesized a series of 2-acylamino-1-(3- or 4-carboxyphenyl)benzamides 8–19 bearing C6, C8, C10, C12, C14, and C16 acyl chains at the 2-amino position of 2-aminobenzoic acid. Enzyme inhibition of these compounds was investigated using in vitro PCAF HAT assays. The inhibitory activities of compounds 8–10, 16, and 19 were similar to that of anacardic acid, and 17 was found to be more active than anacardic acid at 100 μM. Compounds 11–15 showed the low inhibitory activity on PCAF HAT. The cytotoxicity of the synthesized compounds was evaluated by SRB (sulforhodamine B) assay against seven human cancer cell lines: HT-29 (colon), HCT-116 (colon), MDA-231 (breast), A549 (lung), Hep3B (hepatoma), HeLa (cervical) and Caki (kidney) and one normal cell line (HSF). Compound 17 was more active than anacardic acid against human colon cancer (HCT 116, IC(50): 29.17 μM), human lung cancer (A549, IC₅₀: 32.09 μM) cell lines. 18 was more active than anacardic acid against human colon cancer (HT-29, IC₅₀: 35.49 μM and HCT 116, IC₅₀: 27.56 μM), human lung cancer (A549, IC₅₀: 30.69 μM), and human cervical cancer (HeLa, IC₅₀: 34.41 μM) cell lines. The apparent permeability coefficient (P(app), cm/s) values of two compounds (16 and 17) were evaluated as 68.21 and 71.48 × 10⁻⁶ cm/s by Caco-2 cell permeability assay.

  3. Trophoblast cell fusion and differentiation are mediated by both the protein kinase C and a pathways.

    Directory of Open Access Journals (Sweden)

    Waka Omata

    Full Text Available The syncytiotrophoblast of the human placenta is an epithelial barrier that interacts with maternal blood and is a key for the transfer of nutrients and other solutes to the developing fetus. The syncytiotrophoblast is a true syncytium and fusion of progenitor cytotrophoblasts is the cardinal event leading to the formation of this layer. BeWo cells are often used as a surrogate for cytotrophoblasts, since they can be induced to fuse, and then express certain differentiation markers associated with trophoblast syncytialization. Dysferlin, a syncytiotrophoblast membrane repair protein, is up-regulated in BeWo cells induced to fuse by treatment with forskolin; this fusion is thought to occur through cAMP/protein kinase A-dependent mechanisms. We hypothesized that dysferlin may also be up-regulated in response to fusion through other pathways. Here, we show that BeWo cells can also be induced to fuse by treatment with an activator of protein kinase C, and that this fusion is accompanied by increased expression of dysferlin. Moreover, a dramatic synergistic increase in dysferlin expression is observed when both the protein kinase A and protein kinase C pathways are activated in BeWo cells. This synergy in fusion is also accompanied by dramatic increases in mRNA for the placental fusion proteins syncytin 1, syncytin 2, as well as dysferlin. Dysferlin, however, was shown to be dispensable for stimulus-induced BeWo cell syncytialization, since dysferlin knockdown lines fused to the same extent as control cells. The classical trophoblast differentiation marker human chorionic gonadotropin was also monitored and changes in the expression closely parallel that of dysferlin in all of the experimental conditions employed. Thus different biochemical markers of trophoblast fusion behave in concert supporting the hypothesis that activation of both protein kinase C and A pathways lead to trophoblastic differentiation.

  4. NMR studies of human blood cells in health and disease. I. Alterations of the plasma membrane water permeability of erythrocytes

    International Nuclear Information System (INIS)

    Katona, Eva; Doaga, I. O.; Radulet, Diana; Caplanusi, A.; Negreanu, Cezarina; Mihele, Denisa


    Alterations in pathological cases of the human erythrocyte membrane water permeability were investigated by using a Mn 2+ -doping 1 H nuclear magnetic resonance (NMR) technique. The temperature dependence of the apparent water diffusional exchange through erythrocyte membranes in chronic hepatitis, diabetes, dyslipidemia and essential hypertension was measured and compared to healthy controls. Using moderate manganese concentrations (9-18 mM) and Carr-Purcell-Meiboom-Gill pulse sequences with a large number of refocusing π pulses and short interpulse delay (100 μs) our values of the water exchange times (τ e ) across erythrocyte membranes, obtained within a 10 min time period following the moment of doping, were independent of the actual manganese concentration and the Arrhenius plot for water exchange was linear over the range of 22-42 deg C. A marked increase of the water exchange times values was observed in all studied disease states. In case of chronic hepatitis, diabetes and dyslipidemia the changes observed in transmembrane water exchange time were associated with significant increase in the apparent activation energy of the diffusional water permeability thus, pointing out alterations in the function of the erythrocyte water channel. (author)

  5. NMR studies of human blood cells in health and disease. I. Alterations of the plasma membrane water permeability of erythrocytes

    Energy Technology Data Exchange (ETDEWEB)

    Katona, Eva; Doaga, I O; Radulet, Diana [Department of Biophysics, Carol Davila University of Medicine and Pharmaceutics, 8 Blvd. Eroilor Sanitari, POB 15-205, RO-76241 Bucharest (Romania); Caplanusi, A [Medical Biochemistry Department, Carol Davila University of Medicine and Pharmaceutics, 8 Blvd. Eroilor Sanitari, POB 15-205, RO-76241 Bucharest (Romania); Negreanu, Cezarina [Division of New Energy Conversion Methods, Institute of Research and Design for Thermoenergetic Equipment, ICPET-CERCETARE, Bucharest (Romania); Mihele, Denisa [Clinical Laboratory Department, Carol Davila University of Medicine and Pharmaceutics, 8 Blvd. Eroilor Sanitari, POB 15-205, RO-76241 Bucharest (Romania)


    Alterations in pathological cases of the human erythrocyte membrane water permeability were investigated by using a Mn{sup 2+}-doping {sup 1}H nuclear magnetic resonance (NMR) technique. The temperature dependence of the apparent water diffusional exchange through erythrocyte membranes in chronic hepatitis, diabetes, dyslipidemia and essential hypertension was measured and compared to healthy controls. Using moderate manganese concentrations (9-18 mM) and Carr-Purcell-Meiboom-Gill pulse sequences with a large number of refocusing {pi} pulses and short interpulse delay (100 {mu}s) our values of the water exchange times ({tau}{sub e}) across erythrocyte membranes, obtained within a 10 min time period following the moment of doping, were independent of the actual manganese concentration and the Arrhenius plot for water exchange was linear over the range of 22-42 deg C. A marked increase of the water exchange times values was observed in all studied disease states. In case of chronic hepatitis, diabetes and dyslipidemia the changes observed in transmembrane water exchange time were associated with significant increase in the apparent activation energy of the diffusional water permeability thus, pointing out alterations in the function of the erythrocyte water channel. (author)

  6. Intact plant MRI for the study of cell water relations, membrane permeability, cell-to-cell and long distance water transport

    NARCIS (Netherlands)

    As, van H.


    Water content and hydraulic conductivity, including transport within cells, over membranes, cell-to-cell, and long-distance xylem and phloem transport, are strongly affected by plant water stress. By being able to measure these transport processes non-invasely in the intact plant situation in

  7. Effect of GAPDH-derived antimicrobial peptides on sensitive yeasts cells: membrane permeability, intracellular pH and H+-influx/-efflux rates. (United States)

    Branco, Patrícia; Albergaria, Helena; Arneborg, Nils; Prista, Catarina


    Saccharomyces cerevisiae secretes antimicrobial peptides (AMPs) derived from glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which induce the death of several non-Saccharomyces yeasts. Previously, we demonstrated that the naturally secreted GAPDH-derived AMPs (i.e. saccharomycin) caused a loss of culturability and decreased the intracellular pH (pHi) of Hanseniaspora guilliermondii cells. In this study, we show that chemically synthesised analogues of saccharomycin also induce a pHi drop and loss of culturability in H. guilliermondii, although to a lesser extent than saccharomycin. To assess the underlying causes of the pHi drop, we evaluated the membrane permeability to H+ cations of H. guilliermondii cells, after being exposed to saccharomycin or its synthetic analogues. Results showed that the H+-efflux decreased by 75.6% and the H+-influx increased by 66.5% in cells exposed to saccharomycin at pH 3.5. Since H+-efflux via H+-ATPase is energy dependent, reduced glucose consumption would decrease ATP production and consequently H+-ATPase activity. However, glucose uptake rates were not affected, suggesting that the AMPs rather than affecting glucose transporters may affect directly the plasma membrane H+-ATPase or increase ATP leakage due to cell membrane disturbance. Thus, our study revealed that both saccharomycin and its synthetic analogues induced cell death of H. guilliermondii by increasing the proton influx and inhibiting the proton efflux.

  8. Surface binding and uptake of cadmium (Cd2+) by LLC-PK1 cells on permeable membrane supports

    International Nuclear Information System (INIS)

    Prozialeck, W.C.; Lamar, P.C.


    Recent studies have shown that Cd 2+ has relatively specific damaging effects on cell-cell junctions in the renal epithelial cell line, LLC-PK 1 . The objective of the present studies was to examine the surface binding and uptake of Cd 2+ by LLC-PK 1 cells in relation to the disruption of cell-cell junctions. LLC-PK 1 cells on Falcon Cell Culture Inserts were exposed to CdCl 2 containing trace amounts of 109 Cd 2+ from either the apical or the basolateral compartments, and the accumulation of 109 Cd 2+ was monitored for up to 8 h. The integrity of cell-cell junctions was assessed by monitoring the transepithelial electrical resistance. The results showed that the cells accumulated 3-4 times more Cd 2+ from the basolateral compartment than from the apical compartment. The accumulation of Cd 2+ from the basolateral compartment occurred in two phases: a rapid, exponential phase that occurred in 1-2 h and coincided with a decrease in transepithelial resistance, and a slower, linear phase that continued for 6-8 h. The Cd 2+ that accumulated during the rapid phase was easily removed by washing the cells in EGTA, indicating that most of it was bound to sites on the cell surface. By contrast, most of the Cd 2+ that accumulated during the slower phase could not be removed by EGTA, indicating that it had been taken up by the cells. Additional studies showed that the rapid phase of Cd 2+ accumulation was enhanced when Ca 2+ was present at low concentrations (0.1 mM), and was greatly reduced when Ca 2+ was present at high concentrations (10 mM). These results suggest that ld 2+ damages the junctions between LLC-PK 1 cells by interacting with Ca 2+ -sensitive sites on the basolateral cell surface. (orig.)

  9. TRPP2 and TRPV4 form an EGF-activated calcium permeable channel at the apical membrane of renal collecting duct cells.

    Directory of Open Access Journals (Sweden)

    Zhi-Ren Zhang

    Full Text Available Regulation of apical calcium entry is important for the function of principal cells of the collecting duct. However, the molecular identity and the regulators of the transporter/channel, which is responsible for apical calcium entry and what factors regulate the calcium conduction remain unclear.We report that endogenous TRPP2 and TRPV4 assemble to form a 23-pS divalent cation-permeable non-selective ion channel at the apical membrane of renal principal cells of the collecting duct. TRPP2\\TRPV4 channel complex was identified by patch-clamp, immunofluorescence and co-immunprecipitation studies in both principal cells that either possess normal cilia (cilia (+ or in which cilia are absent (cilia (-. This channel has distinct biophysical and pharmacological and regulatory profiles compared to either TRPP2 or TRPV4 channels. The rate of occurrence detected by patch clamp was higher in cilia (- compared to cilia (+ cells. In addition, shRNA knockdown of TRPP2 increased the prevalence of TRPV4 channel activity while knockdown of TRPV4 resulted in TRPP2 activity and knockdown of both proteins vastly decreased the 23-pS channel activity. Epidermal growth factor (EGF stimulated TRPP2\\TRPV4 channel through the EGF receptor (EGFR tyrosine kinase-dependent signaling. With loss of cilia, apical EGF treatment resulted in 64-fold increase in channel activity in cilia (- but not cilia (+ cells. In addition EGF increased cell proliferation in cilia (- cell that was dependent upon TRPP2\\TRPV4 channel mediated increase in intracellular calcium.We conclude that in the absence of cilia, an EGF activated TRPP2\\TRPV4 channel may play an important role in increased cell proliferation and cystogenesis.

  10. Accelerated production of antigen-specific T-cells for pre-clinical and clinical applications using Gas-permeable Rapid Expansion cultureware (G-Rex) (United States)

    Vera, Juan F.; Brenner, Lara J.; Gerdemann, Ulrike; Ngo, Minhtran C.; Sili, Uluhan; Liu, Hao; Wilson, John; Dotti, Gianpietro; Heslop, Helen E.; Leen, Ann M.; Rooney, Cliona M.


    The clinical manufacture of antigen-specific cytotoxic T lymphocytes (CTL) for adoptive immunotherapy is limited by the complexity and time required to produce large numbers with the desired function and specificity. The culture conditions required are rigorous, and in some cases only achieved in 2cm2 wells in which cell growth is limited by gas exchange, nutrients and waste accumulation. Bioreactors developed to overcome these issues tend to be complex, expensive and not always conducive to CTL growth. We observed that antigen-specific CTL undergo seven to ten divisions post-stimulation. However the expected CTL numbers were achieved only in the first week of culture. By recreating the culture conditions present during this first week - low frequency of antigen-specific T-cells and high frequency of feeder cells - we were able to increase CTL expansion to expected levels which could be sustained for several weeks without affecting phenotype or function. However, the number of 24-well plates needed was excessive and cultures required frequent media changes, increasing complexity and manufacturing costs. Therefore, we evaluated novel gas-permeable culture devices (G-Rex) with a silicone membrane at the base allowing gas exchange to occur uninhibited by depth of medium above. This system effectively supports the expansion of CTL and actually increases output by up to 20-fold while decreasing required technician time. Importantly, this amplified cell expansion is not due to more cell divisions but to reduced cell death. This bioprocess optimization increased T-cell output while decreasing the complexity and cost of CTL manufacture, making cell therapy more accessible. PMID:20445351

  11. Bog bilberry (Vaccinium uliginosum L.) extract reduces cultured Hep-G2, Caco-2, and 3T3-L1 cell viability, affects cell cycle progression, and has variable effects on membrane permeability. (United States)

    Liu, Jia; Zhang, Wei; Jing, Hao; Popovich, David G


    Bog bilberry (Vaccinium uliginosum L.) is a blue-pigmented edible berry related to bilberry (Vaccinium myrtillus L.) and the common blueberry (Vaccinium corymbosum). The objective of this study was to investigate the effect of a bog bilberry anthocyanin extract (BBAE) on cell growth, membrane permeability, and cell cycle of 2 malignant cancer cell lines, Caco-2 and Hep-G2, and a nonmalignant murine 3T3-L1 cell line. BBAE contained 3 identified anthocyanins. The most abundant anthocyanin was cyanidin-3-glucoside (140.9 +/- 2.6 microg/mg of dry weight), followed by malvidin-3-glucoside (10.3 +/- 0.3 microg/mg) and malvidin-3-galactoside (8.1 +/- 0.4 microg/mg). Hep-G2 LC50 was calculated to be 0.563 +/- 0.04 mg/mL, Caco-2 LC50 was 0.390 +/- 0.30 mg/mL and 0.214 +/- 0.02 mg/mL for 3T3-L1 cells. LDH release, a marker of membrane permeability, was significantly increased in Hep-G2 cells and Caco-2 cells after 48 and 72 h compared to 24 h. The increase was 21% at 48 h and 57% at 72 h in Caco-2 cells and 66% and 139% in Hep-G2 cells compared to 24 h. However, 3T3-L1 cells showed an unexpected significant lower LDH activity (P < or = 0.05) after 72 h of exposure corresponding to a 21% reduction in LDH release. BBAE treatment increased sub-G1 in all 3 cell lines without influencing cells in the G2/M phase. BBAE treatment reduced the growth and increased the accumulation of sub-G1 cells in 2 malignant and 1 nonmalignant cell line; however, the effect on membrane permeability differs considerably between the malignant and nonmalignant cells and may in part be due to differences in cellular membrane composition.

  12. Permeability prediction in chalks

    DEFF Research Database (Denmark)

    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 p...... significantly using the effective specific surface as the fluid-flow concept. The FZI unit is appropriate for highly permeable sedimentary rocks such as sandstones and limestones that have small surface areas....

  13. Critical role of free cytosolic calcium, but not uncoupling, in mitochondrial permeability transition and cell death induced by diclofenac oxidative metabolites in immortalized human hepatocytes

    International Nuclear Information System (INIS)

    Lim, M.S.; Lim, Priscilla L.K.; Gupta, Rashi; Boelsterli, Urs A.


    Diclofenac is a widely used nonsteroidal anti-inflammatory drug that has been associated with rare but serious hepatotoxicity. Experimental evidence indicates that diclofenac targets mitochondria and induces the permeability transition (mPT) which leads to apoptotic cell death in hepatocytes. While the downstream effector mechanisms have been well characterized, the more proximal pathways leading to the mPT are not known. The purpose of this study was to explore the role of free cytosolic calcium (Ca 2+ c ) in diclofenac-induced cell injury in immortalized human hepatocytes. We show that exposure to diclofenac caused time- and concentration-dependent cell injury, which was prevented by the specific mPT inhibitor cyclosporin A (CsA, 5 μM). At 8 h, diclofenac caused increases in [Ca 2+ ] c (Fluo-4 fluorescence), which was unaffected by CsA. Combined exposure to diclofenac/BAPTA (Ca 2+ chelator) inhibited cell injury, indicating that Ca 2+ plays a critical role in precipitating mPT. Diclofenac decreased the mitochondrial membrane potential, ΔΨ m (JC-1 fluorescence), even in the presence of CsA or BAPTA, indicating that mitochondrial depolarization was not a consequence of the mPT or elevated [Ca 2+ ] c . The CYP2C9 inhibitor sulphaphenazole (10 μM) protected from diclofenac-induced cell injury and prevented increases in [Ca 2+ ] c , while it had no effect on the dissipation of the ΔΨ m . Finally, diclofenac exposure greatly increased the mitochondria-selective superoxide levels secondary to the increases in [Ca 2+ ] c . In conclusion, these data demonstrate that diclofenac has direct depolarizing effects on mitochondria which does not lead to cell injury, while CYP2C9-mediated bioactivation causes increases in [Ca 2+ ] c , triggering the mPT and precipitating cell death

  14. Adenovirus-dependent changes in cell membrane permeability: role of Na/sup +/, K/sup +/-ATPase

    Energy Technology Data Exchange (ETDEWEB)

    Seth, P.; Pastan, I.; Willingham, M.C.


    Adenovirus-dependent release of choline phosphate from KB cells at pH 6.0 was partially blocked by ouabain. In K/sup +/-containing medium, maximum inhibition of release was obtained by 10/sup -5/ M ouabain and half-maximal inhibition was achieved by about 0.5 x 10/sup -6/ M ouabain. Ouabain did not block either the binding or the uptake of adenovirus by KB cells. Without K/sup +/, about 25% of cell-associated choline phosphate was released by adenovirus, whereas with 1 mM K/sup +/ about 50% was released. This activation by K/sup +/ was blocked by 0.1 mM ouabain. HeLa cells behaved like KB cells, but a mutant of HeLa cells resistant to ouabain (D98-OR) released much lower amounts of choline phosphate in response to human adenovirus type 2 (Ad2). Wild-type D98-OR cells bound nearly the same amount of adenovirus as did normal HeLa cells. Ad2 also increased the activity of Na/sup +/, K/sup +/-ATPase in KB cells, with maximum activation at of Ad2 per ml. In D98-OR cells, Ad2 failed to activate Na/sup +/, K/sup +/, ATPase activity. Ad2-dependent lysis of endocytic vesicles (receptosomes) was assayed by measuring Ad2-dependent enhancement of epidermal growth factor-Pseudomonas exotoxin toxicity. This action of adenovirus was increased when K/sup +/ was present in the medium. Under the conditions used, K/sup +/ had no effect on the amount of Ad2 or epidermal growth factor taken up by the cells. On the basis of these results, it is suggested that Ad2-dependent cellular efflux of choline phosphate and adenovirus-dependent lysis of receptosomes may require Na/sup +/, K/sup +/-ATPase activity.

  15. Dansyl chloride labeling of Pseudomonas aeruginosa treated with pyocin R1: change in permeability of the cell envelope. (United States)

    Uratani, Y


    Pyocin R1, a bacteriocin of Pseudomonas aeruginosa, caused an increase in binding of fluorescent label, 1-dimethylaminonaphthalene-5-sulfonyl chloride (dansyl chloride), to sensitive cells. In pyocin R1-treated cells, cytoplasmic soluble proteins and crude ribosomes as well as cell envelopes were labeled by dansyl chloride. The amount of bound dye was proportional to the multiplicity of pyocin R1 and reached a maximal level at high multiplicity. In addition, pyocin R1 rapidly caused an increase in fluorescence intensity of the hydrophobic probes N-phenyl-1-naphthylamine, pyrene, and perylene, which were mixed with cells. These results show that pyocin R1 damages locally a cell envelope barrier to hydrophobic solutes and allows dyes to penetrate into the intracellular space across the barrier. PMID:6799489

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

    Directory of Open Access Journals (Sweden)

    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

  17. Silver ions increase plasma membrane permeability through modulation of intracellular calcium levels in tobacco BY-2 cells

    Czech Academy of Sciences Publication Activity Database

    Klíma, Petr; Laňková, Martina; Vandenbussche, F.; Van Der Straeten, D.; Petrášek, Jan


    Roč. 37, č. 5 (2018), s. 809-818 ISSN 0721-7714 R&D Projects: GA ČR GA16-10948S Grant - others:OPPK(XE) CZ.2.16/3.1.00/21519 Institutional support: RVO:61389030 Keywords : Auxin * Calcium * Ethylene * Silver ions * Tobacco BY-2 cells * Transmembrane transport Subject RIV: ED - Physiology OBOR OECD: Cell biology Impact factor: 2.869, year: 2016

  18. Intestinal permeability study of minoxidil: assessment of minoxidil as a high permeability reference drug for biopharmaceutics classification. (United States)

    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

  19. CDPKs CPK6 and CPK3 function in ABA regulation of guard cell S-type anion- and Ca(2+-permeable channels and stomatal closure.

    Directory of Open Access Journals (Sweden)

    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.

  20. Leflunomide or A77 1726 protect from acetaminophen-induced cell injury through inhibition of JNK-mediated mitochondrial permeability transition in immortalized human hepatocytes

    International Nuclear Information System (INIS)

    Latchoumycandane, Calivarathan; Seah, Quee Ming; Tan, Rachel C.H.; Sattabongkot, Jetsumon; Beerheide, Walter; Boelsterli, Urs A.


    Leflunomide, a disease-modifying anti-rheumatic drug, protects against T-cell-mediated liver injury by poorly understood mechanisms. The active metabolite of leflunomide, A77 1726 (teriflunomide) has been shown to inhibit stress-activated protein kinases (JNK pathway), which are key regulators of mitochondria-mediated cell death. Therefore, we hypothesized that leflunomide may protect from drugs that induce the mitochondrial permeability transition (mPT) by blocking the JNK signaling pathway. To this end, we exposed cultured immortalized human hepatocytes (HC-04) to the standard protoxicant drug acetaminophen (APAP), which induces CsA-sensitive mPT-mediated cell death. We determined the effects of leflunomide on the extent of APAP-induced hepatocyte injury and the upstream JNK-mediated mitochondrial signaling pathways. We found that leflunomide or A77 1726 concentration-dependently protected hepatocytes from APAP (1 mM)-induced mitochondrial permeabilization and lethal cell injury. This was not due to proximal inhibition of CYP-catalyzed APAP bioactivation to its thiol-reactive metabolite. Instead, we demonstrate that leflunomide (20 μM) inhibited the APAP-induced early (3 h) activation (phosphorylation) of JNK1/2, thus inhibiting phosphorylation of the anti-apoptotic protein Bcl-2 and preventing P-Bcl-2-mediated induction of the mPT. This greatly attenuated mitochondrial cytochrome c release, which we used as a marker for mitochondrial permeabilization. The specific JNK2 inhibitor SP600125 similarly protected from APAP-induced cell death. In conclusion, these findings are consistent with our hypothesis that leflunomide protects from protoxicant-induced hepatocyte injury by inhibiting JNK signaling and preventing mPT induction

  1. TRPV4 calcium-permeable channel is a novel regulator of oxidized LDL-induced macrophage foam cell formation. (United States)

    Goswami, Rishov; Merth, Michael; Sharma, Shweta; Alharbi, Mazen O; Aranda-Espinoza, Helim; Zhu, Xiaoping; Rahaman, Shaik O


    Cardiovascular disease is the number one cause of death in United States, and atherosclerosis, a chronic inflammatory arterial disease, is the most dominant underlying pathology. Macrophages are thought to orchestrate atherosclerosis by generating lipid-laden foam cells and by secreting inflammatory mediators. Emerging data support a role for a mechanical factor, e.g., matrix stiffness, in regulation of macrophage function, vascular elasticity, and atherogenesis. However, the identity of the plasma membrane mechanosensor and the mechanisms by which pro-atherogenic signals are transduced/maintained are unknown. We have obtained evidence that TRPV4, an ion channel in the transient receptor potential vanilloid family and a known mechanosensor, is the likely mediator of oxidized low-density lipoprotein (oxLDL)-dependent macrophage foam cell formation, a critical process in atherogenesis. Specifically, we found that: i) genetic ablation of TRPV4 or pharmacologic inhibition of TRPV4 activity by a specific antagonist blocked oxLDL-induced macrophage foam cell formation, and ii) TRPV4 deficiency prevented pathophysiological range matrix stiffness or scratch-induced exacerbation of oxLDL-induced foam cell formation. Mechanistically, we found that: i) plasma membrane localization of TRPV4 was sensitized to the increasing level of matrix stiffness, ii) lack of foam cell formation in TRPV4 null cells was not due to lack of expression of CD36, a major receptor for oxLDL, and iii) TRPV4 channel activity regulated oxLDL uptake but not its binding on macrophages. Altogether, these findings identify a novel role for TRPV4 in regulating macrophage foam cell formation by modulating uptake of oxLDL. These findings suggest that therapeutic targeting of TRPV4 may provide a selective approach to the treatment of atherosclerosis. Copyright © 2017. Published by Elsevier Inc.

  2. Permeability and toxicity characteristics of L-cysteine and 2-methyl-thiazolidine-4-carboxylic acid in Caco-2 cells. (United States)

    Kartal-Hodzic, Alma; Marvola, Tuuli; Schmitt, Mechthild; Harju, Kirsi; Peltoniemi, Marikki; Sivén, Mia


    Acetaldehyde is a known mutagenic substance and has been classified as a group-one carcinogen by the WHO. It is possible to bind acetaldehyde locally in the gastrointestinal (GI) tract with the semi-essential amino acid l-cysteine, which reacts covalently with acetaldehyde and forms compound 2-methyl-thiozolidine-4-carboxylic acid (MTCA). The Caco-2 cell line was used to determine the permeation of l-cysteine and MTCA, as well as the possible cell toxicity of both substances. Neither of the substances permeated through the Caco-2 cells at the concentrations used in this study, and only the highest concentration of MTCA affected the viability of the cells in the MTT (3-[4,5-dimethylthiazol-2yl]-2,5-diphenyltetrazolium bromide) test. These results showed that when l-cysteine is administered in formulations releasing it locally in the lower parts of GI tract, it is not absorbed but can react with acetaldehyde, and that neither l-cysteine nor MTCA is harmful to the cells when present locally in the upper parts of GI tract. This study also shows that MTCA is sensitive at a lower pH of 5.5. Since stable MTCA is desired in different parts of the GI tract, this observation raises concern over the influence of lower pH on l-cysteine-containing product ability to bind and eliminate carcinogenic acetaldehyde.

  3. Modulation of Intestinal Epithelial Permeability in Differentiated Caco-2 Cells Exposed to Aflatoxin M1 and Ochratoxin A Individually or Collectively

    Directory of Open Access Journals (Sweden)

    Yanan Gao


    Full Text Available Aflatoxin M1 (AFM1 and ochratoxin A (OTA are mycotoxins commonly found in milk; however, their effects on intestinal epithelial cells have not been reported. In the present study, we show that AFM1 (0.12 and 12 μM and OTA (0.2 and 20 μM individually or collectively increased the paracellular flux of lucifer yellow and fluorescein isothiocyanate (FITC-dextrans (4 and 40 kDa and decreased transepithelial electrical resistance values in differentiated Caco-2 cells after 48 h of exposure, indicating increased epithelial permeability. Immunoblotting and immunofluorescent analysis revealed that AFM1, OTA, and their combination decreased the expression levels of tight junction (TJ proteins and disrupted their structures, namely, claudin-3, claudin-4, occludin, and zonula occludens-1 (ZO-1, and p44/42 mitogen-activated protein kinase (MAPK partially involved in the mycotoxins-induced disruption of intestinal barrier. The effects of a combination of AFM1 and OTA on intestinal barrier function were more significant (p < 0.05 than those of AFM1 and OTA alone, yielding additive or synergistic effects. The additive or synergistic effects of AFM1 and OTA on intestinal barrier function might affect human health, especially in children, and toxin risks should be considered.

  4. Andrographolide-loaded nanoparticles for brain delivery: Formulation, characterisation and in vitro permeability using hCMEC/D3 cell line. (United States)

    Guccione, Clizia; Oufir, Mouhssin; Piazzini, Vieri; Eigenmann, Daniela Elisabeth; Jähne, Evelyn Andrea; Zabela, Volha; Faleschini, Maria Teresa; Bergonzi, Maria Camilla; Smiesko, Martin; Hamburger, Matthias; Bilia, Anna Rita


    Andrographolide (AG) is a major diterpenoid of the Asian medicinal plant Andrographis paniculata which has shown exciting pharmacological potential for the treatment of inflammation-related pathologies including neurodegenerative disorders. Conversely, the low bioavailability of AG still represents a limiting factor for its use. To overcome these limitations, AG was loaded into human serum albumin based nanoparticles (HSA NPs) and poly ethylcyanoacrylate nanoparticles (PECA NPs). HSA NPs were prepared by thermal (HSAT AG NPs) and chemical cross-linking (HSAC AG NPs), while PECA AG NPs were produced by emulsion-polymerization. NPs were characterized in terms of size, zeta (ζ)-potential, polydispersity, and release studies of AG. In addition, the ability of free AG and AG-loaded in PECA and HSAT NPs to cross the blood-brain barrier (BBB) was assessed using an in vitro BBB model based on human cerebral microvascular endothelial cell line (hCMEC/D3). For BBB drug permeability assays, a quantitative UPLC-MS/MS method for AG in Ringer HEPES buffer was developed and validated according to international regulatory guidelines for industry. Free AG did not permeate the BBB model, as also predicted by in silico studies. HSAT NPs improved by two-fold the permeation of AG while maintaining the integrity of the cell layer, while PECA NPs temporarily disrupted BBB integrity. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Inhibition of NF-κB by a cell permeable form of IκBα induces apoptosis in eosinophils

    International Nuclear Information System (INIS)

    Fujihara, Satoko; Jaffray, Ellis; Farrow, Stuart N.; Rossi, Adriano G.; Haslett, Christopher; Hay, Ronald T.


    An 11 amino acid HIV-TAT peptide can deliver target proteins into a variety of cells in a receptor-independent manner. To generate a highly specific inhibitor of the transcription factor NF-κB, we have fused the TAT-peptide to a version of IκBα that is resistant to signal-induced degradation. TAT-IκBα(S32A, S36A) inhibited NF-κB-dependent transcription in HeLa and A549 cells by retaining NF-κB p65 in the cytoplasm. Introduction of TAT-IκBα(S32A, S36A) into human eosinophils inhibited the nuclear translocation of NF-κB and induced apoptosis. Thus, continuous NF-κB-dependent transcription is important for eosinophil survival. While eosinophils are normally refractive to standard methods of gene delivery, the ability of TAT fusion proteins to be taken up by these cells should enable a detailed molecular analysis of survival pathways in these cells

  6. cGMP and nitric oxide modulate thrombin-induced endothelial permeability : Regulation via different pathways in human aortic and umbilical vein endothelial cells

    NARCIS (Netherlands)

    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

  7. Plasma Membrane Ca2+-Permeable Channels are Differentially Regulated by Ethylene and Hydrogen Peroxide to Generate Persistent Plumes of Elevated Cytosolic Ca2+ During Transfer Cell Trans-Differentiation. (United States)

    Zhang, Hui-ming; van Helden, Dirk F; McCurdy, David W; Offler, Christina E; Patrick, John W


    The enhanced transport capability of transfer cells (TCs) arises from their ingrowth wall architecture comprised of a uniform wall on which wall ingrowths are deposited. The wall ingrowth papillae provide scaffolds to amplify plasma membranes that are enriched in nutrient transporters. Using Vicia faba cotyledons, whose adaxial epidermal cells spontaneously and rapidly (hours) undergo a synchronous TC trans-differentiation upon transfer to culture, has led to the discovery of a cascade of inductive signals orchestrating deposition of ingrowth wall papillae. Auxin-induced ethylene biosynthesis initiates the cascade. This in turn drives a burst in extracellular H2O2 production that triggers uniform wall deposition. Thereafter, a persistent and elevated cytosolic Ca(2+) concentration, resulting from Ca(2+) influx through plasma membrane Ca(2+)-permeable channels, generates a Ca(2+) signal that directs formation of wall ingrowth papillae to specific loci. We now report how these Ca(2+)-permeable channels are regulated using the proportionate responses in cytosolic Ca(2+) concentration as a proxy measure of their transport activity. Culturing cotyledons on various combinations of pharmacological agents allowed the regulatory influence of each upstream signal on Ca(2+) channel activity to be evaluated. The findings demonstrated that Ca(2+)-permeable channel activity was insensitive to auxin, but up-regulated by ethylene through two independent routes. In one route ethylene acts directly on Ca(2+)-permeable channel activity at the transcriptional and post-translational levels, through an ethylene receptor-dependent pathway. The other route is mediated by an ethylene-induced production of extracellular H2O2 which then acts translationally and post-translationally to up-regulate Ca(2+)-permeable channel activity. A model describing the differential regulation of Ca(2+)-permeable channel activity is presented. © The Author 2015. Published by Oxford University Press on

  8. Engineering of a novel tri-functional enzyme with MnSOD, catalase and cell-permeable activities. (United States)

    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. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Permeability of porour rhyolite (United States)

    Cashman, K.; Rust, A.; Wright, H.; Roberge, J.


    The development of permeability in bubble-bearing magmas determines the efficiency of volatile escape during their ascent through volcanic conduits, which, in turn, controls their explosive potential. As permeability requires bubble connectivity, relationships between permeability and porosity in silicic magmas must be controlled by the formation, growth, deformation and coalescence of their constituent bubbles. Although permeability data on porous volcanic pyroclasts are limited, the database can be greatly extended by including data for ceramic and metallic foams1. Several studies indicate that a single number does not adequately describe the permeability of a foam because inertial effects, which predominate at high flow rates, cause deviations from Darcy's law. These studies suggest that permeability is best modeled using the Forschheimer equation to determine both the Darcy permeability (k1) and the non-Darcian (k2) permeability. Importantly, at the high porosities of ceramic foams (75-95%), both k1 and k2 are strongly dependent on pore size and geometry, suggesting that measurement of these parameters provides important information on foam structure. We determined both the connected porosity (by He-pycnometry) and the permeability (k1 and k2) of rhyolitic samples having a wide range in porosity (22-85%) and vesicle textures. In general, these data support previous observations of a power law relationship between connected porosity and Darcy permeability2. In detail, variations in k1 increase at higher porosities. Similarly, k2 generally increases in both mean and standard deviation with increasing porosity. Measurements made on three mutually perpendicular cores from individual pumice clasts suggest that some of the variability can be explained by anisotropy in the vesicle structure. By comparison with ceramic foams, we suggest that the remaining variability results from differences either in average vesicle size or, more likely, in the size of apertures

  10. Progranulin shows cytoprotective effects on trophoblast cells in vitro but does not antagonize TNF-α-induced apoptosis. (United States)

    Stubert, Johannes; Waldmann, Kathrin; Dieterich, Max; Richter, Dagmar-Ulrike; Briese, Volker


    The glycoprotein progranulin directly binds to TNF-receptors and thereby can antagonize the inflammatory effects of TNF-α. Here we analyzed the impact of both cytokines on cytotoxicity and viability of trophoblast cells. Isolated villous first trimester human trophoblast cells and the human choriocarcinoma cell line BeWo were treated with recombinant human progranulin and TNF-α. Analyses were performed by LDH- and MTT-assay and measurement of caspase-8-activity. Progranulin treatment showed some cytoprotective effects on isolated trophoblast cells. However, TNF-α-induced apoptosis was not antagonized by addition of progranulin. Effects were similar, but more pronounced in BeWo cells. The cytoprotective activity of progranulin on trophoblast cells in vitro was only weak and of doubtful biologic relevance. It was not able to antagonize TNF-α. Future studies should focus on possible paracrine activities of progranulin.

  11. Astragaloside IV Inhibits Oxidative Stress-Induced Mitochondrial Permeability Transition Pore Opening by Inactivating GSK-3β via Nitric Oxide in H9c2 Cardiac Cells

    Directory of Open Access Journals (Sweden)

    Yonggui He


    Full Text Available Objective. This study aimed to investigate whether astragaloside IV modulates the mitochondrial permeability transition pore (mPTP opening through glycogen synthase kinase 3β (GSK-3β in H9c2 cells. Methods. H9c2 cells were exposed to astragaloside IV for 20 min. GSK-3β (Ser9, Akt (Ser473, and VASP (Ser239 activities were determined with western blot. The mPTP opening was evaluated by measuring mitochondrial membrane potential (ΔΨm. Nitric oxide (NO generation was measured by 4-amino-5-methylamino-2′, 7′-difluorofluorescein (DAF-FM diacetate. Fluorescence images were obtained with confocal microscopy. Results. Astragaloside IV significantly enhanced GSK-3β phosphorylation and prevented H2O2-induced loss of ΔΨm. These effects of astragaloside IV were reversed by the phosphatidylinositol 3-kinase (PI3K inhibitor LY294002, the NO sensitive guanylyl cyclase selective inhibitor ODQ, and the PKG inhibitor KT5823. Astragaloside IV activated Akt and PKG. Astragaloside IV was also shown to increase NO production, an effect that was reversed by L-NAME and LY294002. Astragaloside IV applied at reperfusion reduced cell death caused by simulated ischemia/reperfusion, indicating that astragaloside IV can prevent reperfusion injury. Conclusions. These data suggest that astragaloside IV prevents the mPTP opening and reperfusion injury by inactivating GSK-3β through the NO/cGMP/PKG signaling pathway. NOS is responsible for NO generation and is activated by the PI3K/Akt pathway.

  12. Soils - Mean Permeability (United States)

    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...

  13. Hydrogen permeability through metals

    International Nuclear Information System (INIS)

    Pisarev, A.A.; Tsvetkov, I.V.; Marenkov, E.D.; Yarko, S.S.


    The mechanisms of hydrogen permeability through one-layer and multi-layer membranes are considered. The effect of surface roughness, crystal defects, cracks and pores is described. Mathematical description of the processes is given [ru

  14. Permeable pavement study (Edison) (United States)

    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...

  15. A synergistic effect of artocarpanone from Artocarpus heterophyllus L. (Moraceae) on the antibacterial activity of selected antibiotics and cell membrane permeability. (United States)

    Septama, Abdi Wira; Xiao, Jianbo; Panichayupakaranant, Pharkphoom


    Artocarpanone isolated from Artocarpus heterophyllus L. (Moraceae) exhibits antibacterial activity. The present study investigated synergistic activity between artocarpanone and tetracycline, ampicillin, and norfloxacin, respectively, against methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa , and Escherichia coli . A broth microdilution method was used for evaluating antibacterial susceptibility. Synergistic effects were identified using a checkerboard method, and a bacterial cell membrane disruption was investigated by assay of released 260 nm absorbing materials following bacteriolysis. Artocarpanone exhibited weak antibacterial activity against MRSA and P. aeruginosa with minimum inhibitory concentrations values of 125 and 500 μg/mL, respectively. However, the compound showed strong antibacterial activity against E. coli (7.8 μg/mL). The interaction between artocarpanone and all tested antibiotics revealed indifference and additive effects against P. aeruginosa and E. coli (fractional inhibitory concentration index [FICI] values of 0.75-1.25). The combination of artocarpanone (31.2 μg/mL) and norfloxacin (3.9 μg/mL) resulted in synergistic antibacterial activity against MRSA, with an FICI of 0.28, while the interaction between artocarpanone and tetracycline, and ampicillin showed an additive effect, with an FICI value of 0.5. A time-kill assay also indicated that artocarpanone had a synergistic effect on the antibacterial activity of norfloxacin. In addition, the combination of artocarpanone and norfloxacin altered the membrane permeability of MRSA. These findings suggest that artocarpanone may be used to enhance the antibacterial activity of norfloxacin against MRSA.

  16. Intestinal Permeability: The Basics

    Directory of Open Access Journals (Sweden)

    Ingvar Bjarnason


    Full Text Available The authors review some of the more fundamental principles underlying the noninvasive assessment of intestinal permeability in humans, the choice of test markers and their analyses, and the practical aspects of test dose composition and how these can be changed to allow the specific assessment of regional permeability changes and other intestinal functions. The implications of increased intestinal permeability in the pathogenesis of human disease is discussed in relation to findings in patients with Crohn’s disease. A common feature of increased intestinal permeability is the development of a low grade enteropathy, and while quantitatively similar changes may be found in Crohn’s disease these seem to predict relapse of disease. Moreover, factors associated with relapse of Crohn’s disease have in common an action to increase intestinal permeability. While increased intestinal permeability does not seem to be important in the etiology of Crohn’s disease it may be a central mechanism in the clinical relapse of disease.

  17. Water permeability in human airway epithelium

    DEFF Research Database (Denmark)

    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 ...

  18. Upscaling of permeability field of fractured rock system: Numerical examples

    KAUST Repository

    Bao, K.; Salama, Amgad; Sun, S.


    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.

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

    Directory of Open Access Journals (Sweden)

    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

  20. DNA excision repair in permeable human fibroblasts

    International Nuclear Information System (INIS)

    Kaufmann, W.K.; Bodell, W.J.; Cleaver, J.E.


    U.v. irradiation of confluent human fibroblasts activated DNA repair, aspects of which were characterized in the cells after they were permeabilized. Incubation of intact cells for 20 min between irradiation and harvesting was necessary to obtain a maximum rate of reparative DNA synthesis. Cells harvested immediately after irradiation before repair was initiated displayed only a small stimulation of DNA synthesis, indicating that permeable cells have a reduced capacity to recognize pyrimidine dimers and activate repair. The distribution of sizes of DNA strands labeled during 10 min of reparative DNA synthesis resembled that of parental DNA. However, during a 60-min incubation of permeable cells at 37 degrees C, parental DNA and DNA labeled by reparative DNA synthesis were both cleaved to smaller sizes. Cleavage also occurred in unirradiated cells, indicating that endogenous nuclease was active during incubation. Repair patches synthesized in permeable cells displayed increased sensitivity to digestion by micrococcal nuclease. However, the change in sensitivity during a chase with unlabeled DNA precursors was small, suggesting that reassembly of nucleosome structure at sites of repair was impaired. To examine whether this deficiency was due to a preponderance of incomplete or unligated repair patches, 3H-labeled (repaired) DNA was purified, then digested with exonuclease III and nuclease S1 to probe for free 3' ends and single-stranded regions. About 85% of the [3H]DNA synthesized during a 10-min pulse resisted digestion, suggesting that a major fraction of the repair patches that were filled were also ligated. U.v. light-activated DNA synthesis in permeable cells, therefore, appears to represent the continuation of reparative gap-filling at sites of excision repair activated within intact cells. Gap-filling and ligation were comparatively efficient processes in permeable cells

  1. Ammonia and urea permeability of mammalian aquaporins

    DEFF Research Database (Denmark)

    Litman, Thomas; Søgaard, Rikke; Zeuthen, Thomas


    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...... groups differ in the amino acid composition of their aromatic/arginine regions. The location of the ammonia-permeable aquaporins in the body parallels that of the Rh proteins. This applies to erythrocytes and to cells associated with nitrogen homeostasis and high rates of anabolism. In the liver, AQPs 8...

  2. Accurate control testing for clay liner permeability

    Energy Technology Data Exchange (ETDEWEB)

    Mitchell, R J


    Two series of centrifuge tests were carried out to evaluate the use of centrifuge modelling as a method of accurate control testing of clay liner permeability. The first series used a large 3 m radius geotechnical centrifuge and the second series a small 0.5 m radius machine built specifically for research on clay liners. Two permeability cells were fabricated in order to provide direct data comparisons between the two methods of permeability testing. In both cases, the centrifuge method proved to be effective and efficient, and was found to be free of both the technical difficulties and leakage risks normally associated with laboratory permeability testing of fine grained soils. Two materials were tested, a consolidated kaolin clay having an average permeability coefficient of 1.2{times}10{sup -9} m/s and a compacted illite clay having a permeability coefficient of 2.0{times}10{sup -11} m/s. Four additional tests were carried out to demonstrate that the 0.5 m radius centrifuge could be used for linear performance modelling to evaluate factors such as volumetric water content, compaction method and density, leachate compatibility and other construction effects on liner leakage. The main advantages of centrifuge testing of clay liners are rapid and accurate evaluation of hydraulic properties and realistic stress modelling for performance evaluations. 8 refs., 12 figs., 7 tabs.

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

    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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. Biostable glucose permeable polymer

    DEFF Research Database (Denmark)


    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...

  5. Exosomes from metastatic cancer cells transfer amoeboid phenotype to non-metastatic cells and increase endothelial permeability: their emerging role in tumor heterogeneity. (United States)

    Schillaci, Odessa; Fontana, Simona; Monteleone, Francesca; Taverna, Simona; Di Bella, Maria Antonietta; Di Vizio, Dolores; Alessandro, Riccardo


    The goal of this study was to understand if exosomes derived from high-metastatic cells may influence the behavior of less aggressive cancer cells and the properties of the endothelium. We found that metastatic colon cancer cells are able to transfer their amoeboid phenotype to isogenic primary cancer cells through exosomes, and that this morphological transition is associated with the acquisition of a more aggressive behavior. Moreover, exosomes from the metastatic line (SW620Exos) exhibited higher ability to cause endothelial hyperpermeability than exosomes from the non metastatic line (SW480Exos). SWATH-based quantitative proteomic analysis highlighted that SW620Exos are significantly enriched in cytoskeletal-associated proteins including proteins activating the RhoA/ROCK pathway, known to induce amoeboid properties and destabilization of endothelial junctions. In particular, thrombin was identified as a key mediator of the effects induced by SW620Exos in target cells, in which we also found a significant increase of RhoA activity. Overall, our results demonstrate that in a heterogeneous context exosomes released by aggressive sub-clones can contribute to accelerate tumor progression by spreading malignant properties that affect both the tumor cell plasticity and the endothelial cell behavior.

  6. The super sickling haemoglobin HbS-Oman: a study of red cell sickling, K+ permeability and associations with disease severity in patients heterozygous for HbA and HbS-Oman (HbA/S-Oman genotype). (United States)

    Al Balushi, Halima W M; Wali, Yasser; Al Awadi, Maha; Al-Subhi, Taimoora; Rees, David C; Brewin, John N; Hannemann, Anke; Gibson, John S


    Studying different sickle cell genotypes may throw light on the pathogenesis of sickle cell disease (SCD). Here, the clinical profile, red cell sickling and K + permeability in 29 SCD patients (15 patients with severe disease and 14 with a milder form) of HbA/S-Oman genotype were analysed. The super sickling nature of this Hb variant was confirmed. The red cell membrane permeability to K + was markedly abnormal with elevated activities of P sickle , Gardos channel and KCl cotransporter (KCC). Results were consistent with Ca 2+ entry and Mg 2+ loss via P sickle stimulating Gardos channel and KCC activities. The abnormal red cell behaviour was similar to that in the commonest genotype of SCD, HbSS, in which the level of mutated Hb is considerably higher. Although activities of all three K + transporters also correlated with the level of HbS-Oman, there was no association between transport phenotype and disease severity. The super sickling behaviour of HbS-Oman may obviate the need for solute loss and red cell dehydration to encourage Hb polymerisation, required in other SCD genotypes. Disease severity was reduced by concurrent α thalassaemia, as observed in other SCD genotypes, and represents an obvious genetic marker for prognostic tests of severity in young SCD patients of the HbA/S-Oman genotype. © 2017 John Wiley & Sons Ltd.

  7. The Mechanism by Which MYCN Amplification Confers an Enhanced Sensitivity to a PCNA-Derived Cell Permeable Peptide in Neuroblastoma Cells

    Directory of Open Access Journals (Sweden)

    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.

  8. Increasing the permeability of Escherichia coli using MAC13243

    DEFF Research Database (Denmark)

    Muheim, Claudio; Götzke, Hansjörg; Eriksson, Anna U.


    molecules that make the outer membrane of Escherichia coli more permeable. We identified MAC13243, an inhibitor of the periplasmic chaperone LolA that traffics lipoproteins from the inner to the outer membrane. We observed that cells were (1) more permeable to the fluorescent probe 1-N...

  9. Catalase and superoxide dismutase conjugated with platelet-endothelial cell adhesion molecule antibody distinctly alleviate abnormal endothelial permeability caused by exogenous reactive oxygen species and vascular endothelial growth factor. (United States)

    Han, Jingyan; Shuvaev, Vladimir V; Muzykantov, Vladimir R


    Reactive oxygen species (ROS) superoxide anion (O(2)()) and hydrogen peroxide (H(2)O(2)) produced by activated leukocytes and endothelial cells in sites of inflammation or ischemia cause endothelial barrier dysfunction that may lead to tissue edema. Antioxidant enzymes (AOEs) catalase and superoxide dismutase (SOD) conjugated with antibodies to platelet-endothelial cell adhesion molecule-1 (PECAM-1) specifically bind to endothelium, quench the corresponding ROS, and alleviate vascular oxidative stress and inflammation. In the present work, we studied the effects of anti-PECAM/catalase and anti-PECAM/SOD conjugates on the abnormal permeability manifested by transendothelial electrical resistance decline, increased fluorescein isothiocyanate-dextran influx, and redistribution of vascular endothelial-cadherin in human umbilical vein endothelial cell (HUVEC) monolayers. Anti-PECAM/catalase protected HUVEC monolayers against H(2)O(2)-induced endothelial barrier dysfunction. Polyethylene glycol-conjugated catalase exerted orders of magnitude lower endothelial uptake and no protective effect, similarly to IgG/catalase. Anti-PECAM/catalase, but not anti-PECAM/SOD, alleviated endothelial hyperpermeability caused by exposure to hypoxanthine/xanthine oxidase, implicating primarily H(2)O(2) in the disruption of the endothelial barrier in this model. Thrombin-induced endothelial permeability was not affected by treatment with anti-PECAM/AOEs or the NADPH oxidase inhibitor apocynin or overexpression of AOEs, indicating that the endogenous ROS play no key role in thrombin-mediated endothelial barrier dysfunction. In contrast, anti-PECAM/SOD, but not anti-PECAM/catalase, inhibited a vascular endothelial growth factor (VEGF)-induced increase in endothelial permeability, identifying a key role of endogenous O(2)() in the VEGF-mediated regulation of endothelial barrier function. Therefore, AOEs targeted to endothelial cells provide versatile molecular tools for testing the roles of

  10. In vivo human buccal permeability of nicotine

    DEFF Research Database (Denmark)

    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...

  11. West Nile virus-induced cell adhesion molecules on human brain microvascular endothelial cells regulate leukocyte adhesion and modulate permeability of the in vitro blood-brain barrier model.

    Directory of Open Access Journals (Sweden)

    Kelsey Roe

    Full Text Available Characterizing the mechanisms by which West Nile virus (WNV causes blood-brain barrier (BBB disruption, leukocyte infiltration into the brain and neuroinflammation is important to understand the pathogenesis of WNV encephalitis. Here, we examined the role of endothelial cell adhesion molecules (CAMs in mediating the adhesion and transendothelial migration of leukocytes across human brain microvascular endothelial cells (HBMVE. Infection with WNV (NY99 strain significantly induced ICAM-1, VCAM-1, and E-selectin in human endothelial cells and infected mice brain, although the levels of their ligands on leukocytes (VLA-4, LFA-1and MAC-1 did not alter. The permeability of the in vitro BBB model increased dramatically following the transmigration of monocytes and lymphocytes across the models infected with WNV, which was reversed in the presence of a cocktail of blocking antibodies against ICAM-1, VCAM-1, and E-selectin. Further, WNV infection of HBMVE significantly increased leukocyte adhesion to the HBMVE monolayer and transmigration across the infected BBB model. The blockade of these CAMs reduced the adhesion and transmigration of leukocytes across the infected BBB model. Further, comparison of infection with highly neuroinvasive NY99 and non-lethal (Eg101 strain of WNV demonstrated similar level of virus replication and fold-increase of CAMs in HBMVE cells suggesting that the non-neuropathogenic response of Eg101 is not because of its inability to infect HBMVE cells. Collectively, these results suggest that increased expression of specific CAMs is a pathological event associated with WNV infection and may contribute to leukocyte infiltration and BBB disruption in vivo. Our data further implicate that strategies to block CAMs to reduce BBB disruption may limit neuroinflammation and virus-CNS entry via 'Trojan horse' route, and improve WNV disease outcome.

  12. 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. (United States)

    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. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Permeability measuremens of brazilian Eucalyptus

    Directory of Open Access Journals (Sweden)

    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.

  14. Permeability of cork for water and ethanol. (United States)

    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.

  15. Electrokinetic effects and fluid permeability

    International Nuclear Information System (INIS)

    Berryman, J.G.


    Fluid permeability of porous media depends mainly on connectivity of the pore space and two physical parameters: porosity and a pertinent length-scale parameter. Electrical imaging methods typically establish connectivity and directly measure electrical conductivity, which can then often be related to porosity by Archie's law. When electrical phase measurements are made in addition to the amplitude measurements, information about the pertinent length scale can then be obtained. Since fluid permeability controls the ability to flush unwanted fluid contaminants from the subsurface, inexpensive maps of permeability could improve planning strategies for remediation efforts. Detailed knowledge of fluid permeability is also important for oil field exploitation, where knowledge of permeability distribution in three dimensions is a common requirement for petroleum reservoir simulation and analysis, as well as for estimates on the economics of recovery

  16. Arabidopsis SNAREs SYP61 and SYP121 coordinate the trafficking of plasma membrane aquaporin PIP2;7 to modulate the cell membrane water permeability. (United States)

    Hachez, Charles; Laloux, Timothée; Reinhardt, Hagen; Cavez, Damien; Degand, Hervé; Grefen, Christopher; De Rycke, Riet; Inzé, Dirk; Blatt, Michael R; Russinova, Eugenia; Chaumont, François


    Plant plasma membrane intrinsic proteins (PIPs) are aquaporins that facilitate the passive movement of water and small neutral solutes through biological membranes. Here, we report that post-Golgi trafficking of PIP2;7 in Arabidopsis thaliana involves specific interactions with two syntaxin proteins, namely, the Qc-SNARE SYP61 and the Qa-SNARE SYP121, that the proper delivery of PIP2;7 to the plasma membrane depends on the activity of the two SNAREs, and that the SNAREs colocalize and physically interact. These findings are indicative of an important role for SYP61 and SYP121, possibly forming a SNARE complex. Our data support a model in which direct interactions between specific SNARE proteins and PIP aquaporins modulate their post-Golgi trafficking and thus contribute to the fine-tuning of the water permeability of the plasma membrane. © 2014 American Society of Plant Biologists. All rights reserved.

  17. Effect of permeability enhancers on paracellular permeability of acyclovir. (United States)

    Ates, Muge; Kaynak, Mustafa Sinan; Sahin, Selma


    According to Biopharmaceutics Classification System (BCS), acyclovir is a class III (high solubility, low permeability) compound, and it is transported through paracellular route by passive diffusion. The aim of this study was to investigate the effect of various pharmaceutical excipients on the intestinal permeability of acyclovir. The single-pass in-situ intestinal perfusion (SPIP) method was used to estimate the permeability values of acyclovir and metoprolol across different intestinal segments (jejunum, ileum and colon). Permeability coefficient (Peff ) of acyclovir was determined in the absence and presence of a permeation enhancer such as dimethyl β-cyclodextrin (DM-β-CD), sodium lauryl sulfate (SLS), sodium caprate (Cap-Na) and chitosan chloride. All enhancers increased the permeability of paracellularly transported acyclovir. Although Cap-Na has the highest permeability-enhancing effect in all segments, permeation-enhancing effect of chitosan and SLS was only significant in ileum. On the other hand, DM-β-CD slightly decreased the permeability in all intestinal segments. These findings have potential implication concerning the enhancement of absorption of paracellularly transported compounds with limited oral bioavailability. In the case of acyclovir, Cap-Na either alone or in combination with SLS or chitosan has the potential to improve its absorption and bioavailability and has yet to be explored. © 2016 Royal Pharmaceutical Society.

  18. Role of different biodegradable polymers on the permeability of ciprofloxacin


    Chakraborti, Chandra Kanti; Sahoo, Subhashree; Behera, Pradipta Kumar


    Since permeability across biological membranes is a key factor in the absorption and distribution of drugs, drug permeation characteristics of three oral suspensions of ciprofloxacin were designed and compared. The three suspensions of ciprofloxacin were prepared by taking biodegradable polymers such as carbopol 934, carbopol 940, and hydroxypropyl methylcellulose (HPMC). The permeability study was performed by using a Franz diffusion cell through both synthetic cellulose acetate membrane and...

  19. Relative Permeability of Fractured Rock

    Energy Technology Data Exchange (ETDEWEB)

    Mark D. Habana


    Contemporary understanding of multiphase flow through fractures is limited. Different studies using synthetic fractures and various fluids have yielded different relative permeability-saturation relations. This study aimed to extend the understanding of multiphase flow by conducting nitrogen-water relative permeability experiments on a naturally-fractured rock from The Geysers geothermal field. The steady-state approach was used. However, steady state was achieved only at the endpoint saturations. Several difficulties were encountered that are attributed to phase interference and changes in fracture aperture and surface roughness, along with fracture propagation/initiation. Absolute permeabilities were determined using nitrogen and water. The permeability values obtained change with the number of load cycles. Determining the absolute permeability of a core is especially important in a fractured rock. The rock may change as asperities are destroyed and fractures propagate or st rain harden as the net stresses vary. Pressure spikes occurred in water a solute permeability experiments. Conceptual models of an elastic fracture network can explain the pressure spike behavior. At the endpoint saturations the water relative permeabilities obtained are much less than the nitrogen gas relative permeabilities. Saturations were determined by weighing and by resistivity calculations. The resistivity-saturation relationship developed for the core gave saturation values that differ by 5% from the value determined by weighing. Further work is required to complete the relative permeability curve. The steady-state experimental approach encountered difficulties due to phase interference and fracture change. Steady state may not be reached until an impractical length of time. Thus, unsteady-state methods should be pursued. In unsteady-state experiments the challenge will be in quantifying rock fracture change in addition to fluid flow changes.

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

    DEFF Research Database (Denmark)

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


    activation of basolaterally cocultured human mononuclear leukocytes were measured. DESIGN: In a coculture model, a layer of differentiated, confluent Caco-2 cells was apically stimulated with growth-arrested, nonpathogenic Escherichia coli. SETTING: Basic human cell culture laboratory. INTERVENTIONS...

  1. Sicilian pistachio (Pistacia vera L.) nut inhibits expression and release of inflammatory mediators and reverts the increase of paracellular permeability in IL-1β-exposed human intestinal epithelial cells. (United States)

    Gentile, C; Perrone, A; Attanzio, A; Tesoriere, L; Livrea, M A


    Dietary approaches to control inflammatory bowel diseases (IBD) may include proanthocyanidin-rich foods. Our previous research showed that a hydrophilic extract from Sicilian pistachio nut (HPE) contains substantial amounts of proanthocyanidins and possesses anti-inflammatory activities. We studied the effects of HPE and of its polymeric proanthocyanidin fraction (PPF) in a cell model that simulated some conditions of IBD, consisting of interleukin (IL)-1β-stimulated Caco-2 cells. HPE was prepared by Pistacia vera L. nuts, and PPF was isolated from HPE by adsorbance chromatography. Proanthocyanidins were quantified as anthocyanidins after acidic hydrolysis. Differentiated Caco-2 cells were pre-incubated with HPE or PPF and then were exposed to IL-1β. Cell viability and parameters associated with nuclear factor-κB (NF-κB) activation were assayed. Adsorption of polymeric proanthocyanidins to the cell membrane was investigated by transepithelial electrical resistance (TEER) measurements. HPE decreased prostaglandin (PG)E2 production, IL-6 and IL-8 release, and cyclooxygenase (COX)-2 expression. HPE also inhibited the increase in paracellular permeability and reduced NF-κB activation. Polymeric proanthocyanidins, tested at a concentration comparable with their content in HPE, produced effects comparable to HPE. Finally, cell exposure to PPF increases TEER of the epithelial monolayers. Our results provide evidence that pistachio nut components inhibit inflammatory response of intestinal epithelial cells in vitro and indicate polymeric proanthocyanidins as the major bioactive nut components. The protection implies inhibition of NF-κB activation and occurs in parallel with the adsorption of polymeric proanthocyanidins to cell membrane. Our findings suggest that intake of small amounts of pistachio nut can exert beneficial effects to gastrointestinal pathophysiology.

  2. Geothermal Permeability Enhancement - Final Report

    Energy Technology Data Exchange (ETDEWEB)

    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.

  3. 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. (United States)

    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.

  4. Aminopurvalanol A, a Potent, Selective, and Cell Permeable Inhibitor of Cyclins/Cdk Complexes, Causes the Reduction of in Vitro Fertilizing Ability of Boar Spermatozoa, by Negatively Affecting the Capacitation-Dependent Actin Polymerization

    Directory of Open Access Journals (Sweden)

    Nicola Bernabò


    Full Text Available The adoption of high-througput technologies demonstrated that in mature spermatozoa are present proteins that are thought to be not present or active in sperm cells, such as those involved in control of cell cycle. Here, by using an in silico approach based on the application of networks theory, we found that Cyclins/Cdk complexes could play a central role in signal transduction active during capacitation. Then, we tested this hypothesis in the vitro model. With this approach, spermatozoa were incubated under capacitating conditions in control conditions (CTRL or in the presence of Aminopurvalanol A a potent, selective and cell permeable inhibitor of Cyclins/Cdk complexes at different concentrations (2, 10, and 20 μM. We found that this treatment caused dose-dependent inhibition of sperm fertilizing ability. We attribute this event to the loss of acrosome integrity due to the inhibition of physiological capacitation-dependent actin polymerization, rather than to a detrimental effect on membrane lipid remodeling or on other signaling pathways such as tubulin reorganization or MAPKs activation. In our opinion, these data could revamp the knowledge on biochemistry of sperm capacitation and could suggest new perspectives in studying male infertility.

  5. Permeability model of sintered porous media: analysis and experiments (United States)

    Flórez Mera, Juan Pablo; Chiamulera, Maria E.; Mantelli, Marcia B. H.


    In this paper, the permeability of porous media fabricated from copper powder sintering process was modeled and measured, aiming the use of the porosity as input parameter for the prediction of the permeability of sintering porous media. An expression relating the powder particle mean diameter with the permeability was obtained, based on an elementary porous media cell, which is physically represented by a duct formed by the arrangement of spherical particles forming a simple or orthorhombic packing. A circular duct with variable section was used to model the fluid flow within the porous media, where the concept of the hydraulic diameter was applied. Thus, the porous is modeled as a converging-diverging duct. The electrical circuit analogy was employed to determine two hydraulic resistances of the cell: based on the Navier-Stokes equation and on the Darcýs law. The hydraulic resistances are compared between themselves and an expression to determine the permeability as function of average particle diameter is obtained. The atomized copper powder was sifted to reduce the size dispersion of the particles. The porosities and permeabilities of sintered media fabricated from powders with particle mean diameters ranging from 20 to 200 microns were measured, by means of the image analysis method and using an experimental apparatus. The permeability data of a porous media, made of copper powder and saturated with distilled water, was used to compare with the permeability model. Permeability literature models, which considers that powder particles have the same diameter and include porosity data as input parameter, were compared with the present model and experimental data. This comparison showed to be quite good.

  6. Effects of saponins and glycoalkaloids on the permeability and viability of mammalian intestinal cells and on the integrity of tissue preparations in vitro

    NARCIS (Netherlands)

    Gee, J.M.; Wortley, G.M.; Johnson, I.T.; Price, K.R.; Rutten, A.A.J.J.L.; Houben, G.F.; Penninks, A.H.


    The effects of potato and tomato glycoalkaloids and a saponin mixture from Gypsophila were investigated in cytotoxicity studies (neutral red uptake, mitochondrial MTT reduction and release of lactate dehydrogenase), using cultured cell lines of rat and human intestinal mucosal epithelium.

  7. Permeability changes and incorporation of labelled thymidine into DNA and whole cells of the fibroblast culture of Chinese hamsters affected by MEA and low temperature

    International Nuclear Information System (INIS)

    Ermekova, V.M.; Kondakova, N.V.; Levitman, M.Kh.; Saugabaeva, K.M.; Ehjdus, L.Kh.


    Action of MEA and low temperature (20degC) on the incorporation of labelled thymidine into DNA and whole cells of the fibroblast culture of chinese hamsters has been studied. It has been found that each of the above-mentioned factors equally decreases the label uptake into the cell and DNA. It is concluded that MEA and low temperature do not substantially influence the rate of DNA synthesis

  8. Ultrastructure, inferred porosity, and gram-staining character of Methanospirillum hungatei filament termini describe a unique cell permeability for this archaeobacterium

    International Nuclear Information System (INIS)

    Beveridge, T.J.; Sprott, G.D.; Whippey, P.


    By light microscopy, Methanospirillum hungatei GP1 stains gram positive at the terminal ends of each multicellular filament and gram negative at all regions in between. This phenomenon was studied further by electron microscopy and energy-dispersive X-ray spectroscopy of Gram-stained cells, using a platinum compound to replace Gram's iodine. Crystal violet-platinum precipitates could be found only in the terminal cells of each filament, which suggested that the multilamellar plugs at the filament ends were involved with stain penetration. When sheaths were isolated by sodium dodecyl sulfate-dithiothreitol treatment, the end plugs could be ejected and their layers could be separated from one another by 0.1 M NaOH treatment. Each plug consisted of at least three individual layers; two were particulate and possessed 14.0-nm particles hexagonally arranged on their surfaces with a spacing of a = b = 18.0 nm, whereas the other was a netting of 12.5-nm holes with spacings and symmetry identical to those of the particulate layers. Optical diffraction and computer image reconstruction were used to clarify the structures of each layer in an intact plug and to provide a high-resolution image of their interdigitated structures. The holes through this composite were three to six times larger than those through the sheath. Accordingly, we propose that the terminal plugs of M. hungatei allow the access of larger solutes than does the sheath and that this is the reason why the end cells of each filament stain gram positive whereas more internal cells are gram negative. Intuitively, since the cell spacers which partition the cells from one another along the filament contain plugs identical in structure to terminal plugs, the diffusion of large solutes for these cells would be unidirectional along the filament-cell axis

  9. Bentonite Permeability at Elevated Temperature

    Directory of Open Access Journals (Sweden)

    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.

  10. Human trophoblast-derived hydrogen sulfide stimulates placental artery endothelial cell angiogenesis. (United States)

    Chen, Dong-Bao; Feng, Lin; Hodges, Jennifer K; Lechuga, Thomas J; Zhang, Honghai


    Endogenous hydrogen sulfide (H2S), mainly synthesized by cystathionine β-synthase (CBS) and cystathionine γ-lyase (CTH), has been implicated in regulating placental angiogenesis; however, the underlying mechanisms are unknown. This study was to test a hypothesis that trophoblasts synthesize H2S to promote placental angiogenesis. Human choriocarcinoma-derived BeWo cells expressed both CBS and CTH proteins, while the first trimester villous trophoblast-originated HTR-8/SVneo cells expressed CTH protein only. The H2S producing ability of BeWo cells was significantly inhibited by either inhibitors of CBS (carboxymethyl hydroxylamine hemihydrochloride, CHH) or CTH (β-cyano-L-alanine, BCA) and that in HTR-8/SVneo cells was inhibited by CHH only. H2S donors stimulated cell proliferation, migration, and tube formation in ovine placental artery endothelial cells (oFPAECs) as effectively as vascular endothelial growth factor. Co-culture with BeWo and HTR-8/SVneo cells stimulated oFPAEC migration, which was inhibited by CHH or BCA in BeWo but CHH only in HTR-8/SVneo cells. Primary human villous trophoblasts (HVT) were more potent than trophoblast cell lines in stimulating oFPAEC migration that was inhibited by CHH and CHH/BCA combination in accordance with its H2S synthesizing activity linked to CBS and CTH expression patterns. H2S donors activated endothelial nitric oxide synthase (NOS3), v-AKT murine thymoma viral oncogene homolog 1 (AKT1), and extracellular signal-activated kinase 1/2 (mitogen-activated protein kinase 3/1, MAPK3/1) in oFPAECs. H2S donor-induced NOS3 activation was blocked by AKT1 but not MAPK3/1 inhibition. In keeping with our previous studies showing a crucial role of AKT1, MAPK3/1, and NOS3/NO in placental angiogenesis, these data show that trophoblast-derived endogenous H2S stimulates placental angiogenesis, involving activation of AKT1, NOS3/NO, and MAPK3/1. © The Authors 2017. Published by Oxford University Press on behalf of Society for the Study

  11. Permeability enhancement by shock cooling (United States)

    Griffiths, Luke; Heap, Michael; Reuschlé, Thierry; Baud, Patrick; Schmittbuhl, Jean


    The permeability of an efficient reservoir, e.g. a geothermal reservoir, should be sufficient to permit the circulation of fluids. Generally speaking, permeability decreases over the life cycle of the geothermal system. As a result, is usually necessary to artificially maintain and enhance the natural permeability of these systems. One of the methods of enhancement -- studied here -- is thermal stimulation (injecting cold water at low pressure). This goal of this method is to encourage new thermal cracks within the reservoir host rocks, thereby increasing reservoir permeability. To investigate the development of thermal microcracking in the laboratory we selected two granites: a fine-grained (Garibaldi Grey granite, grain size = 0.5 mm) and a course-grained granite (Lanhelin granite, grain size = 2 mm). Both granites have an initial porosity of about 1%. Our samples were heated to a range of temperatures (100-1000 °C) and were either cooled slowly (1 °C/min) or shock cooled (100 °C/s). A systematic microstructural (2D crack area density, using standard stereological techniques, and 3D BET specific surface area measurements) and rock physical property (porosity, P-wave velocity, uniaxial compressive strength, and permeability) analysis was undertaken to understand the influence of slow and shock cooling on our reservoir granites. Microstructurally, we observe that the 2D crack surface area per unit volume and the specific surface area increase as a result of thermal stressing, and, for the same maximum temperature, crack surface area is higher in the shock cooled samples. This observation is echoed by our rock physical property measurements: we see greater changes for the shock cooled samples. We can conclude that shock cooling is an extremely efficient method of generating thermal microcracks and modifying rock physical properties. Our study highlights that thermal treatments are likely to be an efficient method for the "matrix" permeability enhancement of

  12. Permeability and Dispersion Coefficients in Rocks with Fracture Network - 12140

    Energy Technology Data Exchange (ETDEWEB)

    Lee, C.K.; Htway, M.Z. [Handong Global University, 3 Namsong-ri, Heunghae-eub, Buk-gu, Pohang, Kyungbuk, 791-708 (Korea, Republic of); Yim, S.P. [Korea Atomic Energy Research Institute, P.O.Box 150, Yusong, Daejon, 305-600 (Korea, Republic of)


    Fluid flow and solute transport are considered for a rock medium with a fracture network with regard to the effective permeability and the dispersion coefficients. To investigate the effects of individual fractures a three-fracture system is chosen in which two are parallel and the third one connects the two at different angles. Specifically the micro-cell boundary-value problems(defined through multiple scale analysis) are solved numerically by using finite elements to calculate the permeability and dispersion coefficients. It is shown that the permeability depends significantly on the pattern of the fracture distribution and the dispersion coefficient is influenced by both the externally imposed pressure gradient (which also reflects the flow field) and the direction of the gradient of solute concentration on the macro-scale. From the calculations of the permeability and dispersion coefficients for solute in a rock medium with a fracture network the following conclusions are drawn. 1. The permeability of fractured medium depends on the primary orientation of the fracture network and is influenced by the connecting fractures in the medium. 2. The cross permeability, e.g., permeability in the direction normal to the direction of the external pressure gradient is rather insensitive to the orientation of the fracture network. 3. Calculation of permeability is most efficiently achieved with optimal discretization across individual fractures and is rather insensitive to the discretization along the fracture.. 4. The longitudinal dispersion coefficient Dxx of a fractured medium depends on both the macro-scale concentration gradient and the direction of the flow (pressure gradient). Hence both features must be considered when investigating solute transport in a fractured medium. (authors)

  13. Mechanism of mucosal permeability enhancement of CriticalSorb® (Solutol® HS15) investigated In Vitro in cell cultures.


    Shubber, Saif; Vllasaliu, Driton; Rauch, Cyril; Jordan, Faron; Illum, Lisbeth; Stolnik, Snjezana


    Purpose CriticalSorb?, with the principal component Solutol? HS15, is a novel mucosal drug delivery system demonstrated to improve the bioavailability of selected biotherapeutics. The intention of this study is to elucidate mechanism(s) responsible for the enhancement of trans-mucosal absorption of biological drugs by Solutol? HS15. Methods Micelle size and CMC of Solutol? HS15 were determined in biologically relevant media. Polarised airway Calu-3 cell layers were used to measure the permeab...

  14. The human leukocyte antigen G promotes trophoblast fusion and β-hCG production through the Erk1/2 pathway in human choriocarcinoma cell lines

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Ji-meng [School of Medicine, Nankai University, Tianjin 300071 (China); State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101 (China); Zhao, Hong-xi [Department of Obstetrics and Gynecology, Tangdu Hospital, Fourth Military Medical University, Xi’an 710038 (China); Wang, Li [Department of Obstetrics and Gynecology, General Hospital of Chinese People’s Liberation Army, Beijing 100853 (China); Gao, Zhi-ying, E-mail: [Department of Obstetrics and Gynecology, General Hospital of Chinese People’s Liberation Army, Beijing 100853 (China); Yao, Yuan-qing, E-mail: [Department of Obstetrics and Gynecology, General Hospital of Chinese People’s Liberation Army, Beijing 100853 (China)


    Highlights: •HLA-G expression promotes BeWo cells fusion and fusogenic gene expression. •HLA-G is capable of inducing β-hCG production in human choriocarcinoma cell lines. •Up-regulation of β-hCG production by HLA-G is mediated via the Erk1/2 pathway. -- Abstract: The human leukocyte antigen G (HLA-G) is expressed on the fetal–maternal interface and plays a role in protecting fetal-derived trophoblasts from the maternal immune response, allowing trophoblasts to invade the uterus. However, HLA-G also possesses immune suppressing-independent functions. We found that HLA-G expressing BeWo choriocarcinoma cells increased cell–cell fusion compared to control BeWo cells under forskolin treatment. Regardless of forskolin treatment, the expression of fusogenic gene mRNAs, including syncytin-1, the transcription factor glial cell missing 1 (Gcm1), and beta human chorionic gonadotropin (β-hCG) were elevated. HLA-G up-regulates β-hCG production in human choriocarcinoma cells because HLA-G knockdown in JEG-3 cells induces a dramatic decrease in β-hCG compared with control cells. The defect in β-hCG production in HLA-G knocked-down cells could not be completely overcome by stimulating hCG production through increasing intracellular cAMP levels. HLA-G expressing cells have increased phosphorylation levels for extracellular signal-regulated kinase1/2 (Erk1/2) in BeWo cells. The Erk1/2 pathway is inactivated after the inhibition of HLA-G expression in JEG-3 cells. Finally, Erk1/2 inhibition was able to suppress the increased hCG production induced by HLA-G expression. Together, these data suggest novel roles for HLA-G in regulating β-hCG production via the modulation of the Erk1/2 pathway and by inducing trophoblast cell fusion.

  15. 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 (United States)

    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.

  16. A Chitin-binding Protein Purified from Moringa oleifera Seeds Presents Anticandidal Activity by Increasing Cell Membrane Permeability and Reactive Oxygen Species Production

    Directory of Open Access Journals (Sweden)

    João X.S. Neto


    Full Text Available Candida species are opportunistic pathogens that infect immunocompromised and/or immunosuppressed patients, particularly in hospital facilities, that besides representing a significant threat to health increase the risk of mortality. Apart from echinocandins and triazoles, which are well tolerated, most of the antifungal drugs used for candidiasis treatment can cause side effects and lead to the development of resistant strains. A promising alternative to the conventional treatments is the use of plant proteins. M. oleifera Lam. is a plant with valuable medicinal properties, including antimicrobial activity. This work aimed to purify a chitin-binding protein from M. oleifera seeds and to evaluate its antifungal properties against Candida species. The purified protein, named Mo-CBP2, represented about 0.2% of the total seed protein and appeared as a single band on native PAGE. By mass spectrometry, Mo-CBP2 presented 13,309 Da. However, by SDS-PAGE, Mo-CBP2 migrated as a single band with an apparent molecular mass of 23,400 Da. Tricine-SDS-PAGE of Mo-CBP2 under reduced conditions revealed two protein bands with apparent molecular masses of 7,900 and 4,600 Da. Altogether, these results suggest that Mo-CBP2 exists in different oligomeric forms. Moreover, Mo-CBP2 is a basic glycoprotein (pI 10.9 with 4.1% (m/m sugar and it did not display hemagglutinating and hemolytic activities upon rabbit and human erythrocytes. A comparative analysis of the sequence of triptic peptides from Mo-CBP2 in solution, after LC-ESI-MS/MS, revealed similarity with other M. oleifera proteins, as the 2S albumin Mo-CBP3 and flocculating proteins, and 2S albumins from different species. Mo-CBP2 possesses in vitro antifungal activity against Candida albicans, C. parapsilosis, C. krusei, and C. tropicalis, with MIC50 and MIC90 values ranging between 9.45–37.90 and 155.84–260.29 μM, respectively. In addition, Mo-CBP2 (18.90 μM increased the cell membrane permeabilization

  17. Effect of Salt Forms of Chitosan on In Vitro Permeability ...

    African Journals Online (AJOL)

    Purpose: To investigate the effect of chitosan (CS) salt forms and pH condition on the transepithelial electrical resistance (TEER) of Caco-2 cell monolayer for enhanced permeability. Methods: Solutions (2 %w/v) of four different salt forms of CS-aspartate (CS-A), CS-ethylene diamine tetraacetate (CS-EDTA), ...

  18. Quantifying Evaporation in a Permeable Pavement System (United States)

    Studies quantifying evaporation from permeable pavement systems are limited to a few laboratory studies and one field application. This research quantifies evaporation for a larger-scale field application by measuring the water balance from lined permeable pavement sections. Th...

  19. Permeable Pavement Research - Edison, New Jersey (United States)

    This presentation provides the background and summary of results collected at the permeable pavement parking lot monitored at the EPA facility in Edison, NJ. This parking lot is surfaced with permeable interlocking concrete pavers (PICP), pervious concrete, and porous asphalt. ...

  20. Simultaneous determination of intestinal permeability and potential drug interactions of complex mixtures using Caco-2 cells and high-resolution mass spectrometry: Studies with Rauwolfia serpentina extract. (United States)

    Flynn, Thomas J; Vohra, Sanah N


    Caco-2 cells are a commonly used model for estimating the intestinal bioavailability of single chemical entity pharmaceuticals. Caco-2 cells, when induced with calcitriol, also express other biological functions such as phase I (CYP) and phase II (glucuronosyltransferases) drug metabolizing enzymes which are relevant to drug-supplement interactions. Intestinal bioavailability is an important factor in the overall safety assessment of products consumed orally. Foods, including herbal dietary supplements, are complex substances with multiple chemical components. Because of potential interactions between components of complex mixtures, more reliable safety assessments can be obtained by studying the commercial products "as consumed" rather than by testing individual chemical components one at a time. The present study evaluated the apparent intestinal permeability (P app ) of a model herbal extract, Rauwolfia serpentina, using both whole plant extracts and the individual purified Rauwolfia alkaloids. All test compounds, endpoint substrates, and their metabolites were quantified using liquid chromatography and high-resolution mass spectrometry. The P app values for individual Rauwolfia alkaloids were comparable whether measured individually or as components of the complete extract. Both Rauwolfia extract and all individual Rauwolfia alkaloids except yohimbine inhibited CYP3A4 activity (midazolam 1'-hydroxylation). Both Rauwolfia extract and all individual Rauwolfia alkaloids except corynanthine and reserpic acid significantly increased glucuronosyltransferase activity (glucuronidation of 4-methylumbelliferone). The positive control, ketoconazole, significantly inhibited both CYP3A4 and glucuronosyltransferase activities. These findings suggest that the Caco-2 assay is capable of simultaneously identifying both bioavailability and potentially hazardous intestinal drug-supplement interactions in complex mixtures. Published by Elsevier B.V.

  1. Cell-to-Cell Contact Results in a Selective Translocation of Maternal Human Immunodeficiency Virus Type 1 Quasispecies across a Trophoblastic Barrier by both Transcytosis and Infection (United States)

    Lagaye, S.; Derrien, M.; Menu, E.; Coïto, C.; Tresoldi, E.; Mauclère, P.; Scarlatti, G.; Chaouat, G.; Barré-Sinoussi, F.; Bomsel, M.


    Mother-to-child transmission can occur in utero, mainly intrapartum and postpartum in case of breastfeeding. In utero transmission is highly restricted and results in selection of viral variant from the mother to the child. We have developed an in vitro system that mimics the interaction between viruses, infected cells present in maternal blood, and the trophoblast, the first barrier protecting the fetus. Trophoblastic BeWo cells were grown as a tight polarized monolayer in a two-chamber system. Cell-free virions applied to the apical pole neither crossed the barrier nor productively infected BeWo cells. In contrast, apical contact with human immunodeficiency virus (HIV)-infected peripheral blood mononuclear cells (PBMCs) resulted in transcytosis of infectious virus across the trophoblastic monolayer and in productive infection correlating with the fusion of HIV-infected PBMCs with trophoblasts. We showed that viral variants are selected during these two steps and that in one case of in utero transmission, the predominant maternal viral variant characterized after transcytosis was phylogenetically indistinguishable from the predominant child's virus. Hence, the first steps of transmission of HIV-1 in utero appear to involve the interaction between HIV type 1-infected cells and the trophoblastic layer, resulting in the passage of infectious HIV by transcytosis and by fusion/infection, both leading to a selection of virus quasispecies. PMID:11312350

  2. Quantifying porosity, compressibility and permeability in Shale

    DEFF Research Database (Denmark)

    Mbia, Ernest Ncha; Fabricius, Ida Lykke; Frykman, Peter

    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...... permeability estimate comparable to the measured one for shale rich in smectite. This is probably because Yang and Aplin model was calibrated in London clay which is rich in smectite....

  3. Viscous fingering with permeability heterogeneity

    International Nuclear Information System (INIS)

    Tan, C.; Homsy, G.M.


    Viscous fingering in miscible displacements in the presence of permeability heterogeneities is studied using two-dimensional simulations. The heterogeneities are modeled as stationary random functions of space with finite correlation scale. Both the variance and scale of the heterogeneities are varied over modest ranges. It is found that the fingered zone grows linearly in time in a fashion analogous to that found in homogeneous media by Tan and Homsy [Phys. Fluids 31, 1330 (1988)], indicating a close coupling between viscous fingering on the one hand and flow through preferentially more permeable paths on the other. The growth rate of the mixing zone increases monotonically with the variance of the heterogeneity, as expected, but shows a maximum as the correlation scale is varied. The latter is explained as a ''resonance'' between the natural scale of fingers in homogeneous media and the correlation scale

  4. Permeability of hydrogen isotopes through Pd-Ag membrane

    International Nuclear Information System (INIS)

    Yoshida, Hiroshi


    This paper represents the permeabilities, diffusion coefficients and isotope effects for hydrogen and deuterium through Pd-25 wt.% Ag alloy tubes The feed gas H 2 or D 2 flowing into the permeation cell was preheated before it reached to the outer surface of the permeation tube made of palladium-silver alloy. Permeation time lag method could be successfully carried out with the present apparatus to measure both permeability and diffusion coefficient. The square-root pressure dependence for the permeation of hydrogen isotopes was observed. The observed systematic temperature dependence indicates that the approximation of the Arrhenius' relation was effective within this experimental conditions. Some tendency of permeation fluxes in relation to the reciprocal temperature, 1/T, was seen. The permeability ratio was larger than the square root of isotopic mass ratio, and it decreased with temperature rise. On the contrary, the diffusion coefficient ratio was much smaller than the square root of isotopic mass ratio. (Kato, T.)

  5. Transparent anodes for polymer photovoltaics: Oxygen permeability of PEDOT

    DEFF Research Database (Denmark)

    Andersen, M.; Carlé, Jon Eggert; Cruys-Bagger, N.


    The oxygen permeability of the transparent organic anode poly(3,4,-ethylene dioxythiophene) with paratoluenesulphonate as the anion (PEDOT:pTS) was determined to be 2.5 +/- 0.7 x 10(-15) cm(3) (STP) CM cm(-2) S-1 Pa-1, and is thus comparable in magnitude to the oxygen permeability of polyethylene......The oxygen permeability of the transparent organic anode poly(3,4,-ethylene dioxythiophene) with paratoluenesulphonate as the anion (PEDOT:pTS) was determined to be 2.5 +/- 0.7 x 10(-15) cm(3) (STP) CM cm(-2) S-1 Pa-1, and is thus comparable in magnitude to the oxygen permeability...... of polyethyleneterephthalate (PET). The oxygen diffusion through bilayers of polyethylene (PE) and PEDOT:pTS and bilayers of PET and PEDOT:pTS was established. The bilayer structures were applied as the carrier substrate and the transparent anode in polymer-based photovoltaic devices employing a mixture of poly(1-methoxy-4......-(2-ethylhexyloxy)-p-phenylenevinylene) (MEH-PPV) and [6,6]-phenyt-C-61-butanoicacidmethylester (PCBM) as the active layer and aluminium as the cathode. The oxygen permeability of the layers and the aluminium cathode was correlated with the lifetime of the solar cell devices. It was found that the performance...

  6. Gap junction protein connexin43 exacerbates lung vascular permeability.

    Directory of Open Access Journals (Sweden)

    James J O'Donnell

    Full Text Available Increased vascular permeability causes pulmonary edema that impairs arterial oxygenation and thus contributes to morbidity and mortality associated with Acute Respiratory Distress Syndrome and sepsis. Although components of intercellular adhesive and tight junctions are critical for maintaining the endothelial barrier, there has been limited study of the roles of gap junctions and their component proteins (connexins. Since connexins can modulate inflammatory signaling in other systems, we hypothesized that connexins may also regulate pulmonary endothelial permeability. The relationships between connexins and the permeability response to inflammatory stimuli were studied in cultured human pulmonary endothelial cells. Prolonged treatment with thrombin, lipopolysaccharide, or pathological cyclic stretch increased levels of mRNA and protein for the major connexin, connexin43 (Cx43. Thrombin and lipopolysaccharide both increased intercellular communication assayed by transfer of microinjected Lucifer yellow. Although thrombin decreased transendothelial resistance in these cells, the response was attenuated by pretreatment with the connexin inhibitor carbenoxolone. Additionally, the decreases of transendothelial resistance produced by either thrombin or lipopolysaccharide were attenuated by reducing Cx43 expression by siRNA knockdown. Both carbenoxolone and Cx43 knockdown also abrogated thrombin-induced phosphorylation of myosin light chain. Taken together, these data suggest that increased lung vascular permeability induced by inflammatory conditions may be amplified via increased expression of Cx43 and intercellular communication among pulmonary endothelial cells.

  7. A fractal model for predicting permeability and liquid water relative permeability in the gas diffusion layer (GDL) of PEMFCs (United States)

    He, Guangli; Zhao, Zongchang; Ming, Pingwen; Abuliti, Abudula; Yin, Caoyong

    In this study, a fractal model is developed to predict the permeability and liquid water relative permeability of the GDL (TGP-H-120 carbon paper) in proton exchange membrane fuel cells (PEMFCs), based on the micrographs (by SEM, i.e. scanning electron microscope) of the TGP-H-120. Pore size distribution (PSD), maximum pore size, porosity, diameter of the carbon fiber, pore tortuosity, area dimension, hydrophilicity or hydrophobicity, the thickness of GDL and saturation are involved in this model. The model was validated by comparison between the predicted results and experimental data. The results indicate that the water relative permeability in the hydrophobicity case is much higher than in the hydrophilicity case. So, a hydrophobic carbon paper is preferred for efficient removal of liquid water from the cathode of PEMFCs.

  8. Do the recommended standards for in vitro biopharmaceutic classification of drug permeability meet the "passive transport" criterion for biowaivers? (United States)

    Žakelj, Simon; Berginc, Katja; Roškar, Robert; Kraljič, Bor; Kristl, Albin


    BCS based biowaivers are recognized by major regulatory agencies. An application for a biowaiver can be supported by or even based on "in vitro" measurements of drug permeability. However, guidelines limit the application of biowaivers to drug substances that are transported only by passive mechanisms. Regarding published permeability data as well as measurements obtained in our institution, one can rarely observe drug substances that conform to this very strict criterion. Therefore, we measured the apparent permeability coefficients of 13 drugs recommended by FDA's Guidance to be used as standards for "in vitro" permeability classification. The asymmetry of permeability data determined for both directions (mucosal-to-serosal and serosalto- mucosal) through the rat small intestine revealed significant active transport for four out of the nine high-permeability standards and for all four low-permeability standard drugs. As could be expected, this asymmetry was abolished at 4°C on rat intestine. The permeability of all nine high-permeability, but none of the low permeability standards, was also much lower when measured with intestinal tissue, Caco-2 cell monolayers or artificial membranes at 4°C compared to standard conditions (37°C). Additionally, concurrent testing of several standard drugs revealed that membrane transport can be affected by the use of internal permeability standards. The implications of the results are discussed regarding the regulatory aspects of biopharmaceutical classification, good practice in drug permeability evaluation and regarding the general relevance of transport proteins with broad specificity in drug absorption.

  9. Effect of aggregate grain size distribution on properties of permeable ...

    African Journals Online (AJOL)

    ) ratio on the mechanical properties of permeable concrete is investigated. The aim of this study is to prepare permeable concrete mixture with optimum properties in terms of strength and permeability. For this purpose, five different permeable ...

  10. Cell-permeable Circadian Clock Proteins

    National Research Council Canada - National Science Library

    Johnson, Carl


    .... These 'biological clocks' are important to human physiology. For example, psychiatric and medical studies have shown that circadian rhythmicity is involved in some forms of depressive illness, 'jet lag', drug tolerance/efficacy, memory, and insomnia...

  11. Involvement of the putative Ca²⁺-permeable mechanosensitive channels, NtMCA1 and NtMCA2, in Ca²⁺ uptake, Ca²⁺-dependent cell proliferation and mechanical stress-induced gene expression in tobacco (Nicotiana tabacum) BY-2 cells. (United States)

    Kurusu, Takamitsu; Yamanaka, Takuya; Nakano, Masataka; Takiguchi, Akiko; Ogasawara, Yoko; Hayashi, Teruyuki; Iida, Kazuko; Hanamata, Shigeru; Shinozaki, Kazuo; Iida, Hidetoshi; Kuchitsu, Kazuyuki


    To gain insight into the cellular functions of the mid1-complementing activity (MCA) family proteins, encoding putative Ca²⁺-permeable mechanosensitive channels, we isolated two MCA homologs of tobacco (Nicotiana tabacum) BY-2 cells, named NtMCA1 and NtMCA2. NtMCA1 and NtMCA2 partially complemented the lethality and Ca²⁺ uptake defects of yeast mutants lacking mechanosensitive Ca²⁺ channel components. Furthermore, in yeast cells overexpressing NtMCA1 and NtMCA2, the hypo-osmotic shock-induced Ca²⁺ influx was enhanced. Overexpression of NtMCA1 or NtMCA2 in BY-2 cells enhanced Ca²⁺ uptake, and significantly alleviated growth inhibition under Ca²⁺ limitation. NtMCA1-overexpressing BY-2 cells showed higher sensitivity to hypo-osmotic shock than control cells, and induced the expression of the touch-inducible gene, NtERF4. We found that both NtMCA1-GFP and NtMCA2-GFP were localized at the plasma membrane and its interface with the cell wall, Hechtian strands, and at the cell plate and perinuclear vesicles of dividing cells. NtMCA2 transcript levels fluctuated during the cell cycle and were highest at the G1 phase. These results suggest that NtMCA1 and NtMCA2 play roles in Ca²⁺-dependent cell proliferation and mechanical stress-induced gene expression in BY-2 cells, by regulating the Ca²⁺ influx through the plasma membrane.

  12. Permeability of highly compacted bentonite

    International Nuclear Information System (INIS)

    Pusch, R.


    The object of the study was the water flow through the bentonite which is caused by hydraulic gradients. The study comprised laboratory tests and theoretical considerations. It was found that high bulk densities reduced the permeability to very low values. It was concluded that practically impervious conditions prevail when the gradients are low. Thus with a regional gradient of 10 -2 and a premeability of 10 -13 m/s the flow rate will not be higher than approximately 1 mm in 30 000 years. (G.B.)

  13. The Effect of Excipients on the Permeability of BCS Class III Compounds and Implications for Biowaivers. (United States)

    Parr, Alan; Hidalgo, Ismael J; Bode, Chris; Brown, William; Yazdanian, Mehran; Gonzalez, Mario A; Sagawa, Kazuko; Miller, Kevin; Jiang, Wenlei; Stippler, Erika S


    Currently, the FDA allows biowaivers for Class I (high solubility and high permeability) and Class III (high solubility and low permeability) compounds of the Biopharmaceutics Classification System (BCS). Scientific evidence should be provided to support biowaivers for BCS Class I and Class III (high solubility and low permeability) compounds. Data on the effects of excipients on drug permeability are needed to demonstrate that commonly used excipients do not affect the permeability of BCS Class III compounds, which would support the application of biowaivers to Class III compounds. This study was designed to generate such data by assessing the permeability of four BCS Class III compounds and one Class I compound in the presence and absence of five commonly used excipients. The permeability of each of the compounds was assessed, at three to five concentrations, with each excipient in two different models: Caco-2 cell monolayers, and in situ rat intestinal perfusion. No substantial increases in the permeability of any of the compounds were observed in the presence of any of the tested excipients in either of the models, with the exception of disruption of Caco-2 cell monolayer integrity by sodium lauryl sulfate at 0.1 mg/ml and higher. The results suggest that the absorption of these four BCS Class III compounds would not be greatly affected by the tested excipients. This may have implications in supporting biowaivers for BCS Class III compounds in general.

  14. Live cell imaging of in vitro human trophoblast syncytialization. (United States)

    Wang, Rui; Dang, Yan-Li; Zheng, Ru; Li, Yue; Li, Weiwei; Lu, Xiaoyin; Wang, Li-Juan; Zhu, Cheng; Lin, Hai-Yan; Wang, Hongmei


    Human trophoblast syncytialization, a process of cell-cell fusion, is one of the most important yet least understood events during placental development. Investigating the fusion process in a placenta in vivo is very challenging given the complexity of this process. Application of primary cultured cytotrophoblast cells isolated from term placentas and BeWo cells derived from human choriocarcinoma formulates a biphasic strategy to achieve the mechanism of trophoblast cell fusion, as the former can spontaneously fuse to form the multinucleated syncytium and the latter is capable of fusing under the treatment of forskolin (FSK). Live-cell imaging is a powerful tool that is widely used to investigate many physiological or pathological processes in various animal models or humans; however, to our knowledge, the mechanism of trophoblast cell fusion has not been reported using a live- cell imaging manner. In this study, a live-cell imaging system was used to delineate the fusion process of primary term cytotrophoblast cells and BeWo cells. By using live staining with Hoechst 33342 or cytoplasmic dyes or by stably transfecting enhanced green fluorescent protein (EGFP) and DsRed2-Nuc reporter plasmids, we observed finger-like protrusions on the cell membranes of fusion partners before fusion and the exchange of cytoplasmic contents during fusion. In summary, this study provides the first video recording of the process of trophoblast syncytialization. Furthermore, the various live-cell imaging systems used in this study will help to yield molecular insights into the syncytialization process during placental development. © 2014 by the Society for the Study of Reproduction, Inc.

  15. Steam-water relative permeability

    Energy Technology Data Exchange (ETDEWEB)

    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.

  16. Clogging in permeable concrete: A review. (United States)

    Kia, Alalea; Wong, Hong S; Cheeseman, Christopher R


    Permeable concrete (or "pervious concrete" in North America) is used to reduce local flooding in urban areas and is an important sustainable urban drainage system. However, permeable concrete exhibits reduction in permeability due to clogging by particulates, which severely limits service life. This paper reviews the clogging mechanism and current mitigating strategies in order to inform future research needs. The pore structure of permeable concrete and characteristics of flowing particulates influence clogging, which occurs when particles build-up and block connected porosity. Permeable concrete requires regular maintenance by vacuum sweeping and pressure washing, but the effectiveness and viability of these methods is questionable. The potential for clogging is related to the tortuosity of the connected porosity, with greater tortuosity resulting in increased potential for clogging. Research is required to develop permeable concrete that can be poured on-site, which produces a pore structure with significantly reduced tortuosity. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Different Methods of Predicting Permeability in Shale

    DEFF Research Database (Denmark)

    Mbia, Ernest Ncha; Fabricius, Ida Lykke; Krogsbøll, Anette

    by two to five orders of magnitudes at lower vertical effective stress below 40 MPa as the content of clay minerals increases causing heterogeneity in shale material. Indirect permeability from consolidation can give maximum and minimum values of shale permeability needed in simulating fluid flow......Permeability is often very difficult to measure or predict in shale lithology. In this work we are determining shale permeability from consolidation tests data using Wissa et al., (1971) approach and comparing the results with predicted permeability from Kozeny’s model. Core and cuttings materials...... effective stress to 9 μD at high vertical effective stress of 100 MPa. The indirect permeability calculated from consolidation tests falls in the same magnitude at higher vertical effective stress, above 40 MPa, as that of the Kozeny model for shale samples with high non-clay content ≥ 70% but are higher...

  18. Analytical Estimation of Water-Oil Relative Permeabilities through Fractures

    Directory of Open Access Journals (Sweden)

    Saboorian-Jooybari Hadi


    Full Text Available Modeling multiphase flow through fractures is a key issue for understanding flow mechanism and performance prediction of fractured petroleum reservoirs, geothermal reservoirs, underground aquifers and carbon-dioxide sequestration. One of the most challenging subjects in modeling of fractured petroleum reservoirs is quantifying fluids competition for flow in fracture network (relative permeability curves. Unfortunately, there is no standard technique for experimental measurement of relative permeabilities through fractures and the existing methods are very expensive, time consuming and erroneous. Although, several formulations were presented to calculate fracture relative permeability curves in the form of linear and power functions of flowing fluids saturation, it is still unclear what form of relative permeability curves must be used for proper modeling of flow through fractures and consequently accurate reservoir simulation. Basically, the classic linear relative permeability (X-type curves are used in almost all of reservoir simulators. In this work, basic fluid flow equations are combined to develop a new simple analytical model for water-oil two phase flow in a single fracture. The model gives rise to simple analytic formulations for fracture relative permeabilities. The model explicitly proves that water-oil relative permeabilities in fracture network are functions of fluids saturation, viscosity ratio, fluids density, inclination of fracture plane from horizon, pressure gradient along fracture and rock matrix wettability, however they were considered to be only functions of saturations in the classic X-type and power (Corey [35] and Honarpour et al. [28, 29] models. Eventually, validity of the proposed formulations is checked against literature experimental data. The proposed fracture relative permeability functions have several advantages over the existing ones. Firstly, they are explicit functions of the parameters which are known for

  19. Low Permeability Polyimide Insulation, Phase I (United States)

    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...

  20. Modelling of water permeability in cementitious materials

    DEFF Research Database (Denmark)

    Guang, Ye; Lura, Pietro; van Breugel, K.


    This paper presents a network model to predict the permeability of cement paste from a numerical simulation of its microstructure. Based on a linked list pore network structure, the effective hydraulic conductivity is estimated and the fluid flow is calculated according to the Hagen-Poiseuille law....... The pressure gradient at all nodes is calculated with the Gauss elimination method and the absolute permeability of the pore network is calculated directly from Darcy's law. Finally, the permeability model is validated by comparison with direct water permeability measurements. According to this model...

  1. Transcytosis Involvement in Transport System and Endothelial Permeability of Vascular Leakage during Dengue Virus Infection

    Directory of Open Access Journals (Sweden)

    Chanettee Chanthick


    Full Text Available The major role of endothelial cells is to maintain homeostasis of vascular permeability and to preserve the integrity of vascular vessels to prevent fluid leakage. Properly functioning endothelial cells promote physiological balance and stability for blood circulation and fluid components. A monolayer of endothelial cells has the ability to regulate paracellular and transcellular pathways for transport proteins, solutes, and fluid. In addition to the paracellular pathway, the transcellular pathway is another route of endothelial permeability that mediates vascular permeability under physiologic conditions. The transcellular pathway was found to be associated with an assortment of disease pathogeneses. The clinical manifestation of severe dengue infection in humans is vascular leakage and hemorrhagic diatheses. This review explores and describes the transcellular pathway, which is an alternate route of vascular permeability during dengue infection that corresponds with the pathologic finding of intact tight junction. This pathway may be the route of albumin transport that causes endothelial dysfunction during dengue virus infection.

  2. Permeability of Aluminium Foams Produced by Replication Casting

    Directory of Open Access Journals (Sweden)

    Maxim L. Cherny


    Full Text Available The replication casting process is used for manufacturing open-pore aluminum foams with advanced performances, such as stability and repeatability of foam structure with porosity over 60%. A simple foam structure model based on the interaction between sodium chloride solid particles poorly wetted by melted aluminum, which leads to the formation of air pockets (or “air collars”, is proposed for the permeability of porous material. The equation for the minimum pore radius of replicated aluminum foam is derived. According to the proposed model, the main assumption of the permeability model consists in a concentration of flow resistance in a circular aperture of radius rmin. The permeability of aluminum open-pore foams is measured using transformer oil as the fluid, changing the fractions of initial sodium chloride. Measured values of minimum pore size are close to theoretically predicted ones regardless of the particle shape. The expression for the permeability of replicated aluminum foam derived on the basis of the “bottleneck” model of porous media agrees well with the experimental data. The obtained data can be applied for commercial filter cells and pneumatic silencers.

  3. Crustal permeability: Introduction to the special issue (United States)

    Ingebritsen, Steven E.; Gleeson, Tom


    The topic of crustal permeability is of broad interest in light of the controlling effect of permeability on diverse geologic processes and also timely in light of the practical challenges associated with emerging technologies such as hydraulic fracturing for oil and gas production (‘fracking’), enhanced geothermal systems, and geologic carbon sequestration. This special issue of Geofluids is also motivated by the historical dichotomy between the hydrogeologic concept of permeability as a static material property that exerts control on fluid flow and the perspective of economic geologists, geophysicists, and crustal petrologists who have long recognized permeability as a dynamic parameter that changes in response to tectonism, fluid production, and geochemical reactions. Issues associated with fracking, enhanced geothermal systems, and geologic carbon sequestration have already begun to promote a constructive dialog between the static and dynamic views of permeability, and here we have made a conscious effort to include both viewpoints. This special issue also focuses on the quantification of permeability, encompassing both direct measurement of permeability in the uppermost crust and inferential permeability estimates, mainly for the deeper crust.

  4. Effect of temperature on sandstone permeability

    DEFF Research Database (Denmark)

    Rosenbrand, Esther; Kjøller, Claus

    Hot water injection in geothermal sandstone aquifers is considered for seasonal energy storage in Denmark. However, an increase in the aquifer temperature might reduce permeability, and thereby increase production costs. An understanding of the factors that control permeability is required in order...... and the Klinkenberg procedure showed the expected correlation between the two measures, however, differences could be around one order of magnitude. In tight gas sandstones, permeability is often sensitive to net stress, which might change due to the pore pressure change in the Klinkenberg procedure. Besides...... affecting the Klinkenberg procedure, the combined effect of slip and changes in permeability would affect production during pressure depletion in tight gas sandstone reservoirs; therefore effects of gas slip and net stress on permeability were combined in a model based on the Klinkenberg equation. A lower...

  5. Microorganism Removal in Permeable Pavement Parking Lots ... (United States)

    Three types of permeable pavements (pervious concrete, permeable interlocking concrete pavers, and porous asphalt) were monitored at the Edison Environmental Center in Edison, New Jersey for indicator organisms such as fecal coliform, enterococci, and E. coli. Results showed that porous asphalt had much lower concentration in monitored infiltrate compared to pervious concrete and permeable interlocking concrete pavers. Concentrations of monitored organisms in infiltrate from porous asphalt were consistently below the bathing water quality standard. Fecal coliform and enterococci exceeded bathing water quality standards more than 72% and 34% of the time for permeable interlocking concrete pavers and pervious concrete, respectively. Purpose is to evaluate the performance of permeable pavement in removing indicator organisms from infiltrating stormwater runoff.

  6. Compact rock material gas permeability properties

    Energy Technology Data Exchange (ETDEWEB)

    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.

  7. Gas and Water Permeability of Concrete

    Energy Technology Data Exchange (ETDEWEB)

    Villar, M. V.; Martin, P. L.; Romero, F. J.; Gutierrez-Rodirgo, V.; Barcala, J. M.


    The gas pressure of concrete samples was measured in an unsteady-state equipment working under low injection pressures and in a newly fine tuned steady-state setup working under different pressures. These measurements allowed the estimation of the intrinsic and relative gas permeability of the concrete and of the effect of boundary conditions on them. Permeability decreased with water content, but it was also greatly affected by the hydraulic history of concrete, i.e. if it had been previously dried or wetted. In particular, and for a given degree of saturation, the gas permeability of concrete previously saturated was lower than if the concrete had been just air dried or saturated after air drying. In any case, the gas permeability was about two orders of magnitude higher than the liquid water permeability (10-16 vs. 10-18 m2), probably due to the chemical reactions taking place during saturation (carbonation). The relative gas permeability of concrete increased sharply for water degrees of saturation smaller than 50%. The boundary conditions also affected the gas permeability, which seemed to be mostly conditioned by the back pressure and the confining pressure, increasing as the former increased and decreasing as the latter increased, i.e. decreasing as the effective pressure increased. Overall the increase of pressure head or injection pressure implied a decrease in gas permeability. External,microcracking during air-drying could not be ruled out as responsible for the decrease of permeability with confining pressure. The apparent permeability obtained applying the Klinkenberg method for a given effective pressure was only slightly smaller than the average of all the values measured for the same confining pressure range. For this reason it is considered that the Klinkenberg effect was not relevant in the range of pressures applied. (Author) 37 refs.

  8. Permeability During Magma Expansion and Compaction (United States)

    Gonnermann, Helge. M.; Giachetti, Thomas; Fliedner, Céline; Nguyen, Chinh T.; Houghton, Bruce F.; Crozier, Joshua A.; Carey, Rebecca J.


    Plinian lapilli from the 1060 Common Era Glass Mountain rhyolitic eruption of Medicine Lake Volcano, California, were collected and analyzed for vesicularity and permeability. A subset of the samples were deformed at a temperature of 975°, under shear and normal stress, and postdeformation porosities and permeabilities were measured. Almost all undeformed samples fall within a narrow range of vesicularity (0.7-0.9), encompassing permeabilities between approximately 10-15 m2 and 10-10 m2. A percolation threshold of approximately 0.7 is required to fit the data by a power law, whereas a percolation threshold of approximately 0.5 is estimated by fitting connected and total vesicularity using percolation modeling. The Glass Mountain samples completely overlap with a range of explosively erupted silicic samples, and it remains unclear whether the erupting magmas became permeable at porosities of approximately 0.7 or at lower values. Sample deformation resulted in compaction and vesicle connectivity either increased or decreased. At small strains permeability of some samples increased, but at higher strains permeability decreased. Samples remain permeable down to vesicularities of less than 0.2, consistent with a potential hysteresis in permeability-porosity between expansion (vesiculation) and compaction (outgassing). We attribute this to retention of vesicle interconnectivity, albeit at reduced vesicle size, as well as bubble coalescence during shear deformation. We provide an equation that approximates the change in permeability during compaction. Based on a comparison with data from effusively erupted silicic samples, we propose that this equation can be used to model the change in permeability during compaction of effusively erupting magmas.

  9. Role of different biodegradable polymers on the permeability of ciprofloxacin

    Directory of Open Access Journals (Sweden)

    Chandra Kanti Chakraborti


    Full Text Available Since permeability across biological membranes is a key factor in the absorption and distribution of drugs, drug permeation characteristics of three oral suspensions of ciprofloxacin were designed and compared. The three suspensions of ciprofloxacin were prepared by taking biodegradable polymers such as carbopol 934, carbopol 940, and hydroxypropyl methylcellulose (HPMC. The permeability study was performed by using a Franz diffusion cell through both synthetic cellulose acetate membrane and excised goat gastrointestinal membranes in acidic as well as alkaline pH. To know the permeability of drug from control/formulations through different membranes in acidic/alkaline pH, cumulative percentage drug permeation, apparent permeability (Papp, flux, and enhancement ratio (ER were calculated. Considering Papp and flux values of all formulations, it is evident that formulation containing HPMC was the most beneficial for improving permeation and diffusivity of ciprofloxacin even after 16 h. Hence, this preparation may be considered as the most suitable formulation to obtain prolonged release action of the drug. The ER values of all formulations, through excised goat intestinal mucosal membrane in alkaline pH, were higher than those formulations through goat stomach mucosal membrane in acidic pH. Enhancement ratio values of those formulations indicate that the permeability of the drug was more enhanced by the polymers in the intestinal part, leading to more bioavailability and prolonged action in that portion of the gastrointestinal tract. It may also be concluded from our results that HPMC containing formulation was the best suspension, which may show effective controlled release action. Even carbopol containing formulations might also produce controlled release action.

  10. Endothelial glycocalyx dysfunction in disease: albuminuria and increased microvascular permeability. (United States)

    Salmon, Andrew H J; Satchell, Simon C


    Appreciation of the glomerular microcirculation as a specialized microcirculatory bed, rather than as an entirely separate entity, affords important insights into both glomerular and systemic microvascular pathophysiology. In this review we compare regulation of permeability in systemic and glomerular microcirculations, focusing particularly on the role of the endothelial glycocalyx, and consider the implications for disease processes. The luminal surface of vascular endothelium throughout the body is covered with endothelial glycocalyx, comprising surface-anchored proteoglycans, supplemented with adsorbed soluble proteoglycans, glycosaminoglycans and plasma constituents. In both continuous and fenestrated microvessels, this endothelial glycocalyx provides resistance to the transcapillary escape of water and macromolecules, acting as an integral component of the multilayered barrier provided by the walls of these microvessels (ie acting in concert with clefts or fenestrae across endothelial cell layers, basement membranes and pericytes). Dysfunction of any of these capillary wall components, including the endothelial glycocalyx, can disrupt normal microvascular permeability. Because of its ubiquitous nature, damage to the endothelial glycocalyx alters the permeability of multiple capillary beds: in the glomerulus this is clinically apparent as albuminuria. Generalized damage to the endothelial glycocalyx can therefore manifest as both albuminuria and increased systemic microvascular permeability. This triad of altered endothelial glycocalyx, albuminuria and increased systemic microvascular permeability occurs in a number of important diseases, such as diabetes, with accumulating evidence for a similar phenomenon in ischaemia-reperfusion injury and infectious disease. The detection of albuminuria therefore has implications for the function of the microcirculation as a whole. The importance of the endothelial glycocalyx for other aspects of vascular function

  11. Suitability of Torrent Permeability Tester to measure air-permeability of covercrete

    Energy Technology Data Exchange (ETDEWEB)

    Andrade, C.; Gonzales-Gasca, C. [Institute of Construction Sciences ' Eduardo Torroja' , Madrid (Spain); Torrent, R. [Portland Cement Institute, (Argentina)


    Suitability of the Torrent Permeability Tester (TPT) to measure the permeability of covercrete to air, both in the laboratory and the field, is investigated, and test results obtained in laboratory studies are discussed. The tests performed included the determination of air permeability (TPT method), oxygen permeability (Cembureau method) and capillary suction, rapid chloride permeability test (ASTM C 1202), as well as a one-year carbonation depth test. Concrete specimens of various compositions and curing regimes were used in the tests; the gas-permeability tests were repeated on the same specimens after 28 days, than again at 6 months and 12 months. Test results confirmed the suitability of the TPT as a useful tool in the characterization of the quality the of concrete cover. It was found to be sensitive to changes in concrete quality; repeatable for sensitive properties such as gas permeability ; also, it was found to correlate well with other durability-related properties. 10 refs., 8 tabs., 8 figs.

  12. Comparative study of four immortalized human brain capillary endothelial cell lines, hCMEC/D3, hBMEC, TY10, and BB19, and optimization of culture conditions, for an in vitro blood-brain barrier model for drug permeability studies. (United States)

    Eigenmann, Daniela E; Xue, Gongda; Kim, Kwang S; Moses, Ashlee V; Hamburger, Matthias; Oufir, Mouhssin


    Reliable human in vitro blood-brain barrier (BBB) models suitable for high-throughput screening are urgently needed in early drug discovery and development for assessing the ability of promising bioactive compounds to overcome the BBB. To establish an improved human in vitro BBB model, we compared four currently available and well characterized immortalized human brain capillary endothelial cell lines, hCMEC/D3, hBMEC, TY10, and BB19, with respect to barrier tightness and paracellular permeability. Co-culture systems using immortalized human astrocytes (SVG-A cell line) and immortalized human pericytes (HBPCT cell line) were designed with the aim of positively influencing barrier tightness. Tight junction (TJ) formation was assessed by transendothelial electrical resistance (TEER) measurements using a conventional epithelial voltohmmeter (EVOM) and an automated CellZscope system which records TEER and cell layer capacitance (CCL) in real-time.Paracellular permeability was assessed using two fluorescent marker compounds with low BBB penetration (sodium fluorescein (Na-F) and lucifer yellow (LY)). Conditions were optimized for each endothelial cell line by screening a series of 24-well tissue culture inserts from different providers. For hBMEC cells, further optimization was carried out by varying coating material, coating procedure, cell seeding density, and growth media composition. Biochemical characterization of cell type-specific transmembrane adherens junction protein VE-cadherin and of TJ proteins ZO-1 and claudin-5 were carried out for each endothelial cell line. In addition, immunostaining for ZO-1 in hBMEC cell line was performed. The four cell lines all expressed the endothelial cell type-specific adherens junction protein VE-cadherin. The TJ protein ZO-1 was expressed in hCMEC/D3 and in hBMEC cells. ZO-1 expression could be confirmed in hBMEC cells by immunocytochemical staining. Claudin-5 expression was detected in hCMEC/D3, TY10, and at a very low level

  13. Permeability Barrier Generation in the Martian Lithosphere (United States)

    Schools, Joe; Montési, Laurent


    Permeability barriers develop when a magma produced in the interior of a planet rises into the cooler lithosphere and crystallizes more rapidly than the lithosphere can deform (Sparks and Parmentier, 1991). Crystallization products may then clog the porous network in which melt is propagating, reducing the permeability to almost zero, i.e., forming a permeability barrier. Subsequent melts cannot cross the barrier. Permeability barriers have been useful to explain variations in crustal thickness at mid-ocean ridges on Earth (Magde et al., 1997; Hebert and Montési, 2011; Montési et al., 2011). We explore here under what conditions permeability barriers may form on Mars.We use the MELTS thermodynamic calculator (Ghiorso and Sack, 1995; Ghiorso et al., 2002; Asimow et al., 2004) in conjunction with estimated Martian mantle compositions (Morgan and Anders, 1979; Wänke and Dreibus, 1994; Lodders and Fegley, 1997; Sanloup et al., 1999; Taylor 2013) to model the formation of permeability barriers in the lithosphere of Mars. In order to represent potential past and present conditions of Mars, we vary the lithospheric thickness, mantle potential temperature (heat flux), oxygen fugacity, and water content.Our results show that permeability layers can develop in the thermal boundary layer of the simulated Martian lithosphere if the mantle potential temperature is higher than ~1500°C. The various Martian mantle compositions yield barriers in the same locations, under matching variable conditions. There is no significant difference in barrier location over the range of accepted Martian oxygen fugacity values. Water content is the most significant influence on barrier development as it reduces the temperature of crystallization, allowing melt to rise further into the lithosphere. Our lower temperature and thicker lithosphere model runs, which are likely the most similar to modern Mars, show no permeability barrier generation. Losing the possibility of having a permeability

  14. Permeability Tests on Silkeborg Sand No. 0000

    DEFF Research Database (Denmark)

    Lund, Willy; Jakobsen, Kim Parsberg

    on the characteristics of the soil matrix, the permeability is determined for different void ratios. All tests are performed on reconstituted specimens of Silkeborg Sand No. 0000. The permeability is determined by use of a falling head apparatus. The apparatus, test procedures and the analysis method are described......The flow through porous media plays an important role in various engineering disciplines, as for example in ground water hydrology and soil mechanics. In the present study the permeability is determined for a fine, saturated sand. As the flow through a porous media strongly depends...

  15. Permeability Tests on Eastern Scheldt Sand

    DEFF Research Database (Denmark)

    Jakobsen, Kim Parsberg

    on the characteristics of the soil matrix, the permeability is determined for different void ratios. All tests are performed on reconstituted specimens of Eastern Scheldt Sand. The permeability is determined by use of a falling head apparatus. Finally the test results are briefly summarised and a relationship between......The flow through porous media plays an important role in various engineering disciplines, as for example in ground water hydrology and soil mechanics. In the present study the permeability is determined for a fine, saturated sand. As the flow through a porous media strongly depends...

  16. Predicting skin permeability from complex chemical mixtures

    International Nuclear Information System (INIS)

    Riviere, Jim E.; Brooks, James D.


    Occupational and environmental exposure to topical chemicals is usually in the form of complex chemical mixtures, yet risk assessment is based on experimentally derived data from individual chemical exposures from a single, usually aqueous vehicle, or from computed physiochemical properties. We present an approach using hybrid quantitative structure permeation relationships (QSPeR) models where absorption through porcine skin flow-through diffusion cells is well predicted using a QSPeR model describing the individual penetrants, coupled with a mixture factor (MF) that accounts for physicochemical properties of the vehicle/mixture components. The baseline equation is log k p = c + mMF + aΣα 2 H + bΣβ 2 H + sπ 2 H + rR 2 + vV x where Σα 2 H is the hydrogen-bond donor acidity, Σβ 2 H is the hydrogen-bond acceptor basicity, π 2 H is the dipolarity/polarizability, R 2 represents the excess molar refractivity, and V x is the McGowan volume of the penetrants of interest; c, m, a, b, s, r, and v are strength coefficients coupling these descriptors to skin permeability (k p ) of 12 penetrants (atrazine, chlorpyrifos, ethylparathion, fenthion, methylparathion, nonylphenol, ρ-nitrophenol, pentachlorophenol, phenol, propazine, simazine, and triazine) in 24 mixtures. Mixtures consisted of full factorial combinations of vehicles (water, ethanol, propylene glycol) and additives (sodium lauryl sulfate, methyl nicotinate). An additional set of 4 penetrants (DEET, SDS, permethrin, ricinoleic acid) in different mixtures were included to assess applicability of this approach. This resulted in a dataset of 16 compounds administered in 344 treatment combinations. Across all exposures with no MF, R 2 for absorption was 0.62. With the MF, correlations increased up to 0.78. Parameters correlated to the MF include refractive index, polarizability and log (1/Henry's Law Constant) of the mixture components. These factors should not be considered final as the focus of these studies

  17. Cocrystals of Hydrochlorothiazide: Solubility and Diffusion/Permeability Enhancements through Drug-Coformer Interactions. (United States)

    Sanphui, Palash; Devi, V Kusum; Clara, Deepa; Malviya, Nidhi; Ganguly, Somnath; Desiraju, Gautam R


    Hydrochlorothiazide (HCT) is a diuretic and a BCS class IV drug with low solubility and low permeability, exhibiting poor oral absorption. The present study attempts to improve the physicochemical properties of the drug using a crystal engineering approach with cocrystals. Such multicomponent crystals of HCT with nicotinic acid (NIC), nicotinamide (NCT), 4-aminobenzoic acid (PABA), succinamide (SAM), and resorcinol (RES) were prepared using liquid-assisted grinding, and their solubilities in pH 7.4 buffer were evaluated. Diffusion and membrane permeability were studied using a Franz diffusion cell. Except for the SAM and NIC cocrystals, all other binary systems exhibited improved solubility. All of the cocrystals showed improved diffusion/membrane permeability compared to that of HCT with the exception of the SAM cocrystal. When the solubility was high, as in the case of PABA, NCT, and RES cocrystals, the flux/permeability dropped slightly. This is in agreement with the expected interplay between solubility and permeability. Improved solubility/permeability is attributed to new drug-coformer interactions. Cocrystals of SAM, however, showed poor solubility and flux. This cocrystal contains a primary sulfonamide dimer synthon similar to that of HCT polymorphs, which may be a reason for its unusual behavior. Hirshfeld surface analysis was carried out in all cases to determine whether a correlation exists between cocrystal permeability and drug-coformer interactions.

  18. Borehole stoneley waves and permeability: Laboratory results

    International Nuclear Information System (INIS)

    Winkler, K.W.; Plona, T.J.; Froelich, B.; Liu, H.L.


    Recent interest in full waveform sonic logging has created the need for full waveform laboratory experiments on model boreholes. Of particular interest is the investigation of Stoneley waves and their interaction with permeable formations. The authors describe experimental results that show how Stoneley wave slowness and attenuation are affected by formation permeability. Both slowness and attenuation (1/Q) are observed to increase with formation permeability. This increase is frequency dependent, being greatest at low frequencies. The presence of simulated mudcakes on the borehole wall reduces the permeability effect on Stoneley waves, but does not eliminate it. The mudcake effect is frequency dependent, being greatest at low frequencies. In our experiments on rocks, the laboratory data is in qualitative agreement with theoretical predictions. In a very well characterized synthetic porous material, theory and experiment are in good quantitative agreement

  19. Octopus microvasculature: permeability to ferritin and carbon. (United States)

    Browning, J


    The permeability of Octopus microvasculature was investigated by intravascular injection of carbon and ferritin. Vessels were tight to carbon while ferritin penetrated the pericyte junction, and was found extravascularly 1-2 min after its introduction. Vesicles occurred rarely in pericytes; fenestrae were absent. The discontinuous endothelial layer did not consitute a permeability barrier. The basement membrane, although retarding the movement of ferritin, was permeable to it; carbon did not penetrate the basement membrane. Evidence indicated that ferritin, and thus similarly sized and smaller water soluble materials, traverse the pericyte junction as a result of bulk fluid flow. Comparisons are made with the convective (or junctional) and slower, diffusive (or vesicular) passage of materials known to occur across the endothelium of continuous capillaries in mammals. Previous macrophysiological determinations concerning the permeability of Octopus vessels are questioned in view of these findings. Possible reasons for some major structural differences in the microcirculatory systems of cephalopods and vertebrates are briefly discussed.

  20. Dentin Permeability of Carious Primary Teeth

    African Journals Online (AJOL)

    primary dental pulp make it difficult to determine which modality offers the best ... The most common pathology of the dentine is dental caries. ... to evaluate dentine permeability is to calculate its hydraulic conductance (Lp) using fluid filtration ...

  1. Permeability of gypsum samples dehydrated in air (United States)

    Milsch, Harald; Priegnitz, Mike; Blöcher, Guido


    We report on changes in rock permeability induced by devolatilization reactions using gypsum as a reference analog material. Cylindrical samples of natural alabaster were dehydrated in air (dry) for up to 800 h at ambient pressure and temperatures between 378 and 423 K. Subsequently, the reaction kinetics, so induced changes in porosity, and the concurrent evolution of sample permeability were constrained. Weighing the heated samples in predefined time intervals yielded the reaction progress where the stoichiometric mass balance indicated an ultimate and complete dehydration to anhydrite regardless of temperature. Porosity showed to continuously increase with reaction progress from approximately 2% to 30%, whilst the initial bulk volume remained unchanged. Within these limits permeability significantly increased with porosity by almost three orders of magnitude from approximately 7 × 10-19 m2 to 3 × 10-16 m2. We show that - when mechanical and hydraulic feedbacks can be excluded - permeability, reaction progress, and porosity are related unequivocally.

  2. Permeability After Impact Testing of Composite Laminates (United States)

    Nettles, Alan T.


    Since composite laminates are beginning to be identified for use in reusable launch vehicle propulsion systems, an understanding of their permeance is needed. A foreign object impact event can cause a localized area of permeability (leakage) in a polymer matrix composite and it is the aim of this study to assess a method of quantifying permeability-after-impact results. A simple test apparatus is presented and variables that could affect the measured values of permeability-after-impact were assessed. Once it was determined that valid numbers were being measured, a fiber/resin system was impacted at various impact levels and the resulting permeability measured, first with a leak check solution (qualitative) then using the new apparatus (quantitative). The results showed that as the impact level increased, so did the measured leakage. As the pressure to the specimen was increased, the leak rate was seen to increase in a non-linear fashion for almost all of the specimens tested.

  3. The effect of novel surfactants and Solutol HS 15 on paclitaxel aqueous solubility and permeability across a Caco-2 monolayer. (United States)

    Alani, Adam W G; Rao, Deepa A; Seidel, Ron; Wang, Jian; Jiao, Jim; Kwon, Glen S


    The effect of novel surfactants on the aqueous solubility and the permeability of paclitaxel across a Caco-2 cell monolayer were examined in this work. The solubility and permeability of paclitaxel was evaluated in the presence of four soft surfactants (SS) KXN441, KXN424, KXN437, and KXN 337 and Solutol HS15. All surfactants increased the aqueous solubility of paclitaxel. Caco-2 cell membrane integrity in the presence of SS and Solutol HS15 was assessed by mannitol permeability and LDH release. All surfactants were tested at 0.5x CMC, 5x CMC and 1.5 mM concentrations. The effect of SSs on paclitaxel permeability was concentration dependent. At all concentrations tested, KXN 441 and Solutol HS 15 showed partially inhibition of drug efflux with no discernable change in mannitol permeability or cytotoxicity as observed with LDH release. At these concentrations, other SSs exhibited some partial efflux inhibition along with compromised membrane integrity and increasing mannitol permeability. In conclusion, all SSs were able to increase the aqueous solubility and permeability of paclitaxel across Caco-2 cells monolayer. However, KXN441 and Solutol HS15 were able to enhance paclitaxel permeability across Caco-2 monolayer without cytotoxicity. (c) 2010 Wiley-Liss, Inc. and the American Pharmacists Association

  4. Surface sedimentation at permeable pavement systems

    DEFF Research Database (Denmark)

    Støvring, Jan; Dam, Torben; Jensen, Marina Bergen


    Newly installed permeable pavement (PP) systems provide high surface infiltration capacity, but the accumulation of sediments causes a decrease in capacity over time, eventually leading to surface clogging. With the aim of investigating local sedimentation processes and the importance of restorat......Newly installed permeable pavement (PP) systems provide high surface infiltration capacity, but the accumulation of sediments causes a decrease in capacity over time, eventually leading to surface clogging. With the aim of investigating local sedimentation processes and the importance...

  5. Negative permeability from random particle composites

    Energy Technology Data Exchange (ETDEWEB)

    Hussain, Shahid, E-mail:


    Artificial media, such as those composed of periodically-spaced wires for negative permittivity and split ring resonators for negative permeability have been extensively investigated for negative refractive index (NRI) applications (Smith et al., 2004; Pendry et al., 1999) [1,2]. This paper presents an alternative method for producing negative permeability: granular (or particulate) composites incorporating magnetic fillers. Artificial media, such as split-ring resonators, are designed to produce a magnetic resonance feature, which results in negative permeability over a narrow frequency range about the resonance frequency. The position of the feature is dependent upon the size of the inclusion. The material in this case is anisotropic, such that the feature is only observable when the materials are orientated in a specific direction relative to the applied field. A similar resonance can be generated in magnetic granular (particulate) materials: ferromagnetic resonance from the natural spin resonance of particles. Although the theoretical resonance profiles in granular composites shows the permeability dipping to negative values, this is rarely observed experimentally due to resonance damping effects. Results are presented for iron in spherical form and in flake form, dispersed in insulating host matrices. The two particle shapes show different permeability performance, with the magnetic flakes producing a negative contribution. This is attributed to the stronger coupling with the magnetic field resulting from the high aspect ratio of the flakes. The accompanying ferromagnetic resonance is strong enough to overcome the effects of damping and produce negative permeability. The size of random particle composites is not dictated by the wavelength of the applied field, so the materials are potentially much thinner than other, more traditional artificial composites at microwave frequencies. - Highlights: • Negative permeability from random particle composites is

  6. Sphingosine kinase inhibition alleviates endothelial permeability induced by thrombin and activated neutrophils. (United States)

    Itagaki, Kiyoshi; Zhang, Qin; Hauser, Carl J


    Inflammation and microvascular thrombosis are interrelated causes of acute lung injury in the systemic inflammatory response syndrome. Neutrophils (polymorphonuclear neutrophil [PMN]) and endothelial cells (EC) activated by systemic inflammatory response syndrome interact to increase pulmonary vascular permeability, but the interactions between PMN and EC are difficult to study. Recently, we reported that sphingosine 1-phosphate is a second messenger eliciting store-operated calcium entry (SOCE) in response to inflammatory agonists in both PMN and EC. Store-operated calcium entry is therefore a target mechanism for the therapeutic modulation of inflammatory PMN-EC interactions. Here, we isolated, modeled, and studied the effects of pharmacologic SOCE inhibition using real-time systems to monitor EC permeability after exposure to activated PMN. We created systems to continuously assess permeability of human pulmonary artery endothelial cells and human microvascular endothelial cells from lung. Endothelial cells show increased permeability after challenge by activated PMN. Such permeability increases can be attenuated by exposure of the cocultures to sphingosine kinase (SK) inhibitors (SKI-2, N,N-dimethylsphingosine [DMS]) or Ca2+ entry inhibitors (Gd3+, MRS-1845). Human microvascular endothelial cells from lung pretreated with SKI-2 or DMS showed decreased permeability when later exposed to activated PMN. Likewise, when PMNs were activated with thapsigargin (TG) in the presence of SKI-2, DMS, Gd, or MRS-1845, their ability to cause EC permeability subsequently was reduced. SKI-2 also inhibited the activation of human pulmonary artery ECs by thrombin. These studies will provide a firm mechanistic foundation for understanding how systemic SOCE inhibition may be used to prevent acute lung injury in vivo.

  7. Transformable ferroelectric control of dynamic magnetic permeability (United States)

    Jiang, Changjun; Jia, Chenglong; Wang, Fenglong; Zhou, Cai; Xue, Desheng


    Magnetic permeability, which measures the response of a material to an applied magnetic field, is crucial to the performance of magnetic devices and related technologies. Its dynamic value is usually a complex number with real and imaginary parts that describe, respectively, how much magnetic power can be stored and lost in the material. Control of permeability is therefore closely related to energy redistribution within a magnetic system or energy exchange between magnetic and other degrees of freedom via certain spin-dependent interactions. To avoid a high power consumption, direct manipulation of the permeability with an electric field through magnetoelectric coupling leads to high efficiency and simple operation, but remains a big challenge in both the fundamental physics and material science. Here we report unambiguous evidence of ferroelectric control of dynamic magnetic permeability in a Co /Pb (Mg1/3Nb2/3) 0.7Ti0.3O3 (Co/PMN-PT) heterostructure, in which the ferroelectric PMN-PT acts as an energy source for the ferromagnetic Co film via an interfacial linear magnetoelectric interaction. The electric field tuning of the magnitude and line shape of the permeability offers a highly localized means of controlling magnetization with ultralow power consumption. Additionally, the emergence of negative permeability promises a new way of realizing functional nanoscale metamaterials with adjustable refraction index.

  8. Defining clogging potential for permeable concrete. (United States)

    Kia, Alalea; Wong, Hong S; Cheeseman, Christopher R


    Permeable concrete is used to reduce urban flooding as it allows water to flow through normally impermeable infrastructure. It is prone to clogging by particulate matter and predicting the long-term performance of permeable concrete is challenging as there is currently no reliable means of characterising clogging potential. This paper reports on the performance of a range of laboratory-prepared and commercial permeable concretes, close packed glass spheres and aggregate particles of varying size, exposed to different clogging methods to understand this phenomena. New methods were developed to study clogging and define clogging potential. The tests involved applying flowing water containing sand and/or clay in cycles, and measuring the change in permeability. Substantial permeability reductions were observed in all samples, particularly when exposed to sand and clay simultaneously. Three methods were used to define clogging potential based on measuring the initial permeability decay, half-life cycle and number of cycles to full clogging. We show for the first time strong linear correlations between these parameters for a wide range of samples, indicating their use for service-life prediction. Copyright © 2018 Elsevier Ltd. All rights reserved.

  9. Comparative field permeability measurement of permeable pavements using ASTM C1701 and NCAT permeameter methods. (United States)

    Li, Hui; Kayhanian, Masoud; Harvey, John T


    Fully permeable pavement is gradually gaining support as an alternative best management practice (BMP) for stormwater runoff management. As the use of these pavements increases, a definitive test method is needed to measure hydraulic performance and to evaluate clogging, both for performance studies and for assessment of permeability for construction quality assurance and maintenance needs assessment. Two of the most commonly used permeability measurement tests for porous asphalt and pervious concrete are the National Center for Asphalt Technology (NCAT) permeameter and ASTM C1701, respectively. This study was undertaken to compare measured values for both methods in the field on a variety of permeable pavements used in current practice. The field measurements were performed using six experimental section designs with different permeable pavement surface types including pervious concrete, porous asphalt and permeable interlocking concrete pavers. Multiple measurements were performed at five locations on each pavement test section. The results showed that: (i) silicone gel is a superior sealing material to prevent water leakage compared with conventional plumbing putty; (ii) both methods (NCAT and ASTM) can effectively be used to measure the permeability of all pavement types and the surface material type will not impact the measurement precision; (iii) the permeability values measured with the ASTM method were 50-90% (75% on average) lower than those measured with the NCAT method; (iv) the larger permeameter cylinder diameter used in the ASTM method improved the reliability and reduced the variability of the measured permeability. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. Expression pattern of matrix metalloproteinases in human gynecological cancer cell lines

    International Nuclear Information System (INIS)

    Schröpfer, Andrea; Kammerer, Ulrike; Kapp, Michaela; Dietl, Johannes; Feix, Sonja; Anacker, Jelena


    Matrix metalloproteinases (MMPs) are involved in the degradation of protein components of the extracellular matrix and thus play an important role in tumor invasion and metastasis. Their expression is related to the progression of gynecological cancers (e.g. endometrial, cervical or ovarian carcinoma). In this study we investigated the expression pattern of the 23 MMPs, currently known in humans, in different gynecological cancer cell lines. In total, cell lines from three endometrium carcinomas (Ishikawa, HEC-1-A, AN3 CA), three cervical carcinomas (HeLa, Caski, SiHa), three chorioncarcinomas (JEG, JAR, BeWo), two ovarian cancers (BG-1, OAW-42) and one teratocarcinoma (PA-1) were examined. The expression of MMPs was analyzed by RT-PCR, Western blot and gelatin zymography. We demonstrated that the cell lines examined can constitutively express a wide variety of MMPs on mRNA and protein level. While MMP-2, -11, -14 and -24 were widely expressed, no expression was seen for MMP-12, -16, -20, -25, -26, -27 in any of the cell lines. A broad range of 16 MMPs could be found in the PA1 cells and thus this cell line could be used as a positive control for general MMP experiments. While the three cervical cancer cell lines expressed 10-14 different MMPs, the median expression in endometrial and choriocarcinoma cells was 7 different enzymes. The two investigated ovarian cancer cell lines showed a distinctive difference in the number of expressed MMPs (2 vs. 10). Ishikawa, Caski, OAW-42 and BeWo cell lines could be the best choice for all future experiments on MMP regulation and their role in endometrial, cervical, ovarian or choriocarcinoma development, whereas the teratocarcinoma cell line PA1 could be used as a positive control for general MMP experiments

  11. 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

    DEFF Research Database (Denmark)

    Saaby, Lasse; Tfelt-Hansen, Peer; Brodin, Birger


    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......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...

  12. Relative permeability of the endothelium and epithelium of rabbit lungs

    International Nuclear Information System (INIS)

    Effros, R.M.; Mason, G.R.; Silverman, P.; Hukkanen, J.


    Electron micrographic studies of lungs suggest that the epithelial cells are more tightly joined than the underlying endothelium, and macromolecules penetrate the endothelium more readily than the epithelium. Comparisons of epithelial and endothelial permeability to small molecules have been based upon the relative rates at which solutes traverse the alveolar-capillary barrier in fluid filled lungs and those at which they equilibrate across the capillaries in air-filled lungs. Because the former process is much slower than the latter, it has been concluded that the epithelium is less permeable to small solutes than the endothelium. However this difference may be related to inadequate access of solutes to airway surfaces. In this study, solute losses from the vascular space were compared to those from the airspace in perfused, fluid-filled rabbit lungs. 36 Cl - and 125 I - were lost from air-spaces almost twice as rapidly as 22 Na + . In contrast, the endothelium is equally permeable to 22 Na + and these anions. Loss of 3 H-mannitol from the perfusate resembled that of 22 Na + for about 30 minutes, after which diffusion of 3 H-mannitol into the tissue nearly ceased. These observations suggest that the epithelium is more permselective than the endothelium. By resisting solute and water transport, the epithelium tends to prevent alveolar flooding and confines edema to the interstitium, where it is less likely to interfere with gas exchange

  13. Permeability response of oil-contaminated compacted clays

    International Nuclear Information System (INIS)

    Silvestri, V.; Mikhail, N.; Soulie, M.


    This paper presents the results of a laboratory investigation on the behavior of motor oil-contaminated, partially saturated compacted clays. For the study, both a natural clay and an artificially purified kaolinite, contaminated with 0 to 8% of motor oil, were firstly compacted following the ASTM standard procedure. Secondly, permeability tests were carried out in a triaxial cell on 10 cm-diameter compacted clay specimens. The results of the investigation indicate that increasing percentages of motor oil decrease both the optimum water content and the optimum dry density of the two clays. However, whereas the optimum water content on the average decreases by about 6% when the percentage contamination increases from 0 to 8%, the corresponding decrease in the optimum dry density is less than 3%. Even though the optimum dry density decreases as the percentage of oil increases from 0 to 8%, there is, however, a range in oil content varying between 2 and 4% for which the optimum dry density is slightly greater than that of the untreated soils. As far as the permeability tests are concerned, the results indicate that as the percentage of oil increases, the coefficient of permeability decreases substantially, especially for clay specimens which were initially compacted on the dry side of optimum

  14. Thermal loading of bentonite. Impact on hydromechanics and permeability

    Energy Technology Data Exchange (ETDEWEB)

    Zihms, Stephanie G.; Harrington, Jon [British Geological Survey, Nickerhill Keyworth (United Kingdom)


    Due to its favorable properties, in particular, low permeability and swelling capacity, bentonite has been favored as an engineered barrier and backfill material for the geological storage of radioactive waste. To ensure safe long-term performance it is important to understand any changes in these properties when the material is subject to heat emitting waste. As such, this study will investigate the hydro-mechanical response of bentonite under multi-step thermal loading subject to a constant volume boundary condition. The experimental set up allows continuous measurements of hydraulic and mechanical response during each phase of the thermal cycle. The constant volume cell was placed inside an oven and connected to a hydraulic system with the water reservoir located externally. A pressure gradient of 4 MPa was placed across the sample for the duration of the test in order to map the evolution of permeability. After initial hydration of the bentonite, in this case signified by reaching the asymptote in total stress, the temperature was raised in 20 C increments from 20 to 80 C followed by a final 10 C step to reach 90 C. Each temperature was held constant for at least 7-10 days to allow the stresses and hydraulic transients to equilibrate. This data set will provide an insight into the hydromechanical behavior of the bentonite and the evolution of its permeability when exposed to elevated temperatures.

  15. Controlling DC permeability in cast steels

    International Nuclear Information System (INIS)

    Sumner, Aaran; Gerada, Chris; Brown, Neil; Clare, Adam


    Annealing (at multiple cooling rates) and quenching (with tempering) was performed on specimens of cast steel of varying composition. The aim was to devise a method for selecting the steel with the highest permeability, from any given range of steels, and then increasing the permeability by heat treatment. Metallographic samples were imaged using optical microscopy to show the effect of the applied heat treatments on the microstructure. Commonly cast steels can have DC permeability altered by the careful selection of a heat treatment. Increases of up to 381% were achieved by annealing using a cooling rate of 6.0 °C/min. Annealing was found to cause the carbon present in the steel to migrate from grain boundaries and from within ferrite crystals into adjacent pearlite crystals. The migration of the carbon resulted in less carbon at grain boundaries and within ferrite crystals reducing the number of pinning sites between magnetic domains. This gives rise to a higher permeability. Quenching then tempering was found to cause the formation of small ferrite crystals with the carbon content of the steel predominately held in the martensitic crystal structures. The results show that with any given range of steel compositions the highest baseline DC permeability will be found with the steel that has the highest iron content and the lowest carbon content. For the samples tested in this paper a cooling rate of 4.5 °C/min resulted in the relative permeability of the sample with the highest baseline permeability, AS4, increasing from 783 to 1479 at 0.5 T. This paper shows how heat treatments commonly applied to hypoeutectoid cast steels, to improve their mechanical performance, can be used to also enhance electromagnetic properties of these alloys. The use of cast steels allows the creation of DC components for electrical machines not possible by the widely used method of stacking of electrical grade sheet steels. - Highlights: • A range of structural steels had their

  16. Controlling DC permeability in cast steels

    Energy Technology Data Exchange (ETDEWEB)

    Sumner, Aaran, E-mail: [University of Nottingham, Nottingham University Park Campus, Nottingham NG7 2RD, England (United Kingdom); Gerada, Chris, E-mail: [Electrical Machines, University of Nottingham, Tower Building, Nottingham NG7 2RD, England (United Kingdom); Brown, Neil, E-mail: [Advanced Electrical Machines Research and Technology at Cummins Power Generation, Peterborough PE2 6FZ, England (United Kingdom); Clare, Adam, E-mail: [Advanced Manufacturing, University of Nottingham, University Park Campus, Nottingham NG7 2RD, England (United Kingdom)


    Annealing (at multiple cooling rates) and quenching (with tempering) was performed on specimens of cast steel of varying composition. The aim was to devise a method for selecting the steel with the highest permeability, from any given range of steels, and then increasing the permeability by heat treatment. Metallographic samples were imaged using optical microscopy to show the effect of the applied heat treatments on the microstructure. Commonly cast steels can have DC permeability altered by the careful selection of a heat treatment. Increases of up to 381% were achieved by annealing using a cooling rate of 6.0 °C/min. Annealing was found to cause the carbon present in the steel to migrate from grain boundaries and from within ferrite crystals into adjacent pearlite crystals. The migration of the carbon resulted in less carbon at grain boundaries and within ferrite crystals reducing the number of pinning sites between magnetic domains. This gives rise to a higher permeability. Quenching then tempering was found to cause the formation of small ferrite crystals with the carbon content of the steel predominately held in the martensitic crystal structures. The results show that with any given range of steel compositions the highest baseline DC permeability will be found with the steel that has the highest iron content and the lowest carbon content. For the samples tested in this paper a cooling rate of 4.5 °C/min resulted in the relative permeability of the sample with the highest baseline permeability, AS4, increasing from 783 to 1479 at 0.5 T. This paper shows how heat treatments commonly applied to hypoeutectoid cast steels, to improve their mechanical performance, can be used to also enhance electromagnetic properties of these alloys. The use of cast steels allows the creation of DC components for electrical machines not possible by the widely used method of stacking of electrical grade sheet steels. - Highlights: • A range of structural steels had their

  17. Permeability and elastic properties of cracked glass under pressure (United States)

    Ougier-Simonin, A.; GuéGuen, Y.; Fortin, J.; Schubnel, A.; Bouyer, F.


    Fluid flow in rocks is allowed through networks of cracks and fractures at all scales. In fact, cracks are of high importance in various applications ranging from rock elastic and transport properties to nuclear waste disposal. The present work aims at investigating thermomechanical cracking effects on elastic wave velocities, mechanical strength, and permeability of cracked glass under pressure. We performed the experiments on a triaxial cell at room temperature which allows for independent controls of the confining pressure, the axial stress, and pore pressure. We produced cracks in original borosilicate glass samples with a reproducible method (thermal treatment with a thermal shock of 300°C). The evolution of the elastic and transport properties have been monitored using elastic wave velocity sensors, strain gage, and flow measurements. The results obtained evidence for (1) a crack family with identified average aspect ratio and crack aperture, (2) a very small permeability which decreases as a power (exponential) function of pressure, and depends on (3) the crack aperture cube. We also show that permeability behavior of a cracked elastic brittle solid is reversible and independent of the fluid nature. Two independent methods (permeability and elastic wave velocity measurements) give these consistent results. This study provides data on the mechanical and transport properties of an almost ideal elastic brittle solid in which a crack population has been introduced. Comparisons with similar data on rocks allow for drawing interesting conclusions. Over the timescale of our experiments, our results do not provide any data on stress corrosion, which should be considered in further study.

  18. Phosphatidylcholine Reverses Ethanol-Induced Increase in Transepithelial Endotoxin Permeability and Abolishes Transepithelial Leukocyte Activation

    DEFF Research Database (Denmark)

    Mitzscherling, Katja; Volynets, Valentina; Parlesak, Alexandr


    Chronic alcohol abuse increases both intestinal bacterial overgrowth and intestinal permeability to macromolecules. Intestinal permeability of endotoxin, a component of the outer cell membrane of Gram-negative bacteria, plays a crucial role in the development of alcohol-induced liver disease (ALD......). As impaired bile flow leads to endotoxemia and the bile component phosphatidylcholine (PC) is therapeutically active in ALD, we tested the hypothesis that conjugated primary bile salts (CPBS) and PC inhibit ethanol-enhanced transepithelial permeability of endotoxin and the subsequent transepithelial...... activation of human leukocytes. For this purpose, we used a model in which intestinal epithelial cells (Caco-2) were basolaterally cocultivated with mononuclear leukocytes. Cells were challenged apically with endotoxin from Escherichia coli K12 and were incubated with or without the addition of CPBS (1.5 m...

  19. Phosphatidylcholine reverses ethanol-induced increase in transepithelial endotoxin permeability and abolishes transepithelial leukocyte activation

    DEFF Research Database (Denmark)

    Mitscherling, K.; Volynets, V.; Parlesak, Alexandr


    BACKGROUND: Chronic alcohol abuse increases both intestinal bacterial overgrowth and intestinal permeability to macromolecules. Intestinal permeability of endotoxin, a component of the outer cell membrane of Gram-negative bacteria, plays a crucial role in the development of alcohol-induced liver...... disease (ALD). As impaired bile flow leads to endotoxemia and the bile component phosphatidylcholine (PC) is therapeutically active in ALD, we tested the hypothesis that conjugated primary bile salts (CPBS) and PC inhibit ethanol-enhanced transepithelial permeability of endotoxin and the subsequent...... transepithelial activation of human leukocytes. METHODS: For this purpose, we used a model in which intestinal epithelial cells (Caco-2) were basolaterally cocultivated with mononuclear leukocytes. Cells were challenged apically with endotoxin from Escherichia coli K12 and were incubated with or without...

  20. Thrombin-induced increase in albumin permeability across the endothelium

    International Nuclear Information System (INIS)

    Garcia, J.G.; Siflinger-Birnboim, A.; Bizios, R.; Del Vecchio, P.J.; Fenton, J.W. II; Malik, A.B.


    We studied the effect of thrombin on albumin permeability across the endothelial monolayer in vitro. Bovine pulmonary artery endothelial cells were grown on micropore membranes. Morphologic analysis confirmed the presence of a confluent monolayer with interendothelial junctions. Albumin permeability was measured by the clearance of 125I-albumin across the endothelial monolayer. The control 125I-albumin clearance was 0.273 +/- 0.02 microliter/min. The native enzyme, alpha-thrombin (10(-6) to 10(-10) M), added to the luminal side of the endothelium produced concentration-dependent increases in albumin clearance (maximum clearance of 0.586 +/- 0.08 microliter/min at 10(-6) M). Gamma (gamma) thrombin (10(-6) M and 10(-8) M), which lacks the fibrinogen recognition site, also produced a concentration-dependent increase in albumin clearance similar to that observed with alpha-thrombin. Moreover, the two proteolytically inactive forms of the native enzyme, i-Pr2 P-alpha-thrombin and D-Phe-Pro-Arg-CH2-alpha-thrombin, increased the 125I-albumin clearance (0.610 +/- 0.09 microliter/min and 0.609 +/- 0.02 microliter/min for i-Pr2 P-alpha-thrombin and D-Phe-Pro-Arg-CH2-alpha-thrombin at 10(-6) M, respectively). Since the modified forms of thrombin lack the fibrinogen recognition and active serine protease sites, the results indicate that neither site is required for increased albumin permeability. The increase in albumin clearance with alpha-thrombin was not secondary to endothelial cell lysis because lactate dehydrogenase concentration in the medium following thrombin was not significantly different from baseline values. There was also no morphological evidence of cell lysis. Moreover, the increase in 125I-albumin clearance induced by alpha-thrombin was reversible by washing thrombin from the endothelium

  1. Effect of desensitizing agents on dentin permeability. (United States)

    Ishihata, Hiroshi; Kanehira, Masafumi; Nagai, Tomoko; Finger, Werner J; Shimauchi, Hidetoshi; Komatsu, Masashi


    To investigate the in vitro efficacy of two dentin desensitizing products at reducing liquid permeability through human dentin discs. The tested hypothesis was that the products, in spite of different chemical mechanisms were not different at reducing or eliminating flow through dentin discs. Dentin slices (1 mm thick) were prepared from 16 extracted human third molars and their permeability was indirectly recorded in a split chamber model, using a chemiluminescence technique, after EDTA treatment (control), after soaking with albumin, and after desensitizer application. Two products were studied: MS Coat, a self-curing resin-containing oxalate product, and Gluma Desensitizer, a glutaraldehyde/HEMA-based agent without initiator. The dentin slices were mounted between an upper chamber, filled with an aqueous solution of 1% potassium ferricyanide and 0.3% hydrogen peroxide, and a lower chamber filled with 1% sodium hydroxide solution and 0.02% luminol. The upper solution was pressurized, and upon contact with the luminol solution a photochemical signal was generated and recorded as a measure of permeability throughout two consecutive pressurizing cycles at 2.5 and 13 kPa (26 and 133 cm H2O), respectively. The permeability of the control and albumin-soaked samples was similarly high. After application of the desensitizing agents, dentin permeability was reduced to virtually zero at both pressure levels (P < 0.001).

  2. Hydrogen concentration control utilizing a hydrogen permeable membrane

    International Nuclear Information System (INIS)

    Keating, S.J. Jr.


    The concentration of hydrogen in a fluid mixture is controlled to a desired concentration by flowing the fluid through one chamber of a diffusion cell separated into two chambers by a hydrogen permeable membrane. A gradient of hydrogen partial pressure is maintained across the membrane to cause diffusion of hydrogen through the membrane to maintain the concentration of hydrogen in the fluid mixture at the predetermined level. The invention has particular utility for the purpose of injecting into and/or separating hydrogen from the reactor coolant of a nuclear reactor system

  3. Myxoviruses do not induce non-specific alterations in membrane permeability early on in infection

    International Nuclear Information System (INIS)

    Foster, K.A.; Micklem, K.J.; Bogomolova, N.N.; Boriskin, Y.S.; Pasternak, C.A.


    The permeability characteristics of cells infected with myxoviruses have been studied by measuring the concentrative uptake of nutrients, the concentration of intracellular K + , and the maintenance of the Na + gradient across the plasma membrane. Cells either show no change at all (Sendai virus-infected BHK cells and measles virus-infected Vero cells) or they show a decreased ability to concentrate nutrients, while intracellular K + and the Na + gradient remain unchanged (Sendai and influenza virus-infected L-1210 cells, measles virus-infected lymphocytes and mumps virus-infected L-41 cells). In no case, therefore, was a change observed that resembles the non-specific increase in membrane permeability induced by haemolytic paramyxoviruses (35, 42) or the non-specific membrane leakiness postulated to take place in infected cells (8, 9). A preliminary account of some of these findings has been presented (39)

  4. Experimental Studies on Permeability of Intact and Singly Jointed Meta-Sedimentary Rocks Under Confining Pressure (United States)

    Wong, Louis Ngai Yuen; Li, Diyuan; Liu, Gang


    Three different types of permeability tests were conducted on 23 intact and singly jointed rock specimens, which were cored from rock blocks collected from a rock cavern under construction in Singapore. The studied rock types belong to inter-bedded meta-sandstone and meta-siltstone with very low porosity and high uniaxial compressive strength. The transient pulse water flow method was employed to measure the permeability of intact meta-sandstone under a confining pressure up to 30 MPa. It showed that the magnitude order of meta-sandstone's intrinsic permeability is about 10-18 m2. The steady-state gas flow method was used to measure the permeability of both intact meta-siltstone and meta-sandstone in a triaxial cell under different confining pressures spanning from 2.5 to 10 MPa. The measured permeability of both rock types ranged from 10-21 to 10-20 m2. The influence of a single natural joint on the permeability of both rock types was studied by using the steady-state water flow method under different confining pressures spanning from 1.25 to 5.0 MPa, including loading and unloading phases. The measured permeability of both jointed rocks ranged from 10-13 to 10-11 m2, where the permeability of jointed meta-siltstone was usually slightly lower than that of jointed meta-sandstone. The permeability of jointed rocks decreases with increasing confining pressure, which can be well fitted by an empirical power law relationship between the permeability and confining pressure or effective pressure. The permeability of partly open cracked specimens is lower than that of open cracked specimens, but it is higher than that of the specimen with a dominant vein for the meta-sandstone under the same confining pressure. The permeability of open cracked rock specimens will partially recover during the unloading confining pressure process. The equivalent crack (joint) aperture is as narrow as a magnitude order of 10-6 m (1 μm) in the rock specimens under confining pressures

  5. Multiple Targets for Novel Therapy of FSGS Associated with Circulating Permeability Factor

    Directory of Open Access Journals (Sweden)

    Virginia J. Savin


    Full Text Available A plasma component is responsible for altered glomerular permeability in patients with focal segmental glomerulosclerosis. Evidence includes recurrence after renal transplantation, remission after plasmapheresis, proteinuria in infants of affected mothers, transfer of proteinuria to experimental animals, and impaired glomerular permeability after exposure to patient plasma. Therapy may include decreasing synthesis of the injurious agent, removing or blocking its interaction with cells, or blocking signaling or enhancing cell defenses to restore the permeability barrier and prevent progression. Agents that may prevent the synthesis of the permeability factor include cytotoxic agents or aggressive chemotherapy. Extracorporeal therapies include plasmapheresis, immunoadsorption with protein A or anti-immunoglobulin, or lipopheresis. Oral or intravenous galactose also decreases Palb activity. Studies of glomeruli have shown that several strategies prevent the action of FSGS sera. These include blocking receptor-ligand interactions, modulating cell reactions using indomethacin or eicosanoids 20-HETE or 8,9-EET, and enhancing cytoskeleton and protein interactions using calcineurin inhibitors, glucocorticoids, or rituximab. We have identified cardiotrophin-like cytokine factor 1 (CLCF-1 as a candidate for the permeability factor. Therapies specific to CLCF-1 include potential use of cytokine receptor-like factor (CRLF-1 and inhibition of Janus kinase 2. Combined therapy using multiple modalities offers therapy to reverse proteinuria and prevent scarring.

  6. In situ permeability testing of rock salt

    International Nuclear Information System (INIS)

    Peterson, E.W.; Lagus, P.L.; Broce, R.D.; Lie, K.


    Storage of transuranic (TRU) wastes in bedded salt formations requires a knowledge of the in situ permeability of SENM rock salt. Since assumptions for safety assessments have been made in which these wastes could generate gas pressures on the order of the lithostatic pressure over geologic time scales, the permeability of the surrounding formation becomes an important parameter for determining the manner in which the gases will be contained or dispersed. This report describes the series of tests conducted in the AEC-7 borehole, located near the WIPP site, to determine the in situ gas flow characteristics of the bedded salt. In these tests, compressed air was injected into the borehole and flow into the surrounding formation measured. These measured flow rates were interpreted in terms of formation permeabilities and porosities which were, in turn, used as modeling parameters for the repository response analysis. Two series of field tests were performed. The first series consisted of a number of whole-hole flow tests conducted to provide preliminary design information required for future operation of a guarded straddle packer system capable of measuring permeabilities > or = 0.1 μdarcy. The second series of tests were conducted using the Systems, Science and Software (S-Cubed) designed guarded straddle packer system. In these interval permeability tests, 100-foot lengths of borehole were isolated and the flow characteristics of the surrounding formation examined. In this report, a complete description of the test procedures, instrumentation, and measurement techniques is first given. The analytical/numerical methods used for data interpretation are then presented, followed by results of the interval and permeability tests. (The whole-hole tests are summarized in Appendix A.) Conclusions are presented in the final section

  7. Role of platelets in maintenance of pulmonary vascular permeability to protein

    International Nuclear Information System (INIS)

    Lo, S.K.; Burhop, K.E.; Kaplan, J.E.; Malik, A.B.


    The authors examined the role of platelets in maintenance of pulmonary vascular integrity by inducing thrombocytopenia in sheep using antiplatelet serum (APS). A causal relationship between thrombocytopenia and increase in pulmonary vascular permeability was established by platelet repletion using platelet-rich plasma (PRP). Sheep were chronically instrumented and lung lymph fistulas prepared to monitor pulmonary lymph flow (Q lym ). A balloon catheter was positioned in the left atrium to assess pulmonary vascular permeability to protein after raising the left atrial pressure (P la ). Thrombocytopenia was maintained for 3 days by daily intramuscular APS injections. In studies using cultured bovine pulmonary artery endothelial monolayers, transendothelia permeability of 125 I-labeled albumin was reduced 50 and 95%, respectively, when 2.5 x 10 7 or 5 x 10 7 platelets were added onto endothelial monolayers. However, addition of 5 x 10 6 platelets or 5 x 10 7 red blood cells did not reduce endothelial monolayer albumin permeability. Results indicate that platelets are required for the maintenance of pulmonary vascular permeability. Reduction in permeability appears to involve an interaction of platelets with the endothelium

  8. Calcium reduces the sodium permeability of luminal membrane vesicles from toad bladder. Studies using a fast-reaction apparatus

    International Nuclear Information System (INIS)

    Chase, H.S. Jr.; Al-Awqati, Q.


    Regulation of the sodium permeability of the luminal membrane is the major mechanism by which the net rate of sodium transport across tight epithelia is varied. Previous evidence has suggested that the permeability of the luminal membrane might be regulated by changes in intracellular sodium or calcium activities. To test this directly, we isolated a fraction of the plasma membrane from the toad urinary bladder, which contains a fast, amiloride-sensitive sodium flux with characteristics similar to those of the native luminal membrane. Using a flow-quench apparatus to measure the initial rate of sodium efflux from these vesicles in the millisecond time range, we have demonstrated that the isotope exchange permeability of these vesicles is very sensitive to calcium. Calcium reduces the sodium permeability, and the half-maximal inhibitory concentration is 0.5 microM, well within the range of calcium activity found in cells. Also, the permeability of the luminal membrane vesicles is little affected by the ambient sodium concentration. These results, when taken together with studies on whole tissue, suggest that cell calcium may be an important regulator of transepithelial sodium transport by its effect on luminal sodium permeability. The effect of cell sodium on permeability may be mediated by calcium rather than by sodium itself

  9. Small-bowel permeability in collagenous colitis

    DEFF Research Database (Denmark)

    Wildt, Signe; Madsen, Jan L; Rumessen, Jüri J


    Collagenous colitis (CC) is a chronic inflammatory bowel disease that affects the colon. However, some patients with CC present with accompanying pathologic small-bowel manifestations such as coeliac disease, defects in bile acid absorption and histopathologic changes in small-intestinal biopsies......, indicating that CC is a pan-intestinal disease. In small-intestinal disease, the intestinal barrier function may be impaired, and the permeability of the small intestine altered. The purpose of this research was to study small-bowel function in patients with CC as expressed by intestinal permeability....

  10. Dopamine enhances duodenal epithelial permeability via the dopamine D5 receptor in rodent. (United States)

    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 D 1 (D 1 R) and D 5 (D 5 R), but whether D1-like receptors influence the duodenal permeability is unclear. FITC-dextran permeability, short-circuit current (I SC ), Western blot, immunohistochemistry and ELISA were used in human D 5 R transgenic mice and hyperendogenous enteric DA (HEnD) rats in this study. Dopamine induced a downward deflection in I SC 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 D 5 R, but not D 1 R, was observed in the duodenum of control rat. In human D 5 R knock-in transgenic mice, duodenal mucosa displayed an increased basal I SC 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. D 5 R knock-down transgenic mice manifested a decreased basal I SC 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 D 5 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.

  11. Acylation of salmon calcitonin modulates in vitro intestinal peptide flux through membrane permeability enhancement

    DEFF Research Database (Denmark)

    Trier, Sofie; Linderoth, Lars; Bjerregaard, Simon


    hypothesize that tailoring the acylation may be used to optimize intestinal translocation. This work aims to characterize acylated analogues of the therapeutic peptide salmon calcitonin (sCT), which lowers blood calcium, by systematically increasing acyl chain length at two positions, in order to elucidate...... to be optimal, as elongating the chain causes greater binding to the cell membrane but similar permeability, and we speculate that increasing the chain length further may decrease the permeability. In conclusion, acylated sCT acts as its own in vitro intestinal permeation enhancer, with reversible effects...... on Caco-2 cells, indicating that acylation of sCT may represent a promising tool to increase intestinal permeability without adding oral permeation enhancers....

  12. Estimation of non-linear effective permeability of magnetic materials with fine structure

    International Nuclear Information System (INIS)

    Waki, H.; Igarashi, H.; Honma, T.


    This paper describes a homogenization method for magnetic materials with fine structure. In this method, the structures of the magnetic materials are assumed to be periodic, and the unit cell is defined. The effective permeability is determined on the basis of magnetic energy balance in the unit cell. This method can be applied not only for linear problems but also for non-linear ones. In this paper, estimation of the effective permeability of non-linear magnetic materials by using the homogenization method is described in detail, and then the validity for the non-liner problems is tested for two-dimensional problems. It is shown that this homogenization method gives accurate non-linear effective permeability

  13. High-permeability criterion for BCS classification: segmental/pH dependent permeability considerations. (United States)

    Dahan, Arik; Miller, Jonathan M; Hilfinger, John M; Yamashita, Shinji; Yu, Lawrence X; Lennernäs, Hans; Amidon, Gordon L


    The FDA classifies a drug substance as high-permeability when the fraction of dose absorbed (F(abs)) in humans is 90% or higher. This direct correlation between human permeability and F(abs) has been recently controversial, since the β-blocker sotalol showed high F(abs) (90%) and low Caco-2 permeability. The purpose of this study was to investigate the scientific basis for this disparity between permeability and F(abs). The effective permeabilities (P(eff)) of sotalol and metoprolol, a FDA standard for the low/high P(eff) class boundary, were investigated in the rat perfusion model, in three different intestinal segments with pHs corresponding to the physiological pH in each region: (1) proximal jejunum, pH 6.5; (2) mid small intestine, pH 7.0; and (3) distal ileum, pH 7.5. Both metoprolol and sotalol showed pH-dependent permeability, with higher P(eff) at higher pH. At any given pH, sotalol showed lower permeability than metoprolol; however, the permeability of sotalol determined at pH 7.5 exceeded/matched metoprolol's at pH 6.5 and 7.0, respectively. Physicochemical analysis based on ionization, pK(a) and partitioning of these drugs predicted the same trend and clarified the mechanism behind these observed results. Experimental octanol-buffer partitioning experiments confirmed the theoretical curves. An oral dose of metoprolol has been reported to be completely absorbed in the upper small intestine; it follows, hence, that metoprolol's P(eff) value at pH 7.5 is not likely physiologically relevant for an immediate release dosage form, and the permeability at pH 6.5 represents the actual relevant value for the low/high permeability class boundary. Although sotalol's permeability is low at pH 6.5 and 7.0, at pH 7.5 it exceeds/matches the threshold of metoprolol at pH 6.5 and 7.0, most likely responsible for its high F(abs). In conclusion, we have shown that, in fact, there is no discrepancy between P(eff) and F(abs) in sotalol's absorption; the data emphasize that

  14. Pulmonary and intestinal permeabilities in alcoholic hepatic cirrhosis

    International Nuclear Information System (INIS)

    De Botton, S.; Huglo, B.; Canva-Delacambre, V.; Colombel, J.F.; Beauchat, V.; Ziegels, P.; Prangere, T.; Steinling, M.; Machandise, X.; Wallaert, B.


    The aim of this prospective study was to evaluate simultaneously the intestinal permeability (IP), usually normal, and the pulmonary permeability, (PP) rather rarely studied, in patients afflicted with hepatic cirrhosis of alcoholic (HCA) origin. Thirty five non-smoker patients, afflicted with HCA, proved by biopsy, without pulmonary pathology and with normal pulmonary scanography were subject to our investigation. The pre-graft hepatic examination contained also respiratory functional explorations as well as bronchi-alveolar clearance (BAC) explorations. After inhalation of the DTPA- 99m Tc aerosols, a 20 min dynamical study in posterior-front condition was achieved. After exponential matching on the activity/time curve of the right lung, the half life (T 1/2 in min) and the Residual Activity at 10 min (RA in %) were calculated. The PI were than estimated and on the basis of urinary activity of EDTA- 51 Cr obtained on 24 h and expressed in % of the uptake activity, according to the Bjarnasson's technique. The results were compared (significant non-parametric tests if p 1/2 and 87.1% ± 6.7 vs 92.8% ± 2.6 (p < 0.002) for RA. It is significantly correlated with the total number of cells (r = -0.379) and with the number of lymphocytes (r = 0.351) in the BAC. For the first time an enhanced PP was observed in HCA, correlated with the increase in the number of cells at BAC

  15. Water permeability of pigmented waterborne coatings

    NARCIS (Netherlands)

    Donkers, P.A.J.; Huinink, H.P.; Erich, S.J.F.; Reuvers, N.J.W.; Adan, O.C.G.


    Coatings are used in a variety of applications. Last decades more and more coating systems are transforming from solvent to waterborne coating systems. In this study the influence of pigments on the water permeability of a waterborne coating system is studied, with special interest in the possible

  16. Foam film permeability: theory and experiment. (United States)

    Farajzadeh, R; Krastev, R; Zitha, Pacelli L J


    The mass transfer of gas through foam films is a prototype of various industrial and biological processes. The aim of this paper is to give a perspective and critical overview of studies carried out to date on the mass transfer of gas through foam films. Contemporary experimental data are summarized, and a comprehensive overview of the theoretical models used to explain the observed effects is given. A detailed description of the processes that occur when a gas molecule passes through each layer that forms a foam film is shown. The permeability of the film-building surfactant monolayers plays an important role for the whole permeability process. It can be successfully described by the models used to explain the permeability of surfactant monolayers on aqueous sub-phase. For this reason, the present paper briefly discusses the surfactant-induced resistance to mass transfer of gases through gas-liquid interface. One part of the paper discusses the experimental and theoretical aspects of the foam film permeability in a train of foam films in a matrix or a cylinder. This special case is important to explain the gas transfer in porous media or in foams. Finally, this paper will highlight the gaps and challenges and sketch possible directions for future research.

  17. Color-magnetic permeability of QCD vacuum

    Energy Technology Data Exchange (ETDEWEB)

    Saito, T [Kyoto Prefectural Univ. of Medicine (Japan); Shigemoto, K


    In the very strong background gauge field the QCD true vacuum has been shown to have lower energy than the ''perturbative vacuum.'' The color-magnetic permeability of the QCD true vacuum is then calculated to be 1/2 within the quark-one-loop approximation.

  18. The Permeability of Rubble Mound Breakwaters

    DEFF Research Database (Denmark)

    Williams, A.F.; Burcharth, H. F.; Adel, H. den


    . 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...

  19. Vascular permeability in cerebral cavernous malformations

    DEFF Research Database (Denmark)

    Mikati, Abdul G; Khanna, Omaditya; Zhang, Lingjiao


    Patients with the familial form of cerebral cavernous malformations (CCMs) are haploinsufficient for the CCM1, CCM2, or CCM3 gene. Loss of corresponding CCM proteins increases RhoA kinase-mediated endothelial permeability in vitro, and in mouse brains in vivo. A prospective case-controlled observ...

  20. Programs for the calculi of blocks permeabilities

    International Nuclear Information System (INIS)

    Gomez Hernandez, J.J.; Sovero Sovero, H.F.


    This report studies the stochastic analysis of radionuclide transport. The permeability values of blocks are necessary to do a numeric model for the flux and transport problems in ground soils. The determination of block value by function on grill value is the objective of this program

  1. Radionuclide assessment of pulmonary microvascular permeability

    Energy Technology Data Exchange (ETDEWEB)

    Groeneveld, A.B.J. [Medical Intensive Care Unit, Department of Internal Medicine, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam (Netherlands)


    The literature has been reviewed to evaluate the technique and clinical value of radionuclide measurements of microvascular permeability and oedema formation in the lungs. Methodology, modelling and interpretation vary widely among studies. Nevertheless, most studies agree on the fact that the measurement of permeability via pulmonary radioactivity measurements of intravenously injected radiolabelled proteins versus that in the blood pool, the so-called pulmonary protein transport rate (PTR), can assist the clinician in discriminating between permeability oedema of the lungs associated with the adult respiratory distress syndrome (ARDS) and oedema caused by an increased filtration pressure, for instance in the course of cardiac disease, i.e. pressure-induced pulmonary oedema. Some of the techniques used to measure PTR are also able to detect subclinical forms of lung microvascular injury not yet complicated by permeability oedema. This may occur after cardiopulmonary bypass and major vascular surgery, for instance. By paralleling the clinical severity and course of the ARDS, the PTR method may also serve as a tool to evaluate new therapies for the syndrome. Taken together, the currently available radionuclide methods, which are applicable at the bedside in the intensive care unit, may provide a gold standard for detecting minor and major forms of acute microvascular lung injury, and for evaluating the severity, course and response to treatment. (orig.). With 2 tabs.

  2. Thromboxane A2 increases endothelial permeability through upregulation of interleukin-8

    International Nuclear Information System (INIS)

    Kim, Su-Ryun; Bae, Soo-Kyung; Park, Hyun-Joo; Kim, Mi-Kyoung; Kim, Koanhoi; Park, Shi-Young; Jang, Hye-Ock; Yun, Il; Kim, Yung-Jin; Yoo, Mi-Ae; Bae, Moon-Kyoung


    Thromboxane A 2 (TXA 2 ), a major prostanoid formed from prostaglandin H 2 by thromboxane synthase, is involved in the pathogenesis of a variety of vascular diseases. In this study, we report that TXA 2 mimetic U46619 significantly increases the endothelial permeability both in vitro and in vivo. U46619 enhanced the expression and secretion of interleukin-8 (IL-8), a major inducer of vascular permeability, in endothelial cells. Promoter analysis showed that the U46619-induced expression of IL-8 was mainly regulated by nuclear factor-κB (NF-κB). U46619 induced the activation of NF-κB through IκB kinase (IKK) activation, IκB phosphorylation and NF-κB nuclear translocation. Furthermore, the inhibition of IL-8 or blockade of the IL-8 receptor attenuated the U46619-induced endothelial cell permeability by modulating the cell-cell junctions. Overall, these results suggest that U46619 promotes vascular permeability through the production of IL-8 via NF-κB activation in endothelial cells.

  3. A new water permeability measurement method for unsaturated tight materials using saline solutions

    International Nuclear Information System (INIS)

    Malinsky, Laurent; Talandier, Jean


    Document available in extended abstract form only. Relative water permeability of material in a radioactive waste disposal is a key parameter to simulate and predict saturation state evolution. In this paper we present a new measurement method and the results obtained for Callovo-Oxfordian (Cox) clay-stone, host rock of the underground Andra laboratory at Bure (Meuse/Haute-Marne). Relative water permeability of such a low permeability rock as Cox clay-stone has been measured up to now by an indirect method. It consists in submitting a rock sample to successive relative humidity steps imposed by saline solutions. The transient mass variation during each step and the mass at hydric equilibrium are interpreted generally by using an inverse analysis method. The water relative permeability function of water saturation is derived from water diffusion coefficient evolution and water retention curve. The proposed new method consists in directly measuring the water flux across a flat cylindrical submitted to a relative humidity gradient. Two special cells have been developed. The tightness of the lateral sample surface is insured by crushing a polyurethane ring surrounding the sample set in an aluminium device placed over a Plexiglas vessel filled with a saline solution. One of the cells is designed to allow humidity measurement in the cell. These cells can also be used to measure the relative humidity produced by a saline solution or by an unsaturated material. During a permeability measurement, the cell with the sample to be tested is continuously weighted in a Plexiglas box in which a saline solution imposes a different relative humidity at the upper sample face. The experimental set-up is shown on Figure 1. The mean permeability of the sample is proportional to the rate of mass variation when steady state is reached. The result of one test is shown on Figure 2(a). Twenty four permeability measurements have been performed on four argillite samples of 15 mm in height and

  4. Astrocyte–endothelial interactions and blood–brain barrier permeability* (United States)

    Abbott, N Joan


    The blood–brain barrier (BBB) is formed by brain endothelial cells lining the cerebral microvasculature, and is an important mechanism for protecting the brain from fluctuations in plasma composition, and from circulating agents such as neurotransmitters and xenobiotics capable of disturbing neural function. The barrier also plays an important role in the homeostatic regulation of the brain microenvironment necessary for the stable and co-ordinated activity of neurones. The BBB phenotype develops under the influence of associated brain cells, especially astrocytic glia, and consists of more complex tight junctions than in other capillary endothelia, and a number of specific transport and enzyme systems which regulate molecular traffic across the endothelial cells. Transporters characteristic of the BBB phenotype include both uptake mechanisms (e.g. GLUT-1 glucose carrier, L1 amino acid transporter) and efflux transporters (e.g. P-glycoprotein). In addition to a role in long-term barrier induction and maintenance, astrocytes and other cells can release chemical factors that modulate endothelial permeability over a time-scale of seconds to minutes. Cell culture models, both primary and cell lines, have been used to investigate aspects of barrier induction and modulation. Conditioned medium taken from growing glial cells can reproduce some of the inductive effects, evidence for involvement of diffusible factors. However, for some features of endothelial differentiation and induction, the extracellular matrix plays an important role. Several candidate molecules have been identified, capable of mimicking aspects of glial-mediated barrier induction of brain endothelium; these include TGFβ, GDNF, bFGF, IL-6 and steroids. In addition, factors secreted by brain endothelial cells including leukaemia inhibitory factor (LIF) have been shown to induce astrocytic differentiation. Thus endothelium and astrocytes are involved in two-way induction. Short-term modulation of brain

  5. Predicting permeability of regular tissue engineering scaffolds: scaling analysis of pore architecture, scaffold length, and fluid flow rate effects. (United States)

    Rahbari, A; Montazerian, H; Davoodi, E; Homayoonfar, S


    The main aim of this research is to numerically obtain the permeability coefficient in the cylindrical scaffolds. For this purpose, a mathematical analysis was performed to derive an equation for desired porosity in terms of morphological parameters. Then, the considered cylindrical geometries were modeled and the permeability coefficient was calculated according to the velocity and pressure drop values based on the Darcy's law. In order to validate the accuracy of the present numerical solution, the obtained permeability coefficient was compared with the published experimental data. It was observed that this model can predict permeability with the utmost accuracy. Then, the effect of geometrical parameters including porosity, scaffold pore structure, unit cell size, and length of the scaffolds as well as entrance mass flow rate on the permeability of porous structures was studied. Furthermore, a parametric study with scaling laws analysis of sample length and mass flow rate effects on the permeability showed good fit to the obtained data. It can be concluded that the sensitivity of permeability is more noticeable at higher porosities. The present approach can be used to characterize and optimize the scaffold microstructure due to the necessity of cell growth and transferring considerations.

  6. Sub-core permeability and relative permeability characterization with Positron Emission Tomography (United States)

    Zahasky, C.; Benson, S. M.


    This study utilizes preclinical micro-Positron Emission Tomography (PET) to image and quantify the transport behavior of pulses of a conservative aqueous radiotracer injected during single and multiphase flow experiments in a Berea sandstone core with axial parallel bedding heterogeneity. The core is discretized into streamtubes, and using the micro-PET data, expressions are derived from spatial moment analysis for calculating sub-core scale tracer flux and pore water velocity. Using the flux and velocity data, it is then possible to calculate porosity and saturation from volumetric flux balance, and calculate permeability and water relative permeability from Darcy's law. Full 3D simulations are then constructed based on this core characterization. Simulation results are compared with experimental results in order to test the assumptions of the simple streamtube model. Errors and limitations of this analysis will be discussed. These new methods of imaging and sub-core permeability and relative permeability measurements enable experimental quantification of transport behavior across scales.

  7. Remote assessment of permeability/thermal diffusivity of consolidated clay sediments

    International Nuclear Information System (INIS)

    Lovell, M.A.; Ogden, P.


    The aim of this project was to examine the feasibility of predicting marine sediment permeability and thermal diffusivity by remote geophysical observations. For this purpose a modified consolidation cell was developed and constructed and tests on deep sea sediment samples carried out. Results and conclusions of a nineteen month programme are presented. (U.K.)

  8. Permeability, diffusivity and solubility of carbon dioxide in fluoropolymers: An experimental and modeling study

    DEFF Research Database (Denmark)

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

    Directory of Open Access Journals (Sweden)

    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.

  10. Solubility and Permeability Studies of Aceclofenac in Different Oils

    African Journals Online (AJOL)

    The solubility and permeability of aceclofenac were compared with the hydroalcoholic solution of ... the use of lipid based systems such as micro- or .... carriers/vehicles for enhanced solubility and permeability ... modifications: A recent review.

  11. Investigation clogging dynamic of permeable pavement systems using embedded sensors (United States)

    Permeable pavement is a stormwater control measure commonly selected in both new and retrofit applications. However, there is limited information about the clogging mechanism of these systems that effects the infiltration. A permeable pavement site located at the Seitz Elementary...

  12. Preliminary study of soil permeability properties using principal component analysis (United States)

    Yulianti, M.; Sudriani, Y.; Rustini, H. A.


    Soil permeability measurement is undoubtedly important in carrying out soil-water research such as rainfall-runoff modelling, irrigation water distribution systems, etc. It is also known that acquiring reliable soil permeability data is rather laborious, time-consuming, and costly. Therefore, it is desirable to develop the prediction model. Several studies of empirical equations for predicting permeability have been undertaken by many researchers. These studies derived the models from areas which soil characteristics are different from Indonesian soil, which suggest a possibility that these permeability models are site-specific. The purpose of this study is to identify which soil parameters correspond strongly to soil permeability and propose a preliminary model for permeability prediction. Principal component analysis (PCA) was applied to 16 parameters analysed from 37 sites consist of 91 samples obtained from Batanghari Watershed. Findings indicated five variables that have strong correlation with soil permeability, and we recommend a preliminary permeability model, which is potential for further development.

  13. Development of calcium-permeable alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors in cultured neocortical neurons visualized by cobalt staining

    DEFF Research Database (Denmark)

    Jensen, J B; Schousboe, A; Pickering, D S


    The developmental expression of calcium (Ca2+)-permeable alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and kainate receptors in cultured neocortical neurons was evaluated by using cobalt uptake, a histochemical method that identifies cells expressing Ca2+-permeable, non-N-methyl-D-aspartate...

  14. Long-term Metal Performance of Three Permeable Pavements (United States)

    EPA constructed a 4,000-m2 parking lot surfaced with three permeable pavements (permeable interlocking concrete pavers, pervious concrete, and porous asphalt) on the Edison Environmental Center in Edison, NJ in 2009. Samples from each permeable pavement infiltrate were collected...

  15. The kinetics of denitrification in permeable sediments

    DEFF Research Database (Denmark)

    Evrard, Victor; Glud, Ronnie N.; Cook, Perran L. M.


    Permeable sediments comprise the majority of shelf sediments, yet the rates of denitrification remain highly uncertain in these environments. Computational models are increasingly being used to understand the dynamics of denitrification in permeable sediments, which are complex environments...... on sediments taken from six shallow coastal sites in Port Phillip Bay, Victoria, Australia. The results showed that denitrification commenced rapidly (within 30 min) after the onset of anoxia and the kinetics could be well described by Michaelis-Menten kinetics with half saturation constants (apparent K...... in cohesive sediments despite organic carbon contents one order of magnitude lower for the sediments studied here. The ratio of sediment O-2 consumption to V-max was in the range of 0.02-0.09, and was on average much lower than the theoretical ratio of 0.8. As a consequence, models implemented...

  16. Nonequilibrium gas absorption in rotating permeable media (United States)

    Baev, V. K.; Bazhaikin, A. N.


    The absorption of ammonia, sulfur dioxide, and carbon dioxide by water and aqueous solutions in rotating permeable media, a cellular porous disk, and a set of spaced-apart thin disks has been considered. The efficiency of cleaning air to remove these impurities is determined, and their anomalously high solubility (higher than equilibrium value) has been discovered. The results demonstrate the feasibility of designing cheap efficient rotor-type absorbers to clean gases of harmful impurities.

  17. A Reconciliation of Packed Column Permeability Data: Column Permeability as a Function of Particle Porosity

    Directory of Open Access Journals (Sweden)

    Hubert M. Quinn


    Full Text Available In his textbook teaching of packed bed permeability, Georges Guiochon uses mobile phase velocity as the fluid velocity term in his elaboration of the Darcy permeability equation. Although this velocity frame makes a lot of sense from a thermodynamic point of view, it is valid only with respect to permeability at a single theoretical boundary condition. In his more recent writings, however, Guiochon has departed from his long-standing mode of discussing permeability in terms of the Darcy equation and has embraced the well-known Kozeny-Blake equation. In this paper, his teaching pertaining to the constant in the Kozeny-Blake equation is examined and, as a result, a new correlation coefficient is identified and defined herein based on the velocity frame used in his teaching. This coefficient correlates pressure drop and fluid velocity as a function of particle porosity. We show that in their experimental protocols, Guiochon et al. have not adhered to a strict material balance of permeability which creates a mismatch of particle porosity and leads to erroneous conclusions regarding the value of the permeability coefficient in the Kozeny-Blake equation. By correcting the experimental data to properly reflect particle porosity we reconcile the experimental results of Guiochon and Giddings, resulting in a permeability reference chart which is presented here for the first time. This reference chart demonstrates that Guiochon’s experimental data, when properly normalized for particle porosity and other related discrepancies, corroborates the value of 267 for the constant in the Kozeny-Blake equation which was derived by Giddings in 1965.

  18. Atrial natriuretic factor increases vascular permeability

    International Nuclear Information System (INIS)

    Lockette, W.; Brennaman, B.


    An increase in central blood volume in microgravity may result in increased plasma levels of atrial natriuretic factor (ANF). Since elevations in plasma ANF are found in clinical syndromes associated with edema, and since space motion sickness induced by microgravity is associated with an increase in central blood volume and facial edema, we determined whether ANF increases capillary permeability to plasma protein. Conscious, bilaterally nephrectomized male rats were infused with either saline, ANF + saline, or hexamethonium + saline over 2 h following bolus injections of 125I-albumin and 14C-dextran of similar molecular size. Blood pressure was monitored and serial determinations of hematocrits were made. Animals infused with 1.0 ANF had significantly higher hematocrits than animals infused with saline vehicle. Infusion of ANF increased the extravasation of 125I-albumin, but not 14C-dextran from the intravascular compartment. ANF also induced a depressor response in rats, but the change in blood pressure did not account for changes in capillary permeability to albumin; similar depressor responses induced by hexamethonium were not accompanied by increased extravasation of albumin from the intravascular compartment. ANF may decrease plasma volume by increasing permeability to albumin, and this effect of ANF may account for some of the signs and symptoms of space motion sickness

  19. Ammonia gas permeability of meat packaging materials. (United States)

    Karim, Faris; Hijaz, Faraj; Kastner, Curtis L; Smith, J Scott


    Meat products are packaged in polymer films designed to protect the product from exterior contaminants such as light, humidity, and harmful chemicals. Unfortunately, there is almost no data on ammonia permeability of packaging films. We investigated ammonia permeability of common meat packaging films: low-density polyethylene (LDPE; 2.2 mil), multilayer polyolefin (MLP; 3 mil), and vacuum (V-PA/PE; 3 mil, 0.6 mil polyamide/2.4 mil polyethylene). The films were fabricated into 10 × 5 cm pouches and filled with 50 mL deionized water. Pouches were placed in a plexiglass enclosure in a freezer and exposed to 50, 100, 250, or 500 ppm ammonia gas for 6, 12, 24, and 48 h at -17 ± 3 °C and 21 ± 3 °C. At freezing temperatures, no ammonia residues were detected and no differences in pH were found in the water. At room temperature, ammonia levels and pH of the water increased significantly (P packaging materials have low ammonia permeability and protect meat products exposed to ammonia leaks during frozen storage.

  20. Permeable treatment wall design and cost analysis

    International Nuclear Information System (INIS)

    Manz, C.; Quinn, K.


    A permeable treatment wall utilizing the funnel and gate technology has been chosen as the final remedial solution for one industrial site, and is being considered at other contaminated sites, such as a closed municipal landfill. Reactive iron gates will be utilized for treatment of chlorinated VOCs identified in the groundwater. Alternatives for the final remedial solution at each site were evaluated to achieve site closure in the most cost effective manner. This paper presents the remedial alternatives and cost analyses for each site. Several options are available at most sites for the design of a permeable treatment wall. Our analysis demonstrates that the major cost factor's for this technology are the design concept, length, thickness, location and construction methods for the reactive wall. Minimizing the amount of iron by placement in the most effective area and construction by the lowest cost method is critical to achieving a low cost alternative. These costs dictate the design of a permeable treatment wall, including selection of a variety of alternatives (e.g., a continuous wall versus a funnel and gate system, fully penetrating gates versus partially penetrating gates, etc.). Selection of the appropriate construction methods and materials for the site can reduce the overall cost of the wall

  1. Permeability of different size waste particles

    Directory of Open Access Journals (Sweden)

    Sabina Gavelytė


    Full Text Available The world and life style is changing, but the most popular disposal route for waste is landfill globally until now. We have to think about waste prevention and preparing for re-use or recycling firstly, according to the waste disposal hierarchy. Disposed waste to the landfill must be the last opportunity. In a landfill, during waste degradation processes leachate is formed that can potentially cause clogging of bottom drainage layers. To ensure stability of a landfill construction, the physical properties of its components have to be controlled. The hydrology of precipitation, evaporation, runoff and the hydraulic performance of the capping and liner materials are important controls of the moisture content. The water balance depends also on the waste characteristics and waste particle size distribution. The aim of this paper is to determine the hydraulic permeability in a landfill depending on the particle size distribution of municipal solid waste disposed. The lab experiment results were compared with the results calculated with DEGAS model. Samples were taken from a landfill operated for five years. The samples particle sizes are: >100 mm, 80 mm, 60 mm, 40 mm, 20 mm, 0.01 mm and <0.01 mm. The permeability test was conducted using the column test. The paper presents the results of experiment and DEGAS model water permeability with waste particle size.

  2. Effect of high density lipoproteins on permeability of rabbit aorta to low density lipoproteins

    International Nuclear Information System (INIS)

    Klimov, A.N.; Popov, V.A.; Nagornev, V.A.; Pleskov, V.M.


    A study was made on the effect of high density lipoproteins (HDL) on the permeability of rabbit aorta to low density lipoproteins (LDL) after intravenous administration of human HDL and human ( 125 I)LDL to normal and hypercholesterolemic rabbits. Evaluation of radioactivity in plasma and aorta has shown that the administration of a large dose of HDL decreased the aorta permeability rate for ( 125 I)LDL on an average by 19% in normal rabbits, and by 45% in rabbits with moderate hypercholesterolemia. A historadiographic study showed that HDL also decreased the vessel wall permeability to ( 125 I)LDL in normal and particularly in hypercholesterolemic animals. The suggestion was made that HDL at very high molar concentration can hamper LDL transportation through the intact endothelial layer into the intima due to the ability of HDL to compete with LDL in sites of low affinity on the surface of endothelial cells. (author)

  3. Enhancing the intestinal membrane permeability of zanamivir: a carrier mediated prodrug approach. (United States)

    Gupta, Sheeba Varghese; Gupta, Deepak; Sun, Jing; Dahan, Arik; Tsume, Yasuhiro; Hilfinger, John; Lee, Kyung-Dall; Amidon, Gordon L


    The purpose of this study was to improve the membrane permeability and oral absorption of the poorly permeable anti-influenza agent, zanamivir. The poor oral bioavailability is attributed to the high polarity (cLogP ∼ -5) resulting from the polar and zwitterionic nature of zanamivir. In order to improve the permeability of zanamivir, prodrugs with amino acids were developed to target the intestinal membrane transporter, hPepT1. Several acyloxy ester prodrugs of zanamivir conjugated with amino acids were synthesized and characterized. The prodrugs were evaluated for their chemical stability in buffers at various pHs and for their transport and tissue activation by enzymes. The acyloxy ester prodrugs of zanamivir were shown to competitively inhibit [(3)H]Gly-Sar uptake in Caco-2 cells (IC(50): 1.19 ± 0.33 mM for L-valyl prodrug of zanamivir). The L-valyl prodrug of zanamivir exhibited ∼3-fold higher uptake in transfected HeLa/hPepT1 cells compared to wild type HeLa cells, suggesting, at least in part, carrier mediated transport by the hPepT1 transporter. Further, enhanced transcellular permeability of prodrugs across Caco-2 monolayer compared to the parent drug (P(app) = 2.24 × 10(-6) ± 1.33 × 10(-7) cm/s for L-valyl prodrug of zanamivir), with only parent zanamivir appearing in the receiver compartment, indicates that the prodrugs exhibited both enhanced transport and activation in intestinal mucosal cells. Most significantly, several of these prodrugs exhibited high intestinal jejunal membrane permeability, similar to metoprolol, in the in situ rat intestinal perfusion system, a system highly correlated with human jejunal permeability. In summary, this mechanistic targeted prodrug strategy, to enhance oral absorption via intestinal membrane carriers such as hPepT1, followed by activation to parent drug (active pharmaceutical ingredient or API) in the mucosal cell, significantly improves the intestinal epithelial cell permeability of zanamivir and has the

  4. Permeability and Microstructure of Suspension Plasma-Sprayed YSZ Electrolytes for SOFCs on Various Substrates (United States)

    Marr, Michael; Kesler, Olivera


    Yttria-stabilized zirconia electrolyte coatings for solid oxide fuel cells were deposited by suspension plasma spraying using a range of spray conditions and a variety of substrates, including finely structured porous stainless steel disks and cathode layers on stainless steel supports. Electrolyte permeability values and trends were found to be highly dependent on which substrate was used. The most gas-tight electrolyte coatings were those deposited directly on the porous metal disks. With this substrate, permeability was reduced by increasing the torch power and reducing the stand-off distance to produce dense coating microstructures. On the substrates with cathodes, electrolyte permeability was reduced by increasing the stand-off distance, which reduced the formation of segmentation cracks and regions of aligned and concentrated porosity. The formation mechanisms of the various permeability-related coating features are discussed and strategies for reducing permeability are presented. The dependences of electrolyte deposition efficiency and surface roughness on process conditions and substrate properties are also presented.

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

    KAUST Repository

    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.

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

    KAUST Repository

    Akhtar, Faheem Hassan; Kumar, Mahendra; Villalobos, Luis Francisco; Shevate, Rahul; Vovusha, Hakkim; Schwingenschlö gl, Udo; Peinemann, Klaus-Viktor


    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. Genetic Manipulation of Outer Membrane Permeability: Generating Porous Heterogeneous Catalyst Analogs in Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Patel, TN; Park, AHA; Bantat, S


    The limited permeability of the E. coli outer membrane can significantly hinder whole-cell biocatalyst performance. In this study, the SARS coronavirus small envelope protein (SCVE) was expressed in E. coli cells previously engineered for periplasmic expression of carbonic anhydrase (CA) activity. This maneuver increased small molecule uptake by the cells, resulting in increased apparent CA activity of the biocatalysts. The enhancements in activity were quantified using methods developed for traditional heterogeneous catalysis. The expression of the SCVE protein was found to significantly reduce the Thiele moduli (phi), as well as increase the effectiveness factors (eta), effective diffusivities (D-e), and permeabilities (P) of the biocatalysts. These catalytic improvements translated into superior performance of the biocatalysts for the precipitation of calcium carbonate from solution which is an attractive strategy for long-term sequestration of captured carbon dioxide. Overall, these results demonstrate that synthetic biology approaches can be used to enhance heterogeneous catalysts incorporated into microbial whole-cell scaffolds.

  8. The use of tracers in the study of the ionic permeability of the nerve

    International Nuclear Information System (INIS)

    Guinnebault, M.


    The use of the radioactive tracers 24 Na and 42 K, and the electrophysiological fine study carried out on the plant Cephalopods nervous cells have made it possible, during ten years, to study the relationships between the electrical phenomena observed during the propagation of the nervous impulse and the ionic movements which occur at the cell membrane between the intra- and extra-cellular media. In the state of rest, the nerve fibres present a certain permeability towards sodium and potassium, a permeability which allows the renewal, from the extra-cellular medium, of the cations contained in the cytoplasm. At the moment of the passage of the nervous impulse, it is possible to demonstrate a temporary increase in the permeability towards the sodium which causes the entry of Na + ions into the nerve fibre, whilst, a few milliseconds later, the increase in the potassium permeability causes the departure of an equivalent quantity of K + ions. A recovery phase follows the passage of the nervous impulse, during which the original ionic conditions are re-established. If the potassium movements can be considered as passive, the expulsion of Na + requires, on the part of the cell, the use of a part of the metabolic energy. (author) [fr

  9. Evolution of permeability in diatomaceous rocks mediated by pressure solution

    International Nuclear Information System (INIS)

    Yasuhara, Hideaki; Kinoshita, Naoki; Kurikami, Hiroshi; Kishida, Kiyoshi


    A conceptual model is presented to follow the evolution of permeability in diatomaceous rocks mediated by pressure solution. The progress of compaction and the evolution of permeability may be followed with time. Specifically, the main minerals of diatomaceous rocks that are quartz, cristobalite, and amorphous silica, are focused to examine differences of the permeability evolutions among them at effective stresses of 5, and 10 MPa, and temperatures of 20 and 90degC. The rates and magnitudes of permeability reduction increase with increase of the dissolution rate constants. Ultimate permeabilities reduce to the order of 90% at the completion of dissolution-mediated compaction. (author)

  10. EDZ and permeability in clayey rocks

    International Nuclear Information System (INIS)

    Levasseur, Severine; Collin, Frederic; Charlier, Robert; Besuelle, Pierre; Chambon, Rene; Viggiani, Cino


    Document available in extended abstract form only. Deep geological layers are being considered as potential host rocks for the high level radioactivity waste disposals. During drilling in host rocks, an excavated damaged zone - EDZ is created. The fluid transmissivity may be modified in this damaged zone. This paper deals with the permeability evolution in relation with diffuse and/or localized crack propagation in the material. We mainly focus on argillaceous rocks and on some underground laboratories: Mol URL in Boom clay, Bure URL in Callovo-Oxfordian clay and Mont-Terri URL in Opalinus clay. First, observations of damage around galleries are summarized. Structure of damage in localized zone or in fracture has been observed at underground gallery scale within the excavation damaged zone (EDZ). The first challenge for a correct understanding of all the processes occurring within the EDZ is the characterization at the laboratory scale of the damage and localization processes. The observation of the initiation and propagation of the localized zones needs for advanced techniques. X-ray tomography is a non-destructive imaging technique that allows quantification of internal features of an object in 3D. If mechanical loading of a specimen is applied inside a X-ray CT apparatus, successive 3D images at different loading steps show the evolution of the specimen. However, in general volumetric strain in a shear band is small compared to the shear strain and, unfortunately, in tomographic images grey level is mainly sensitive to the local mass density field. Such a limitation has been recently overcome by complementing X-ray tomography with 3D Volumetric Digital Image Correlation (V-DIC) which allows the determination of the full strain tensor field. Then it is possible to further explore the progression of localized deformation in the specimen. The second challenge is the robust modelling of the strain localized process. In fact, modelling the damage process with finite

  11. Soluble CD40 ligand directly alters glomerular permeability and may act as a circulating permeability factor in FSGS. (United States)

    Doublier, Sophie; Zennaro, Cristina; Musante, Luca; Spatola, Tiziana; Candiano, Giovanni; Bruschi, Maurizio; Besso, Luca; Cedrino, Massimo; Carraro, Michele; Ghiggeri, Gian Marco; Camussi, Giovanni; Lupia, Enrico


    CD40/CD40 ligand (CD40L) dyad, a co-stimulatory bi-molecular complex involved in the adaptive immune response, has also potent pro-inflammatory actions in haematopoietic and non-haematopoietic cells. We describe here a novel role for soluble CD40L (sCD40L) as modifier of glomerular permselectivity directly acting on glomerular epithelial cells (GECs). We found that stimulation of CD40, constitutively expressed on GEC cell membrane, by the sCD40L rapidly induced redistribution and loss of nephrin in GECs, and increased albumin permeability in isolated rat glomeruli. Pre-treatment with inhibitors of CD40-CD40L interaction completely prevented these effects. Furthermore, in vivo injection of sCD40L induced a significant reduction of nephrin and podocin expression in mouse glomeruli, although no significant increase of urine protein/creatinine ratio was observed after in vivo injection. The same effects were induced by plasma factors partially purified from post-transplant plasma exchange eluates of patients with focal segmental glomerulosclerosis (FSGS), and were blocked by CD40-CD40L inhibitors. Moreover, 17 and 34 kDa sCD40L isoforms were detected in the same plasmapheresis eluates by Western blotting. Finally, the levels of sCD40Lwere significantly increased in serum of children both with steroid-sensitive and steroid-resistant nephrotic syndrome (NS), and in adult patients with biopsy-proven FSGS, compared to healthy subjects, but neither in children with congenital NS nor in patients with membranous nephropathy. Our results demonstrate that sCD40L directly modifies nephrin and podocin distribution in GECs. Moreover, they suggest that sCD40L contained in plasmapheresis eluates from FSGS patients with post-transplant recurrence may contribute, presumably cooperating with other mediators, to FSGS pathogenesis by modulating glomerular permeability.

  12. Soluble CD40 ligand directly alters glomerular permeability and may act as a circulating permeability factor in FSGS.

    Directory of Open Access Journals (Sweden)

    Sophie Doublier

    Full Text Available CD40/CD40 ligand (CD40L dyad, a co-stimulatory bi-molecular complex involved in the adaptive immune response, has also potent pro-inflammatory actions in haematopoietic and non-haematopoietic cells. We describe here a novel role for soluble CD40L (sCD40L as modifier of glomerular permselectivity directly acting on glomerular epithelial cells (GECs. We found that stimulation of CD40, constitutively expressed on GEC cell membrane, by the sCD40L rapidly induced redistribution and loss of nephrin in GECs, and increased albumin permeability in isolated rat glomeruli. Pre-treatment with inhibitors of CD40-CD40L interaction completely prevented these effects. Furthermore, in vivo injection of sCD40L induced a significant reduction of nephrin and podocin expression in mouse glomeruli, although no significant increase of urine protein/creatinine ratio was observed after in vivo injection. The same effects were induced by plasma factors partially purified from post-transplant plasma exchange eluates of patients with focal segmental glomerulosclerosis (FSGS, and were blocked by CD40-CD40L inhibitors. Moreover, 17 and 34 kDa sCD40L isoforms were detected in the same plasmapheresis eluates by Western blotting. Finally, the levels of sCD40Lwere significantly increased in serum of children both with steroid-sensitive and steroid-resistant nephrotic syndrome (NS, and in adult patients with biopsy-proven FSGS, compared to healthy subjects, but neither in children with congenital NS nor in patients with membranous nephropathy. Our results demonstrate that sCD40L directly modifies nephrin and podocin distribution in GECs. Moreover, they suggest that sCD40L contained in plasmapheresis eluates from FSGS patients with post-transplant recurrence may contribute, presumably cooperating with other mediators, to FSGS pathogenesis by modulating glomerular permeability.

  13. Blood brain barrier permeability of (−-epigallocatechin gallate, its proliferation-enhancing activity of human neuroblastoma SH-SY5Y cells, and its preventive effect on age-related cognitive dysfunction in mice

    Directory of Open Access Journals (Sweden)

    Monira Pervin


    Conclusion: Cognitive dysfunction in mice is suppressed after ingesting GTCs when a low concentration of EGCG is incorporated into the brain parenchyma via the BBB. Nerve cell proliferation/differentiation was enhanced by a low concentration of EGCG. Furthermore, the additive effect of EGC and GA suggests that EGCG sustains a preventive effect after the hydrolysis to EGC and GA.

  14. Towards cavitation-enhanced permeability in blood vessel on a chip (United States)

    De Luca, R.; Silvani, G.; Scognamiglio, C.; Sinibaldi, G.; Peruzzi, G.; Chinappi, M.; Kiani, M. F.; Casciola, C. M.


    The development of targeted delivery systems releasing pharmaceutical agents directly at the desired site of action may improve their therapeutic efficiency while minimizing damage to healthy tissues, toxicity to the patient and drug waste. In this context, we have developed a bio-inspired microdevice mimicking the tumour microvasculature which represents a valuable tool for assessing the enhancement of blood vessel permeability due to cavitation. This novel system allows us to investigate the effects of ultrasound-driven microbubbles that temporarily open the endothelial intercellular junctions allowing drug to extravasate blood vessels into tumour tissues. The blood vessel on a chip consists of a tissue chamber and two independent vascular channels (width 200 µm, height 100 µm, length 2762 µm) cultured with endothelial cells placed side-by-side and separated by a series of 3 µm pores. Its geometry and dimensions mimic the three-dimensional morphology, size and flow characteristics of microvessels in vivo. The early stage of this project had a twofold objective: 1. To define the protocol for culturing of Human Umbilical Vein Endothelial Cells (HUVECs) within the vascular channel; 2. To develop a fluorescence based microscopy technique for measuring permeability. We have developed a reliable and reproducible protocol to culture endothelial cells within the artificial vessels in a realistic manner: HUVECs show the typical elongated shape in the direction of flow, exhibit tight junction formation and form a continuous layer with a central lumen that completely covers the channels wall. As expected, the permeability of cell-free device is higher than the one cultured with HUVECs in the vascular channels. The proposed blood vessel on a chip and the permeability measurement protocol have a significant potential to allow for the study of cavitation-enhanced permeability of the endothelium and improve efficiency in screening drug delivery systems.

  15. Studies of cesium permeability of potassium transporter from Arabidopsis

    International Nuclear Information System (INIS)

    Kobayashi, Daisuke; Yamagami, Mutsumi; Hisamatsu, Shun'ichi; Inaba, Jiro; Uozumi, Nobuyuki; Hazama, Akihiro


    Cs-137 is an important radionuclide for safety assessment of nuclear facilities, and its transfer via plants is an important route from the environment to humans. Studies of Cs uptake mechanisms by plants are essential for understanding 137 Cs movement in soil-to-plant systems and in plants. Since uptake of Cs is considered to be mediated by K transport protein, we investigated Cs + permeability of two Arabidopsis K + transporters, AKT2 and AtHKT1, by using Xenopus oocytes expression systems and two-electrode voltage-clamp technique under various pH conditions. The data showed AKT2 and AtHKT1 did not transport Cs + at external pH in the 5.5-7.5 range. These results indicated that AKT2 and AtHKT1 did not contribute to Cs + influx into cells under physiological conditions in plants. (author)

  16. The Effect of Bacteria Penetration on Chalk Permeability

    DEFF Research Database (Denmark)

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

    number of B. licheniformis was detected on the effluent compared with P. putida. However, in the experiment with B. licheniformis mainly spores were detected in the effluent. The core permeability decreased rapidly during injection of bacteria and a starvation period of 12 days did not allow......Bacteria selective plugging is one of the mechanisms through which microorganisms can be applied for enhanced oil recovery. Bacteria can plug the water-bearing zones of a reservoir, thus altering the flow paths and improving sweep efficiency. It is known that the bacteria can penetrate deeply...... into reservoirs, however, a complete understanding of the penetration behavior of bacteria is lacking, especially in chalk formations where the pore throat sizes are almost comparable with the sizes of bacteria vegetative cells. This study investigates the penetration of bacteria into chalk. Two bacteria types...

  17. Evaluation of permeable fractures in rock aquifers (United States)

    Bok Lee, Hang


    In this study, the practical usefulness and fundamental applicability of a self-potential (SP) method for identifying the permeable fractures were evaluated by a comparison of SP methods with other geophysical logging methods and hydraulic tests. At a 10 m-shallow borehole in the study site, the candidates of permeable fractures crossing the borehole were first determined by conventional geophysical methods such as an acoustic borehole televiwer, temperature, electrical conductivity and gamma-gamma loggings, which was compared to the analysis by the SP method. Constant pressure injection and recovery tests were conducted for verification of the hydraulic properties of the fractures identified by various logging methods. The acoustic borehole televiwer and gamma-gamma loggings detected the open space or weathering zone within the borehole, but they cannot prove the possibility of a groundwater flow through the detected fractures. The temperature and electrical conductivity loggings had limitations to detect the fractured zones where groundwater in the borehole flows out to the surrounding rock aquifers. Comparison of results from different methods showed that there is a best correlation between the distribution of hydraulic conductivity and the variation of the SP signals, and the SP logging can estimate accurately the hydraulic activity as well as the location of permeable fractures. Based on the results, the SP method is recommended for determining the hydraulically-active fractures rather than other conventional geophysical loggings. This self-potential method can be effectively applied in the initial stage of a site investigation which selects the optimal location and evaluates the hydrogeological property of fractures in target sites for the underground structure including the geothermal reservoir and radioactive waste disposal.

  18. Permeability log using new lifetime measurements

    International Nuclear Information System (INIS)

    Dowling, D.J.; Boyd, J.F.; Fuchs, J.A.


    Comparative measurements of thermal neutron decay time are obtained for a formation after irradiation with a pulsed neutron source. Chloride ions in formation fluids are concentrated by the electrosmosis effect using charged poles on a well logging sonde. The formation is irradiated with fast neutrons and a first comparative measure of the thermal neutron decay time or neutron lifetime is taken. The chloride ions are then dispersed by acoustic pumping with a magnetostrictive transducer. The formation is then again irradiated with fast neutrons and a comparative measure of neutron lifetime is taken. The comparison is a function of the variation in chloride concentration between the two measurements which is related to formation permeability

  19. Permeability Evolution and Rock Brittle Failure


    Sun Qiang; Xue Lei; Zhu Shuyun


    This paper reports an experimental study of the evolution of permeability during rock brittle failure and a theoretical analysis of rock critical stress level. It is assumed that the rock is a strain-softening medium whose strength can be described by Weibull’s distribution. Based on the two-dimensional renormalization group theory, it is found that the stress level λ c (the ratio of the stress at the critical point to the peak stress) depends mainly on the homogeneity index or shape paramete...

  20. Nitric oxide turnover in permeable river sediment

    DEFF Research Database (Denmark)

    Schreiber, Frank; Stief, Peter; Kuypers, Marcel M M


    We measured nitric oxide (NO) microprofiles in relation to oxygen (O2) and all major dissolved N-species (ammonium, nitrate, nitrite, and nitrous oxide [N2O]) in a permeable, freshwater sediment (River Weser, Germany). NO reaches peak concentrations of 0.13 μmol L-1 in the oxic zone and is consumed......-nitroso-N-acetylpenicillamine (SNAP) (1) confirmed denitrification as the main NO consumption pathway, with N2O as its major product, (2) showed that denitrification combines one free NO molecule with one NO molecule formed from nitrite to produce N2O, and (3) suggested that NO inhibits N2O reduction....

  1. Gyroid Nanoporous Membranes with Tunable Permeability

    DEFF Research Database (Denmark)

    Li, Li; Schulte, Lars; Clausen, Lydia D.


    -linked 1,2-polybutadiene (1,2-PB) membranes with uniform pores that, if needed, can be rendered hydrophilic. The gyroid porosity has the advantage of isotropic percolation with no need for structure prealignment. Closed (skin) or opened (nonskin) outer surface can be simply realized by altering...... 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...

  2. Development and characterization of a new oral dapsone nanoemulsion system: permeability and in silico bioavailability studies. (United States)

    Monteiro, Lidiane M; Lione, Viviane F; do Carmo, Flavia A; do Amaral, Lilian H; da Silva, Julianna H; Nasciutti, Luiz E; Rodrigues, Carlos R; Castro, Helena C; de Sousa, Valeria P; Cabral, Lucio M


    Dapsone is described as being active against Mycobacterium leprae, hence its role in the treatment of leprosy and related pathologies. Despite its therapeutic potential, the low solubility of dapsone in water results in low bioavailability and high microbial resistance. Nanoemulsions are pharmaceutical delivery systems derived from micellar solutions with a good capacity for improving absorption. The aim of this work was to develop and compare the permeability of a series of dapsone nanoemulsions in Caco-2 cell culture against that of effective permeability in the human body simulated using Gastroplus™ software. The release profiles of the dapsone nanoemulsions using different combinations of surfactants and cosolvent showed a higher dissolution rate in simulated gastric and enteric fluid than did the dispersed dapsone powder. The drug release kinetics were consistent with a Higuchi model. This comparison of dapsone permeability in Caco-2 cells with effective permeability in the human body simulated by Gastroplus showed a good correlation and indicates potential improvement in the biodisponibility of dapsone using this new system.

  3. Endothelial glycocalyx shedding and vascular permeability in severely injured trauma patients

    DEFF Research Database (Denmark)

    Rahbar, Elaheh; Cardenas, Jessica C; Baimukanova, Gyulnar


    of trauma patients. METHODS: Plasma samples were collected from 5 healthy consented volunteers and 22 severely injured trauma patients upon admission to the emergency department. ELISA assays were performed to quantify shed HA, HS, CS and syndecan-1 in plasma. A colloid osmometer and Electric Cell......-substrate Impedance Sensing (ECIS) system were used to measure plasma colloid osmotic pressure (COP) and cell permeability, respectively. Thrombin generation was measured using a calibrated automated thrombogram (CAT). Initial vital signs, routine laboratory values, and injury severity scores (ISS) were recorded. Non......COP (≤16 mmHg) had significantly increased syndecan-1 and HA compared to those with normal COP, which corresponded to increased cell permeability via ECIS. CS and HS did not vary between COP groups. Lastly, patients with low COP displayed reduced peak thrombin...

  4. Permeability-Porosity Relationships of Subduction Zone Sediments (United States)

    Gamage, K.; Screaton, E.; Bekins, B.; Aiello, I.


    Permeability-porosity relationships for sediments from Northern Barbados, Costa Rica, Nankai, and Peru subduction zones were examined based on their sediment type and grain size distribution. Greater correlation was observed between permeability and porosity for siliciclastic sediments, diatom oozes, and nannofossil chalk than for nannofossil oozes. For siliciclastic sediments, grouping of sediments by clay content yields relationships that are generally consistent with results from other marine settings and suggest decreasing permeability for a given porosity as clay content increases. Correction of measured porosities for smectite content generally improves the quality of permeability-porosity relationships. The relationship between permeability and porosity for diatom oozes may be controlled by the amount of clay present in the ooze, causing diatom oozes to behave similarly to siliciclastic sediments. For a given porosity the nannofossil oozes have higher permeability values by 1.5 orders of magnitude than the siliciclastic sediments. However, the use of a permeability-porosity relation may not be appropriate for unconsolidated carbonates such as nannofossil oozes. This study provided insight to the effects of porosity correction for smectite, variations in lithology and grain size in permeability-porosity relationships. However, further progress in delineating controls on permeability will require more careful and better documented permeability tests on characterized samples.

  5. Development and characterization of a new oral dapsone nanoemulsion system: permeability and in silico bioavailability studies

    Directory of Open Access Journals (Sweden)

    Monteiro LM


    Full Text Available Lidiane M Monteiro,1 Viviane F Lione,1 Flavia A do Carmo,1 Lilian H do Amaral,1 Julianna H da Silva,2 Luiz E Nasciutti,2 Carlos R Rodrigues,1 Helena C Castro,3 Valeria P de Sousa,1 Lucio M Cabral11Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, 2Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, 3Instituto de Biologia, Universidade Federal Fluminense, Niterói, Rio de Janeiro, BrasilBackground: Dapsone is described as being active against Mycobacterium leprae, hence its role in the treatment of leprosy and related pathologies. Despite its therapeutic potential, the low solubility of dapsone in water results in low bioavailability and high microbial resistance. Nanoemulsions are pharmaceutical delivery systems derived from micellar solutions with a good capacity for improving absorption. The aim of this work was to develop and compare the permeability of a series of dapsone nanoemulsions in Caco-2 cell culture against that of effective permeability in the human body simulated using GastroplusTM software.Methods and results: The release profiles of the dapsone nanoemulsions using different combinations of surfactants and cosolvent showed a higher dissolution rate in simulated gastric and enteric fluid than did the dispersed dapsone powder. The drug release kinetics were consistent with a Higuchi model.Conclusion: This comparison of dapsone permeability in Caco-2 cells with effective permeability in the human body simulated by Gastroplus showed a good correlation and indicates potential improvement in the biodisponibility of dapsone using this new system.Keywords: dapsone, nanoemulsions, antibacterial, permeability, Caco-2 cell, GastroplusTM

  6. Bistable Bacterial Growth Rate in Response to Antibiotics with Low Membrane Permeability (United States)

    Elf, Johan; Nilsson, Karin; Tenson, Tanel; Ehrenberg, Måns


    We demonstrate that growth rate bistability for bacterial cells growing exponentially at a fixed external antibiotic concentration can emerge when the cell wall permeability for the drug is low and the growth rate sensitivity to the intracellular drug concentration is high. Under such conditions, an initially high growth rate can remain high, due to dilution of the intracellular drug concentration by rapid cell volume increase, while an initially low growth rate can remain low, due to slow cell volume increase and insignificant drug dilution. Our findings have implications for the testing of novel antibiotics on growing bacterial strains.

  7. Stationary and Dynamic Permeability and Coupling Coefficient Measurements in Sintered Glass Bead Systems (United States)

    Gueven, I.; Steeb, H.; Luding, S.


    Electrokinetic waves describe the coupling between seismic and electromagnetic waves that exist in porous media. The coupling between them arise from an electrochemical boundary layer between grain and fluid interface of saturated porous media. Acoustical waves cause a disturbance of the electrical fluid charge within the double layer, which therefore creates an electric streaming current (seismoelectric effect). Inversely, electromagnetic waves can generate mechanical signals (electroseismic effect). Electrokinetic conversion potentially combines high seismic resolution with good electromagnetic hydrocarbon sensitivity. The (stationary and frequency-dependent) streaming potential coefficient is a key property, which gives rise to the coupling between electromagnetic and acoustical waves. It depends strongly on the fluid conductivity, porosity, tortuosity, permeability, pore throat and zeta potential of porous media. We examine experimentally both, the stationary and dynamic permeabilities and coupling coefficients of sintered glass bead systems. For this purpose a multi-purpose measuring cell was developed which allows us to carry out - besides common ultrasound experiments - also to perform stationary and frequency-dependent permeability and coupling coefficient measurements. For the experiments sintered mono- and slightly polydisperse glass bead samples with different glass bead diameters between 0.4 and 8mm and porosities ranging between 21 and 39% were used. The stationary and dynamic permeability and streaming potential measurements are supported by μCT scans which enable us a deeper insight into the porous medium. Based on the μCT scans of the produced sintered glass bead samples essential influence parameters, like tortuosity, porosity, effective particle diameters and pore throats in different regions of the entire scanned region have been analyzed in detail to understand the laboratory experiments, cf. Illustration 1. In addition lattice Boltzmann

  8. Influence of Cholesterol on the Oxygen Permeability of Membranes: Insight from Atomistic Simulations. (United States)

    Dotson, Rachel J; Smith, Casey R; Bueche, Kristina; Angles, Gary; Pias, Sally C


    Cholesterol is widely known to alter the physical properties and permeability of membranes. Several prior works have implicated cell membrane cholesterol as a barrier to tissue oxygenation, yet a good deal remains to be explained with regard to the mechanism and magnitude of the effect. We use molecular dynamics simulations to provide atomic-resolution insight into the influence of cholesterol on oxygen diffusion across and within the membrane. Our simulations show strong overall agreement with published experimental data, reproducing the shapes of experimental oximetry curves with high accuracy. We calculate the upper-limit transmembrane oxygen permeability of a 1-palmitoyl,2-oleoylphosphatidylcholine phospholipid bilayer to be 52 ± 2 cm/s, close to the permeability of a water layer of the same thickness. With addition of cholesterol, the permeability decreases somewhat, reaching 40 ± 2 cm/s at the near-saturating level of 62.5 mol % cholesterol and 10 ± 2 cm/s in a 100% cholesterol mimic of the experimentally observed noncrystalline cholesterol bilayer domain. These reductions in permeability can only be biologically consequential in contexts where the diffusional path of oxygen is not water dominated. In our simulations, cholesterol reduces the overall solubility of oxygen within the membrane but enhances the oxygen transport parameter (solubility-diffusion product) near the membrane center. Given relatively low barriers to passing from membrane to membrane, our findings support hydrophobic channeling within membranes as a means of cellular and tissue-level oxygen transport. In such a membrane-dominated diffusional scheme, the influence of cholesterol on oxygen permeability is large enough to warrant further attention. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  9. 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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  10. Assessing the permeability of engineered capillary networks in a 3D culture.

    Directory of Open Access Journals (Sweden)

    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

  11. Estimation of permeability and permeability anisotropy in horizontal wells through numerical simulation of mud filtrate invasion

    Energy Technology Data Exchange (ETDEWEB)

    Pereira, Nelson [PETROBRAS S.A., Rio de Janeiro, RJ (Brazil). Exploracao e Producao; Altman, Raphael; Rasmus, John; Oliveira, Jansen [Schlumberger Servicos de Petroleo Ltda., Rio de Janeiro, RJ (Brazil)


    This paper describes how permeability and permeability anisotropy is estimated in horizontal wells using LWD (logging-while-drilling) laterolog resistivity data. Laterolog-while-drilling resistivity passes of while-drilling and timelapse (while reaming) were used to capture the invasion process. Radial positions of water based mud invasion fronts were calculated from while-drilling and reaming resistivity data. The invasion process was then recreated by constructing forward models with a fully implicit, near-wellbore numerical simulation such that the invasion front at a given time was consistent with the position of the front predicted by resistivity inversions. The radial position of the invasion front was shown to be sensitive to formation permeability. The while-drilling environment provides a fertile scenario to investigate reservoir dynamic properties because mud cake integrity and growth is not fully developed which means that the position of the invasion front at a particular point in time is more sensitive to formation permeability. The estimation of dynamic formation properties in horizontal wells is of particular value in marginal fields and deep-water offshore developments where running wireline and obtaining core is not always feasible, and where the accuracy of reservoir models can reduce the risk in field development decisions. (author)


    Directory of Open Access Journals (Sweden)

    Helena Vučenović


    Full Text Available Geosynthetic clay liners (GCL are manufactured hydraulic barriers consisting of mineral and geosynthetic components. They belong to a group of geosynthetic products whose primary purpose is to seal and they have been used in many geotechnical and hydrotechnical applications, landfi lls and liquid waste lagoons for quite a while. They are used in landfill final cover systems to prevent the infi ltration of precipitation into the landfi ll body and the penetration of gases and liquids from the landfill into the atmosphere and environment. Laboratory and fi eld research and observations on regulated landfi lls have proven the eff ectiveness of GCL as a barrier for the infi ltration of precipitation into the landfi ll body as well as the drainage of fl uid beneath the landfill. Due to the presence of high concentrations of gases in the landfill body, there is a growing interest in determining the efficiency of GCL as a gas barrier. It was not until the last twenty years that the importance of this topic was recognized. In this article, current GCL gas permeability studies, the testing methods and test results of gas permeability in laboratory conditions are described.

  13. Haemophilia, AIDS and lung epithelial permeability

    Energy Technology Data Exchange (ETDEWEB)

    O' Doherty, M.J.; Page, C.J.; Harrington, C.; Nunan, T.; Savidge, G. (Haemophilia Centre and Coagulation Research Unit, Department of Nuclear Medicine, Rayne Institute, St. Thomas' Hospital, London (United Kingdom))


    Lung {sup 99m}Tc DTPA transfer was measured in HIV antibodypositive haemophiliacs (11 smokers, 26 nonsmokers, 5 patients with Pneumocystis carinii pneumonia (PCP)). Lung {sup 99m}Tc DTPA transfer as a marker of lung epithelial permeability was measured as the half time of transfer (from airspace into blood). This half time was faster in smokers compred to nonsmokers and the transfer curve was monoexponential. In nonsmokers no difference was observed between asymptomatic HIV-positive haemophiliacs and normal subjects, with the exception of the lung bases. At the lung basis in HIV-positive haemophiliac nonsmokers the transfer was faster than in normal individuals, implying increased alveolar permeability. Pneumocystis carinii pneumonia resulted in a rapid transfer of {sup 99m}Tc DTPA (mean T50 of 2 minutes) and the transfer curve was biphasic, confirming previous observations in homosexual HIV antibody-positive patients with PCP. These changes returned to a monoexponential profile by 6 weeks following successful treatment. The DTPA lung transfer study may enable clinicians to instigate therapy for PCP without the need for initial bronchoscopy and provide a noninvasive method for the reassessment of patients should further respiratory signs or symptoms develop. This method is considered to be highly cost-effective in that it obviates the use of factor VIII concentrates required to cover bronchoscopic procedures and, with its early application and ease of use as a follow-up investigation, permits the evaluation of patients on an outpatient basis, thus reducing hospital costs. (au).

  14. Haemophilia, AIDS and lung epithelial permeability

    International Nuclear Information System (INIS)

    O'Doherty, M.J.; Page, C.J.; Harrington, C.; Nunan, T.; Savidge, G.


    Lung 99m Tc DTPA transfer was measured in HIV antibodypositive haemophiliacs (11 smokers, 26 nonsmokers, 5 patients with Pneumocystis carinii pneumonia (PCP)). Lung 99m Tc DTPA transfer as a marker of lung epithelial permeability was measured as the half time of transfer (from airspace into blood). This half time was faster in smokers compred to nonsmokers and the transfer curve was monoexponential. In nonsmokers no difference was observed between asymptomatic HIV-positive haemophiliacs and normal subjects, with the exception of the lung bases. At the lung basis in HIV-positive haemophiliac nonsmokers the transfer was faster than in normal individuals, implying increased alveolar permeability. Pneumocystis carinii pneumonia resulted in a rapid transfer of 99m Tc DTPA (mean T50 of 2 minutes) and the transfer curve was biphasic, confirming previous observations in homosexual HIV antibody-positive patients with PCP. These changes returned to a monoexponential profile by 6 weeks following successful treatment. The DTPA lung transfer study may enable clinicians to instigate therapy for PCP without the need for initial bronchoscopy and provide a noninvasive method for the reassessment of patients should further respiratory signs or symptoms develop. This method is considered to be highly cost-effective in that it obviates the use of factor VIII concentrates required to cover bronchoscopic procedures and, with its early application and ease of use as a follow-up investigation, permits the evaluation of patients on an outpatient basis, thus reducing hospital costs. (au)

  15. Permeability of protective coatings to tritium

    International Nuclear Information System (INIS)

    Braun, J.M.


    The permeability of four protective coatings to tritium gas and tritiated water was investigated. The coatings, including two epoxies, one vinyl and one urethane, were selected for their suitability in CANDU plant service in Ontario Hydro. Sorption rates of tritium gas into the coatings were considerably larger than for tritiated water, by as much as three to four orders of magnitude. However, as a result of the very large solubility of tritiated water in the coatings, the overall permeability to tritium gas and tritiated water are comparable, being somewhat larger for HTO. Marked differences were also evident among the four coatings, the vinyl proving to be unique in behaviour and morphology. Because of a highly porous surface structure water condensation takes place at high relative humidities, leading to an abnormally high retention of free water. Desorption rates from the four coatings were otherwise quite similar. Of practical importance was the observation that more effective desorption of tritiated water could be carried out at relatively high humidities, in this case 60%. It was believed that isotopic exchange was responsible for this phenomenon. It appears that epoxy coatings having a high pigment-to-binder ratio are most suited for coating concrete in tritium handling facilities

  16. Salt-saturated concrete strength and permeability

    International Nuclear Information System (INIS)

    Pfeifle, T.W.; Hansen, F.D.; Knowles, M.K.


    Laboratory-scale experiments applicable to the use of salt-saturated concrete as a seal material for a transuranic waste repository have been completed. Nitrogen gas permeability measurements were made using a flexible-wall permeameter, a confining pressure of 1 MPa, and gas pressure gradients ranging from 0.3 MPa to 0.75 MPa. Results show that salt-saturated concrete has very low intrinsic permeability with values ranging from 9.4 x 10 -22 m 2 to 9.7 x 10 -17 m 2 . Strength and deformation characteristics were investigated under conditions of triaxial compression with confining pressures ranging from 0 to 15 MPa using either axial strain-rate or axial stress-rate control and show that the failure strength of concrete increases with confining pressure which can be adequately described through pressure-sensitive failure criteria. Axial, radial, and volumetric strains were also measured during each test and these data were used to determine elastic properties. Experimental results are applicable in the design and analysis of scale-related functions and apply to other concrete structures subjected to compressive loadings such as dams and prestressed structural members

  17. Effective permeability in micropores from molecular simulations

    International Nuclear Information System (INIS)

    Botan, A.; Vermorel, R.; Brochard, L.; Hantal, G.; Pellenq, R.


    Document available in extended abstract form only. Despite many years' efforts and a large numbers of proposed models, the description of transport properties in clays is still an open question. The reason for this is that structurally clay is an extremely heterogeneous material. The pore size varies from a few to 20 angstroms for interlayer (micro) porosity, from 20 A to 500 A for interparticle (meso) porosity, and 500 A to μm and more for natural (macro) fractures. One further problem with the description of the transport properties is the presence of adsorption/desorption processes onto clay particles, which are coupled with swelling/shrinkage of the particles. Any volumetric changes in the particles affect the meso-pore aperture, and thus, the total permeability of the system. The various processes affecting the permeability occur on different spatial and temporal scales, that requires a multi-scale modeling approach. The most complete model to date is a dual porosity mode. Here the total flow is often written as a sum of the macropore flow and micropore flow. The flow through macro-pores is generally considered to be laminar and obeys Darcy's law, whereas flow through the matrix (micropore flow) may be modeled using Fick's law. The micropore flow involves both Knudsen and surface diffusion mechanisms. An accurate accounting of adsorption-desorption processes or the consideration of binary mixture greatly complicate analytical description. The goal of this study is to improve macro-scale model, the dual porosity model, for the transport properties of fluids in micropores from molecular simulations. The main idea is that we reproduce an experimental set-up used for permeability measurements, as illustrated in Figure 1. High density and low density regions are settled at each end of the membrane that allows to attain a steady flow. The densities in these regions are controlled by Grand Canonical Monte Carlo simulation; the molecular motions are described by

  18. Calculation of large scale relative permeabilities from stochastic properties of the permeability field and fluid properties

    Energy Technology Data Exchange (ETDEWEB)

    Lenormand, R.; Thiele, M.R. [Institut Francais du Petrole, Rueil Malmaison (France)


    The paper describes the method and presents preliminary results for the calculation of homogenized relative permeabilities using stochastic properties of the permeability field. In heterogeneous media, the spreading of an injected fluid is mainly sue to the permeability heterogeneity and viscosity fingering. At large scale, when the heterogeneous medium is replaced by a homogeneous one, we need to introduce a homogenized (or pseudo) relative permeability to obtain the same spreading. Generally, is derived by using fine-grid numerical simulations (Kyte and Berry). However, this operation is time consuming and cannot be performed for all the meshes of the reservoir. We propose an alternate method which uses the information given by the stochastic properties of the field without any numerical simulation. The method is based on recent developments on homogenized transport equations (the {open_quotes}MHD{close_quotes} equation, Lenormand SPE 30797). The MHD equation accounts for the three basic mechanisms of spreading of the injected fluid: (1) Dispersive spreading due to small scale randomness, characterized by a macrodispersion coefficient D. (2) Convective spreading due to large scale heterogeneities (layers) characterized by a heterogeneity factor H. (3) Viscous fingering characterized by an apparent viscosity ration M. In the paper, we first derive the parameters D and H as functions of variance and correlation length of the permeability field. The results are shown to be in good agreement with fine-grid simulations. The are then derived a function of D, H and M. The main result is that this approach lead to a time dependent . Finally, the calculated are compared to the values derived by history matching using fine-grid numerical simulations.

  19. Herbal medicines that benefit epidermal permeability barrier function

    Directory of Open Access Journals (Sweden)

    Lizhi Hu


    Full Text Available Epidermal permeability barrier function plays a critical role in regulating cutaneous functions. Hence, researchers have been searching for effective and affordable regimens to enhance epidermal permeability barrier function. In addition to topical stratum corneum lipids, peroxisome proliferator-activated receptor, and liver X receptor ligands, herbal medicines have been proven to benefit epidermal permeability barrier function in both normal and diseased skin, including atopic dermatitis, glucocorticoid-induced skin damage, and UVB-damaged skin. The potential mechanisms by which herbal medicines improve the permeability barrier include stimulation of epidermal differentiation, lipid production, antimicrobial peptide expression, and antioxidation. Therefore, utilization of herbal medicines could be a valuable alternative approach to enhance epidermal permeability barrier function in order to prevent and/or treat skin disorders associated with permeability barrier abnormalities.

  20. Therapeutic benefits of enhancing permeability barrier for atopic eczema

    Directory of Open Access Journals (Sweden)

    George Man


    Full Text Available The regulatory role of epidermal permeability barrier function in cutaneous inflammation has been well appreciated. While barrier disruption induces cutaneous inflammation, improvement of permeability barrier function alleviates inflammation. Studies have demonstrated that improvement of epidermal permeability barrier function not only prevents the development of atopic eczema, but also delays the relapse of these diseases. Moreover, enhancing the epidermal permeability barrier also alleviates atopic eczema. Furthermore, co-applications of barrier enhancing products with glucocorticoids can increase the therapeutic efficacy and reduce the adverse effects of glucocorticoids in the treatment of atopic eczema. Therefore, utilization of permeability barrier enhancing products alone or in combination with glucocorticoids could be a valuable approach in the treatment of atopic eczema. In this review, we discuss the benefits of improving the epidermal permeability barrier in the management of atopic eczema.

  1. Active intestinal drug absorption and the solubility-permeability interplay. (United States)

    Porat, Daniel; Dahan, Arik


    The solubility-permeability interplay deals with the question: what is the concomitant effect on the drug's apparent permeability when increasing the apparent solubility with a solubility-enabling formulation? The solubility and the permeability are closely related, exhibit certain interplay between them, and ongoing research throughout the past decade shows that treating the one irrespectively of the other may be insufficient. The aim of this article is to provide an overview of the current knowledge on the solubility-permeability interplay when using solubility-enabling formulations for oral lipophilic drugs, highlighting active permeability aspects. A solubility-enabling formulation may affect the permeability in opposite directions; the passive permeability may decrease as a result of the apparent solubility increase, according to the solubility-permeability tradeoff, but at the same time, certain components of the formulation may inhibit/saturate efflux transporters (when relevant), resulting in significant apparent permeability increase. In these cases, excipients with both solubilizing and e.g. P-gp inhibitory properties may lead to concomitant increase of both the solubility and the permeability. Intelligent development of such formulation will account for the simultaneous effects of the excipients' nature/concentrations on the two arms composing the overall permeability: the passive and the active arms. Overall, thorough mechanistic understanding of the various factors involved in the solubility-permeability interplay may allow developing better solubility-enabling formulations, thereby exploiting the advantages analyzed in this article, offering oral delivery solution even for BCS class IV drugs. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Study on Surface Permeability of Concrete under Immersion


    Liu, Jun; Xing, Feng; Dong, Biqin; Ma, Hongyan; Pan, Dong


    In this paper, concrete specimens are immersed in ultrapure water, to study the evolutions of surface permeability, pore structure and paste microstructure following the prolonging of immersion period. According to the results, after 30-day immersion, the surface permeability of concrete becomes higher as compared with the value before immersion. However, further immersion makes the surface permeability decrease, so that the value measured after 150-day immersion is only half that measured af...

  3. Epithelial Permeability Alterations in an In Vitro Air-Liquid Interface Model of Allergic Fungal Rhinosinusitis (United States)

    Den Beste, Kyle A.; Hoddeson, Elizabeth K.; Parkos, Charles A.; Nusrat, Asma; Wise, Sarah K.


    Background Chronic rhinosinusitis (CRS) is an inflammatory upper-airway disease with numerous etiologies. Patients with a characteristic subtype of CRS, allergic fungal rhinosinusitis (AFRS), display increased expression of Th2 cytokines and antigen-specific IgE. Various sinonasal inflammatory conditions are associated with alterations in epithelial barrier function. The aim of this study was to compare epithelial permeability and intercellular junctional protein expression amongst cultured primary sinonasal cells from AFRS patients versus non-inflammatory controls. Methods Epithelial cells isolated from paranasal sinus mucosa of AFRS and non-inflammatory control patients were grown to confluence on permeable supports and transitioned to air-liquid interface (ALI). Trans-epithelial resistance (TER) was measured with a horizontal Ussing chamber to characterize the functional permeability of each cell type. After TER recordings were complete, a panel of intercellular junctional proteins was assessed by Western blot and immunofluorescence labeling followed by confocal microscopy. Results After 12 samples were measured from each group, we observed a 41% mean decrease in TER in AFRS cells (296±89 ohms × cm2) compared to control (503±134 ohms × cm2, P=0.006). TER deficits observed in AFRS were associated with decreased expression of the tight junction proteins occludin and Junctional Adhesion Molecule-A (JAM-A), and increased expression of a leaky tight junction protein claudin-2. Conclusions Cultured sinonasal epithelium from AFRS patients displayed increased epithelial permeability and altered expression of intercellular junctional proteins. Given that these cells were not incubated with inflammatory cytokines in vitro, the cultured AFRS epithelial alterations may represent a retained modification in protein expression from the in vivo phenotype. PMID:22927233

  4. Particulate matter air pollution causes oxidant-mediated increase in gut permeability in mice

    Directory of Open Access Journals (Sweden)

    Keshavarzian Ali


    Full Text Available Abstract Background Exposure to particulate matter (PM air pollution may be an important environmental factor leading to exacerbations of inflammatory illnesses in the GI tract. PM can gain access to the gastrointestinal (GI tract via swallowing of air or secretions from the upper airways or mucociliary clearance of inhaled particles. Methods We measured PM-induced cell death and mitochondrial ROS generation in Caco-2 cells stably expressing oxidant sensitive GFP localized to mitochondria in the absence or presence of an antioxidant. C57BL/6 mice were exposed to a very high dose of urban PM from Washington, DC (200 μg/mouse or saline via gastric gavage and small bowel and colonic tissue were harvested for histologic evaluation, and RNA isolation up to 48 hours. Permeability to 4kD dextran was measured at 48 hours. Results PM induced mitochondrial ROS generation and cell death in Caco-2 cells. PM also caused oxidant-dependent NF-κB activation, disruption of tight junctions and increased permeability of Caco-2 monolayers. Mice exposed to PM had increased intestinal permeability compared with PBS treated mice. In the small bowel, colocalization of the tight junction protein, ZO-1 was lower in the PM treated animals. In the small bowel and colon, PM exposed mice had higher levels of IL-6 mRNA and reduced levels of ZO-1 mRNA. Increased apoptosis was observed in the colon of PM exposed mice. Conclusions Exposure to high doses of urban PM causes oxidant dependent GI epithelial cell death, disruption of tight junction proteins, inflammation and increased permeability in the gut in vitro and in vivo. These PM-induced changes may contribute to exacerbations of inflammatory disorders of the gut.

  5. Study on road surface source pollution controlled by permeable pavement (United States)

    Zheng, Chaocheng


    The increase of impermeable pavement in urban construction not only increases the runoff of the pavement, but also produces a large number of Non-Point Source Pollution. In the process of controlling road surface runoff by permeable pavement, a large number of particulate matter will be withheld when rainwater is being infiltrated, so as to control the source pollution at the source. In this experiment, we determined the effect of permeable road surface to remove heavy pollutants in the laboratory and discussed the related factors that affect the non-point pollution of permeable pavement, so as to provide a theoretical basis for the application of permeable pavement.

  6. Calculation of Permeability inside the Basket including one Fuel Assembly

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Seung Hwan; Bang, Kyung Sik; Lee, Ju an; Choi, Woo Seok [KAERI, Daejeon (Korea, Republic of)


    In general, the porous media model and the effective thermal conductivity were used to simply the fuel assembly. The methods of calculating permeability were compared considering the flow inside a basket which includes a nuclear fuel. Detailed fuel assembly was a computational modeling and the flow characteristics were investigated. The flow inside the basket which included a fuel assembly is analyzed by CFD. As the height of the fuel assembly increases, the pressure drop linearly increased. The inertia resistance could be neglected. Three methods to calculate the permeability were compared. The permeability by the friction factor is 50% less than the permeability by wall shear stress and pressure drop.

  7. Fast Laplace solver approach to pore-scale permeability (United States)

    Arns, C. H.; Adler, P. M.


    We introduce a powerful and easily implemented method to calculate the permeability of porous media at the pore scale using an approximation based on the Poiseulle equation to calculate permeability to fluid flow with a Laplace solver. The method consists of calculating the Euclidean distance map of the fluid phase to assign local conductivities and lends itself naturally to the treatment of multiscale problems. We compare with analytical solutions as well as experimental measurements and lattice Boltzmann calculations of permeability for Fontainebleau sandstone. The solver is significantly more stable than the lattice Boltzmann approach, uses less memory, and is significantly faster. Permeabilities are in excellent agreement over a wide range of porosities.

  8. Frictional stability-permeability relationships for fractures in shales (United States)

    Fang, Yi; Elsworth, Derek; Wang, Chaoyi; Ishibashi, Takuya; Fitts, Jeffrey P.


    There is wide concern that fluid injection in the subsurface, such as for the stimulation of shale reservoirs or for geological CO2 sequestration (GCS), has the potential to induce seismicity that may change reservoir permeability due to fault slip. However, the impact of induced seismicity on fracture permeability evolution remains unclear due to the spectrum of modes of fault reactivation (e.g., stable versus unstable). As seismicity is controlled by the frictional response of fractures, we explore friction-stability-permeability relationships through the concurrent measurement of frictional and hydraulic properties of artificial fractures in Green River shale (GRS) and Opalinus shale (OPS). We observe that carbonate-rich GRS shows higher frictional strength but weak neutral frictional stability. The GRS fracture permeability declines during shearing while an increased sliding velocity reduces the rate of permeability decline. By comparison, the phyllosilicate-rich OPS has lower friction and strong stability while the fracture permeability is reduced due to the swelling behavior that dominates over the shearing induced permeability reduction. Hence, we conclude that the friction-stability-permeability relationship of a fracture is largely controlled by mineral composition and that shale mineral compositions with strong frictional stability may be particularly subject to permanent permeability reduction during fluid infiltration.

  9. In vitro blood-brain barrier permeability predictions for GABAA receptor modulating piperine analogs. (United States)

    Eigenmann, Daniela Elisabeth; Dürig, Carmen; Jähne, Evelyn Andrea; Smieško, Martin; Culot, Maxime; Gosselet, Fabien; Cecchelli, Romeo; Helms, Hans Christian Cederberg; Brodin, Birger; Wimmer, Laurin; Mihovilovic, Marko D; Hamburger, Matthias; Oufir, Mouhssin


    The alkaloid piperine from black pepper (Piper nigrum L.) and several synthetic piperine analogs were recently identified as positive allosteric modulators of γ-aminobutyric acid type A (GABAA) receptors. In order to reach their target sites of action, these compounds need to enter the brain by crossing the blood-brain barrier (BBB). We here evaluated piperine and five selected analogs (SCT-66, SCT-64, SCT-29, LAU397, and LAU399) regarding their BBB permeability. Data were obtained in three in vitro BBB models, namely a recently established human model with immortalized hBMEC cells, a human brain-like endothelial cells (BLEC) model, and a primary animal (bovine endothelial/rat astrocytes co-culture) model. For each compound, quantitative UHPLC-MS/MS methods in the range of 5.00-500ng/mL in the corresponding matrix were developed, and permeability coefficients in the three BBB models were determined. In vitro predictions from the two human BBB models were in good agreement, while permeability data from the animal model differed to some extent, possibly due to protein binding of the screened compounds. In all three BBB models, piperine and SCT-64 displayed the highest BBB permeation potential. This was corroborated by data from in silico prediction. For the other piperine analogs (SCT-66, SCT-29, LAU397, and LAU399), BBB permeability was low to moderate in the two human BBB models, and moderate to high in the animal BBB model. Efflux ratios (ER) calculated from bidirectional permeability experiments indicated that the compounds were likely not substrates of active efflux transporters. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Acoustic--nuclear permeability logging system

    International Nuclear Information System (INIS)

    Dowling, D.J.; Arnold, D.M.


    A down hole logging tool featuring a neutron generator, an acoustic disturbance generator, and a radiation detection system is described. An array of acoustic magnetostriction transducers is arranged about the target of a neutron accelerator. Two gamma ray sensors are separated from the accelerator target by shielding. According to the method of the invention, the underground fluid at the level of a formation is bombarded by neutrons which react with oxygen in the fluid to produce unstable nitrogen 16 particles according to the reaction 16 O(n,p) 16 N. Acoustic pulses are communicated to the fluid, and are incident on the boundary of the borehole at the formation. The resulting net flow of fluid across the boundary is determined from radiation detection measurements of the decaying 16 N particles in the fluid. A measure of the permeability of the formation is obtained from the determination of net fluid flow across the boundary

  11. Treatment for cracked and permeable Houston clay

    International Nuclear Information System (INIS)

    Vipulanandan, C.; Leung, M.


    In this study, the treatability of a field clay (obtained from Houston, Texas) and a clay-sand mixture to reduce their hydraulic conductivity was evaluated. Remolded field clay and clay-sand mixture with and without methanol contamination were treated to reduce their hydraulic conductivity by permeating very dilute grout solutions. The concentration of sodium silicate in the grout solution was 8%, while the solid content in the cement grout was 0.3%. The hydraulic conductivity of permeable Houston clay (hydraulic conductivity >10 -5 cm/sec) could be reduced to less than 10 -7 cm/sec (U.S. EPA limit for soil barriers) by permeating with a selected combination of grout solutions

  12. Migration of particulates in permeable rock columns

    International Nuclear Information System (INIS)

    Cropper, R.L.


    The migration of radioactive material through soil and permeable rock formations have become a major topic of concern due to the interest in the licensing of new radioactive waste disposal sites. Previously, research has been conducted in relation to deep repositories; however, similar situations arise in the vadose zone, where there is a higher probability of naturally-occurring particulates of organic nature and for the incursion of water. Test data has provided information which suggests that particulates will travel through porous media subject to various delay mechnisms and must be included in any consideration of waste migration. Data concerning particulate migration must and should be considered in the future when radioactive waste disposal sites are licensed

  13. Composite binders for concrete with reduced permeability

    International Nuclear Information System (INIS)

    Fediuk, R; Yushin, A


    Composite binder consisting of cement (55%), acid fly ash (40%) and limestone (5%) has been designed. It is obtained by co-milling to a specific surface of 550 kg/m 2 , it has an activity of 77.3 MPa and can produce a more dense cement stone structure. Integrated study revealed that the concrete on the composite binder basis provides an effective diffusion coefficient D. So we can conclude that the concrete layer protects buildings from toxic effects of expanded polystyrene. Low water absorption of the material (2.5% by weight) is due to the structure of its cement stone pore space. Besides lime powder prevents the penetration of moisture, reduces water saturation of the coverage that has a positive effect on useful life period. It also explains rather low water vapor permeability of the material - 0.021 mg/(m- hour-Pa). (paper)

  14. Consolidation and permeability of salt in brine

    International Nuclear Information System (INIS)

    Shor, A.J.; Baes, C.F. Jr.; Canonico, C.M.


    The consolidation and loss of permeability of salt crystal aggregates, important in assessing the effects of water in salt repositories, has been studied as a function of several variables. The kinetic behavior was similar to that often observed in sintering and suggested the following expression for the time dependence of the void fraction: phi(t) = phi(0) - (A/B)ln(1 + Bt/z(0) 3 ), where A and B are rate constants and z(0) is initial average particle size. With brine present, A and phi(0) varied linearly with stress. The initial void fraction was also dependent to some extent on the particle size distribution. The rate of consolidation was most rapid in brine and least rapid in the presence of only air as the fluid. A brine containing 5 m MgCl 2 showed an intermediate rate, presumably because of the greatly reduced solubility of NaCl. A substantial wall effect was indicated by an observed increase in the void fraction of consolidated columns with distance from the top where the stress was applied and by a dependence of consolidation rate on the column height and radius. The distance through which the stress fell by a factor of phi was estimated to change inversely as the fourth power of the column diameter. With increasing temperature (to 85 0 C), consolidation proceeded somewhat more rapidly and the wall effect was reduced. The permeability of the columns dropped rapidly with consolidation, decreasing with about the sixth power of the void fraction. In general, extrapolation of the results to repository conditions confirms the self-sealing properties of bedded salt as a storage medium for radioactive waste

  15. Colloid transport in dual-permeability media (United States)

    Leij, Feike J.; Bradford, Scott A.


    It has been widely reported that colloids can travel faster and over longer distances in natural structured porous media than in uniform structureless media used in laboratory studies. The presence of preferential pathways for colloids in the subsurface environment is of concern because of the increased risks for disease caused by microorganisms and colloid-associated contaminants. This study presents a model for colloid transport in dual-permeability media that includes reversible and irreversible retention of colloids and first-order exchange between the aqueous phases of the two regions. The model may also be used to describe transport of other reactive solutes in dual-permeability media. Analytical solutions for colloid concentrations in aqueous and solid phases were obtained using Laplace transformation and matrix decomposition. The solutions proved convenient to assess the effect of model parameters on the colloid distribution. The analytical model was used to describe effluent concentrations for a bromide tracer and 3.2- or 1-μm-colloids that were observed after transport through a composite 10-cm long porous medium made up of a cylindrical lens or core of sand and a surrounding matrix with sand of a different grain size. The tracer data were described very well and realistic estimates were obtained for the pore-water velocity in the two flow domains. An accurate description was also achieved for most colloid breakthrough curves. Dispersivity and retention parameters were typically greater for the larger 3.2-μm-colloids while both reversible and irreversible retention rates tended to be higher for the finer sands than the coarser sand. The relatively small sample size and the complex flow pattern in the composite medium made it difficult to reach definitive conclusions regarding transport parameters for colloid transport.


    Institute of Scientific and Technical Information of China (English)

    CHENGuo-Shen; GONGSai-Jun; DUJie; MARun-Zhen; ZHOURong-Rong; LIULiang-Chu


    Transdermal permeability of estradiol was carried out by using Valia-Chien double compartment permeation cells for the following regions of intact skin and skin without stratum corncum: chest, abdomen, hip, upper arm, thigh and back. The estradiol permeation rates and accumulative amounts within 72h in vitro were examined by HPLC. The results showed that the permeation rates of intact skin from different regions of the body

  17. Decreased outer membrane permeability in imipenem-resistant mutants of Pseudomonas aeruginosa.


    Trias, J; Dufresne, J; Levesque, R C; Nikaido, H


    The outer membrane of imipenem-resistant mutants of Pseudomonas aeruginosa was shown to have decreased permeability to imipenem but not to cephaloridine. These experiments were performed with intact cells and liposomes containing imipenem-hydrolyzing beta-lactamase derived from Pseudomonas maltophilia, in both cases utilizing an imipenem concentration of 50 microM. In contrast, liposome swelling assays using imipenem at 8 mM detected no significant difference between the imipenem-resistant mu...

  18. Design Guidance for Application of Permeable Barriers to Remediate Dissolved Chlorinated Solvents, (United States)


    fill slurry composed of a reactive medium, such as iron powder and guar gum , can then be injected into the fracture to form a reactive treatment zone...slurry (Owaidat, 1996). The slurry, which is composed of powdered guar bean, acts to maintain the integrity of the trench walls during installation of...the cell. The guar gum will later biodegrade to mostly water after wall completion, and will have minimal effect on the permeability of the trench

  19. Involvement of membrane lipids in radiation damage to potassium-ion permeability of Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Suzuki, S [Tokyo Univ. (Japan). Inst. for Medical Science; Akamatsu, Y


    Radiation damage to K/sup +/ permeability of an unsaturated fatty acid auxotroph of E.coli grown with oleate or linolenate was investigated at different temperatures. A remarkable effect of radiation was observed at 0/sup 0/C with cells that had been grown in linolenate at 42/sup 0/C. This indicates that, besides protein, membrane lipids at least are involved in the radiation damage. The damage also seems to be affected by the fluidity of membrane lipids.

  20. Increased gut permeability in cancer cachexia: mechanisms and clinical relevance. (United States)

    Bindels, Laure B; Neyrinck, Audrey M; Loumaye, Audrey; Catry, Emilie; Walgrave, Hannah; Cherbuy, Claire; Leclercq, Sophie; Van Hul, Matthias; Plovier, Hubert; Pachikian, Barbara; Bermúdez-Humarán, Luis G; Langella, Philippe; Cani, Patrice D; Thissen, Jean-Paul; Delzenne, Nathalie M


    Intestinal disorders often occur in cancer patients, in association with body weight loss, and this alteration is commonly attributed to the chemotherapy. Here, using a mouse model of cancer cachexia induced by ectopic transplantation of C26 cancer cells, we discovered a profound alteration in the gut functions (gut permeability, epithelial turnover, gut immunity, microbial dysbiosis) independently of any chemotherapy. These alterations occurred independently of anorexia and were driven by interleukin 6. Gut dysfunction was found to be resistant to treatments with an anti-inflammatory bacterium ( Faecalibacterium prausnitzii ) or with gut peptides involved in intestinal cell renewal (teduglutide, a glucagon-like peptide 2 analogue). The translational value of our findings was evaluated in 152 colorectal and lung cancer patients with or without cachexia. The serum level of the lipopolysaccharide-binding protein, often presented as a reflection of the bacterial antigen load, was not only increased in cachectic mice and cancer patients, but also strongly correlated with the serum IL-6 level and predictive of death and cachexia occurrence in these patients. Altogether, our data highlight profound alterations of the intestinal homeostasis in cancer cachexia occurring independently of any chemotherapy and food intake reduction, with potential relevance in humans. In addition, we point out the lipopolysaccharide-binding protein as a new biomarker of cancer cachexia related to gut dysbiosis.

  1. Measurement of Interfacial Area Production and Permeability within Porous Media

    International Nuclear Information System (INIS)

    Crandall, Dustin; Ahmadi, Goodarz; Smith, Duane H.


    An understanding of the pore-level interactions that affect multi-phase flow in porous media is important in many subsurface engineering applications, including enhanced oil recovery, remediation of dense non-aqueous liquid contaminated sites, and geologic CO 2 sequestration. Standard models of two-phase flow in porous media have been shown to have several shortcomings, which might partially be overcome using a recently developed model based on thermodynamic principles that includes interfacial area as an additional parameter. A few static experimental studies have been previously performed, which allowed the determination of static parameters of the model, but no information exists concerning the interfacial area dynamic parameters. A new experimental porous flow cell that was constructed using stereolithography for two-phase gas-liquid flow studies was used in conjunction with an in-house analysis code to provide information on dynamic evolution of both fluid phases and gas-liquid interfaces. In this paper, we give a brief introduction to the new generalized model of two-phase flow model and describe how the stereolithography flow cell experimental setup was used to obtain the dynamic parameters for the interfacial area numerical model. In particular, the methods used to determine the interfacial area permeability and production terms are shown.

  2. Caco-2 Permeability Studies and In Vitro hERG Liability Assessment of Tryptanthrin and Indolinone. (United States)

    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. Georg Thieme Verlag KG Stuttgart · New York.

  3. Modeling the Hydrologic Processes of a Permeable Pavement System (United States)

    A permeable pavement system can capture stormwater to reduce runoff volume and flow rate, improve onsite groundwater recharge, and enhance pollutant controls within the site. A new unit process model for evaluating the hydrologic performance of a permeable pavement system has be...

  4. Performances of Metal Concentrations from Three Permeable Pavement Infiltrates (United States)

    The U.S. Environmental Protection Agency designed and constructed a 4000-m2 parking lot in Edison, New Jersey in 2009. The parking lot is surfaced with three permeable pavements: permeable interlocking concrete pavers, pervious concrete, and porous asphalt. Water sampling was con...

  5. Permeable Pavement Research at the Edison Environmental Center (United States)

    There are few detailed studies of full-scale, replicated, actively-used permeable pavement systems. Practitioners need additional studies of permeable pavement systems in its intended application (parking lot, roadway, etc.) across a range of climatic events, daily usage conditio...

  6. Nitrogen Transformations in Three Types of Permeable Pavement (United States)

    In 2009, USEPA constructed a 0.4-ha (1-ac) parking lot at the Edison Environmental Center in Edison, NJ, that incorporated three different permeable pavement types - permeable interlocking concrete pavers (PICP), pervious concrete (PC), and porous asphalt (PA). The driving lanes...

  7. Update to Permeable Pavement Research at the Edison ... (United States)

    The EPA’s Urban Watershed Management Branch (UWMB) has been monitoring the permeable pavement demonstration site at the Edison Environmental Center, NJ since 2010. This site has three different types of permeable pavements including interlocking concrete permeable pavers, pervious concrete, and porous asphalt. The permeable pavements are limited to parking spaces while adjacent driving lanes are impermeable and drain to the permeable surfaces. The parking lot is instrumented for continuous monitoring with thermistors and water content reflectometers that measure moisture as infiltrate passes through the storage gallery beneath the permeable pavements into the underlying native soil. Each permeable surface of the parking lot has four lined sections that capture infiltrate in tanks for water quality analyses; these tanks are capable of holding volumes up to 4.1 m3, which represents up to 38 mm (1.5 in.) for direct rainfall on the porous pavement and runoff from adjacent driving lanes that drain into the permeable surface.Previous technical releases concerning the demonstration site focused on monitoring techniques, observed chloride and nutrient concentrations, surface hydrology, and infiltration and evaporation rates. This presentation summarizes these past findings and addresses current water quality efforts including pH, solids analysis, total organic carbon, and chemical oxygen demand. Stormwater runoff continues to be a major cause of water pollution in

  8. Permeability dependence of streaming potential coefficient in porous media

    NARCIS (Netherlands)

    Thanh, L.D.; Sprik, R.


    In theory, the streaming potential coefficient depends not only on the zeta potential but also on the permeability of the rocks that partially determines the surface conductivity of the rocks. However, in practice, it is hard to show the permeability dependence of streaming potential coefficients

  9. A drainage data-based calculation method for coalbed permeability

    International Nuclear Information System (INIS)

    Lai, Feng-peng; Li, Zhi-ping; Fu, Ying-kun; Yang, Zhi-hao


    This paper establishes a drainage data-based calculation method for coalbed permeability. The method combines material balance and production equations. We use a material balance equation to derive the average pressure of the coalbed in the production process. The dimensionless water production index is introduced into the production equation for the water production stage. In the subsequent stage, which uses both gas and water, the gas and water production ratio is introduced to eliminate the effect of flush-flow radius, skin factor, and other uncertain factors in the calculation of coalbed methane permeability. The relationship between permeability and surface cumulative liquid production can be described as a single-variable cubic equation by derivation. The trend shows that the permeability initially declines and then increases after ten wells in the southern Qinshui coalbed methane field. The results show an exponential relationship between permeability and cumulative water production. The relationship between permeability and cumulative gas production is represented by a linear curve and that between permeability and surface cumulative liquid production is represented by a cubic polynomial curve. The regression result of the permeability and surface cumulative liquid production agrees with the theoretical mathematical relationship. (paper)

  10. Permeability of crust is key to crispness retention

    NARCIS (Netherlands)

    Hirte, A.; Hamer, R.J.; Meinders, M.B.J.; Primo-Martin, C.


    Bread loses crispness rapidly after baking because water originating from the wet crumb accumulates in the dry crust. This water accumulation might be increased by the dense and low permeable character of the bread crust. Our objective was to investigate the influence of permeability of the crust on

  11. Principal permeability determination from multiple horizontal well tests

    Energy Technology Data Exchange (ETDEWEB)

    Economides, M. [Texas A and M Univ., TX (United States); Munoz, A.; Ehlig-Economides, C.


    A method for obtaining principal permeability magnitudes and direction that requires only the linear flow regime from transient tests in three horizontal wells oriented in three distinct and arbitrary directions, is described. Well design optimization strategies require knowledge of both the principal permeability orientation as well as the horizontal permeability magnitudes. When the degree of horizontal permeability anisotropy (i.e. permeability in the bedding plane with respect to direction) is significant, the productivity of a long horizontal well will depend greatly on its direction, especially when the well is first brought into production. Productivities have been found to deviate substantially among wells in the same reservoir and this deviation has been attributed to differences in well orientation. In view of this fact, measuring permeability anisotropy becomes a compelling necessity. The success of the proposed method is illustrated by a case study in which the principal permeability magnitudes and direction from three wells were used to predict the productivity of a fourth well within 10 per cent. Use of the computed principal permeabilities from the case study, it was possible to forecast the cumulative production to show the significance of well trajectory optimization on the discounted cash flow and the net present value. 20 refs., 3 figs.

  12. Determination of hydrogen permeability in uncoated and coated superalloys (United States)

    Bhattacharyya, S.; Vesely, E. J., Jr.; Hill, V. L.


    Hydrogen permeability, diffusivity, and solubility data were obtained for eight wrought and cast high temperature alloys over the range 650 to 815 C. Data were obtained for both uncoated alloys and wrought alloys coated with four commercially available coatings. Activation energies for permeability, diffusivity and solubility were calculated.

  13. Unsaturated and Saturated Permeabilities of Fiber Reinforcement: Critics and Suggestions

    Directory of Open Access Journals (Sweden)

    Chung Hae ePARK


    Full Text Available In general, permeability measurement results show a strong scattering according to the measurement method, the type of test fluid and the fluid injection condition, even though permeability is regarded as a unique property of porous medium. In particular, the discrepancy between the unsaturated and saturated permeabilities for the same fabric has been widely reported. In the literature, relative permeability has been adopted to model the unsaturated flow. This approach has some limits in the modeling of double-scale porosity medium. We address this issue of permeability measurement by rigorously examining the mass conservation condition. Finally, we identify that the pressure gradient is non-linear with positive curvature in the unsaturated flow and a misinterpretation of pressure gradient is the main reason for the difference between the saturated and unsaturated permeabilities of the same fiber reinforcement. We propose to use a fixed value of permeability and to modify the mass conservation equation if there are air voids which are entrapped inside the fiber tow. Finally, we also suggest some guidelines and future perspectives to obtain more consistent permeability measurement results.

  14. The long noncoding RNA TUG1 regulates blood-tumor barrier permeability by targeting miR-144. (United States)

    Cai, Heng; Xue, Yixue; Wang, Ping; Wang, Zhenhua; Li, Zhen; Hu, Yi; Li, Zhiqing; Shang, Xiuli; Liu, Yunhui


    Blood-tumor barrier (BTB) limits the delivery of chemotherapeutic agent to brain tumor tissues. Long non-coding RNAs (lncRNAs) have been shown to play critical regulatory roles in various biologic processes of tumors. However, the role of lncRNAs in BTB permeability is unclear. LncRNA TUG1 (taurine upregulated gene 1) was highly expressed in glioma vascular endothelial cells from glioma tissues. It also upregulated in glioma co-cultured endothelial cells (GEC) from BTB model in vitro. Knockdown of TUG1 increased BTB permeability, and meanwhile down-regulated the expression of the tight junction proteins ZO-1, occludin, and claudin-5. Both bioinformatics and luciferase reporter assays demonstrated that TUG1 influenced BTB permeability via binding to miR-144. Furthermore, Knockdown of TUG1 also down-regulated Heat shock transcription factor 2 (HSF2), a transcription factor of the heat shock transcription factor family, which was defined as a direct and functional downstream target of miR-144. HSF2 up-regulated the promoter activities and interacted with the promoters of ZO-1, occludin, and claudin-5 in GECs. In conclusion, our results indicate that knockdown of TUG1 increased BTB permeability via binding to miR-144 and then reducing EC tight junction protein expression by targeting HSF2. Thus, TUG1 may represent a useful future therapeutic target for enhancing BTB permeability.

  15. Prediction of permeability changes in an excavation response zone

    International Nuclear Information System (INIS)

    Kinoshita, Naoto; Ishii, Takashi; Kuroda, Hidetaka; Tada, Hiroyuki


    In geologic disposal of radioactive wastes, stress changes due to cavern excavation may expand the existing fractures and create possible bypasses for groundwater. This paper proposes a simple method for predicting permeability changes in the excavation response zones. Numerical analyses using this method predict that the response zones created by cavern excavation would differ greatly in thickness and permeability depending on the depth of the cavern site and the initial in-situ stress, that when the cavern site is deeper, response zones would expand more and permeability would increases more, and that if the ratio of horizontal to vertical in-situ stress is small, extensive permeable zones at the crown and the bottom would occur, whereas if the ratio is large, extensive permeable zones would occur in the side walls. (orig.)

  16. Effect Of Hot Water Injection On Sandstone Permeability

    DEFF Research Database (Denmark)

    Rosenbrand, Esther; Fabricius, Ida Lykke


    of published results regarding the effect of temperature on sandstone permeability. These tests are performed with mineral oil, nitrogen gas, distilled water and solutions of NaCl, KCl, CaCl2 as well as brines that contain a mixture of salts. Thirteen sandstone formations, ranging from quartz arenites...... to formations with a significant fraction of fine particles including clay minerals are investigated. The porosities range from 0.10 to 0.30 and permeabilities span the range from 1 to 1000 md. To compare different rock types, specific surface is determined from permeability and porosity using Kozeny’s equation...... not account for all the permeability reductions observed. Permeablity reduction occurs both when distilled water is the saturating fluid as well as in tests with NaCl, KCl or CaCl2 solutions, however, this is not the case in tests with mineral oil or nitrogen gas. The formation of a filter cake or influx...

  17. A theoretical model for gas permeability in a composite membrane

    International Nuclear Information System (INIS)

    Serrano, D. A


    We present in this work an analytical expression for permeability in a two-layer composite membrane, which was derived assuming the same hypothesis as those of Adzumi model for permeability in a homogeneous membrane. Whereas in Adzumi model permeability shows a linear dependence on the mean pressure, our model for a composite membrane related permeability to pressure through a rather complex expression, which covers the whole range of flow, from molecular-Knudsen to viscous-Poiseuille regimes. The expression obtained for permeability contained information of membrane structural properties as pore size, porosity and thickness of each layer, as well as gas nature and operational conditions. Our two-layer-model expression turns into Adzumi formula when the structure of the layers approach to each other. [es

  18. Steady flow in voids and closed cracks in permeable media

    International Nuclear Information System (INIS)

    Rae, J.


    This paper considers what happens when a steady flow in a permeable medium meets two concentric spheres which have different permeabilities. This can form a first stage model for water flow near an engineered cavity in rock or a concreted waste package placed in filler material as in a nuclear waste repository. Results are obtained in terms of the simplest spherical harmonics, which lets them be used easily. Included are the well-known result that a highly permeable sphere will see only a few times the flux which would occur if it had the permeability of its surroundings, and the less well-known result, though unsurprising, that a spherical region surrounded by a highly permeable shell will see almost no flow, as it will almost all by-pass. A companion paper will include more geometrical effects by replacing the spheres by ellipsoids. (author)

  19. Ozone alteration of membrane permeability in Chlorella. I. Permeability of potassium ion as measured by 86Rubidium tracer

    International Nuclear Information System (INIS)

    Heath, R.L.; Frederick, P.E.


    The addition of ozone to a suspension of Chlorella sorokiniana causes a rapid loss of K + , as measured by efflux of 86 Rb from prelabeled cells. The efflux of the tracer is stimulated some 15 to 20 times over that of the control. For about 100 microliters per liter ozone, about 25 minutes (6 x 10 -8 moles O 3 delivered per minute) of exposure are required for a 50% depletion of the intracellular K + . The stimulation of K + efflux is nearly linearly dependent upon the amount of ozone delivered into the solution. Following short pulses of ozone (lasting 1 to 5 minutes), efflux rates return to the control level but only after about 15 minutes. While influx of K + is ultimately inhibited by ozone, at low concentrations or for short exposure times the tracer influx is stimulated 100 to 200%. Ozone stimulation of an active pump mechanism is unlikely in view of a concomitant decrease in respiration. Thus, this influx may represent movement of K + along its electrochemical gradient. Assuming that influx and efflux are in steady-state according to the Goldman equation, it was calculated that the membrane potential for K + of -80 to -90 millivolts in control cells drops to -40 millivolts with ozone exposure and is accompanied by a calculated increased permeability to K + of 2- to 3-fold. 25 references, 6 figures

  20. Permeability, storage and hydraulic diffusivity controlled by earthquakes (United States)

    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

  1. In-situ permeability measurements with direct push techniques: Phase II topical report

    International Nuclear Information System (INIS)

    Lowry, W.; Mason, N.; Chipman, V.; Kisiel, K.; Stockton, J.


    This effort designed, fabricated, and field tested the engineering prototype of the Cone Permeametertrademark system. The integrated system includes the instrumented penetrometer probe, air and water pumps, flowrate controls, flow sensors, and a laptop-controlled data system. All of the equipment is portable and can be transported as luggage on airlines. The data system acquired and displays the process measurements (pressures, flows, and downhole temperature) in real time and calculates the resulting permeability. The measurement probe is a 2 inch diameter CPT rod section, incorporating a screened injection zone near the lower end of the rod and multiple sensitive absolute pressure sensors embedded in the probe at varying distances from the injection zone. Laboratory tests in a large test cell demonstrated the system's ability to measure nominally 1 Darcy permeability soil (30 to 40 Darcy material had been successfully measured in the Phase 1 effort). These tests also provided a shakedown of the system and identified minor instrument problems, which were resolved. Supplemental numerical modeling was conducted to evaluate the effects of layered permeability (heterogeneity) and anisotropy on the measurement system's performance. The general results of the analysis were that the Cone Permeameter could measure accurately, in heterogeneous media, the volume represented by the sample port radii if the outer pressure ports were used. Anisotropic permeability, while readily analyzed numerically, is more complicated to resolve with the simple analytical approach of the 1-D model, and will need further work to quantify. This phase culminated in field demonstrations at the DOE Savannah River Site. Saturated hydraulic conductivity measurements were completed at the D-Area Coal Pile Runoff Basin, and air permeability measurements were conducted at the M Area Integrated Demonstration Site and the 321 M area. The saturated hydraulic conductivity measurements were the most

  2. Effect of temperature on damage and permeability of clayey soils and rocks

    International Nuclear Information System (INIS)

    Monfared, M.


    Storage of exothermic radioactive waste in deep low permeability geological formations such as clayey rocks and plastic clays is a solution considered for long term repositories. However the excavation of underground galleries creates a damaged zone (EDZ). The effect of the damage zone on the transport properties of the geological barrier has been widely studied. Within the framework of the TIMODAZ European project, emphasis has been put on the effect of temperature. As a partner of this project, the current work is performed to investigate the coupling effect between temperature, damage and permeability on Boom clay and Opalinus clay through an experimental study. View to the experimental difficulties related to the low permeability materials, a new hollow cylinder triaxial cell with short drainage path specifically designed to study the thermo-hydro-mechanical behaviour of very low permeable materials is developed during this work. The tests and the numerical analysis show that the short sample drainage path reduces significantly the time needed to re-saturate an initially unsaturated sample and it also permits to achieve drained conditions (i.e. negligible excess pore pressure during testing) with a higher loading rate. For Boom clay, the effect of the pore water thermal pressurisation on a sample with a pre-existing shear band is investigated. The undrained heating under shear stress decreases the effective stress on the sample which leads to its failure. An existing failure plane in the sample behaves like a preferential weakness plane which can be reactivated by pore water thermal pressurisation. The estimated shearing resistance along the sheared plane is smaller than that of the intact material. For the Opalinus clay-stone, drained heating on a saturated sample shows that this clay-stone behaves like a slightly over consolidated material (thermo-elasto-plastic behaviour) with transition from expansion to contraction at 65 C. The decrease of the permeability

  3. Small GTP-Binding Protein Rac Is an Essential Mediator of Vascular Endothelial Growth Factor-Induced Endothelial Fenestrations and Vascular Permeability

    DEFF Research Database (Denmark)

    Eriksson, A.; Cao, R.; Tritsaris, K.


    fenestrated endothelium, a feature linked with increased vascular permeability. A cell-permeable Rac antagonist (TAT-RacN17) converted VEGF-induced, leaky vascular plexuses into well-defined vascular networks. In addition, this Rac mutant blocked formation of VEGF-induced endothelial fenestrations...... in mediation of VEGF-induced vascular permeability but less so in neovascularization. This may have conceptual implications for applying Rac antagonists in treatment and prevention of VEGF-induced vascular leakage and edema in connection with ischemic disorders....

  4. Using artificial intelligence to predict permeability from petrographic data

    Energy Technology Data Exchange (ETDEWEB)

    Maqsood Ali; Adwait Chawathe [New Mexico Petroleum Recovery Research Centre (Mexico)


    Petrographic data collected during thin section analysis can be invaluable for understanding the factors that control permeability distribution. Reliable prediction of permeability is important for reservoir characterization. The petrographic elements (mineralogy, porosity types, cements and clays, and pore morphology) interact with each other uniquely to generate a specific permeability distribution. It is difficult to quantify accurately this interaction and its consequent effect on permeability, emphasizing the non-linear nature of the process. To capture these non-linear interactions, neural networks were used to predict permeability from petrographic data. The neural net was used as a multivariate correlative tool because of its ability to learn the non-linear relationships between multiple input and output variables. The study was conducted on the upper Queen formation called the Shattuck Member (Permian age). The Shattuck Member is composed of very fine-grained arkosic sandstone. The core samples were available from the Sulimar Queen and South Lucky Lake fields located in Chaves County, New Mexico. Nineteen petrographic elements were collected for each permeability value using a combined minipermeameter-petrographic technique. In order to reduce noise and overfitting the permeability model, these petrographic elements were screened, and their control (ranking) with respect to permeability was determined using fuzzy logic. Since the fuzzy logic algorithm provides unbiased ranking, it was used to reduce the dimensionality of the input variables. Based on the fuzzy logic ranking, only the most influential petrographic elements were selected as inputs for permeability prediction. The neural net was trained and tested using data from Well 1-16 in the Sulimar Queen field. Relying on the ranking obtained from the fuzzy logic analysis, the net was trained using the most influential three, five, and ten petrographic elements. A fast algorithm (the scaled conjugate

  5. Factors Influencing Stormwater Mitigation in Permeable Pavement

    Directory of Open Access Journals (Sweden)

    Chun Yan Liu


    Full Text Available Permeable pavement (PP is used worldwide to mitigate surface runoff in urban areas. Various studies have examined the factors governing the hydrologic performance of PP. However, relatively little is known about the relative importance of these governing factors and the long-term hydrologic performance of PP. This study applied numerical models—calibrated and validated using existing experimental results—to simulate hundreds of event-based and two long-term rainfall scenarios for two designs of PP. Based on the event-based simulation results, rainfall intensity, rainfall volume, thickness of the storage layer and the hydraulic conductivity of the subgrade were identified as the most influential factors in PP runoff reduction. Over the long term, PP performed significantly better in a relatively drier climate (e.g., New York, reducing nearly 90% of runoff volume compared to 70% in a relatively wetter climate (e.g., Hong Kong. The two designs of PP examined performed differently, and the difference was more apparent in the relatively wetter climate. This study generated insights that will help the design and implementation of PP to mitigate stormwater worldwide.

  6. Electrically Driven Ion Separations in Permeable Membranes

    Energy Technology Data Exchange (ETDEWEB)

    Bruening, Merlin [Michigan State Univ., East Lansing, MI (United States)


    Membranes are attractive for a wide range of separations due to their low energy costs and continuous operation. To achieve practical fluxes, most membranes consist of a thin, selective skin on a highly permeable substrate that provides mechanical strength. Thus, this project focused on creating new methods for forming highly selective ultrathin skins as well as modeling transport through these coatings to better understand their unprecedented selectivities. The research explored both gas and ion separations, and the latter included transport due to concentration, pressure and electrical potential gradients. This report describes a series of highlights of the research and then provides a complete list of publications supported by the grant. These publications have been cited more than 4000 times. Perhaps the most stunning finding is the recent discovery of monovalent/divalent cation and anion selectivities around 1000 when modifying cation- and anion-exchange membranes with polyelectrolyte multilayers (PEMs). This discovery builds on many years of exciting research. (Citation numbers refer to the journal articles in the bibliography.)

  7. Permeability of WIPP Salt During Damage Evolution and Healing

    International Nuclear Information System (INIS)



    The presence of damage in the form of microcracks can increase the permeability of salt. In this paper, an analytical formulation of the permeability of damaged rock salt is presented for both initially intact and porous conditions. The analysis shows that permeability is related to the connected (i.e., gas accessible) volumetric strain and porosity according to two different power-laws, which may be summed to give the overall behavior of a porous salt with damage. This relationship was incorporated into a constitutive model, known as the Multimechanism Deformation Coupled Fracture (MDCF) model, which has been formulated to describe the inelastic flow behavior of rock salt due to coupled creep, damage, and healing. The extended model was used to calculate the permeability of rock salt from the Waste Isolation Pilot Plant (WIPP) site under conditions where damage evolved with stress over a time period. Permeability changes resulting from both damage development under deviatoric stresses and damage healing under hydrostatic pressures were considered. The calculated results were compared against experimental data from the literature, which indicated that permeability in damaged intact WIPP salt depends on the magnitude of the gas accessible volumetric strain and not on the total volumetric strain. Consequently, the permeability of WIPP salt is significantly affected by the kinetics of crack closure, but shows little dependence on the kinetics of crack removal by sintering

  8. Compression characteristics and permeability of saturated Gaomiaozi ca-bentonite

    International Nuclear Information System (INIS)

    Sun Wenjing; Sun De'an; Fang Lei


    The compression characteristics and permeability of compacted Gaomiaozi Ca-bentonite saturated by the water uptake tests are studied by conducting a series of one-dimension compression tests. The permeability coefficient can be calculated by the Terzaghi's one-dimensional consolidation theory after the consolidation coefficient is obtained by the square root of time method. It is found that the compression curves of compacted specimens saturated by the water uptake tests tend to be consistent in the relatively high stress range. The compression indexes show a linear decrease with increasing dry density and the swelling index is a constant. The permeability coefficient decreases with increasing compression stress, and they show the linear relationship in double logarithmic coordinates. Meanwhile, the permeability coefficient shows a linear decrease with decreasing void ratio, which has no relationship with initial states, stress states and stress paths. The permeability coefficient k of GMZ Ca-bentonite at dry density Pd of 1.75 g/cm 3 can be calculated as 2.0 × 10 -11 cm/s by the linear relationship between Pd and log k. It is closed to the permeability coefficient of GMZ Ca-bentonite with the same dry density published in literature, which testifies that the method calculating the permeability coefficient is feasible from the consolidation coefficient obtained by the consolidation test. (authors)

  9. Prediction of Hydrocarbon Reservoirs Permeability Using Support Vector Machine

    Directory of Open Access Journals (Sweden)

    R. Gholami


    Full Text Available Permeability is a key parameter associated with the characterization of any hydrocarbon reservoir. In fact, it is not possible to have accurate solutions to many petroleum engineering problems without having accurate permeability value. The conventional methods for permeability determination are core analysis and well test techniques. These methods are very expensive and time consuming. Therefore, attempts have usually been carried out to use artificial neural network for identification of the relationship between the well log data and core permeability. In this way, recent works on artificial intelligence techniques have led to introduce a robust machine learning methodology called support vector machine. This paper aims to utilize the SVM for predicting the permeability of three gas wells in the Southern Pars field. Obtained results of SVM showed that the correlation coefficient between core and predicted permeability is 0.97 for testing dataset. Comparing the result of SVM with that of a general regression neural network (GRNN revealed that the SVM approach is faster and more accurate than the GRNN in prediction of hydrocarbon reservoirs permeability.

  10. Microfluidic passive permeability assay using nanoliter droplet interface lipid bilayers. (United States)

    Nisisako, Takasi; Portonovo, Shiva A; Schmidt, Jacob J


    Membrane permeability assays play an important role in assessing drug transport activities across biological membranes. However, in conventional parallel artificial membrane permeability assays (PAMPA), the membrane model used is dissimilar to biological membranes physically and chemically. Here, we describe a microfluidic passive permeability assay using droplet interface bilayers (DIBs). In a microfluidic network, nanoliter-sized donor and acceptor aqueous droplets are alternately formed in cross-flowing oil containing phospholipids. Subsequently, selective removal of oil through hydrophobic pseudo-porous sidewalls induces the contact of the lipid monolayers, creating arrayed planar DIBs between the donor and acceptor droplets. Permeation of fluorescein from the donor to the acceptor droplets was fluorometrically measured. From the measured data and a simple diffusion model we calculated the effective permeabilities of 5.1 × 10(-6) cm s(-1), 60.0 × 10(-6) cm s(-1), and 87.6 × 10(-6) cm s(-1) with donor droplets at pH values of 7.5, 6.4 and 5.4, respectively. The intrinsic permeabilities of specific monoanionic and neutral fluorescein species were obtained similarly. We also measured the permeation of caffeine in 10 min using UV microspectroscopy, obtaining a permeability of 20.8 × 10(-6) cm s(-1). With the small solution volumes, short measurement time, and ability to measure a wide range of compounds, this device has considerable potential as a platform for high-throughput drug permeability assays.

  11. Characterization and estimation of permeability correlation structure from performance data

    Energy Technology Data Exchange (ETDEWEB)

    Ershaghi, I.; Al-Qahtani, M. [Univ. of Southern California, Los Angeles, CA (United States)


    In this study, the influence of permeability structure and correlation length on the system effective permeability and recovery factors of 2-D cross-sectional reservoir models, under waterflood, is investigated. Reservoirs with identical statistical representation of permeability attributes are shown to exhibit different system effective permeability and production characteristics which can be expressed by a mean and variance. The mean and variance are shown to be significantly influenced by the correlation length. Detailed quantification of the influence of horizontal and vertical correlation lengths for different permeability distributions is presented. The effect of capillary pressure, P{sub c1} on the production characteristics and saturation profiles at different correlation lengths is also investigated. It is observed that neglecting P{sub c} causes considerable error at large horizontal and short vertical correlation lengths. The effect of using constant as opposed to variable relative permeability attributes is also investigated at different correlation lengths. Next we studied the influence of correlation anisotropy in 2-D reservoir models. For a reservoir under five-spot waterflood pattern, it is shown that the ratios of breakthrough times and recovery factors of the wells in each direction of correlation are greatly influenced by the degree of anisotropy. In fully developed fields, performance data can aid in the recognition of reservoir anisotropy. Finally, a procedure for estimating the spatial correlation length from performance data is presented. Both the production performance data and the system`s effective permeability are required in estimating the correlation length.

  12. Stress dependence of permeability of intact and fractured shale cores. (United States)

    van Noort, Reinier; Yarushina, Viktoriya


    Whether a shale acts as a caprock, source rock, or reservoir, understanding fluid flow through shale is of major importance for understanding fluid flow in geological systems. Because of the low permeability of shale, flow is thought to be largely confined to fractures and similar features. In fracking operations, fractures are induced specifically to allow for hydrocarbon exploration. We have constructed an experimental setup to measure core permeabilities, using constant flow or a transient pulse. In this setup, we have measured the permeability of intact and fractured shale core samples, using either water or supercritical CO2 as the transporting fluid. Our measurements show decreasing permeability with increasing confining pressure, mainly due to time-dependent creep. Furthermore, our measurements show that for a simple splitting fracture, time-dependent creep will also eliminate any significant effect of this fracture on permeability. This effect of confinement on fracture permeability can have important implications regarding the effects of fracturing on shale permeability, and hence for operations depending on that.

  13. Update to Permeable Pavement Research at the Edison Environmental Center - abstract (United States)

    Abstract The EPA’s Urban Watershed Management Branch (UWMB) has been monitoring the permeable pavement demonstration site at the Edison Environmental Center, NJ since 2010. This site has three different types of permeable pavement including: interlocking concrete permeable pavers...

  14. Detection of semi-volatile organic compounds in permeable pavement infiltrate (United States)

    Abstract The Edison Environmental Center (EEC) performs research on green infrastructure (GI) treatment options. One such treatment option is the use of permeable pavements. EEC constructed a parking lot comprised of three different permeable systems: permeable asphalt, porous ...

  15. Update to permeable pavement research at the Edison Environmental Center - slides (United States)

    Abstract: The EPA’s Urban Watershed Management Branch (UWMB) has been monitoring the permeable pavement demonstration site at the Edison Environmental Center, NJ since 2010. This site has three different types of permeable pavement including: interlocking concrete permeable paver...

  16. Cortactin deficiency is associated with reduced neutrophil recruitment but increased vascular permeability in vivo. (United States)

    Schnoor, Michael; Lai, Frank P L; Zarbock, Alexander; Kläver, Ruth; Polaschegg, Christian; Schulte, Dörte; Weich, Herbert A; Oelkers, J Margit; Rottner, Klemens; Vestweber, Dietmar


    Neutrophil extravasation and the regulation of vascular permeability require dynamic actin rearrangements in the endothelium. In this study, we analyzed in vivo whether these processes require the function of the actin nucleation-promoting factor cortactin. Basal vascular permeability for high molecular weight substances was enhanced in cortactin-deficient mice. Despite this leakiness, neutrophil extravasation in the tumor necrosis factor-stimulated cremaster was inhibited by the loss of cortactin. The permeability defect was caused by reduced levels of activated Rap1 (Ras-related protein 1) in endothelial cells and could be rescued by activating Rap1 via the guanosine triphosphatase (GTPase) exchange factor EPAC (exchange protein directly activated by cAMP). The defect in neutrophil extravasation was caused by enhanced rolling velocity and reduced adhesion in postcapillary venules. Impaired rolling interactions were linked to contributions of β(2)-integrin ligands, and firm adhesion was compromised by reduced ICAM-1 (intercellular adhesion molecule 1) clustering around neutrophils. A signaling process known to be critical for the formation of ICAM-1-enriched contact areas and for transendothelial migration, the ICAM-1-mediated activation of the GTPase RhoG was blocked in cortactin-deficient endothelial cells. Our results represent the first physiological evidence that cortactin is crucial for orchestrating the molecular events leading to proper endothelial barrier function and leukocyte recruitment in vivo.

  17. Application of in vitro BBB model to measure permeability of nanoparticles

    International Nuclear Information System (INIS)

    Hanada, S; Kanaya, F; Yamamoto, K; Fujoka, K; Manome, Y; Inoue, Y


    In both pharmaceutical and toxicological fields, one of major issues has been the possibility of nanoparticle uptake to central nerve system. For the safe use of nanoparticles, it is integral to evaluate the permeability of nanoparticles through BBB. In our collaborative research group reported that a few nanoparticles accumulated in brain in animal experiment, as an in vitro model, we applied commercially available cell-based BBB model for establishing evaluation method, which is quick, quantitative and equivalent to in vivo assay. We assayed 30–1500 nm silica and surface charge dependent Qdots. Our results showed the size-dependency and the surface modification dependency. We compared our assay to several animal experiments. There are both equivalence and discrepancy with animal experiments. Our BBB model can be useful tools for evaluating size-dependent permeability, but not for surface modification-dependent permeability. Our BBB assay is non-serum assay and we have not adequately reflected the serum-related interaction between nanoparticles and cell surfaces. To clear up the discrepancy of our BBB model, serum-based assay and low-concentration detection will be needed.

  18. Wave transmission over permeable submerged breakwaters; Transmision del oleaje en rompeolas sumergidos permeables

    Energy Technology Data Exchange (ETDEWEB)

    Ruiz-y-Zurvia-Flores, Jaime Roberto; Fragoso-Sandoval, Lucio [Instituto Politecnico Nacional(Mexico)


    The permeable submerged breakwaters represent a coastal protection alternative, where some degree of wave energy transmission is acceptable. Such would be the case of tourist beach protection in Mexico. In previous researches, like those performed by D'Angremond et al. (1996), Seabrook and Hall (1998), and Briganti et al. (2003), the empirical formulas developed, give only some limited information over the spatial distribution of wave energy over the structure. Therefore, a decision was made to conduct a study on a reduced physical model of a permeable submerged breakwater based on the results presented by those researchers and with possible applications. Therefore this paper presents the development of a study of wave transmission over permeable submerged breakwaters performed in a reduced physical model of different sections of a submerged rockfill breakwater of the trapezoidal type. This was done in a narrow wave flume with a hydraulic irregular wave generator controlled by a computer that was used to generate and to reproduce different types of irregular waves to be used in the tests. It also has a wave meter with four sensors, and they are connected to a computer in order to process the wave data. The main objective of the study was to determine in an experimental way the influence of the several parameters of submerged breakwater over the wave transmission coefficient. Our experimental results were comparable to those obtained by D'Angremond et al. (1996) and Seabrook and Hall (1998). The results show that the sumerged breakwater parameters of most influence over the wave transmission coefficient were relative submergence and the relative width crest of the sumerged breakwater, and that the formula by Seabrook and Hall correlates best with our results. [Spanish] Los rompeolas sumergidos permeables representan actualmente una alternativa de proteccion de costas, donde un cierto grado de transmision de energia del oleaje es aceptable, como seria el

  19. Co-fabrication of chitosan and epoxy photoresist to form microwell arrays with permeable hydrogel bottoms (United States)

    Ornoff, Douglas M.; Wang, Yuli; Proctor, Angela; Shah, Akash S.; Allbritton, Nancy L.


    Microfabrication technology offers the potential to create biological platforms with customizable patterns and surface chemistries, allowing precise control over the biochemical microenvironment to which a cell or group of cells is exposed. However, most microfabricated platforms grow cells on impermeable surfaces. This report describes the co-fabrication of a micropatterned epoxy photoresist film with a chitosan film to create a freestanding array of permeable, hydrogel-bottomed microwells. These films possess optical properties ideal for microscopy applications, and the chitosan layers are semi-permeable with a molecular exclusion of 9.9 ± 2.1 kDa. By seeding cells into the microwells, overlaying inert mineral oil, and supplying media via the bottom surface, this hybrid film permits cells to be physically isolated from one another but maintained in culture for at least 4 days. Arrays co-fabricated using these materials reduce both large-molecular-weight biochemical crosstalk between cells and mixing of different clonal populations, and will enable high-throughput studies of cellular heterogeneity with increased ability to customize dynamic interrogations compared to materials in currently available technologies. PMID:26447557

  20. Fractal Theory for Permeability Prediction, Venezuelan and USA Wells (United States)

    Aldana, Milagrosa; Altamiranda, Dignorah; Cabrera, Ana


    Inferring petrophysical parameters such as permeability, porosity, water saturation, capillary pressure, etc, from the analysis of well logs or other available core data has always been of critical importance in the oil industry. Permeability in particular, which is considered to be a complex parameter, has been inferred using both empirical and theoretical techniques. The main goal of this work is to predict permeability values on different wells using Fractal Theory, based on a method proposed by Pape et al. (1999). This approach uses the relationship between permeability and the geometric form of the pore space of the rock. This method is based on the modified equation of Kozeny-Carman and a fractal pattern, which allows determining permeability as a function of the cementation exponent, porosity and the fractal dimension. Data from wells located in Venezuela and the United States of America are analyzed. Employing data of porosity and permeability obtained from core samples, and applying the Fractal Theory method, we calculated the prediction equations for each well. At the beginning, this was achieved by training with 50% of the data available for each well. Afterwards, these equations were tested inferring over 100% of the data to analyze possible trends in their distribution. This procedure gave excellent results in all the wells in spite of their geographic distance, generating permeability models with the potential to accurately predict permeability logs in the remaining parts of the well for which there are no core samples, using even porority logs. Additionally, empirical models were used to determine permeability and the results were compared with those obtained by applying the fractal method. The results indicated that, although there are empirical equations that give a proper adjustment, the prediction results obtained using fractal theory give a better fit to the core reference data.

  1. Experimental Study of Bacterial Penetration into Chalk Rock: Mechanisms and Effect on Permeability

    DEFF Research Database (Denmark)

    Halim, Amalia Yunita; Shapiro, Alexander; Eliasson Lantz, Anna


    Bacterial selective plugging is one of the mechanisms through which microorganisms can be applied for enhanced oil recovery, as bacteria can plug the water-swept zones of a reservoir, thus altering the flow paths and improving sweep efficiency. However, complete understanding of the penetration...... behavior of bacteria is lacking, especially in chalk formations where characteristic pore throat sizes are comparable with the sizes of bacterial cells. In this study, two bacterial strains, Bacillus licheniformis 421 (spore-forming) and Pseudomonas putida K12 (non-spore forming) were used to investigate...... the penetration of bacteria into chalk and its effect on permeability reduction. The core plugs were produced from Stevns Klint outcrop with low permeability (2–4 mD) and with pore sizes comparable to bacterial sizes. Both types of bacteria were able to penetrate and to be transported through the cores to some...

  2. Fluorescein isothiocyanate (FITC)-Dextran Extravasation as a Measure of Blood-Brain Barrier Permeability (United States)

    Natarajan, Reka; Northrop, Nicole


    The blood-brain barrier (BBB) is formed in part by vascular endothelial cells that constitute the capillaries and microvessels of the brain. The function of this barrier is to maintain homeostasis within the brain microenvironment and buffer the brain from changes in the periphery. A dysfunction of the BBB would permit circulating molecules and pathogens typically restricted to the periphery to enter the brain and interfere with normal brain function. As increased permeability of the BBB is associated with several neuropathologies, it is important to have a reliable and sensitive method that determines BBB permeability and the degree of BBB disruption. A detailed protocol is presented for assessing the integrity of the BBB by transcardial perfusion of a 10,000 Da FITC labeled dextran molecule and its visualization to determine the degree of extravasation from brain microvessels. PMID:28398646

  3. Prediction of the permeability of neutral drugs inferred from their solvation properties

    KAUST Repository

    Milanetti, Edoardo


    Motivation: Determination of drug absorption is an important component of the drug discovery and development process in that it plays a key role in the decision to promote drug candidates to clinical trials. We have developed a method that, on the basis of an analysis of the dynamic distribution of water molecules around a compound obtained by molecular dynamics simulations, can compute a parameter-free value that correlates very well with the compound permeability measured using the human colon adenocarcinoma (Caco-2) cell line assay. Results: The method has been tested on twenty-three neutral drugs for which a consistent set of experimental data is available. We show here that our method reproduces the experimental data better than other existing tools. Furthermore it provides a detailed view of the relationship between the hydration and the permeability properties of molecules.

  4. Multiple Factors Influence Glomerular Albumin Permeability in Rats (United States)

    Sandoval, Ruben M.; Wagner, Mark C.; Patel, Monica; Campos-Bilderback, Silvia B.; Rhodes, George J.; Wang, Exing; Wean, Sarah E.; Clendenon, Sherry S.


    Different laboratories recently reported incongruous results describing the quantification of albumin filtration using two-photon microscopy. We investigated the factors that influence the glomerular sieving coefficient for albumin (GSCA) in an effort to explain these discordant reports and to develop standard operating procedures for determining GSCA. Multiple factors influenced GSCA, including the kidney depth of image acquisition (10–20 μm was appropriate), the selection of fluorophore (probes emitting longer wavelengths were superior), the selection of plasma regions for fluorescence measurements, the size and molecular dispersion characteristics of dextran polymers if used, dietary status, and the genetic strain of rat. Fasting reduced the GSCA in Simonsen Munich Wistar rats from 0.035±0.005 to 0.016±0.004 (Palbumin transcytosis with vesicular and tubular delivery to and fusion with the basolateral membrane in S1 proximal tubule cells. In summary, these results help explain the previously conflicting microscopy and micropuncture data describing albumin filtration and highlight the dynamic nature of glomerular albumin permeability. PMID:22223875

  5. Effects of Atelocollagen Formulation Containing Oligonucleotide on Endothelial Permeability

    Directory of Open Access Journals (Sweden)

    Koji Hanai


    Full Text Available Atelocollagen is a major animal protein that is used as a highly biocompatible biomaterial. To date, atelocollagen has been used as an effective drug delivery technology to sustain the release of antitumor proteins and to enhance the antitumor activity of oligonucleotides in in vivo models. However, the biological effects of this technology are not fully understood. In the present study, we investigated the effects of atelocollagen on endothelial paracellular barrier function. An atelocollagen formulation containing oligonucleotides specifically increased the permeability of two types of endothelial cells, and the change was dependent on the molecular size, structure of the oligonucleotides used and the concentrations of the oligonucleotide and atelocollagen in the formulation. An immunohistochemical examination revealed that the formulation had effects on the cellular skeleton and intercellular structure although it did not affect the expression of adherens junction or tight junction proteins. These changes were induced through p38 MAP kinase signaling. It is important to elucidate the biological functions of atelocollagen in order to be able to exploit its drug delivery properties.

  6. Autoradiographic study of the permeability characteristics of the small intestine

    Energy Technology Data Exchange (ETDEWEB)

    Kingham, J G.C.; Baker, J H; Loehry, C A [Royal Victoria Hospital, Bournemouth (UK)


    This autoradiographic study demonstrates the distribution of a range of small solutes and macromolecules in the mucosa of the guinea-pig small intestine after intracardiac injection. The substances investigated were: /sup 14/C-urea, /sup 3/H-mannose, /sup 3/H-inulin, and /sup 125/I polyvinylpyrrolidone (PVP). Small bowel biopsies were taken at intervals from one to 60 minutes after injection and the tissues processed for autoradiography. Light microscopic examination of the autoradiographs showed that the compartmental distribution depended on the molecular size of the substances being studied. Urea and mannose, as small solutes, were uniformly distributed throughout the intravascular, extravascular, and epithelial compartments. Inulin was evenly distributed in the vessel lumen and extravascular space but there was a considerable drop in concentration in the epithelium. PVP exhibited the most marked gradients, the concentration being greatest in the vascular lumina, lower in the extravascular space, least in the epithelium. Thus there appear to be two barriers to macromolecular passage which are freely permeable to small solutes: the capillary wall and the epithelium. At a light microscopical level it was not possible to observe whether the limiting membrane of each of these barriers is the cell plasmalemmal membrane or the basement membrane. The selectivity of the epithelial barrier was greater than that of the capillary barrier.

  7. Determining Surface Infiltration Rate of Permeable Pavements with Digital Imaging

    Directory of Open Access Journals (Sweden)

    Caterina Valeo


    Full Text Available Cell phone images of pervious pavement surfaces were used to explore relationships between surface infiltration rates (SIR measured using the ASTM C1701 standard test and using a simple falling head test. A fiber-reinforced porous asphalt surface and a highly permeable material comprised of stone, rubber and a polymer binder (Porous Pave were tested. Images taken with a high-resolution cellphone camera were acquired as JPEG files and converted to gray scale images in Matlab® for analysis. The distribution of gray levels was compared to the surface infiltration rates obtained for both pavements with attention given to the mean of the distribution. Investigation into the relationships between mean SIR and parameters determined from the gray level distribution produced in the image analysis revealed that mean SIR measured in both pavements were proportional to the inverse of the mean of the distribution. The relationships produced a coefficient of determination over 85% using both the ASTM and the falling head test in the porous asphalt surface. SIR measurements determined with the ASTM method were highly correlated with the inverse mean of the distribution of gray levels in the Porous Pave material as well, producing coefficients of determination of over 90% and Kendall’s tau-b of roughly 70% for nonparametric data.

  8. Determination of filtrations and permeability of an earth dam

    International Nuclear Information System (INIS)

    Gomez, H.R.; Baro, G.B.; Gillen, Ricardo.


    The aim of this work was to measure with the aid of a radioactive tracer the speed flow of the water filtrating from Sumampa Dam in northeastern Catamarca, while being in operation, and with these data determine if the actual permeability corresponds to the projected one. Iodine-131 was used as tracer and periodical samples were taken from the down stream water in order to determine its activity concentration. In previous perforations ionic interchange resines were used so as to measure simultaneously the fixed Iodine-131. The permeability of the dam was calculated from the obtained speed based on time-concentration curves and applying Darcy formulas for permeability. (author) [es

  9. Electrical conductivity and magnetic permeability measurement of case hardened steels (United States)

    Tian, Yong


    For case carburized steels, electrical conductivity and magnetic permeability profiles are needed to develop model-based case depth characterization techniques for the purpose of nondestructive quality control. To obtain fast and accurate measurement of these material properties, four-point potential drop approaches are applied on circular-shaped discs cut from steel rings with different case depths. First, a direct current potential drop (DCPD) approach is applied to measure electrical conductivity. Subsequently, an alternating current potential drop (ACPD) approach is used to measure magnetic permeability. Practical issues in measurement design and implementation are discussed. Depth profiles of electrical conductivity and magnetic permeability are reported.

  10. Instrumentation for Measurement of Gas Permeability of Polymeric Membranes (United States)

    Upchurch, Billy T.; Wood, George M.; Brown, Kenneth G.; Burns, Karen S.


    A mass spectrometric 'Dynamic Delta' method for the measurement of gas permeability of polymeric membranes has been developed. The method is universally applicable for measurement of the permeability of any gas through polymeric membrane materials. The usual large sample size of more than 100 square centimeters required for other methods is not necessary for this new method which requires a size less than one square centimeter. The new method should fulfill requirements and find applicability for industrial materials such as food packaging, contact lenses and other commercial materials where gas permeability or permselectivity properties are important.

  11. Compressible fluid flow through rocks of variable permeability

    International Nuclear Information System (INIS)

    Lin, W.


    The effectiveness of course-grained igneous rocks as shelters for burying radioactive waste can be assessed by determining the rock permeabilities at their in situ pressures and stresses. Analytical and numerical methods were used to solve differential equations of one-dimensional fluid flow through rocks with permeabilities from 10 4 to 1 nD. In these calculations, upstream and downstream reservoir volumes of 5, 50, and 500 cm 3 were used. The optimal size combinations of the two reservoirs were determined for measurements of permeability, stress, strain, acoustic velocity, and electrical conductivity on low-porosity, coarse-grained igneous rocks

  12. Effect of confining pressure on permeability behavior of Beishan granite

    International Nuclear Information System (INIS)

    Ma Like; Li Yunfeng; Zhao Xingguang; Tan Guohuan


    By using of the Electro-Hydraulic Servo-controlled Rock Mechanics Testing System (MTS 815.04) in the University of Hong Kong, a series of permeability tests were performed on specimens of Beishan granite at different confining pressures. The result indicates that: (1) there is a decrease of permeability due to progressive closure of initial microcracks and the corresponding volumetric strain is compressive when the confining pressures increase from 2.5 MPa to 15 MPa, (2) when the confining pressures decrease from 15 MPa to 2.5 MPa, there is an increase of permeability in this stage in relation with the volumetric dilation. (authors)

  13. Membrane permeability of the human granulocyte to water, dimethyl sulfoxide, glycerol, propylene glycol and ethylene glycol. (United States)

    Vian, Alex M; Higgins, Adam Z


    Granulocytes are currently transfused as soon as possible after collection because they rapidly deteriorate after being removed from the body. This short shelf life complicates the logistics of granulocyte collection, banking, and safety testing. Cryopreservation has the potential to significantly increase shelf life; however, cryopreservation of granulocytes has proven to be difficult. In this study, we investigate the membrane permeability properties of human granulocytes, with the ultimate goal of using membrane transport modeling to facilitate development of improved cryopreservation methods. We first measured the equilibrium volume of human granulocytes in a range of hypo- and hypertonic solutions and fit the resulting data using a Boyle-van't Hoff model. This yielded an isotonic cell volume of 378 μm(3) and an osmotically inactive volume of 165 μm(3). To determine the permeability of the granulocyte membrane to water and cryoprotectant (CPA), cells were injected into well-mixed CPA solution while collecting volume measurements using a Coulter Counter. These experiments were performed at temperatures ranging from 4 to 37°C for exposure to dimethyl sulfoxide, glycerol, ethylene glycol, and propylene glycol. The best-fit water permeability was similar in the presence of all of the CPAs, with an average value at 21°C of 0.18 μmatm(-1)min(-1). The activation energy for water transport ranged from 41 to 61 kJ/mol. The CPA permeability at 21°C was 6.4, 1.0, 8.4, and 4.0 μm/min for dimethyl sulfoxide, glycerol, ethylene glycol, and propylene glycol, respectively, and the activation energy for CPA transport ranged between 59 and 68 kJ/mol. Copyright © 2013 Elsevier Inc. All rights reserved.

  14. Do phosphoinositides regulate membrane water permeability of tobacco protoplasts by enhancing the aquaporin pathway? (United States)

    Ma, Xiaohong; Shatil-Cohen, Arava; Ben-Dor, Shifra; Wigoda, Noa; Perera, Imara Y; Im, Yang Ju; Diminshtein, Sofia; Yu, Ling; Boss, Wendy F; Moshelion, Menachem; Moran, Nava


    Enhancing the membrane content of PtdInsP 2 , the already-recognized protein-regulating lipid, increased the osmotic water permeability of tobacco protoplasts, apparently by increasing the abundance of active aquaporins in their membranes. While phosphoinositides are implicated in cell volume changes and are known to regulate some ion channels, their modulation of aquaporins activity has not yet been reported for any organism. To examine this, we compared the osmotic water permeability (P f) of protoplasts isolated from tobacco (Nicotiana tabacum) cultured cells (NT1) with different (genetically lowered or elevated relative to controls) levels of inositol trisphosphate (InsP3) and phosphatidyl inositol [4,5] bisphosphate (PtdInsP2). To achieve this, the cells were transformed with, respectively, the human InsP3 5-phosphatase ('Ptase cells') or human phosphatidylinositol (4) phosphate 5-kinase ('PIPK cells'). The mean P f of the PIPK cells was several-fold higher relative to that of controls and Ptase cells. Three results favor aquaporins over the membrane matrix as underlying this excessive P f: (1) transient expression of the maize aquaporin ZmPIP2;4 in the PIPK cells increased P f by 12-30 μm s(-1), while in the controls only by 3-4 μm s(-1). (2) Cytosol acidification-known to inhibit aquaporins-lowered the P f in the PIPK cells down to control levels. (3) The transcript of at least one aquaporin was elevated in the PIPK cells. Together, the three results demonstrate the differences between the PIPK cells and their controls, and suggest a hitherto unobserved regulation of aquaporins by phosphoinositides, which could occur through direct interaction or indirect phosphoinositides-dependent cellular effects.

  15. Permeability of skin and oral mucosa to water and horseradish peroxidase as related to the thickness of the permeability barrier

    International Nuclear Information System (INIS)

    Squier, C.A.; Hall, B.K.


    The permeability of porcine skin and keratinized and nonkeratinized oral mucosa to tritium-labeled water and horseradish peroxidase (HRPO) was determined using perfusion chambers. Small blocks from each tissue were also incubated with HRPO and the extent of penetration visualized microscopically; this enabled measurements to be made of the thickness of the permeability barrier to this water-soluble tracer. Results obtained after inverting the oral mucosa in the chambers or adding metabolic inhibitors indicated that both compounds diffuse across the tissue. The permeability constants derived directly in the study showed that skin was less permeable than oral mucosa and that the floor of the mouth was significantly more permeable than all other regions. When these constants were normalized in terms of a standard permeability barrier thickness and the different tissues compared, the values obtained for skin were again less than those of the oral regions but, of these, the buccal mucosa was significantly higher. The difference in permeability between epidermis and keratinized oral epithelium may be due to differences in the volume density of membrane-coating granules known to exist between the tissues; differences between the oral mucosal regions may reflect differences in the nature of the intercellular barrier material

  16. Pannexin 1 channels mediate 'find-me' signal release and membrane permeability during apoptosis. (United States)

    Chekeni, Faraaz B; Elliott, Michael R; Sandilos, Joanna K; Walk, Scott F; Kinchen, Jason M; Lazarowski, Eduardo R; Armstrong, Allison J; Penuela, Silvia; Laird, Dale W; Salvesen, Guy S; Isakson, Brant E; Bayliss, Douglas A; Ravichandran, Kodi S


    Apoptotic cells release 'find-me' signals at the earliest stages of death to recruit phagocytes. The nucleotides ATP and UTP represent one class of find-me signals, but their mechanism of release is not known. Here, we identify the plasma membrane channel pannexin 1 (PANX1) as a mediator of find-me signal/nucleotide release from apoptotic cells. Pharmacological inhibition and siRNA-mediated knockdown of PANX1 led to decreased nucleotide release and monocyte recruitment by apoptotic cells. Conversely, PANX1 overexpression enhanced nucleotide release from apoptotic cells and phagocyte recruitment. Patch-clamp recordings showed that PANX1 was basally inactive, and that induction of PANX1 currents occurred only during apoptosis. Mechanistically, PANX1 itself was a target of effector caspases (caspases 3 and 7), and a specific caspase-cleavage site within PANX1 was essential for PANX1 function during apoptosis. Expression of truncated PANX1 (at the putative caspase cleavage site) resulted in a constitutively open channel. PANX1 was also important for the 'selective' plasma membrane permeability of early apoptotic cells to specific dyes. Collectively, these data identify PANX1 as a plasma membrane channel mediating the regulated release of find-me signals and selective plasma membrane permeability during apoptosis, and a new mechanism of PANX1 activation by caspases.

  17. Flexible control of cellular encapsulation, permeability, and release in a droplet-templated bifunctional copolymer scaffold. (United States)

    Chen, Qiushui; Chen, Dong; Wu, Jing; Lin, Jin-Ming


    Designing cell-compatible, bio-degradable, and stimuli-responsive hydrogels is very important for biomedical applications in cellular delivery and micro-scale tissue engineering. Here, we report achieving flexible control of cellular microencapsulation, permeability, and release by rationally designing a diblock copolymer, alginate-conjugated poly(N-isopropylacrylamide) (Alg-co-PNiPAM). We use the microfluidic technique to fabricate the bifunctional copolymers into thousands of mono-disperse droplet-templated hydrogel microparticles for controlled encapsulation and triggered release of mammalian cells. In particular, the grafting PNiPAM groups in the synthetic cell-laden microgels produce lots of nano-aggregates into hydrogel networks at elevated temperature, thereafter enhancing the permeability of microparticle scaffolds. Importantly, the hydrogel scaffolds are readily fabricated via on-chip quick gelation by triggered release of Ca 2+ from the Ca-EDTA complex; it is also quite exciting that very mild release of microencapsulated cells is achieved via controlled degradation of hydrogel scaffolds through a simple strategy of competitive affinity of Ca 2+ from the Ca-Alginate complex. This finding suggests that we are able to control cellular encapsulation and release through ion-induced gelation and degradation of the hydrogel scaffolds. Subsequently, we demonstrate a high viability of microencapsulated cells in the microgel scaffolds.

  18. Metal concentrations from permeable pavement parking lot in Edison, NJ (United States)

    U.S. Environmental Protection Agency — The U.S. Environmental Protection Agency constructed a 4000-m2 parking lot in Edison, New Jersey in 2009. The parking lot is surfaced with three permeable pavements...

  19. Investigation of negative permeability metamaterials for wireless power transfer

    Directory of Open Access Journals (Sweden)

    Wenhui Xin


    Full Text Available In order to enhance the transmission efficiency of wireless power transfer (WPT, a negative permeability metamaterials (NPM with a structure of honeycomb composed by units of hexagon-shaped spirals copper is proposed in this paper. The unit parameters of the NPM are optimized, to make sure the negative permeability at the special frequency. The S-parameters of the designed NPM are measured by a network analyzer and the permeability is extracted, it shows the honeycomb NPM has a negative permeability at 6.43 MHz. A two-coil WPT is setup and the transmission efficiency of WPT embedded with NPM at the different position and with different structure are investigated. The measured results show that the 2-slab honeycomb NPM have a good perform compared with the 1-slab NPM, and the efficiency can be increased up to 51%. The results show that honeycomb NPM embedded in the WPT help to improve the transmission efficiency remarkable.

  20. Xenon NMR measurements of permeability and tortuosity in reservoir rocks. (United States)

    Wang, Ruopeng; Pavlin, Tina; Rosen, Matthew Scott; Mair, Ross William; Cory, David G; Walsworth, Ronald Lee


    In this work we present measurements of permeability, effective porosity and tortuosity on a variety of rock samples using NMR/MRI of thermal and laser-polarized gas. Permeability and effective porosity are measured simultaneously using MRI to monitor the inflow of laser-polarized xenon into the rock core. Tortuosity is determined from measurements of the time-dependent diffusion coefficient using thermal xenon in sealed samples. The initial results from a limited number of rocks indicate inverse correlations between tortuosity and both effective porosity and permeability. Further studies to widen the number of types of rocks studied may eventually aid in explaining the poorly understood connection between permeability and tortuosity of rock cores.

  1. Investigation of negative permeability metamaterials for wireless power transfer (United States)

    Xin, Wenhui; Mi, Chunting Chris; He, Fei; Jiang, Meng; Hua, Dengxin


    In order to enhance the transmission efficiency of wireless power transfer (WPT), a negative permeability metamaterials (NPM) with a structure of honeycomb composed by units of hexagon-shaped spirals copper is proposed in this paper. The unit parameters of the NPM are optimized, to make sure the negative permeability at the special frequency. The S-parameters of the designed NPM are measured by a network analyzer and the permeability is extracted, it shows the honeycomb NPM has a negative permeability at 6.43 MHz. A two-coil WPT is setup and the transmission efficiency of WPT embedded with NPM at the different position and with different structure are investigated. The measured results show that the 2-slab honeycomb NPM have a good perform compared with the 1-slab NPM, and the efficiency can be increased up to 51%. The results show that honeycomb NPM embedded in the WPT help to improve the transmission efficiency remarkable.

  2. Quantifying tidally driven benthic oxygen exchange across permeable sediments

    DEFF Research Database (Denmark)

    McGinnis, Daniel F.; Sommer, Stefan; Lorke, Andreas


    Continental shelves are predominately (approximate to 70%) covered with permeable, sandy sediments. While identified as critical sites for intense oxygen, carbon, and nutrient turnover, constituent exchange across permeable sediments remains poorly quantified. The central North Sea largely consists...... of permeable sediments and has been identified as increasingly at risk for developing hypoxia. Therefore, we investigate the benthic O-2 exchange across the permeable North Sea sediments using a combination of in situ microprofiles, a benthic chamber, and aquatic eddy correlation. Tidal bottom currents drive...... the variable sediment O-2 penetration depth (from approximate to 3 to 8 mm) and the concurrent turbulence-driven 25-fold variation in the benthic sediment O-2 uptake. The O-2 flux and variability were reproduced using a simple 1-D model linking the benthic turbulence to the sediment pore water exchange...

  3. Investigation clogging dynamic of permeable pavement systems using embeded sensors (United States)

    U.S. Environmental Protection Agency — Permeable pavement is a stormwater control measure commonly selected in both new and retrofit applications. However, there is limited information about the clogging...

  4. Investigation of stormwater quality improvements utilizing permeable friction course (PFC). (United States)


    This report describes research into the water quality and hydraulics of the Permeable Friction Course (PFC). : Water quality monitoring of 3 locations in the Austin area indicates up to a 90 percent reduction in pollutant : discharges from PFC compar...

  5. Wood Permeability in Eucalyptus grandis and Eucalyptus dunnii

    Directory of Open Access Journals (Sweden)

    Raphael Nogueira Rezende


    Full Text Available ABSTRACT The objective of this study was to evaluate the flow of air and water in Eucalyptus grandis and Eucalyptus dunnii wood. Wood was collected from four trees aged 37 years in an experimental plantation of the Federal University of Lavras, Brazil. Planks were cut off the basal logs to produce specimens for air and water permeability testing. Results indicated that the longitudinal permeability to air and water of E. grandis wood were, on average, 5% and 10% higher, respectively, than that of E. dunnii wood. E. grandis and E. dunnii wood showed neither air nor water flow in the test for permeability transversal to the fibers, and longitudinal permeability to air exceeded that to water by approximately 50 fold in both species.

  6. Porosity, permeability, and their relationship in granite, basalt, and tuff

    International Nuclear Information System (INIS)


    This report discusses the porosity, storage, and permeability of fractured (mainly crystalline) rock types proposed as host rock for nuclear waste repositories. The emphasis is on the inter-relationships of these properties, but a number of reported measurements are included as well. The porosity of rock is shown to consist of fracture porosity and matrix porosity; techniques are described for determining the total interconnected porosity through both laboratory and field measurement. Permeability coefficient, as obtained by experiments ranging from laboratory to crustal scale, is discussed. Finally, the problem of determining the relationship between porosity and permeability is discussed. There is no simple, all encompassing relationship that describes the dependence of permeability upon porosity. However, two particular cases have been successfully analyzed: flow through a single rough fracture, and flow through isotropic porous rock. These two cases are discussed in this report

  7. Diagenetic effect on permeabilities of geothermal sandstone reservoirs

    DEFF Research Database (Denmark)

    Weibel, Rikke; Olivarius, Mette; Kristensen, Lars

    The Danish subsurface contains abundant sedimentary deposits, which can be utilized for geothermal heating. The Upper Triassic – Lower Jurassic continental-marine sandstones of the Gassum Formation has been utilised as a geothermal reservoir for the Thisted Geothermal Plant since 1984 extracting...... and permeability is caused by increased diagenetic changes of the sandstones due to increased burial depth and temperatures. Therefore, the highest water temperatures typically correspond with the lowest porosities and permeabilities. Especially the permeability is crucial for the performance of the geothermal......-line fractures. Continuous thin chlorite coatings results in less porosity- and permeability-reduction with burial than the general reduction with burial, unless carbonate cemented. Therefore, localities of sandstones characterized by these continuous chlorite coatings may represent fine geothermal reservoirs...

  8. Study on Surface Permeability of Concrete under Immersion. (United States)

    Liu, Jun; Xing, Feng; Dong, Biqin; Ma, Hongyan; Pan, Dong


    In this paper, concrete specimens are immersed in ultrapure water, to study the evolutions of surface permeability, pore structure and paste microstructure following the prolonging of immersion period. According to the results, after 30-day immersion, the surface permeability of concrete becomes higher as compared with the value before immersion. However, further immersion makes the surface permeability decrease, so that the value measured after 150-day immersion is only half that measured after 30-day immersion. The early increase in surface permeability should be mainly attributed to the leaching of calcium hydroxide, while the later decrease to the refinement of pore structure due to hydration. The two effects work simultaneously and compete throughout the immersion period. The proposed mechanisms get support from microscopic measurements and observations.

  9. Water permeability is a measure of severity in acute appendicitis. (United States)

    Pini, Nicola; Pfeifle, Viktoria A; Kym, Urs; Keck, Simone; Galati, Virginie; Holland-Cunz, Stefan; Gros, Stephanie J


    Acute appendicitis is the most common indication for pediatric abdominal emergency surgery. Determination of the severity of appendicitis on clinical grounds is challenging. Complicated appendicitis presenting with perforation, abscess or diffuse peritonitis is not uncommon. The question remains why and when acute appendicitis progresses to perforation. The aim of this study was to assess the impact of water permeability on the severity of appendicitis. We show that AQP1 expression and water permeability in appendicitis correlate with the stage of inflammation and systemic infection parameters, leading eventually to perforation of the appendix. AQP1 is also expressed within the ganglia of the enteric nervous system and ganglia count increases with inflammation. Severity of appendicitis can be correlated with water permeability measured by AQP1 protein expression and increase of ganglia count in a progressive manner. This introduces the question if regulation of water permeability can present novel curative or ameliorating therapeutic options.

  10. Airspace Analyzer for Assessing Airspace Directional Permeability, Phase I (United States)

    National Aeronautics and Space Administration — We build a software tool which enables the user (airline or Air Traffic Service Provider (ATSP)) the ability to analyze the flight-level-by-flight-level permeability...

  11. Measurement of HTO permeability of materials for protective appliances

    International Nuclear Information System (INIS)

    Yamamoto, H.; Tomooka, M.; Kato, S.; Murata, M.; Kinouchi, N.; Yamamoto, H.


    Tritiated water (HTO) vapor permeabilities were measured for plastic and rubber films used for protective appliances (suits, gloves, wrappings, etc.). The measurement data prove that polyehylene and butyl rubbers are materials suitable for HTO protective appliances with their lower permeability. The data also indicate that desiccating protective appliances before reuse is effective for restoring their original resistances to penetrating HTO vapor when they are repeatedly used. (author)

  12. Transport zonation limits coupled nitrification-denitrification in permeable sediments

    DEFF Research Database (Denmark)

    Kessler, Adam John; Glud, R.N.; Cardenas, M.B.


    - and N-15-N-2 gas. The measured two-dimensional profiles correlate with computational model simulations, showing a deep pool of N-2 gas forming, and being advected to the surface below ripple peaks. Further isotope pairing calculations on these data indicate that coupled nitrification......-denitrification is severely limited in permeable sediments because the flow and transport field limits interaction between oxic and anoxic pore water. The approach allowed for new detailed insight into subsurface denitrification zones in complex permeable sediments....

  13. Quantitation of small intestinal permeability during normal human drug absorption


    Levitt, David G


    Background Understanding the quantitative relationship between a drug?s physical chemical properties and its rate of intestinal absorption (QSAR) is critical for selecting candidate drugs. Because of limited experimental human small intestinal permeability data, approximate surrogates such as the fraction absorbed or Caco-2 permeability are used, both of which have limitations. Methods Given the blood concentration following an oral and intravenous dose, the time course of intestinal absorpti...

  14. Highly permeable, cement-bounded backfilling mortars for SMA repositories

    International Nuclear Information System (INIS)

    Jacobs, F.; Mayer, G.; Wittmann, F.H.


    In low- and intermediate-level waste repositories, gas is produced due e.g. to corrosion. This gas must be able to escape from the repository in order to prevent damage to the repository structure. A cement-based backfill should take over this function. For this purpose, the composition of cement-based materials was varied to study their influence on porosity and permeability. In parallel to this study the behaviour of fresh concrete, the liberation of the heat of hydration and the hardened concrete properties were investigated. To characterize the permeability of cement-based materials the following parameters are important: 1) composition of the material (pore fabric), 2) storage conditions (degree of saturation), 3) degree of hydration (age), 4) measuring fluid. A change in the composition of cement-based materials can vary the permeability by ten orders of magnitude. It is shown that, by using dense aggregates, the transport of the fluid takes place through the matrix and along the aggregate/matrix interface. By using porous aggregates the permeability can be increased by two orders of magnitude. In the case of a dense matrix, porous aggregates do not alter the permeability. Increasing the matrix content or interface content increases permeability. Hence light weight mortars are an obvious choice. Like-grained mixes showed higher permeabilities in combination with better mechanical properties but, in comparison to normal mixes, they showed worse flow properties. With the composition cement-: water-: aggregate content 1:0.4:5.33 the likegrained mix with aggregates ranging from 2 to 3 mm proved to be a suitable material. With a low compaction after 28 days this mix reaches a permeability of 4.10 -12 m 2 and an uniaxial cylinder compressive strength of 16 N/mm 2 . (author) 58 figs., 23 tabs., refs

  15. Experimental Validation of a Permeability Model for Enrichment Membranes

    International Nuclear Information System (INIS)

    Orellano, Pablo; Brasnarof, Daniel; Florido Pablo


    An experimental loop with a real scale diffuser, in a single enrichment-stage configuration, was operated with air at different process conditions, in order to characterize the membrane permeability.Using these experimental data, an analytical geometric-and-morphologic-based model was validated.It is conclude that a new set of independent measurements, i.e. enrichment, is necessary in order to fully characterize diffusers, because of its internal parameters are not univocally determinated with permeability experimental data only

  16. Improvement of air permeability of Bubbfil nanofiber membrane

    Directory of Open Access Journals (Sweden)

    Wang Fei-Yan


    Full Text Available Nanofiber membranes always have extremely high filter efficiency and remarkably low pressure drop. In order to further improve air permeability of bubbfil nanofiber membranes, the plasma technology is used for surface treatment in this paper. The results show that plasma treatment can improve air permeability by 4.45%. Under higher power plasma treatment, earthworm like etchings are produced on the membrane surface with fractal dimensions of about 1.138.

  17. Permeability, zonulin production, and enteropathy in dermatitis herpetiformis. (United States)

    Smecuol, Edgardo; Sugai, Emilia; Niveloni, Sonia; Vázquez, Horacio; Pedreira, Silvia; Mazure, Roberto; Moreno, María Laura; Label, Marcelo; Mauriño, Eduardo; Fasano, Alessio; Meddings, Jon; Bai, Julio César


    Dermatitis herpetiformis (DH) is characterized by variable degrees of enteropathy and increased intestinal permeability. Zonulin, a regulator of tight junctions, seems to play a key role in the altered intestinal permeability that characterizes the early phase of celiac disease. Our aim was to assess both intestinal permeability and serum zonulin levels in a group of patients with DH having variable grades of enteropathy. We studied 18 DH patients diagnosed on the basis of characteristic immunoglobulin (Ig)A granular deposits in the dermal papillae of noninvolved skin. Results were compared with those of classic celiac patients, patients with linear IgA dermatosis, and healthy controls. According to Marsh's classification, 5 patients had no evidence of enteropathy (type 0), 4 patients had type II, 2 patients had type IIIb damage, and 7 patients had a more severe lesion (type IIIc). Intestinal permeability (lactulose/mannitol ratio [lac/man]) was abnormal in all patients with DH. Patients with more severe enteropathy had significantly greater permeability ( P zonulin concentration (enzyme-linked immunosorbent assay) for patients with DH was 2.1 +/- .3 ng/mg with 14 of 16 (87.5%) patients having abnormally increased values. In contrast, patients with linear IgA dermatosis had normal histology, normal intestinal permeability, and negative celiac serology. Increased intestinal permeability and zonulin up-regulation are common and concomitant findings among patients with DH, likely involved in pathogenesis. Increased permeability can be observed even in patients with no evidence of histologic damage in biopsy specimens. Patients with linear IgA dermatosis appear to be a distinct population with no evidence of gluten sensitivity.

  18. Evaluation of permeability of compacted bentonite ground considering heterogeneity by geostatistics

    International Nuclear Information System (INIS)

    Tanaka, Yukihisa; Nakamura, Kunihiko; Kudo, Kohji; Hironaga, Michihiko; Nakagami, Motonori; Niwase, Kazuhito; Komatsu, Shin-ichi


    The permeability of the bentonite ground as an engineered barrier is possibly designed to the value which is lower than that determined in terms of required performance because of heterogeneous distribution of permeability in the ground, which might be considerable when the ground is created by the compaction method. The effect of heterogeneity in the ground on the permeability of the bentonite ground should be evaluated by overall permeability of the ground, whereas in practice, the effect is evaluated by the distribution of permeability in the ground. Thus, in this study, overall permeability of the bentonite ground is evaluated from the permeability of the bentonite ground is evaluated from the permeability distribution determined using the geostatistical method with the dry density data as well as permeability data of the undisturbed sample recovered from the bentonite ground. Consequently, it was proved through this study that possibility of overestimation of permeability of the bentonite ground can be reduced if the overall permeability is used. (author)

  19. Hybrid green permeable pave with hexagonal modular pavement systems

    International Nuclear Information System (INIS)

    Rashid, M A; Abustan, I; Hamzah, M O


    Modular permeable pavements are alternatives to the traditional impervious asphalt and concrete pavements. Pervious pore spaces in the surface allow for water to infiltrate into the pavement during rainfall events. As of their ability to allow water to quickly infiltrate through the surface, modular permeable pavements allow for reductions in runoff quantity and peak runoff rates. Even in areas where the underlying soil is not ideal for modular permeable pavements, the installation of under drains has still been shown to reflect these reductions. Modular permeable pavements have been regarded as an effective tool in helping with stormwater control. It also affects the water quality of stormwater runoff. Places using modular permeable pavement has been shown to cause a significant decrease in several heavy metal concentrations as well as suspended solids. Removal rates are dependent upon the material used for the pavers and sub-base material, as well as the surface void space. Most heavy metals are captured in the top layers of the void space fill media. Permeable pavements are now considered an effective BMP for reducing stormwater runoff volume and peak flow. This study examines the extent to which such combined pavement systems are capable of handling load from the vehicles. Experimental investigation were undertaken to quantify the compressive characteristics of the modular. Results shows impressive results of achieving high safety factor for daily life vehicles.

  20. Detection of semi-volatile organic compounds in permeable ... (United States)

    Abstract The Edison Environmental Center (EEC) has a research and demonstration permeable parking lot comprised of three different permeable systems: permeable asphalt, porous concrete and interlocking concrete permeable pavers. Water quality and quantity analysis has been ongoing since January, 2010. This paper describes a subset of the water quality analysis, analysis of semivolatile organic compounds (SVOCs) to determine if hydrocarbons were in water infiltrated through the permeable surfaces. SVOCs were analyzed in samples collected from 11 dates over a 3 year period, from 2/8/2010 to 4/1/2013.Results are broadly divided into three categories: 42 chemicals were never detected; 12 chemicals (11 chemical test) were detected at a rate of less than 10% or less; and 22 chemicals were detected at a frequency of 10% or greater (ranging from 10% to 66.5% detections). Fundamental and exploratory statistical analyses were performed on these latter analyses results by grouping results by surface type. The statistical analyses were limited due to low frequency of detections and dilutions of samples which impacted detection limits. The infiltrate data through three permeable surfaces were analyzed as non-parametric data by the Kaplan-Meier estimation method for fundamental statistics; there were some statistically observable difference in concentration between pavement types when using Tarone-Ware Comparison Hypothesis Test. Additionally Spearman Rank order non-parame

  1. Long-term bioventing performance in low-permeability soils

    International Nuclear Information System (INIS)

    Phelps, M.B.; Stanin, F.T.; Downey, D.C.


    Short-term and long-term bioventing treatability testing has shown that in situ air injection and extraction is a practical method for sustaining increased oxygen levels and enhancing aerobic biodegradation of petroleum hydrocarbons in low-permeability soils. At several test sites, initial physical parameter analysis of soils and air permeability tests indicated that impacted soils (fine sandy silts and clays) had low air permeabilities. Measurements of depleted soil-gas oxygen levels and increased soil-gas carbon dioxide levels indicated that the natural process of aerobic biodegradation of petroleum hydrocarbons was oxygen-limited. Initial treatability testing consisted of air permeability tests to measure the