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Sample records for brush border membranes

  1. Taurine transport in renal brush-border-membrane vesicles.

    PubMed Central

    Rozen, R; Tenenhouse, H S; Scriver, C R

    1979-01-01

    Taurine transport in isolated brush-border-membrane vesicles from rat kidney is concentrative and it is driven by the Na+ gradient and transmembrane potential difference; binding is not a significant component of net uptake. The Na+-dependent component of net uptake is saturable with an apparent Km of 17 microM. The taurine-transport mechanism is selective for beta-amino compounds. PMID:486101

  2. Isolation of renal brush borders.

    PubMed

    Morré, D James; Hammond, Timothy

    2007-03-01

    Methods are described to isolate intact brush borders and brush border membranes from renal cell homogenates. A rapid method yields sealed vesicles that reconstitute renal brush border transport. In one variation of this protocol, 10 to 20 mM CaCl2 or MgCl2 is added to aggregate non-brush border structures for subsequent removal by centrifugation. For analytical studies, guidance is provided for subsequent purification steps including preparative free-flow and aqueous two-phase partition. Marker enzymes and morphological parameters are included for assessment of yield and fraction purity. PMID:18228514

  3. The pathogenic antigen of Heymann nephritis is a membrane glycoprotein of the renal proximal tubule brush border.

    PubMed Central

    Kerjaschki, D; Farquhar, M G

    1982-01-01

    Purified brush border fractions prepared from rat kidneys were solubilized in detergent, iodinated, and subjected to immunoprecipitation to identify the pathogenic antigen present in brush border membranes that is responsible for the production of Heymann nephritis (HN). Purified IgG prepared from the sera of rabbits or rats immunized with a crude cortical preparation, known as Fx1A, precipitated multiple peptides, whereas IgG eluted from glomeruli of rats with active or passive HN specifically immunoprecipitated a single large glycoprotein (Mr = 330,000). This protein (gp330) was subsequently purified by gel filtration and lentil lectin affinity chromatography from detergent-solubilized brush border membranes. When rats were immunized with purified gp330, they developed anti-brush border antibodies and active HN. IgG prepared from the serum of rats with active HN caused passive HN when injected into normal recipients. Rats immunized against brush border membrane proteins depleted of gp330 developed anti-brush border antibodies but did not develop HN. Further analysis of gp330 indicated that it is solubilized by detergent treatment of isolated brush border microvilli, and its antigenic component is released from intact microvilli by trypsin. By immunoperoxidase staining it was localized to the luminal side of the brush border membranes. These results indicate that (i) gp330 is the pathogenic antigen of HN; (ii) the antigen is a glycoprotein of the brush border membrane; and (iii) it is disposed with its pathogenic domain(s) facing the tubule lumen. Images PMID:6752952

  4. Membrane stress causes inhibition of water channels in brush border membrane vesicles from kidney proximal tubule.

    PubMed

    Soveral, G; Macey, R I; Moura, T F

    1997-08-01

    Brush border membrane vesicles (BBMV) from rabbit kidney proximal tubule cells, prepared with different internal solute concentrations (cellobiose buffer 13, 18 or 85 mosM) developed an hydrostatic pressure difference across the membrane of 18.7 mosM, that causes a membrane tension close to 5 x 10(-5) N cm-1. When subjected to several hypertonic osmotic shocks an initial delay of osmotic shrinkage (a lag time), corresponding to a very small change in initial volume was apparent. This initial osmotic response, which is significantly retarded, was correlated with the initial period of elevated membrane tension, suggesting that the water permeability coefficient is inhibited by membrane stress. We speculate that this inhibition may serve to regulate cell volume in the proximal tubule. PMID:9468597

  5. Calcium uptake by brush-border and basolateral membrane vesicles in chick duodenum

    SciTech Connect

    Takito, J.; Shinki, T.; Sasaki, T.; Suda, T. )

    1990-01-01

    Calcium uptake was compared between duodenal brush-border membrane vesicles (BBMV) and basolateral membrane vesicles (BLMV) isolated from vitamin D-deficient chicks and those injected with 625 ng of 1 alpha,25-dihydroxyvitamin D3 (1 alpha,25(OH)2D3). The uptake by BBMV in the 1 alpha,25-(OH)2D3-treated birds attained a maximum (280% of the control) at 12 h and was maintained at an elevated level (210%) at 24 h after the injection of the vitamin. In contrast, ATP-dependent calcium uptake by BLMV reached a maximum (185% of the control) at 6 h and decreased to the control level at 24 h. The kinetic analysis revealed that 1 alpha,25(OH)2D3 increased Vmax values without any changes in apparent Km values in both BBMV and BLMV. The activity of ATP-dependent calcium uptake was localized exclusively in the basolateral membrane, and the activity was inhibited by vanadate (IC50, 1 microM), but not by oligomycin, theophylline, calmodulin, trifluoperazine, or calbindin D28K. These results indicate that calcium transport through both the brush-border and basolateral membranes is involved in the 1 alpha,25(OH)2D3-dependent intestinal calcium absorption. The initiation of calcium absorption by 1 alpha,25(OH)2D3 appears to be due to an increase in the rate of calcium efflux at the basolateral membrane rather than the rate at the brush-border membrane.

  6. Lipid rafts in epithelial brush borders: atypical membrane microdomains with specialized functions.

    PubMed

    Danielsen, E Michael; Hansen, Gert H

    2003-10-31

    Epithelial cells that fulfil high-throughput digestive/absorptive functions, such as small intestinal enterocytes and kidney proximal tubule cells, are endowed with a dense apical brush border. It has long been recognized that the microvillar surface of the brush border is organized in cholesterol/sphingolipid-enriched membrane microdomains commonly known as lipid rafts. More recent studies indicate that microvillar rafts, in particular those of enterocytes, have some unusual properties in comparison with rafts present on the surface of other cell types. Thus, microvillar rafts are stable rather than transient/dynamic, and their core components include glycolipids and the divalent lectin galectin-4, which together can be isolated as "superrafts", i.e., membrane microdomains resisting solubilization with Triton X-100 at physiological temperature. These glycolipid/lectin-based rafts serve as platforms for recruitment of GPI-linked and transmembrane digestive enzymes, most likely as an economizing effort to secure and prolong their digestive capability at the microvillar surface. However, in addition to microvilli, the brush border surface also consists of membrane invaginations between adjacent microvilli, which are the only part of the apical surface sterically accessible for membrane fusion/budding events. Many of these invaginations appear as pleiomorphic, deep apical tubules that extend up to 0.5-1 microm into the underlying terminal web region. Their sensitivity to methyl-beta-cyclodextrin suggests them to contain cholesterol-dependent lipid rafts of a different type from the glycolipid-based rafts at the microvillar surface. The brush border is thus an example of a complex membrane system that harbours at least two different types of lipid raft microdomains, each suited to fulfil specialized functions. This conclusion is in line with an emerging, more varied view of lipid rafts being pluripotent microdomains capable of adapting in size, shape, and content to

  7. Proteomic analysis of the mosquito Aedes aegypti midgut brush border membrane vesicles

    PubMed Central

    Popova-Butler, Alexandra; Dean, Donald H.

    2009-01-01

    We analyzed brush border membrane vesicle proteins from isolated midguts of the mosquito Aedes aegypti, by two proteomic methods: two-dimensional gel electrophoresis (isoelectric focusing and SDS-PAGE) and a shotgun two-dimensional liquid chromatographic (LS/LS) approach based on multidimensional protein identification technology (MudPIT). We were interested in the most abundant proteins of the apical brush border midgut membrane. About 400 spots were detected on 2D gels and 39 spots were cored and identified by mass spectrometry. 86 proteins were identified by MudPIT. Three proteins, arginine kinase, putative allergen and actin are shown to be the most predominant proteins in the sample. The total number of 36 proteins detected by both methods represents the most abundant proteins in the BBMV. PMID:19133270

  8. Specific binding of lactoferrin to brush-border membrane: Ontogeny and effect of glycan chain

    SciTech Connect

    Davidson, L.A.; Loennerdal, B. )

    1988-04-01

    Bioavailability of iron from human milk is exceptionally high. It has been suggested that lactoferrin, the major iron-binding protein in human milk, may participate in this high iron bioavailability from milk. The authors examined the interaction of lactoferrin with the intestinal brush-border membrane using the rhesus monkeys as a model. Brush-border membrane vesicles were prepared from monkeys of various ages. Binding studies with {sup 59}Fe-labeled human and monkey lactoferrin were performed to examine interaction of lactoferrin with the brush-border membrane. Specific saturable binding of lactoferrin was found at all ages studied. The dissociation constant for lactoferrin-receptor binding was 9 {times} 10{sup {minus}6} M. In contrast, no binding of serum transferrin or bovine lactoferrin occurred. Removal of fucose from the lactoferrin glycans resulted in a significant decrease in binding. It was concluded that lactoferrin in milk may function in the process of iron absorption through interaction with a small intestinal receptor and that fucosylated glycans on the carbohydrate chain of lactoferrin are necessary for receptor recognition.

  9. Light-chain binding sites on renal brush-border membranes

    SciTech Connect

    Batuman, V.; Dreisbach, A.W.; Cyran, J.

    1990-05-01

    Immunoglobulin light chains are low-molecular-weight proteins filtered at the renal glomerulus and catabolized within the proximal tubular epithelium. Excessive production and urinary excretion of light chains are associated with renal dysfunction. They also interfere with proximal renal tubule epithelial functions in vitro. We studied the binding of 125I-labeled kappa- and lambda-light chains, obtained from the urine of multiple myeloma patients, to rat and human renal proximal tubular brush-border membranes. Light-chain binding to brush borders was also demonstrated immunologically by flow cytometry. Computer analysis of binding data was consistent with presence of a single class of low-affinity, high-capacity, non-cooperative binding sites with relative selectivity for light chains on both rat and human kidney brush-border membranes. The dissociation constants of light chains ranged from 1.6 X 10(-5) to 1.2 X 10(-4) M, and maximum binding capacity ranged from 4.7 +/- 1.3 X 10(-8) to 8.0 +/- 0.9 X 10(-8) (SD) mol/mg protein at 25 degrees C. Kappa- and lambda-light chains competed with each other for binding with comparable affinity constants. Competition by albumin and beta-lactoglobulin, however, was much weaker, suggesting relative site selectivity for light chains. These binding sites probably function as endocytotic receptors for light chains and possibly other low-molecular-weight proteins.

  10. "Nonclassical" secretion of annexin A2 to the lumenal side of the enterocyte brush border membrane.

    PubMed

    Danielsen, E Michael; van Deurs, Bo; Hansen, Gert H

    2003-12-16

    Annexin A2 is a member of the annexin family of Ca(2+)-dependent lipid binding proteins and believed to be engaged in membrane transport processes in a number of cell types. In small intestinal enterocytes, we localized annexin A2 to the brush border region, where it was found mainly on the lumenal side of the microvilli, showing an apical secretion by a "nonclassical" mechanism. In addition, annexin A2 was associated with surface-connected, deep apical tubules in the apical terminal web region and with an underlying pleiomorphic, tubulo-vesicular compartment (subapical compartment/multivesicular bodies). By subcellular fractionation, the 36 kDa full-length form of annexin A2 was approximately equally distributed between the Mg(2+)-precipitated fraction (containing intracellular and basolateral membranes) and the microvillar membrane fraction. In addition, a 33 kDa molecular form of annexin A2 was seen in the latter fraction that could be generated from the full-length annexin A2 by digestion with trypsin. Taken together, the results suggest that annexin A2 acts in exocytic apical membrane trafficking and is proteolytically cleaved in situ by pancreatic proteinases once it has become externalized to the lumenal side of the brush border membrane. On the basis of its well-known membrane fusogenic properties, we propose a model for the nonclassical membrane translocation of annexin A2. PMID:14661980

  11. Enzymatic and transport characteristics of isolated snake renal brush-border membranes

    SciTech Connect

    Benyajati, S.; Dantzler, W.H. )

    1988-07-01

    Brush-border membranes (BBM) or proximal tubules were isolated from the kidney of the garter snake (Thamnophis sirtalis) by a procedure involving hypotonic lysis, Ca precipitation, and differential centrifugation. The isolated membranes were enriched 15-fold in brush-border enzyme activities (alkaline phosphatase, {gamma}-glutamyl transpeptidase) compared with whole kidney homogenates and were substantially free of other contaminating membranes. The yield of the BBM preparation was 40%. The BBM vesicular transport of several organic solutes was characterized by a rapid filtration technique at 25{degree}C. D-glucose, p-aminohippurate (PAH), and urate entered the same osmotically active space and binding was minimal. An uptake overshoot for 3-O-methyl-D-glucose by reptilian BBM was observed only in the presence of an inwardly directed NaCl gradient and was abolished by 0.1 mM phlorizin. Reptilian BBM exhibited Na-gradient-stimulated uptake of PAH (90 {mu}M) with an overshoot that was inhibited by other organic acids and by 4-acetamido-4{prime}-isothiocyanostilbene-2,2{prime}-disulfonic acid (SITS). In contrast, urate uptake appeared to be Na independent and not appreciably affected by other organic anions or SITS. The presence of specific transport systems for organic solutes in the isolated membrane preparation distinctly characterizes the BBM of reptilian kidney.

  12. Thyroid hormones increase Na -H exchange activity in renal brush border membranes

    SciTech Connect

    Kinsella, J.; Sacktor, B.

    1985-06-01

    Na -H exchange activity, i.e., amiloride-sensitive Na and H flux, in renal proximal tubule brush border (luminal) membrane vesicles was increased in the hyperthyroid rat and decreased in the hypothyroid rat, relative to the euthyroid animal. A positive correlation was found between Na -H exchange activity and serum concentrations of thyroxine (T4) and triiodothyronine (T3). The thyroid status of the animal did not alter amiloride-insensitive Na uptake. The rate of passive pH gradient dissipation was higher in membrane vesicles from hyperthyroid rats compared to the rate in vesicles from hypothyroid animals, a result which would tend to limit the increase in Na uptake in vesicles from hyperthyroid animals. Na -dependent phosphate uptake was increased in membrane vesicles from hyperthyroid rats; Na -dependent D-glucose and L-proline uptakes were not changed by the thyroid status of the animal. The effect of thyroid hormones in increasing the uptake of Na in the brush border membrane vesicle is consistent with the action of the hormones in enhancing renal Na reabsorption.

  13. Detection of renal brush border membrane enzymes for evaluation of renal injury in neonatal scleredema

    PubMed Central

    Ren, Qing; Zhang, Yongjun; Yang, Jinying; Wei, Lixia; Zhao, Lili; Yang, Qiaozhi

    2015-01-01

    Objective: To evaluate renal brush border membrane enzymes in urine as an indicator for renal injury in neonatal scleredema (NS). Methods: Sixty nine NS patients in our hospital were enrolled and divided into mild group and moderate/severe group. Patients were further randomly divided into therapy and control subgroups for 7 days ligustrazine administration. Urine samples were collected to detect renal brush border membrane enzymes (RBBME) by ELISA and β2-microglobulin (β2-MG) by radioimmunoassay (RIA). The results were compared with those of 30 normal neonates. Data were statistically analyzed using SPSS13.0 software. Results: Both RBBME and β2-MG were found to be higher in urine in NS patients than normal controls (P < 0.01). Level of RBBME increased with the severity of NS (P <0.05), while urinary β2-MG did not (P >0.05). After being treated with ligustrazine, a medicine for renal function recovery, both RBBME and β2-MG were similarly significantly decreased comparing to untreated groups (P < 0.05). 79.7% of NS patients showed abnormal RBBME while only 52.2% had an abnormal urinary β2-MG (χ2=11.65,P < 0.01). Conclusion: RBBME was more sensitive than β2-MG in reflecting the renal injury in NS. Examination of RBBME effectively reflected the recovery of renal injury after treatment with ligustrazine. PMID:25878616

  14. Ontogeny of Na/H antiporter activity in rabbit renal brush border membrane vesicles.

    PubMed Central

    Beck, J C; Lipkowitz, M S; Abramson, R G

    1991-01-01

    The development of the Na/H antiporter was studied in renal brush border membrane vesicles (BBMV) from fetal and adult rabbits using isotopic and fluorescent techniques. The kinetics of the antiporter studied by 22Na+ uptake revealed that the Vmax was only 25% of that in the adult; however, the Km's for Na+ were not significantly different. These data were confirmed by a fluorescent assay using the pH-sensitive probe, acridine orange: the Vmax was significantly lower in the fetal BBMV. Conductive Na+ movement was estimated from amiloride-insensitive 22Na+ uptake and the rate of alkalinization induced by K+, an ion whose relative conductance was found to be similar to that of Na+. Although relative Na+ conductance was significantly greater in fetal BBMV, the lower Vmax in fetal vesicles could not be ascribed to this factor. Maternal administration of betamethasone (50 micrograms/kg intramuscularly) for 2 d before delivery significantly increased the Vmax of the antiporter to levels observed in the adult; Km was unaffected. Na/K ATPase activity increased fourfold after betamethasone, but the specific activities of four brush border marker enzymes and the kinetics of Na(+)-glucose cotransport were unchanged. These data indicate that there is a developmental increase in brush border Na/H exchange which is the result of an increase in the number and/or the turnover number of the carriers. Further, these data suggest that the postnatal increase in antiporter activity may be related to the surge in glucocorticoid concentration that occurs perinatally. Images PMID:1645751

  15. Differences in neutral amino acid and glucose transport between brush border and basolateral plasma membrane of intestinal epithelial cells.

    PubMed

    Hopfer, U; Sigrist-Nelson, K; Ammann, E; Murer, H

    1976-12-01

    A comparison of L-valine and D-glucose transport was carried out with vesicles of plasma membrane isolated either from the luminal (brush border) or from the contra-luminal (basolateral) region of small intestinal epithelial cells. The existence of transport systems for both non-electrolytes was demonstrated by stereospecificity and saturability of uptake, as well as tracer coupling. Transport of L-valine and D-glucose differs markedly in the two types of plasma membrane with respect to stimulation by Na+. The presence of Na+ stimulated initial L-valine and D-glucose uptake in brush border, but not in basolateral membrane. Moreover, an electro-chemical Na+ gradient, oriented with the lower potential on the inside, supported accumulation of the non-electrolytes above medium concentration only in the brush border membrane. L-Valine and D-glucose transport also were saturated at lower concentrations in brush border (10-20 mM) than in basolateral plasma membranes (30-50 mM). A third difference between the two membranes was found in the effectiveness of known inhibitors of D-glucose transport. In brush border membranes phlorizin was more potent than phloretin and 2', 3', 4'-trihydroxy-4-methoxy chalcone and cytochalasin B did not inhibit at all. In contrast, with the basolateral plasma membranes the order of potency was changed to phloretin = 2',3',4'-trihydroxy-4-methoxy chalcone greater than cytochalasin B greater than phlorizin. These results indicate the presence of different types of transport systems for monosaccharides and neutral amino acids in the luminal and contra-luminal region of the plasma membrane. Active transepithelial transport can be explained on the basis of the different properties of the non-electrolyte transport systems in the two cellular regions and an electro-chemical Na+ gradient that is dependent on cellular metabolism. PMID:137908

  16. Association and dissociation of Escherichia coli heat-stable enterotoxin from rat brush border membrane receptors

    SciTech Connect

    Cohen, M.B.; Thompson, M.R.; Overmann, G.J.; Giannella, R.A.

    1987-02-01

    Escherichia coli heat-stable enterotoxin (ST) binds to receptors on rat intestinal cells and brush border membranes (BBM). We devised experiments to examine the reversibility of ST binding. We found that both /sup 125/I-labeled ST and native ST were spontaneously dissociable from the BBM receptor. Radiolabeled ST bound to BBM was also dissociated by the addition of avid goat anti-ST antiserum. Furthermore, using a computer program for analysis of ligand binding, we calculated an apparent Ka of 10(8) liters/mol from competitive inhibition and saturation-binding data. This is significantly lower than the value previously reported by others. Our findings, of a lower Ka and a reversible ST-binding process, suggest that a therapeutic strategy of removing bound ST from its receptor or competing with the enterocyte receptor for unbound ST might be successful in terminating ST-induced secretion.

  17. H+ V-ATPase-Energized Transporters in Brush Border Membrane Vesicles from Whole Larvae of Aedes Aegypti

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Brush Border Membrane vesicles (BBMVs) from Whole larvae of Aedes aegypti (AeBBMVWs ) contain an H+ V-ATPase (V), a Na+/H+ antiporter, NHA1 (A) and a Na+-coupled, nutrient amino acid transporter, NAT8 (N), VAN for short. All V-ATPase subunits are present in the Ae. aegypti genome and in the vesicles...

  18. Characteristics of glutamine transport in dog jejunal brush-border membrane vesicles.

    PubMed

    Bulus, N M; Abumrad, N N; Ghishan, F K

    1989-07-01

    The present study characterizes glutamine transport across brush-border membrane vesicles (BBMV) prepared from dog jejunum. The purity of these vesicles was demonstrated by a 20-fold enrichment of leucine aminopeptidase, a marker for BBM. Glutamine uptake was found to occur into an osmotically active space with no membrane binding and to exhibit temperature and pH dependence (optimal uptake at pH 7-7.5). Glutamine uptake was driven by an inwardly directed Na+ gradient with a distinct overshoot not observed under K+ gradient. Lithium could not substitute for Na+ as a stimulator of glutamine uptake. Na+-dependent glutamine uptake was not inhibited by methylaminoisobutyric acid, a typical substrate for system A, and was found to be electrogenic and saturable with a Km of 0.97 +/- 0.58 mM and a Vmax of 3.93 +/- 0.99 nmol.mg protein-1.10 s-1. A Na+-glutamine coupling ratio of 1:1 could be demonstrated by a plot of Hill transformation. Na+-independent glutamine uptake was found to be electroneutral and saturable with a Km of 3.70 +/- 0.66 mM and a Vmax of 2.70 +/- 1.55 nmol.mg protein-1.10 s-1. Inhibition studies confirmed the presence of a Na+-dependent as well as a Na+-independent carrier for glutamine uptake. We conclude that glutamine uptake across dog BBMV occurs via two transport systems: a Na+-dependent high-affinity system similar to the neutral brush-border system and a Na+-independent lower-affinity system similar to system L. PMID:2750912

  19. Effects of dimethylsulfoxide and mercurial sulfhydryl reagents on water and solute permeability of rat kidney brush border membranes.

    PubMed

    van Hoek, A N; de Jong, M D; van Os, C H

    1990-12-14

    The effects of dimethylsulfoxide, DMSO, and mercurial sulfhydryl reagents have been studied on water and small solute permeability of rat renal brush border membrane vesicles. Water and solute permeability was measured by mixing membrane vesicles with hypertonic solutions in a stopped-flow apparatus and following osmotically-induced changes in vesicular volume via changes in scattered light intensity. The rate constant of the fast osmotic shrinkage is proportional to the osmotic water permeability, while the rate constant of the slow reswelling phase is proportional to the solute permeability. Using mannitol as the osmotic agent, the osmotic shrinkage of rat renal brush border membrane vesicles followed a biphasic time course. 80% of the vesicles shrunk with a rate constant of approx. 50 s-1 and 20% with a rate constant of approx. 2 s-1. DMSO decreased dose-dependently the amplitude of the fast osmotic shrinkage, without affecting its rate constant. In contrast to DMSO, HgCl2 decreased the rate constant but not the amplitude of the fast osmotic shrinkage of renal brush border vesicles. Between 40-50 microM HgCl2, the inhibition of the fast osmotic shrinkage was completed. DMSO and HgCl2 increase the activation energy of water permeation in renal membranes from 3 to 12-15 kcal/mol. DMSO and HgCl2 did not affect the rate constant of the slow osmotic shrinkage of renal membrane vesicles and were also without effect on osmotic shrinkage of small intestinal brush border and pure phospholipid vesicles. In renal brush border membranes, HgCl2 at low concentrations (less than 10 microM) increased by 15-fold the permeability to NaCl and urea but not to mannitol, an effect which precedes the inhibition of water permeability at higher HgCl2 concentrations. The increase in small solute permeability was irreversible while the inhibition of water permeability could be reversed with cysteine and dithiothreitol. We conclude that water and small solute pathways in rat renal brush

  20. Sulfate uptake by crustacean hepatopancreatic brush border membrane vesicles. [Homarus americanus

    SciTech Connect

    Gerencser, G.A.; Cattey, M.A; Ahearn, G.A. Univ. of Hawaii, Honolulu )

    1990-02-26

    Purified brush border membrane vesicles (BBMV) were prepared from Atlantic lobster (Homarus americanus) hepatopancreas using differential centrifugation and Mg{sup +2} precipitation techniques. Uptake of 0.1 mM {sup 35}SO{sub 4}{sup {minus}2} was stimulated by pre-loading vesicles with Cl{sup {minus}} leading to a transient accumulation of isotope more than twice that at equilibrium. Pre-loading with HCO{sub 3}{sup {minus}} or gluconate had no effect on sulfate uptake. No stimulation of {sup 35}SO{sub 4}{sup {minus}2} was observed in the presence of inwardly directed Na{sup +} or tetramethylammonium{sup +} gradients. Uptake of the divalent anion was strongly stimulated by inwardly directed proton gradients (pH{sub o} < pH{sub i}) and markedly inhibited by outwardly directed proton gradients (pH{sub o} > pH{sub i}). {sup 35}SO{sub 4}{sup {minus}2}/Cl{sup {minus}} exchange was enhanced by imposing a transmembrane inside positive K{sup +} diffusion potential and inhibited by a membrane potential of the opposite polarity (K{sup +}/valinomycin). Results suggest the presence of a proton-dependent, electrogenic anion antiport mechanism in BBMV isolated from the crustacean hepatopancreas.

  1. Heparin blocks /sup 125/I-calmodulin internalization by isolated rat renal brush border membrane vesicles

    SciTech Connect

    Meezan, E.; Elgavish, A.; Roden, L.; Wallace, R.W.

    1986-03-05

    /sup 125/I-Calmodulin is internalized by isolated rat renal brush border membrane vesicles (BBV) in a time, temperature and calcium dependent manner. Internalization of /sup 125/I-calmodulin into the osmotically sensitive space of BBV was distinguished from binding of the ligand to the outer BBV surface by examining the interaction of ligand and BBV at different medium osmolarities (300-1100 mosm), uptake was inversely proportional to medium osmolarity. Internalized /sup 125/I-calmodulin was intact and Western blots of solubilized BBV with /sup 125/I-calmodulin demonstrated the presence of several calmodulin-binding proteins of 143, 118, 50, 47.5, 46.5 and 35 kilodaltons which could represent potential intravesicular binding sites for the ligand. Heparin and the related glycosaminoglycan heparin sulfate both showed a dose-dependent inhibition (0.5-50 ..mu..g/ml) of /sup 125/I-calmodulin uptake by BBV, but other sulfated and nonsulfated glycosaminoglycans including chondroitin sulfates, keratan sulfate and hyaluronic acid showed little or no inhibitory effect. Desulfation of heparin virtually abolished the inhibition of uptake while depolymerization reduced it. Heparin did not block the binding of /sup 125/I-calmodulin to BBV proteins as assessed by Western blotting technique suggesting its effect was on internalization of the ligand rather than on its association with internal membrane proteins.

  2. Improved stability of rabbit and rat intestinal brush border membrane vesicles using phospholipase inhibitors.

    PubMed

    Maenz, D D; Chenu, C; Bellemare, F; Berteloot, A

    1991-11-01

    The initial rates of Na(+)-dependent D-aspartate and D-glucose uptakes were shown to decline from the time of resuspension of brush border membrane vesicles isolated from rabbit and rat jejunum by standard divalent cation precipitation procedures. The former were however more stable than the latter and followed quite closely the decrease in the intravesicular volume, thus suggesting that the loss of transport activity may involve both nonspecific opening of the vesicles and either direct or indirect specific inactivation of the transporters. Uptake rates for both substrates did tend to stabilize at 6-24 h from resuspension, however this final 'next day' uptake activity was too low to be of practical use in kinetic studies. Freezing aliquots of rabbit jejunal vesicles in liquid N2 until the time of assay resulted in complete stabilization of D-glucose uptake. A modified homogenate buffer designed to inhibit a broad spectrum of phospholipase activities resulted in a partial stabilization of glucose transport by rabbit jejunal vesicles with, on average, an over 6-fold enrichment in the 'next day' stable specific activity of uptake as compared to unfrozen vesicles. The modified homogenate buffer also improved the stability and the 'next day' specific activities of D-glucose uptake in rat jejunal brush border vesicles and D-aspartic acid uptake in rabbit jejunal vesicles. It also completely stabilized the intravesicular volume in the latter preparation. An evaluation of the kinetic parameters of Na(+)-dependent D-glucose transport in rabbit vesicles prepared from either the standard homogenate media and frozen in liquid N2 or the modified media and allowed to stabilize overnight, revealed a single transport system with a Km of 0.31-0.32 mM as the best model to fit the data. As such the modifications to the homogenate media do not appear to effect the functional properties of D-glucose transport in the membrane. While being less efficient in stabilizing the vesicles than

  3. Acute and chronic exposure to ethanol and the electrophysiology of the brush border membrane of rat small intestine.

    PubMed Central

    al-Balool, F; Debnam, E S; Mazzanti, R

    1989-01-01

    In this study we have investigated the effects of (a) chronic ethanol intake on glucose and galactose absorption across the rat jejunum in vivo and on the potential difference across the isolated brush border membrane (Vm) and (b) acute exposure to ethanol (4% or 8%) and acetaldehyde (0.25%) on changes in Vm associated with Na(+)-dependent galactose absorption across the jejunum and ileum. Chronic ethanol intake was associated with hyperpolarization of Vm and an enhanced galactose but not glucose transport. Acute ethanol and acetaldehyde were without effect on Vm whether or not galactose was present. We conclude that while a greater electrochemical gradient across the brush border membrane is a likely explanation for the stimulation of galactose absorption induced by ethanol feeding, factors other than changes in Vm are responsible for the inhibitory effects of acute ethanol. PMID:2612984

  4. Direct effect of vanadium on citrate uptake by rat renal brush border membrane vesicles (BBMV).

    PubMed

    Sato, Kazuhiro; Kusaka, Yukinori; Akino, Hironobu; Kanamaru, Hiroshi; Okada, Kenichiro

    2002-07-01

    Vanadium pentoxide is used as a catalyst and a ferrovanadium alloy ingredient in automotive steels and in jet engines and airframes. In addition, vanadium is found in fuel oils. Thus, occupational exposures to vanadium pentoxide and trioxide may occur during the cleaning of oil-fired ship boilers, and from oil-fired power station boilers. Occupational exposure to vanadium pentoxide induces green tongue, asthmatic symptoms and albuminuria with cast. Urinary citrate is freely filtered at the glomerulus, and its reabsorption in the proximal tubule is the major determinant of the rate of renal excretion. In this study, we exposed rat renal brush border membrane vesicles (BBMV) to vanadium pentoxide and examined their citrate uptake characteristics. The preincubation of BBMV with 1 mM V2O5 for 8 hours significantly inhibited citrate uptake compared with that of BBMV without V2O5, preincubation. These findings indicate that the preincubation of BBMV with vanadium pentoxide results in a time-dependent inhibition of citrate uptake by BBMV. These findings might contribute to nephrotoxicity in vanadium exposure. PMID:12141377

  5. Sulfate inhibits ( sup 14 C)phosphonoformic acid binding to renal brush-border membranes

    SciTech Connect

    Tenenhouse, H.S.; Lee, J. )

    1990-08-01

    To examine the specificity of the phosphonoformic acid (PFA) interaction with the Na(+)-dependent phosphate transporter of mouse renal brush-border membrane vesicles, we compared the effects of anions on Na(+)-dependent (14C)PFA binding and Na(+)-dependent phosphate transport. Inhibition of PFA binding was achieved by PFA (% control = 0 +/- 1), sulfate (15 +/- 2), arsenate (35 +/- 1), phosphate (59 +/- 2), and nitrate (68 +/- 4), whereas inhibition of phosphate transport was only apparent with phosphate (0 +/- 1), PFA (22 +/- 4), and arsenate (37 +/- 5). Succinate and gluconate had no effect on either Na(+)-dependent process. Under conditions where Na(+)-dependent PFA binding was maximally inhibited by phosphate (% control = 65 +/- 4), further inhibition could be achieved by sulfate (26 +/- 5%). Na(+)-dependent PFA binding was competitively inhibited by phosphate (apparent Ki = 8.9 +/- 1.2 mM) and noncompetitively inhibited by sulfate (apparent Ki = 2.6 +/- 0.5 mM). We found that PFA inhibited Na(+)-dependent sulfate transport (50% inhibition at 9 mM PFA) as well as Na(+)-dependent phosphate transport (50% inhibition at 0.5 mM PFA). We also examined the pH dependence of Na(+)-dependent PFA binding and Na(+)-dependent phosphate and sulfate transport. PFA binding was optimal at pH = 7.4, whereas phosphate transport increased with increasing pH, and sulfate transport increased with decreasing pH.

  6. Effect of chronic (4 weeks) ingestion of ethanol on transport of proline into intestinal brush border membrane vesicles

    SciTech Connect

    Beesley, R.C.; Jones, T.D.

    1986-03-01

    Hamsters were separated into two groups and fed liquid diets ad lib. Controls were given a diet similar to that described by DeCarli and Lieber while alcoholics received the same diet containing 5% ethanol isocalorically substituted for sucrose. The volume of diet consumed daily and the gain in body weights of alcoholics were not significantly different from those of controls. After four weeks the animals were sacrificed and the upper third of the small intestine was used to prepare brush border membrane vesicles. In the presence of a Na/sup +/ gradient, uptake of proline into vesicles prepared from both groups was rapid, reaching a maximum accumulation in 1 to 2 min and then decreasing to the equilibrium level. To normalize the results, the amount of proline take up at each time point was divided by the amount present at equilibrium. From the normalized data it was concluded that both the rate of uptake and the maximum accumulation of proline into brush border membrane vesicles isolated from alcoholics were significantly less than those obtained with vesicles from controls. These results suggest that chronic ingestion of ethanol results in a reduction in Na/sup +/-dependent transport of proline across the brush border membrane and, thus, may contribute to the malnutrition which is frequently associated with chronic alcoholism.

  7. Characterization of tryptophan transport in human placental brush-border membrane vesicles.

    PubMed Central

    Ganapathy, M E; Leibach, F H; Mahesh, V B; Howard, J C; Devoe, L D; Ganapathy, V

    1986-01-01

    The characteristics of tryptophan uptake in isolated human placental brush-border membrane vesicles were investigated. Tryptophan uptake in these vesicles was predominantly Na+-independent. Uptake of tryptophan as measured with short incubations occurred exclusively by a carrier-mediated process, but significant binding of this amino acid to the membrane vesicles was observed with longer incubations. The carrier-mediated system obeyed Michaelis-Menten kinetics, with an apparent affinity constant of 12.7 +/- 1.0 microM and a maximal velocity of 91 +/- 5 pmol/15 s per mg of protein. The kinetic constants were similar in the presence and absence of a Na+ gradient. Competition experiments showed that tryptophan uptake was effectively inhibited by many neutral amino acids except proline, hydroxyproline and 2-(methylamino)isobutyric acid. The inhibitory amino acids included aromatic amino acids as well as other system-1-specific amino acids (system 1 refers to the classical L system, according to the most recent nomenclature of amino acid transport systems). The transport system showed very low affinity for D-isomers, was not affected by phloretin or glucose but was inhibited by p-azidophenylalanine and N-ethylmaleimide. The uptake rates were only minimally affected by change in pH over the range 4.5-8.0. Tryptophan uptake markedly responded to trans-stimulation, and the amino acids capable of causing trans-stimulation included all amino acids with system-1-specificity. The patterns of inhibition of uptake of tryptophan and leucine by various amino acids were very similar. We conclude that system t, which is specific for aromatic amino acids, is absent from human placenta and that tryptophan transport in this tissue occurs via system 1, which has very broad specificity. PMID:3800932

  8. Ezetimibe Promotes Brush Border Membrane-to-Lumen Cholesterol Efflux in the Small Intestine.

    PubMed

    Nakano, Takanari; Inoue, Ikuo; Takenaka, Yasuhiro; Ono, Hiraku; Katayama, Shigehiro; Awata, Takuya; Murakoshi, Takayuki

    2016-01-01

    Ezetimibe inhibits Niemann-Pick C1-like 1 (NPC1L1), an apical membrane cholesterol transporter of enterocytes, thereby reduces intestinal cholesterol absorption. This treatment also increases extrahepatic reverse cholesterol transport via an undefined mechanism. To explore this, we employed a trans-intestinal cholesterol efflux (TICE) assay, which directly detects circulation-to-intestinal lumen 3H-cholesterol transit in a cannulated jejunal segment, and found an increase of TICE by 45%. To examine whether such increase in efflux occurs at the intestinal brush border membrane(BBM)-level, we performed luminal perfusion assays, similar to TICE but the jejunal wall was labelled with orally-given 3H-cholesterol, and determined elevated BBM-to-lumen cholesterol efflux by 3.5-fold with ezetimibe. Such increased efflux probably promotes circulation-to-lumen cholesterol transit eventually; thus increases TICE. Next, we wondered how inhibition of NPC1L1, an influx transporter, resulted in increased efflux. When we traced orally-given 3H-cholesterol in mice, we found that lumen-to-BBM 3H-cholesterol transit was rapid and less sensitive to ezetimibe treatment. Comparison of the efflux and fractional cholesterol absorption revealed an inverse correlation, indicating the efflux as an opposite-regulatory factor for cholesterol absorption efficiency and counteracting to the naturally-occurring rapid cholesterol influx to the BBM. These suggest that the ezetimibe-stimulated increased efflux is crucial in reducing cholesterol absorption. Ezetimibe-induced increase in cholesterol efflux was approximately 2.5-fold greater in mice having endogenous ATP-binding cassette G5/G8 heterodimer, the major sterol efflux transporter of enterocytes, than the knockout counterparts, suggesting that the heterodimer confers additional rapid BBM-to-lumen cholesterol efflux in response to NPC1L1 inhibition. The observed framework for intestinal cholesterol fluxes may provide ways to modulate the flux

  9. Ezetimibe Promotes Brush Border Membrane-to-Lumen Cholesterol Efflux in the Small Intestine

    PubMed Central

    Nakano, Takanari; Inoue, Ikuo; Takenaka, Yasuhiro; Ono, Hiraku; Katayama, Shigehiro; Awata, Takuya; Murakoshi, Takayuki

    2016-01-01

    Ezetimibe inhibits Niemann-Pick C1-like 1 (NPC1L1), an apical membrane cholesterol transporter of enterocytes, thereby reduces intestinal cholesterol absorption. This treatment also increases extrahepatic reverse cholesterol transport via an undefined mechanism. To explore this, we employed a trans-intestinal cholesterol efflux (TICE) assay, which directly detects circulation-to-intestinal lumen 3H-cholesterol transit in a cannulated jejunal segment, and found an increase of TICE by 45%. To examine whether such increase in efflux occurs at the intestinal brush border membrane(BBM)-level, we performed luminal perfusion assays, similar to TICE but the jejunal wall was labelled with orally-given 3H-cholesterol, and determined elevated BBM-to-lumen cholesterol efflux by 3.5-fold with ezetimibe. Such increased efflux probably promotes circulation-to-lumen cholesterol transit eventually; thus increases TICE. Next, we wondered how inhibition of NPC1L1, an influx transporter, resulted in increased efflux. When we traced orally-given 3H-cholesterol in mice, we found that lumen-to-BBM 3H-cholesterol transit was rapid and less sensitive to ezetimibe treatment. Comparison of the efflux and fractional cholesterol absorption revealed an inverse correlation, indicating the efflux as an opposite-regulatory factor for cholesterol absorption efficiency and counteracting to the naturally-occurring rapid cholesterol influx to the BBM. These suggest that the ezetimibe-stimulated increased efflux is crucial in reducing cholesterol absorption. Ezetimibe-induced increase in cholesterol efflux was approximately 2.5-fold greater in mice having endogenous ATP-binding cassette G5/G8 heterodimer, the major sterol efflux transporter of enterocytes, than the knockout counterparts, suggesting that the heterodimer confers additional rapid BBM-to-lumen cholesterol efflux in response to NPC1L1 inhibition. The observed framework for intestinal cholesterol fluxes may provide ways to modulate the flux

  10. Proline transport by brush-border membrane vesicles of lobster antennal glands

    SciTech Connect

    Behnke, R.D.; Wong, R.K.; Huse, S.M.; Reshkin, S.J.; Ahearn, G.A. )

    1990-02-01

    Purified brush-border membrane vesicles (BBMV) of lobster antennal gland labyrinth and bladder were separately formed by a magnesium precipitation technique. L-(3H)proline uptake was stimulated by a transmembrane NaCl gradient (outside (o) greater than inside (i)) to a greater extent in BBMV from labyrinth than those from the bladder. Detailed study of the labyrinth proline-transport processes revealed a specific dependence on NaCl, with negligible stimulatory effects by NaSCN, Na-gluconate, or KCl. A transmembrane proton gradient (o greater than i) was without stimulatory effect on proline transport. A transmembrane potential difference alone, in the presence of equilibrated NaCl and L-(3H)proline, led to net influx of the labeled amino acid, suggesting that the uptake process was electrogenic and capable of bringing about the net transfer of positive charge to the vesicle interior. Although a transmembrane Na gradient alone, in the presence of equilibrated Cl and L-(3H)proline, was able to bring about the net influx of the amino acid, a transmembrane Cl gradient alone under Na- and L-(3H)proline-equilibrated conditions was not, suggesting that only the Na gradient could energize the carrier process through cotransport, while the anion served an essential activating role. Proline influx by these vesicles occurred by the combination of at least one saturable Michaelis-Menten carrier system (apparent Kt = 0.37 mM; apparent JM = 1.19 nmol.mg protein-1.10 s-1) and apparent diffusion (P = 0.33 nmol.mg protein-1.10 s-1.mM-1). Static head analysis of the transport process suggested a cotransport stoichiometry of 2 Na:1 proline with essential activation by Cl ion.

  11. Bacillus thuringiensis delta-endotoxin binding to brush border membrane vesicles of rice stem borers.

    PubMed

    Alcantara, Edwin P; Aguda, Remedios M; Curtiss, April; Dean, Donald H; Cohen, Michael B

    2004-04-01

    The receptor binding step in the molecular mode of action of five delta-endotoxins (Cry1Ab, Cry1Ac, Cry1C, Cry2A, and Cry9C) from Bacillus thuringiensis was examined to find toxins with different receptor sites in the midgut of the striped stem borer (SSB) Chilo suppressalis (Walker) and yellow stem borer (YSB) Scirpophaga incertulas (Walker) (Lepidoptera: Pyralidae). Homologous competition assays were used to estimate binding affinities (K(com)) of (125)I-labelled toxins to brush border membrane vesicles (BBMV). The SSB BBMV affinities in decreasing order was: Cry1Ab = Cry1Ac > Cry9C > Cry2A > Cry1C. In YSB, the order of decreasing affinities was: Cry1Ac > Cry1Ab > Cry9C = Cry2A > Cry1C. The number of binding sites (B(max)) estimated by homologous competition binding among the Cry toxins did not affect toxin binding affinity (K(com)) to both insect midgut BBMVs. Results of the heterologous competition binding assays suggest that Cry1Ab and Cry1Ac compete for the same binding sites in SSB and YSB. Other toxins bind with weak (Cry1C, Cry2A) or no affinity (Cry9C) to Cry1Ab and Cry1Ac binding sites in both species. Cry2A had the lowest toxicity to 10-day-old SSB and Cry1Ab and Cry1Ac were the most toxic. Taken together, the results of this study show that Cry1Ab or Cry1Ac could be combined with either Cry1C, Cry2A, or Cry9C for more durable resistance in transgenic rice. Cry1Ab should not be used together with Cry1Ac because a mutation in one receptor site could diminish binding of both toxins. PMID:15027071

  12. Uptake of /sup 75/Se-selenite by brush border membrane vesicles from chick duodenum stimulated by vitamin D

    SciTech Connect

    Mykkanen, H.M.; Wasserman, R.H.

    1989-02-01

    Brush border membrane vesicles were isolated from mucosal homogenates of duodena from normal, rachitic and vitamin D-treated rachitic chicks using a discontinuous sucrose gradient, and further purified by glycerol gradient centrifugation. In vitro uptake of 75Se-selenite by purified brush border membrane vesicles was studied using a rapid filtration technique. The time course of 75Se uptake was non-linear; rapid initial binding was followed by a gradual decrease in the rate of uptake until an equilibrium value was reached at 60-120 min. The initial binding at 36 s was not affected by selenite concentration in the incubation buffer, while the fractional rate of uptake between the 36 s and 2 min time periods was clearly lower with 1 mM Se than with 4-100 microM Se. 75Se uptake did not show any dependency on the external Na-gradient, nor could it be inhibited by other anions (arsenate, phosphate). Treatment of rachitic chicks either with cholecalciferol (500 Iu, 72 h) or with 1,25(OH)2-cholecalciferol (0.5 microgram given 16 h prior to isolation of the vesicles) significantly enhanced 75Se uptake. A threefold excess of mannitol in the outside buffer reduced 75Se uptake by vesicles from vitamin D-deficient and D-treated chicks 60% and 35% respectively, but had no effect on vesicles from vitamin D-treated chicks preloaded with 75Se. Neither saponin treatment nor excess cold selenite could release the label from the vesicles preloaded with 75Se. These data are compatible with the hypothesis that selenite easily crosses the brush border membrane into the intravesicular space and, once inside, is tightly bound by the membrane.

  13. A protein complex in the brush-border membrane explains a Hartnup disorder allele.

    PubMed

    Kowalczuk, Sonja; Bröer, Angelika; Tietze, Nadine; Vanslambrouck, Jessica M; Rasko, John E J; Bröer, Stefan

    2008-08-01

    Protein absorption in the intestine is mediated by proteases and brush-border peptidases together with peptide and amino acid transporters. Neutral amino acids are generated by a variety of aminopeptidases and carboxypeptidases and are subsequently taken up by the amino acid transporter B(0)AT1 (SLC6A19), which is mutated in Hartnup disorder. Coexpression of B(0)AT1 together with the brush-border carboxypeptidase angiotensin-converting enzyme 2 (ACE2) in Xenopus laevis oocytes led to a dramatic increase of transporter expression at the oocyte surface. Other members of the SLC6 family were not stimulated by coexpression with ACE2. Addition of a peptide containing a carboxyterminal leucine residue to ACE2- and B(0)AT1-coexpressing oocytes caused inward currents due to Na(+)-leucine cotransport, demonstrating the formation of a metabolic complex. Coexpression of the Hartnup disorder causing mutation B(0)AT1(R240Q) showed reduced interaction with ACE2 and its renal paralogue collectrin. This would result in reduced surface expression in both kidney and intestine, thereby explaining the onset of the disorder in individuals carrying this mutation. PMID:18424768

  14. Microbial sphingomyelinase induces RhoA-mediated reorganization of the apical brush border membrane and is protective against invasion.

    PubMed

    Saslowsky, David E; Thiagarajah, Jay R; McCormick, Beth A; Lee, Jean C; Lencer, Wayne I

    2016-04-01

    The apical brush border membrane (BBM) of intestinal epithelial cells forms a highly structured and dynamic environmental interface that serves to regulate cellular physiology and block invasion by intestinal microbes and their products. How the BBM dynamically responds to pathogenic and commensal bacterial signals can define intestinal homeostasis and immune function. We previously found that in model intestinal epithelium, the conversion of apical membrane sphingomyelin to ceramide by exogenous bacterial sphingomyelinase (SMase) protected against the endocytosis and toxicity of cholera toxin. Here we elucidate a mechanism of action by showing that SMase induces a dramatic, reversible, RhoA-dependent alteration of the apical cortical F-actin network. Accumulation of apical membrane ceramide is necessary and sufficient to induce the actin phenotype, and this coincides with altered membrane structure and augmented innate immune function as evidenced by resistance to invasion by Salmonella. PMID:26864627

  15. Microbial sphingomyelinase induces RhoA-mediated reorganization of the apical brush border membrane and is protective against invasion

    PubMed Central

    Saslowsky, David E.; Thiagarajah, Jay R.; McCormick, Beth A.; Lee, Jean C.; Lencer, Wayne I.

    2016-01-01

    The apical brush border membrane (BBM) of intestinal epithelial cells forms a highly structured and dynamic environmental interface that serves to regulate cellular physiology and block invasion by intestinal microbes and their products. How the BBM dynamically responds to pathogenic and commensal bacterial signals can define intestinal homeostasis and immune function. We previously found that in model intestinal epithelium, the conversion of apical membrane sphingomyelin to ceramide by exogenous bacterial sphingomyelinase (SMase) protected against the endocytosis and toxicity of cholera toxin. Here we elucidate a mechanism of action by showing that SMase induces a dramatic, reversible, RhoA-dependent alteration of the apical cortical F-actin network. Accumulation of apical membrane ceramide is necessary and sufficient to induce the actin phenotype, and this coincides with altered membrane structure and augmented innate immune function as evidenced by resistance to invasion by Salmonella. PMID:26864627

  16. Glucose transport by brush-border membrane vesicles after proximal resection or ileo-jejunal transposition in the rat.

    PubMed Central

    Menge, H; Murer, H; Robinson, J W

    1978-01-01

    1. The functional properties of the ileal mucosa following jejunal resection or interposition in the jejunum were studied with the help of membrane vesicles. 2. Despite the development of functional and morphological features in the mucosa which are more generally associated with the jejunum than with the ileum, examination of brush-border vesicles derived from ileal and jejunal enterocytes from resected or transposed intestines reveals that the functional characteristics of apical membranes isolated from the different regions of the intestine are maintained after these surgical manoeuvres. Vesicles from control ileum develop an 'overshoot' uptake of glucose that is 3-4 times smaller than that of jejunal vesicles, a difference that is still observed in membranes prepared from mucosa of ileal loops following proximal resection or interposition in the jejunum. Images Fig. 3 PMID:625015

  17. High affinity 3H-Phe uptake by brush border membrane vesicles from whole larvae of Aedes aegypti (AaBMVw)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Brush border membrane vesicles from whole Aedes aegypti larvae (AaBBMVw) are confirmed to be valid preparations for membrane transport studies. The Abdul-Rauf and Ellar method was used to isolate AaBBMVw that were frozen, stored for several months, transported to a distant site, thawed and used to s...

  18. Phosphorylation of adenosine in renal brush-border membrane vesicles by an exchange reaction catalysed by adenosine kinase.

    PubMed Central

    Sayós, J; Solsona, C; Mallol, J; Lluis, C; Franco, R

    1994-01-01

    Uptake of [3H]adenosine in brush-border membrane (BBM) vesicles from either rat or pig kidney leads to an accumulation of intravesicular [3H]AMP. The lack of significant levels of ATP and the presence of AMP in BBM indicated that a phosphotransfer between [3H]adenosine and AMP occurs. The phosphotransfer activity is inhibited by iodotubercidin, which suggests that it is performed by adenosine kinase acting in an ATP-independent manner. The existence of a similar phosphotransferase activity was demonstrated in membrane-free extracts from pig kidney. From the compounds tested it was shown that a variety of mononucleotides could act as phosphate donors. The results suggest that phosphotransfer reactions may be physiologically relevant in kidney. PMID:8110185

  19. Influence of dietary partially hydrogenated fat high in trans fatty acids on lipid composition and function of intestinal brush border membrane in rats.

    PubMed

    Ghafoorunissa, S A.I.

    2001-02-01

    The effect of dietary hydrogenated fat (Indian vanaspati) high in trans fatty acids (6 en%) on lipid composition, fluidity and function of rat intestinal brush border membrane was studied at 2 and 8 en% of linoleic acid. Three groups of weanling rats were fed rice-pulse based diet containing 10% fat over a ten week period: Group I (groundnut oil), Group II (vanaspati), Group III (vanaspati + safflower oil). The functionality of the brush border membrane was assessed by the activity of membrane bound enzymes and transport of D-glucose and L-leucine. The levels of total cholesterol and phospholipids were similar in all groups. The data on fatty acid composition of membrane phospholipids showed that, at 2 en% of linoleic acid in the diet, trans fatty acids lowered arachidonic acid and increased linoleic acid contents indicating altered polyunsaturated fatty acid metabolism. Alkaline phosphatase activity was increased while the activities of sucrase, gamma-glutamyl transpeptidase and transport of D-glucose and L-leucine were not altered by dietary trans fatty acids. However at higher intake of linoleic acid in the diet, trans fatty acids have no effect on polyunsaturated fatty acid composition and alkaline phosphatase activity of intestinal brush border membrane. These data suggest that feeding dietary fat high in trans fatty acids is associated with alteration in intestinal brush border membrane polyunsaturated fatty acid composition and alkaline phosphatase activity only when the dietary linoleic acid is low. PMID:11182555

  20. Binding Analyses of Bacillus thuringiensis Cry δ-Endotoxins Using Brush Border Membrane Vesicles of Ostrinia nubilalis

    PubMed Central

    Hua, Gang; Masson, Luke; Jurat-Fuentes, Juan Luis; Schwab, George; Adang, Michael J.

    2001-01-01

    Transgenic corn expressing the Bacillus thuringiensis Cry1Ab gene is highly insecticidal to Ostrinia nubilalis (European corn borer) larvae. We ascertained whether Cry1F, Cry9C, or Cry9E recognizes the Cry1Ab binding site on the O. nubilalis brush border by three approaches. An optical biosensor technology based on surface plasmon resonance measured binding of brush border membrane vesicles (BBMV) injected over a surface of immobilized Cry toxin. Preincubation with Cry1Ab reduced BBMV binding to immobilized Cry1Ab, whereas preincubation with Cry1F, Cry9C, or Cry9E did not inhibit BBMV binding. BBMV binding to a Cry1F-coated surface was reduced when vesicles were preincubated in Cry1F or Cry1Ab but not Cry9C or Cry9E. A radioligand approach measured 125I-Cry1Ab toxin binding to BBMV in the presence of homologous (Cry1Ab) and heterologous (Cry1Ac, Cry1F, Cry9C, or Cry9E) toxins. Unlabeled Cry1Ac effectively competed for 125I-Cry1Ab binding in a manner comparable to Cry1Ab itself. Unlabeled Cry9C and Cry9E toxins did not inhibit 125I-Cry1Ab binding to BBMV. Cry1F inhibited 125I-Cry1Ab binding at concentrations greater than 500 nM. Cry1F had low-level affinity for the Cry1Ab binding site. Ligand blot analysis identified Cry1Ab, Cry1Ac, and Cry1F binding proteins in BBMV. The major Cry1Ab signals on ligand blots were at 145 kDa and 154 kDa, but a strong signal was present at 220 kDa and a weak signal was present at 167 kDa. Cry1Ac and Cry1F binding proteins were detected at 220 and 154 kDa. Anti-Manduca sexta aminopeptidase serum recognized proteins of 145, 154, and 167 kDa, and anti-cadherin serum recognized the 220 kDa protein. We speculate that isoforms of aminopeptidase and cadherin in the brush border membrane serve as Cry1Ab, Cry1Ac, and Cry1F binding proteins. PMID:11157257

  1. Effects of different Brush Border Membrane Vesicle isolation protocols on proteomic analysis of Cry1Ac binding proteins from the midgut of Helicoverpa armigera

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The brush border membrane vesicles (BBMV) isolated from midgut cells of insect have been widely used for studying of the binding receptors and action mode of Cry proteins produced by Bacillus thuringiensis (Bt). There are several methods for isolating insect BBMV used in one-dimension electrophoresi...

  2. Binding of E. coli heat-stable enterotoxin to rat intestinal brush borders and to basolateral membranes

    SciTech Connect

    Guarino, A.; Cohen, M.B.; Overmann, G.; Thompson, M.R.; Giannella, R.A.

    1987-09-01

    We studied the binding of E. coli heat-stable enterotoxin (STa) to rat brush borders (BB) and to basolateral membranes (BLM) using a biologically active monoiodinated radioligand (( /sup 125/I)STa) and highly enriched BB and BLM preparations free of other significant organelle contamination. Binding of (/sup 125/I)STa to BB was specific; time-, temperature-, and pH-dependent; saturable; and partially reversible. Nonlabeled toxin competitively inhibited the binding of radioligand to BB in a dose-related manner. Scatchard analysis revealed a single class of receptors with an apparent affinity constant of 8.7 +/- 1.5 X 10(8) l/mol. Binding was not affected by amino acids, sugars, and lectins. Proteolytic enzymes significantly decreased binding, although several did so by modifying the radioligand. Trypsin inhibited binding without modifying the radioligand thus supporting the proteinaceous nature of the receptor. Since the enrichment in binding activity in the BB over the homogenate was significantly lower than the enrichment in sucrase activity, we concluded that binding activity is probably associated with other membranous domains, but direct examination revealed no binding activity on basolateral membranes.

  3. Restoring effect of selenium on the molecular content, structure and fluidity of diabetic rat kidney brush border cell membrane.

    PubMed

    Gurbanov, Rafig; Bilgin, Mehmet; Severcan, Feride

    2016-04-01

    Diabetic kidney disease (DKD) is a dominant factor standing for kidney impairments during diabetes. In this study, attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy was used to disclose the diabetes-induced structural changes in the kidney and evaluate the effects of selenium on diabetes. The increase in the area of the olefinic band indicated increased amount of lipid peroxidation end products in diabetic kidney brush border cell membrane. Moreover, saturated lipid content of this cell membrane considerably diminished. DKD was found to disrupt lipid order and cause a decrease in membrane dynamics. However, the administration of selenium at low and medium doses was shown to improve these conditions by changing the lipid contents toward control values, restoring the ordered structure of the lipids and membrane dynamics. Curve-fitting and artificial neural network (ANN) analyses of secondary structures of proteins demonstrated a relative increase in α-helix and reduction in the β-sheet during diabetes in comparison to the control group, which were ameliorated following selenium treatment at low and medium doses. These findings were further confirmed by applying hierarchical cluster analysis (HCA) and principal component analysis (PCA). A clear separation of the experimental groups was obtained with high heterogeneity in the lipid and protein regions. These chemometric analyses showed that the low and medium doses of selenium-treated diabetic groups are successfully segregated from the diabetic group and clustered closer to the control. The study suggests that medium and, more predominantly, low-dose selenium treatment can be efficient in eliminating diabetes-induced structural alterations. PMID:26850735

  4. Exploring the Midgut Transcriptome and Brush Border Membrane Vesicle Proteome of the Rice Stem Borer, Chilo suppressalis (Walker)

    PubMed Central

    Peng, Chuanhua; Wang, Xiaoping; Li, Fei; Lin, Yongjun

    2012-01-01

    The rice stem borer, Chilo suppressalis (Walker) (Lepidoptera: Pyralidae), is one of the most detrimental pests affecting rice crops. The use of Bacillus thuringiensis (Bt) toxins has been explored as a means to control this pest, but the potential for C. suppressalis to develop resistance to Bt toxins makes this approach problematic. Few C. suppressalis gene sequences are known, which makes in-depth study of gene function difficult. Herein, we sequenced the midgut transcriptome of the rice stem borer. In total, 37,040 contigs were obtained, with a mean size of 497 bp. As expected, the transcripts of C. suppressalis shared high similarity with arthropod genes. Gene ontology and KEGG analysis were used to classify the gene functions in C. suppressalis. Using the midgut transcriptome data, we conducted a proteome analysis to identify proteins expressed abundantly in the brush border membrane vesicles (BBMV). Of the 100 top abundant proteins that were excised and subjected to mass spectrometry analysis, 74 share high similarity with known proteins. Among these proteins, Western blot analysis showed that Aminopeptidase N and EH domain-containing protein have the binding activities with Bt-toxin Cry1Ac. These data provide invaluable information about the gene sequences of C. suppressalis and the proteins that bind with Cry1Ac. PMID:22666467

  5. Myosin Ia is required for CFTR brush border membrane trafficking and ion transport in the mouse small intestine.

    PubMed

    Kravtsov, Dmitri V; Caputo, Christina; Collaco, Anne; Hoekstra, Nadia; Egan, Marie E; Mooseker, Mark S; Ameen, Nadia A

    2012-08-01

    In enterocytes of the small intestine, endocytic trafficking of CFTR channels from the brush border membrane (BBM) to the subapical endosomes requires the minus-end motor, myosin VI (Myo6). The subapical localization of Myo6 is dependent on myosin Ia (Myo1a) the major plus-end motor associated with the BBM, suggestive of functional synergy between these two motors. In villus enterocytes of the Myo1a KO mouse small intestine, CFTR accumulated in syntaxin-3 positive subapical endosomes, redistributed to the basolateral domain and was absent from the BBM. In colon, where villi are absent and Myo1a expression is low, CFTR exhibited normal localization to the BBM in the Myo1a KO similar to WT. cAMP-stimulated CFTR anion transport in the small intestine was reduced by 58% in the KO, while anion transport in the colon was comparable to WT. Co-immunoprecipitation confirmed the association of CFTR with Myo1a. These data indicate that Myo1a is an important regulator of CFTR traffic and anion transport in the BBM of villus enterocytes and suggest that Myo1a may power apical CFTR movement into the BBM from subapical endosomes. Alternatively, it may anchor CFTR channels in the BBM of villus enterocytes as was proposed for Myo1a's role in BBM localization of sucrase-isomaltase. PMID:22510086

  6. Binding kinetics of Clostridium difficile toxins A and B to intestinal brush border membranes from infant and adult hamsters

    SciTech Connect

    Rolfe, R.D. )

    1991-04-01

    This study was undertaken to determine if the relative resistance of neonates and infants to Clostridium difficile-associated intestinal disease can be related to age-dependent differences in intestinal receptors for C. difficile toxins A and B. Brush border membranes (BBMs) from the small intestines of adult and infant hamsters were examined for their ability to bind radiolabeled toxins A and B. (125I)toxin A bound to both infant and adult hamster BBMs at physiological temperature, whereas (125I)toxin B did not bind to the BBMs under any of the conditions examined. The number of (125I)toxin A molecules bound at saturation was approximately 4 x 10(10) per micrograms of membrane protein for adult BBMs and 1 x 10(11) per micrograms of membrane protein for infant BBMs. Scatchard plot analysis suggested the presence of a single class of toxin A binding sites on both infant and adult hamster BBMs. Maximal binding capacity and Kd values were 0.63 pmol/mg of protein and 66.7 nM, respectively, for the infant BBMs, and 0.24 pmol/mg of protein and 27 nM, respectively, for the adult BBMs. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analyses of extracted BBM proteins revealed differences in the proteins of infant and adult BBMs. However, there were not any detectable differences in the protein bands which bound (125I)toxin A between infant and adult hamsters. The results from these investigations indicate that differences in the binding kinetics of toxins A and/or B to infant and adult hamster BBMs do not account for the observed differences in their susceptibility to C. difficile-associated intestinal disease.

  7. Psychological stress impairs Na+-dependent glucose absorption and increases GLUT2 expression in the rat jejunal brush-border membrane.

    PubMed

    Boudry, Gaëlle; Cheeseman, Christopher I; Perdue, Mary H

    2007-02-01

    Chronic psychological stress impacts many functions of the gastrointestinal tract. However, the effect of stress on nutrient absorption is poorly documented. This study was designed to investigate glucose transporters in rats submitted to different periods of water-avoidance stress (WAS). Rats were subjected to WAS (1 h/day) for 1, 5, or 10 consecutive days. Four hours after the last WAS session, rats were killed and segments of jejunum were mounted in Ussing chambers to study electrophysiological properties of the jejunum and Na+-dependent glucose absorption kinetics. Mucosa was obtained to prepare brush-border membrane vesicles (BBMV) used to measure [14C]fructose uptake as well as sodium-glucose transporter 1 (SGLT-1) and GLUT2 expression by Western blot analysis. Exposure of animals to WAS induced a decrease in Na+-dependent glucose absorption Vmax after 1, 5, and 10 days without any change in SGLT-1 expression. Potential difference across the jejunum was decreased for all stressed groups. Furthermore, we observed an increase in phloretin-sensitive uptake of [14C]fructose by BBMV after 1, 5, or 10 days of WAS, which was not present in control animals. This suggested the abnormal appearance of GLUT2 in the brush border, which was confirmed by Western blot analysis. We concluded that psychological stress induces major changes in glucose transport with a decrease in Na+-dependent glucose absorption and an increase in GLUT2 expression at the brush-border membrane level. PMID:17053095

  8. Membrane insertion of the Bacillus thuringiensis Cry1Ab toxin: single mutation in domain II block partitioning of the toxin into the brush border membrane.

    PubMed

    Nair, Manoj S; Liu, Xinyan Sylvia; Dean, Donald H

    2008-05-27

    The umbrella and penknife models hypothesize that insecticidal Bacillus thuringiensis Cry toxins partition into the apical membrane of the insect midgut by insertion of only two alpha-helices from domain I of the protein, alpha-helices 4 and 5 in the case of the umbrella model and alpha-helices 5 and 6 in the case of the penknife model. Neither model envisages membrane partitioning by domains II and III. In this study, we present data suggesting that mutations in the domain II residue, F371, affect insertion of the whole toxin into Manduca sexta brush border membrane vesicles (BBMVs). Using steady state fluorescence measurements combined with a proteinase K protection assay, we show that mutants of F371 have lost their ability to insert into the BBMV, even though binding to cadherin is almost unaffected. The study also identifies a difference in partitioning of toxins into artificial lipid vesicles (SUVs) as opposed to native BBMVs. While the F371 mutations block insertion of domains I and II into BBMVs, they only block domain II insertion into SUVs. Bioassay and voltage clamping of midguts also confirm the fluorescence data that the noninserting mutants are nontoxic. Our study leads us to propose that, in contrast to previous models of individual free helices inserting into the membrane, the toxin enters into the membrane as a whole molecule or oligomers of the molecule, wherein the domain II residue F371 has a vital role to play in membrane insertion. PMID:18457427

  9. Angiotensin 2 directly increases rabbit renal brush-border membrane sodium transport: Presence of local signal transduction system

    SciTech Connect

    Morduchowicz, G.A.; Sheikh-Hamad, D.; Dwyer, B.E.; Stern, N.; Jo, O.D.; Yanagawa, N. )

    1991-05-01

    In the present study, the authors have examined the direct actions of angiotensin II (AII) in rabbit renal brush border membrane (BBM) where binding sites for AII exist. Addition of AII (10(-11)-10(-7) M) was found to stimulate 22Na+ uptake by the isolated BBM vesicles directly. All did not affect the Na(+)-dependent BBM glucose uptake, and the effect of AII on BBM 22Na+ uptake was inhibited by amiloride, suggesting the involvement of Na+/H+ exchange mechanism. BBM proton permeability as assessed by acridine orange quenching was not affected by AII, indicating the direct effect of AII on Na+/H+ antiport system. In search of the signal transduction mechanism, it was found that AII activated BBM phospholipase A2 (PLA) and that BBM contains a 42-kDa guanine nucleotide-binding regulatory protein (G-protein) that underwent pertussis toxin (PTX)-catalyzed ADP-ribosylation. Addition of GTP potentiated, while GDP-beta S or PTX abolished, the effects of AII on BBM PLA and 22Na+ uptake, suggesting the involvement of G-protein in AII's actions. On the other hand, inhibition of PLA by mepacrine prevented AII's effect on BBM 22Na+ uptake, and activation of PLA by mellitin or addition of arachidonic acid similarly enhanced BBM 22Na+ uptake, suggesting the role of PLA activation in mediating AII's effect on BBM 22Na+ uptake. In summary, results of the present study show a direct stimulatory effect of AII on BBM Na+/H+ antiport system, and suggest the presence of a local signal transduction system involving G-protein mediated PLA activation.

  10. Binding of isolectins from red kidney bean (Phaseolus vulgaris) to purified rat brush-border membranes.

    PubMed

    Boldt, D H; Banwell, J G

    1985-12-13

    Ingestion of red kidney bean phytohemagglutinin causes impaired growth and intestinal malabsorption, and facilitates bacterial colonization in the small intestine of weanling rats. We have studied interactions of the highly purified phytohemagglutinin erythroagglutinating (E4) and mitogenic (L4) isolectins with microvillous membrane vesicles prepared from rat small intestines. E4 and L4 were radioiodinated with 125I by the chloramine-T technique. E4 and L4 isolectins both bound to microvillous membrane vesicles. Binding was saturable and reversible. Each mg of membrane protein bound 744 +/- 86 micrograms E4 and 213 +/- 21 micrograms L4. The apparent Ka for E4 and L4 binding was 2.5 x 10(-6) and 13.0 x 10(-6) M-1, respectively. Binding of each 125I-labelled isolectin was abolished by 100-fold excess of unlabelled isolectin. In each case binding also was inhibited by appropriate oligosaccharide inhibitors, indicating that isolectin-microvillous membrane interactions were mediated by carbohydrate recognition. Patterns of saccharide inhibition of isolectin binding were different for E4 and L4. Competitive binding experiments demonstrated mutual noncompetitive inhibition of E4 and L4 binding consistent with steric hindrance. Therefore, E4 and L4 each bound to its own set of receptors. Based on the known saccharide specificities of E4 and L4, these data indicate that there are differences in expression of complex asparagine-linked biantennary and tri- or tetraantennary oligosaccharides at the microvillous surface. The data also provide the possibility that direct interactions of one or more phytohemagglutinin isolectins with intestinal mucosa in vivo may contribute to the antinutritional effects associated with ingestion of crude red kidney beans. PMID:4063394

  11. Hepatocyte growth factor activates phosphoinositide 3-kinase C2 beta in renal brush-border plasma membranes.

    PubMed Central

    Crljen, Vladiana; Volinia, Stefano; Banfic, Hrvoje

    2002-01-01

    Upon stimulation of renal cortical slices with hepatocyte growth factor (HGF), inositol lipid metabolism was studied in basal-lateral plasma membranes (BLM) and brush-border plasma membranes (BBM). Whereas in BLM rapid increases in 1,2-diacylglycerol, PtdIns(3,4,5)P(3) and PtdIns(3,4)P(2) were observed, suggesting that in BLM HGF activates both phospholipase C (PLC) and phosphoinositide 3-kinase (PI3K), in BBM only HGF-induced transient accumulation of PtdIns3P was seen, which was temporarily delayed from signalling events in BLM and could be blocked by the PtdIns-specific-PLC inhibitor ET-18-OCH(3) and the calpain inhibitor calpeptin, suggesting that 3-kinase activation in BBM lies downstream of PLC activation in BLM and is a calpain-mediated event. Moreover, the increase in immunoprecipitable PI3K-C2 beta activity, which is sensitive to wortmannin (10 nM) and shows strong preference for PtdIns over PtdIns4P as a substrate, was observed only in BBM upon stimulation of renal cortical slices with HGF and could be mimicked by the Ca(2+) ionophore A23187 and blocked by the cell-penetrant Ca(2+) chelator BAPTA-AM [1,2-bis-(2-aminophenoxy)ethane-N,N,N',N'-tetra-acetic acid tetrakis(acetoxymethyl ester)]. On Western blots PI3K-C2 beta revealed a single immunoreactive band of 180 kDa in BLM and BBM, while after stimulation with HGF a gel shift of 18 kDa was noticed only in BBM, suggesting that the observed enzyme activation is achieved by proteolysis. When BBM were subjected to short-term (15 min) exposure to mu-calpain, a similar gel shift together with an increase in PI3K-C2 beta activity was observed, when compared with the BBM harvested after HGF stimulation. The above-mentioned gel shift and increase in PI3K-C2 beta activity could be prevented by the calpain inhibitor calpeptin. The data presented in this report show that in renal cells there is a spatial separation of the inositol lipid signalling system between BLM and BBM, and that HGF causes activation of PLC and

  12. Heterogeneity of L-alanine transport systems in brush-border membrane vesicles from rat placenta during late gestation.

    PubMed Central

    Alonso-Torre, S R; Serrano, M A; Medina, J M; Alvarado, F

    1992-01-01

    The placental uptake of L-alanine was studied by using purified brush-border membrane vesicles from rat trophoblasts. Saturation curves were carried out at 37 degrees C in buffers containing 100 mM (zero-trans)-NaSCN, -NaCl, -KSCN, -KCl, or -N-methyl-D-glucamine gluconate. The uncorrected uptake results were fitted by non-linear regression analysis to an equation involving one diffusional component either one or two saturable Michaelian transport terms. In the presence of NaCl, two distinct L-alanine transport systems were distinguished, named respectively System 1 (S-1; Vm1 about 760 pmol/s per mg of protein; KT1 = 0.5 mM) and System 2 (S-2; Vm2 about 1700 pmol/s per mg; KT2 = 9 mM). In contrast, in the presence of K+ (KCl = KSCN) or in the absence of any alkali-metal ions (N-methyl-D-glucamine gluconate), only one saturable system was apparent, which we identify as S-2. When Na+ is present, S-1, but not S-2, appears to be rheogenic, since its maximal transport capacity significantly increases in the presence of an inside-negative membrane potential, created either by replacing Cl- with the permeant anion thiocyanate (NaSCN > NaCl) or by applying an appropriate K+ gradient and valinomycin. alpha-(Methylamino)isobutyrate (methyl-AIB) appears to be a substrate of S-1, but not of S-2. For reasons that remain to be explained, however, methyl-AIB inhibits S-2. We conclude that S-1 represents a truly Na(+)-dependent mechanism, where Na+ behaves as an obligatory activator, whereas S-2 cannot discriminate between Na+ and K+, although its activity is higher in the presence of alkali-metal ions than in their absence (Na+ = K+ > N-methyl-D-glucammonium ion). S-2 appears to be fully developed 2 days before birth, whereas S-1 undergoes a capacity-type activation between days 19.5 and 21.5 of gestation, i.e. its apparent Vmax. nearly doubles, whereas its KT remains constant. PMID:1445280

  13. Isolation and characterization of rabbit kidney brush borders

    PubMed Central

    Quirk, S. J.; Robinson, G. B.

    1972-01-01

    1. Brush borders were isolated from rabbit kidney-cortex homogenates by rate-zonal centrifugation through a sucrose density gradient in a B-XIV zonal rotor, followed by differential centrifugation. 2. The method of preparation gave brush borders of high purity with a reasonable yield. The morphological appearance supported the evidence from enzymic and chemical investigations, that the brush borders were only slightly contaminated with endoplasmic reticulum, mitochondria, lysosomes and nuclei. 3. The molar ratio of cholesterol to phospholipid lay within the range found in other plasma membranes, but the carbohydrate content was double that found in liver plasma membranes. 4. Alkaline phosphatase, maltase, trehalase and aminopeptidase were major enzymic constituents of the brush borders, and had an approximately equal yield and enrichment, but none of these enzymes fulfilled the criteria for marker enzymes. 5. Mg2+-dependent and Na+,K+-dependent adenosine triphosphatases, although found in brush borders, had low yields and low enrichments. ImagesPLATE 2PLATE 1 PMID:4264701

  14. Transport of L-leucine hydroxy analogue and L-lactate in rabbit small-intestinal brush-border membrane vesicles.

    PubMed

    Friedrich, M; Murer, H; Berger, E G

    1991-05-01

    Substitution of the alpha-amino group of amino acids by hydroxyl groups yields hydroxy analogues (HA), which have been ascribed beneficial effects in nitrogen-sparing diets for uremic patients. In this study, intestinal uptake of L-leucine HA (L-LeuHA) and L-lactate into rabbit jejunal brush-border membrane vesicles was investigated. An inward-directed H+ or Na+ gradient stimulated uptake of both labelled substrates in a voltage-clamped assay. The H+ gradient was the major driving force of uptake as compared with the Na+ gradient, and it led to a transient accumulation of both L-LeuHA and L-lactate. The proton ionophore carbonylcyanide p-trifluoromethoxyphenylhydrazone (FCCP) reduced the initial H(+)-gradient-driven uptake rates of both substrates, but was without effect on Na(+)-gradient-driven uptakes. The H(+)-gradient-driven L-LeuHA uptake was saturable (apparent Kt = 15.4 mM). Alpha-HA of L-leucine, L-isoleucine, L-valine, D-leucine, D-valine or L-lactate inhibited the H(+)-gradient-driven L-LeuHA or L-lactate uptakes whereas free branched-chain amino acids had no effect. Preloading the vesicles with one of the L- or D-HA of branched-chain amino acids or with L-lactate stimulated tracer L-LeuHA and also tracer L-lactate uptakes in the presence of a H+ gradient. It is concluded that H(+)-gradient-driven transport of L- and D-stereoisomeric HA of branched-chain amino acids as well as of L-lactate across rabbit intestinal brush-border membranes is mediated by the same carrier. Furthermore, there exists a Na+ gradient-driven L-lactate transport system in the rabbit intestinal brush-border membrane. PMID:1876483

  15. Dexamethasone modulates rat renal brush border membrane phosphate transporter mRNA and protein abundance and glycosphingolipid composition.

    PubMed Central

    Levi, M; Shayman, J A; Abe, A; Gross, S K; McCluer, R H; Biber, J; Murer, H; Lötscher, M; Cronin, R E

    1995-01-01

    Glucocorticoids are important regulators of renal phosphate transport. This study investigates the role of alterations in renal brush border membrane (BBM) sodium gradient-dependent phosphate transport (Na-Pi cotransporter) mRNA and protein abundance in the dexamethasone induced inhibition of Na-Pi cotransport in the rat. Dexamethasone administration for 4 d caused a 1.5-fold increase in the Vmax of Na-Pi cotransport (1785 +/- 119 vs. 2759 +/- 375 pmol/5 s per mg BBM protein in control, P < 0.01), which was paralleled by a 2.5-fold decrease in the abundance of Na-Pi mRNA and Na-Pi protein. There was also a 1.7-fold increase in BBM glucosylceramide content (528 +/- 63 vs. 312 +/- 41 ng/mg BBM protein in control, P < 0.02). To determine whether the alteration in glucosylceramide content per se played a functional role in the decrease in Na-Pi cotransport, control rats were treated with the glucosylceramide synthase inhibitor, D-threo-1-phenyl-2-decanoyl-amino-3-morpholino-1-propanol (PDMP). The resultant 1.5-fold decrease in BBM glucosylceramide content (199 +/- 19 vs. 312 +/- 41 ng/mg BBM protein in control, P < 0.02) was associated with a 1.4-fold increase in Na-Pi cotransport activity (1422 +/- 73 vs. 1048 +/- 85 pmol/5 s per mg BBM protein in control, P < 0.01), and a 1.5-fold increase in BBM Na-Pi protein abundance. Thus, dexamethasone-induced inhibition of Na-Pi cotransport is associated with a decrease in BBM Na-Pi cotransporter abundance, and an increase in glucosylceramide. Since primary alteration in BBM glucosylceramide content per se directly and selectively modulates BBM Na-Pi cotransport activity and Na-Pi protein abundance, we propose that the increase in BBM glucosylceramide content plays an important role in mediating the inhibitory effect of dexamethasone on Na-Pi cotransport activity. Images PMID:7615789

  16. Staphylococcus aureus enterotoxins A- and B: binding to the enterocyte brush border and uptake by perturbation of the apical endocytic membrane traffic.

    PubMed

    Danielsen, E Michael; Hansen, Gert H; Karlsdóttir, Edda

    2013-04-01

    Enterotoxins of Staphylococcus aureus are among the most common causes of food poisoning. Acting as superantigens they intoxicate the organism by causing a massive uncontrolled T cell activation that ultimately may lead to toxic shock and death. In contrast to our detailed knowledge regarding their interaction with the immune system, little is known about how they penetrate the epithelial barrier to gain access to their targets. We therefore studied the uptake of two staphylococcal enterotoxins (SEs), SEA and SEB, using organ cultured porcine jejunal explants as model system. Attachment of both toxins to the villus surface was scarce and patchy compared with that of cholera toxin B (CTB). SEA and SEB also bound to microvillus membrane vesicles in vitro, but less efficiently than CTB, and the binding was sensitive to treatment with endoglycoceramidase II, indicating that a glycolipid, possibly digalactosylceramide, acts as cell surface receptor at the brush border. Both SEs partitioned poorly with detergent resistant membranes (DRMs) of the microvillus, suggesting a weak association with lipid raft microdomains. Where attachment occurred, cellular uptake of SEA and SEB was also observed. In enterocytes, constitutive apical endocytosis normally proceeds only to subapical early endosomes present in the actomyosin-rich "terminal web" region. But, like CTB, both SEA and SEB penetrated deep into the cytoplasm. In conclusion, the data show that after binding to the enterocyte brush border SEA and SEB perturb the apical membrane trafficking, enabling them to engage in transcytosis to reach their target cells in the subepithelial lamina propria. PMID:23180309

  17. Specificity of Bacillus thuringiensis endotoxins is correlated with the presence of high-affinity binding sites in the brush border membrane of target insect midguts

    SciTech Connect

    Hofmann, C.; Vanderbruggen, H.; Hoefte, H.; Van Rie, J.; Jansens, S.; Van Mellaert, H. )

    1988-11-01

    Binding studies were performed with two {sup 125}I-labeled Bacillus thuringiensis {delta}-endotoxins on brush border membrane vesicles prepared from the larval midgut of the tobacco hornworm Manduca sexta or the cabbage butterfly Pieris brassicae. One {delta}-endotoxin, Bt2-protoxin, is a 130-kDa recombinant crystalline protein from B. thuringiensis subsp. berliner. It kills larvae of both insect species. The active Bt2-toxin is a 60-kDa proteolytic fragment of the Bt2-protoxin. It binds saturably and with high affinity to brush border membrane vesicles from the midgut of both species. The other {delta}-endotoxin, Bt4412-protoxin, is a 136-kDa crystalline protein from B. thuringiensis subsp. thuringiensis, which is highly toxic for P. brassicae, but not for M. sexta larvae. Bt4412-toxin, obtained after proteolytic activation of Bt4412-protoxin, shows high-affinity saturable binding to P. brassicae vesicles but not to M. sexta vesicles. The correlation between toxicity and specific binding is further strengthened by competition studies. Other B. thuringiensis {delta}-endotoxins active against M. sexta compete for binding of {sup 125}I-labeled Bt2-toxin to M. sexta vesicles, whereas toxins active against dipteran or coleopteran larvae do not compete. Bt2-toxin and Bt4412-toxin bind to different sites on P. brassicae vesicles.

  18. Competitive, uncompetitive, and mixed inhibitors of the alkaline phosphatase activity associated with the isolated brush border membrane of the tapeworm Hymenolepis diminuta.

    PubMed

    Pappas, P W; Leiby, D A

    1989-06-01

    Several compounds were tested as inhibitors of the alkaline phosphatase (AlkPase) activity associated with the isolated brush border membrane of the tapeworm, Hymenolepis diminuta. Molybdate, arsenate, arsenite and beta-glycerophosphate (BGP) were competitive inhibitors of the hydrolysis of p-nitrophenyl phosphate, while levamisole and clorsulon were uncompetitive and mixed inhibitors, respectively. Molybdate was also a competitive inhibitor of the hydrolysis of BGP and 5'-adenosine monophosphate, and levamisole was an uncompetitive inhibitor of BGP hydrolysis. The apparent inhibitor constants (Ki') for molybdate and levamisole were virtually identical regardless of the substrate, and these data support the hypothesis that the AlkPase activity is represented by a single membrane-bound enzyme with low substrate specificity. Quinacrine, Hg2+, and ethylenediaminetetraacetate were also potent inhibitors of the AlkPase activity, but the mechanisms by which these latter three inhibitors function were not clear. PMID:2768348

  19. Receptors on the brush border membrane of the insect midgut as determinants of the specificity of Bacillus thuringiensis delta-endotoxins.

    PubMed Central

    Van Rie, J; Jansens, S; Höfte, H; Degheele, D; Van Mellaert, H

    1990-01-01

    To investigate the biochemical basis of the differences in the insecticidal spectrum of Bacillus thuringiensis insecticidal crystal proteins (ICPs), we performed membrane binding and toxicity assays with three different ICPs and three lepidopteran species. The three ICPs have different toxicity patterns in the three selected target species. Binding studies with these 125I-labeled ICPs revealed high-affinity saturable binding to brush border membrane vesicles of the sensitive species. ICPs with no toxicity against a given species did not bind saturably to vesicles of that species. Together with previous data that showed a correlation between toxicity and ICP binding, our data support the statement that differences in midgut ICP receptors are a major determinant of differences in the insecticidal spectrum of the entire lepidopteran-specific ICP family. Receptor site heterogeneity in the insect midgut occurs frequently and results in sensitivity to more than one type of ICP. Images PMID:2339890

  20. Dietary protein reduction in sheep and goats: different effects on L-alanine and L-leucine transport across the brush-border membrane of jejunal enterocytes.

    PubMed

    Schröder, B; Schöneberger, M; Rodehutscord, M; Pfeffer, E; Breves, G

    2003-08-01

    It was the aim of this study to examine the potential regulatory effects of a long-term low dietary protein supply on the transport capacity of the jejunal brush-border membrane for amino acids. For this purpose, we used the neutral amino acids L-alanine (representative for nonessential amino acids) and L-leucine (representative for essential amino acids) as model substances. Ten sheep lambs, 8 weeks of age and 19-27 kg body weight, were allotted to two dietary regimes with either adequate or reduced protein supply which was achieved by 17.9% and 9.7% of crude protein in the concentrated feed, respectively. The feeding periods were 4-6 weeks in length. Similarly, eight goat kids of 5-7 weeks of age and 8-14 kg body weight were allotted to either adequate (crude protein 20.1%, feeding period 9-12 weeks) or reduced protein supply (10.1%, feeding period 17-18 weeks). Dietary protein reduction in lambs caused a significant body weight loss of 0.6 +/- 0.7 kg, whereas the body weight in control animals increased by 1.9 +/- 0.7 kg (P<0.05). Plasma urea concentrations decreased significantly by 60% (low protein 2.3 +/- 0.1 versus control 5.7 +/- 0.2 mmol l(-1), P<0.001). In kids, reduction of dietary protein intake led to significant decreases of the daily weight gain by 48% from 181 +/- 8 g to 94 +/- 3 g (P<0.001) and daily dry matter intake by 27% from 568 +/- 13 g to 417 +/- 6 g (P<0.01). Respective urea concentrations in plasma were reduced by 77% from 5.2 +/- 0.4 to 1.2 +/- 0.2 mmol l(-1) (P<0.01). Kinetic analyses of the initial rates of alanine uptake into isolated jejunal brush-border membrane vesicles from sheep and goats as affected by low dietary protein supply yielded that the apparent Km was neither significantly different between the species nor significantly affected by the feeding regime thus ranging between 0.12 and 0.16 mmol.l(-1). Reduction of dietary protein, however, resulted in significantly decreased Vmax values of the transport system by 25

  1. Brush border membrane vesicle and Caco-2 cell line: Two experimental models for evaluation of absorption enhancing effects of saponins, bile salts, and some synthetic surfactants

    PubMed Central

    Moghimipour, Eskandar; Tabassi, Sayyed Abolghassem Sajadi; Ramezani, Mohammad; Handali, Somayeh; Löbenberg, Raimar

    2016-01-01

    The aim of this study was to investigate the influence of absorption enhancers in the uptake of hydrophilic compounds. The permeation of the two hydrophilic drug models gentamicin and 5 (6)-carboxyfluorescein (CF) across the brush border membrane vesicles and Caco-2 cell lines were evaluated using total saponins of Acanthophyllum squarrosum, Quillaja saponaria, sodium lauryl sulfate, sodium glycocholate, sodium taurodeoxycholate, and Tween 20 as absorption enhancers. Transepithelial electrical resistance (TEER) measurement was utilized to assess the paracellular permeability of cell lines. Confocal laser scanning microscopy (CLSM) was performed to obtain images of the distribution of CF in Caco-2 cells. These compounds were able to loosen tight junctions, thus increasing paracellular permeability. CLSM confirmed the effect of these absorption enhancers on CF transport across Caco-2 lines and increased the Caco-2 permeability via transcellular route. It was also confirmed that the decrease in TEER was transient and reversible after removal of permeation enhancers. PMID:27429925

  2. Brush border membrane vesicle and Caco-2 cell line: Two experimental models for evaluation of absorption enhancing effects of saponins, bile salts, and some synthetic surfactants.

    PubMed

    Moghimipour, Eskandar; Tabassi, Sayyed Abolghassem Sajadi; Ramezani, Mohammad; Handali, Somayeh; Löbenberg, Raimar

    2016-01-01

    The aim of this study was to investigate the influence of absorption enhancers in the uptake of hydrophilic compounds. The permeation of the two hydrophilic drug models gentamicin and 5 (6)-carboxyfluorescein (CF) across the brush border membrane vesicles and Caco-2 cell lines were evaluated using total saponins of Acanthophyllum squarrosum, Quillaja saponaria, sodium lauryl sulfate, sodium glycocholate, sodium taurodeoxycholate, and Tween 20 as absorption enhancers. Transepithelial electrical resistance (TEER) measurement was utilized to assess the paracellular permeability of cell lines. Confocal laser scanning microscopy (CLSM) was performed to obtain images of the distribution of CF in Caco-2 cells. These compounds were able to loosen tight junctions, thus increasing paracellular permeability. CLSM confirmed the effect of these absorption enhancers on CF transport across Caco-2 lines and increased the Caco-2 permeability via transcellular route. It was also confirmed that the decrease in TEER was transient and reversible after removal of permeation enhancers. PMID:27429925

  3. Performance, organ zinc concentration, jejunal brush border membrane enzyme activities and mRNA expression in piglets fed with different levels of dietary zinc.

    PubMed

    Martin, Lena; Pieper, Robert; Schunter, Nadine; Vahjen, Wilfried; Zentek, Jürgen

    2013-06-01

    This study aimed at investigating the effect of dietary zinc on performance, jejunal brush border membrane enzyme activities and mRNA levels of enzymes and two zinc transporters in piglets. A total of 126 piglets were weaned at 26 ±1 days of age and randomly allocated into three groups fed with diets 50, 150 and 2500 mg zinc/kg. Performance was recorded and at weekly intervals, eight piglets per group were killed. The activities of isolated brush border membrane enzymes including lactase, maltase, sucrase, aminopeptidase-N and intestinal alkaline phosphatase (IAP), and the relative transcript abundance of aminopeptidase-N (APN), sucrase-isomaltase (SUC), IAP and the two zinc transporters SLC39A4 (ZIP4) and SLC30A1 (ZnT1) were investigated in the jejunum. Feeding pharmacological zinc levels increased weight gain (p < 0.001) during the first week, but performance was lower (p < 0.05) in the third week. Organ zinc concentrations were increased by high dietary zinc level. The activity of IAP was higher (p < 0.05) with the highest dietary zinc level, no effects were determined for other enzymes. Dietary zinc level had no effect on transcript abundance of digestive enzymes. The mRNA levels decreased (p < 0.001) for ZIP4, and increased for ZnT1 (p < 0.05) with pharmacological zinc levels. In conclusion, pharmacological zinc levels improved performance in the short-term. Intestinal mRNA level of zinc transporters changed with high zinc supply, but this did not prevent zinc accumulation in tissues, suggesting hampered homoeostatic regulation. This might cause impaired performance during longer supply. PMID:23742645

  4. Oxalate transport by anion exchange across rabbit ileal brush border.

    PubMed Central

    Knickelbein, R G; Aronson, P S; Dobbins, J W

    1986-01-01

    This study demonstrates the presence of oxalate transporters on the brush border membrane of rabbit ileum. We found that an inside alkaline (pH = 8.5 inside, 6.5 outside) pH gradient stimulated [14C]oxalate uptake 10-fold at 1 min with a fourfold accumulation above equilibrated uptake at 5 min. 1 mM 4,4'-diisothiocyanostilbene-2,2'-disulfonate (disodium salt; DIDS) profoundly inhibited the pH-gradient stimulated oxalate uptake. Using an inwardly directed K+ gradient and valinomycin, we found no evidence for potential sensitive oxalate uptake. In contrast to Cl:HCO3 exchange, HCO3 did not stimulate oxalate uptake more than was seen with a pH gradient in the absence of HCO3. An outwardly directed Cl gradient (50 mM inside, 5 mM outside) stimulated oxalate uptake 10-fold at 1 min with a fivefold accumulation above equilibrated uptake. Cl-stimulated oxalate uptake was largely inhibited by DIDS. Addition of K+ and nigericin only slightly decreased the Cl gradient-stimulated oxalate uptake, which indicates that this stimulation was not primarily due to the Cl gradient generating an inside alkaline pH gradient via Cl:OH exchange. Further, an outwardly directed oxalate gradient stimulated 36Cl uptake. These results suggested that both oxalate:OH and oxalate:Cl exchange occur on the brush border membrane. To determine if one or both of these exchanges were on contaminating basolateral membrane, the vesicle preparation was further fractionated into a brush border and basolateral component using sucrose density gradient centrifugation. Both exchangers localized to the brush border component. A number of organic anions were examined (outwardly directed gradient) to determine if they could stimulate oxalate and Cl uptake. Only formate and oxaloacetate were found to stimulate oxalate and Cl uptake. An inwardly directed Na gradient only slightly stimulated oxalate uptake, which was inhibited by DIDS. PMID:3003149

  5. Glucose and fructose uptake by Limulus polyphemus hepatopancreatic brush border and basolateral membrane vesicles: evidence for Na+-dependent sugar transport activity.

    PubMed

    Sterling, Kenneth M; Ahearn, Gregory A

    2011-05-01

    [(3)H]-fructose and [(3)H]-glucose transport activities were determined in brush border membrane vesicles (BBMV) and basolateral membrane vesicles (BLMV) from Limulus polyphemus (horseshoe crab) hepatopancreas. Glucose transport was equilibrative in the absence of sodium and sodium dependent in the presence of sodium in BBMV, suggesting GLUT-like and SGLT-like transport activity. Glucose transport by BLMV was equilibrative and sodium independent. Fructose uptake by BBMV and BLMV was equilibrative in the absence of sodium and sodium dependent in the presence of sodium. Western blot analysis using a rabbit anti-mouse SGLT-1 polyclonal antibody indicated the presence of a cross-reacting horseshoe crab BBMV protein of similar molecular weight to the mammalian SGLT1. Sequence alignment of the mouse SGLT-4 and SGLT1 with a translated, horseshoe crab-expressed sequence tag also indicated significant identity between species. Fructose and glucose uptake in the absence and presence of sodium by hepatopancreas BBMV and BLMV indicated the presence of sodium-dependent transport activity for each sugar that may result from the presence of transporters similar to those described for other species. PMID:21184084

  6. Experimental type II diabetes and related models of impaired glucose metabolism differentially regulate glucose transporters at the proximal tubule brush border membrane.

    PubMed

    Chichger, Havovi; Cleasby, Mark E; Srai, Surjit K; Unwin, Robert J; Debnam, Edward S; Marks, Joanne

    2016-06-01

    What is the central question of this study? Although SGLT2 inhibitors represent a promising treatment for patients suffering from diabetic nephropathy, the influence of metabolic disruption on the expression and function of glucose transporters is largely unknown. What is the main finding and its importance? In vivo models of metabolic disruption (Goto-Kakizaki type II diabetic rat and junk-food diet) demonstrate increased expression of SGLT1, SGLT2 and GLUT2 in the proximal tubule brush border. In the type II diabetic model, this is accompanied by increased SGLT- and GLUT-mediated glucose uptake. A fasted model of metabolic disruption (high-fat diet) demonstrated increased GLUT2 expression only. The differential alterations of glucose transporters in response to varying metabolic stress offer insight into the therapeutic value of inhibitors. SGLT2 inhibitors are now in clinical use to reduce hyperglycaemia in type II diabetes. However, renal glucose reabsorption across the brush border membrane (BBM) is not completely understood in diabetes. Increased consumption of a Western diet is strongly linked to type II diabetes. This study aimed to investigate the adaptations that occur in renal glucose transporters in response to experimental models of diet-induced insulin resistance. The study used Goto-Kakizaki type II diabetic rats and normal rats rendered insulin resistant using junk-food or high-fat diets. Levels of protein kinase C-βI (PKC-βI), GLUT2, SGLT1 and SGLT2 were determined by Western blotting of purified renal BBM. GLUT- and SGLT-mediated d-[(3) H]glucose uptake by BBM vesicles was measured in the presence and absence of the SGLT inhibitor phlorizin. GLUT- and SGLT-mediated glucose transport was elevated in type II diabetic rats, accompanied by increased expression of GLUT2, its upstream regulator PKC-βI and SGLT1 protein. Junk-food and high-fat diet feeding also caused higher membrane expression of GLUT2 and its upstream regulator PKC

  7. Studies on the effect of sodium arsenate on the enzymes of carbohydrate metabolism, brush border membrane, and oxidative stress in the rat kidney.

    PubMed

    Shahid, Faaiza; Rizwan, Sana; Khan, Md Wasim; Khan, Sara Anees; Naqshbandi, Ashreeb; Yusufi, Ahad Noor Khan

    2014-03-01

    Arsenic is an environmental pollutant and its contamination in drinking water poses serious world wide environmental health threats. It produces multiple adverse effects in various tissues, including the kidney. However, biochemical mechanism and renal response to its toxic insult are not completely elucidated. We hypothesized that sodium arsenate (ARS) induces oxidative stress and alters the structure and metabolic functions of kidney. Male Wistar rats were administered ARS (10 mg/kg body weight/day), intraperitoneally daily for 10 days. ARS administration increased blood urea nitrogen, serum creatinine, cholesterol, glucose, and phospholipids but decreased inorganic phosphate, indicating kidney toxicity. The activity of brush border membrane (BBM) enzymes significantly lowered in both cortex and medulla. Activity of hexokinase, lactate dehydrogenase, glucose-6-phosphate dehydrogenases, and NADP-malic enzyme significantly increased whereas malate dehydrogenase, glucose-6-phosphatase, and fructose 1,6 bis phosphatase decreased by ARS exposure. The activity of superoxide dismutase, GSH-peroxidase, and catalase were selectively altered in renal tissues along with an increase in lipid peroxidation. The present results indicated that ARS induced oxidative stress caused severe renal damage that resulted in altered levels of carbohydrate metabolism and BBM enzymes. PMID:24562057

  8. Characterization of a protein cofactor that mediates protein kinase A regulation of the renal brush border membrane Na(+)-H+ exchanger.

    PubMed Central

    Weinman, E J; Steplock, D; Wang, Y; Shenolikar, S

    1995-01-01

    Activation of cAMP-dependent protein kinase A inhibits the renal proximal tubule brush border membrane Na(+)-H+ exchanger by a process involving participation of a regulatory cofactor (NHE-RF) that is distinct from the transporter itself. Recent studies from this laboratory reported a partial amino acid sequence of this putative cofactor (Weinman, E. J., D. H. Steplock, and S. Shenolikar. 1993. J. Clin. Invest. 92:1781-1786). The present experiments detail the structure of the NHE-RF protein as determined from molecular cloning studies. A codon-biased oligonucleotide probe to a portion of the amino acid sequence of the putative cofactor was used to isolate a 1.9-kb cDNA from a rabbit renal library. The encoded protein is 358 amino acids in length and is rich in proline residues. Search of existing data bases indicates that NHE-RF is a unique protein. Using a reticulocyte lysate, the cDNA translated a product of approximately 44 kD, which was recognized by an affinity-purified polyclonal antibody to NHE-RF. Potential phosphorylation sites for protein kinase A are present. The mRNA for the protein is expressed in kidney, proximal small intestine, and liver. Reverse transcription/PCR studies in the kidney indicate the presence of mRNA for NHE-RF in several distinct nephron segments including the proximal tubule. Images PMID:7738182

  9. High affinity (3)H-phenylalanine uptake by brush border membrane vesicles from whole larvae of Aedes aegypti (AaBBMVw).

    PubMed

    Sterling, Kenneth M; Okech, Bernard A; Xiang, Minghui A; Linser, Paul J; Price, David A; Vanekeris, Leslie; Becnel, James J; Harvey, William R

    2012-04-01

    Brush border membrane vesicles from whole Aedes aegypti larvae (AaBBMVw) are confirmed to be valid preparations for membrane transport studies. The Abdul-Rauf and Ellar method was used to isolate AaBBMVw that were frozen, stored for several months, transported to a distant site, thawed and used to study Na(+)-coupled, (3)H-labeled, phenylalanine (Phe) uptake. The affinity for all components of the uptake was very high with half maximal values in the sub-micromolar range. By contrast a K(0.5)(Phe) of 0.2mM and a K(0.5)(Na) of 26 mM were calculated from Phe-induced electrical currents in Xenopus oocytes that were heterologously expressing the Anopheles gambiae symporter (co-transporter), AgNAT8, in a buffer with 98 mM Na(+). What accounts for the >1000-fold discrepancy in affinity for substrates between the BBMV and oocyte experiments? Is it because Ae. aegypti were used to isolate BBMVw whereas An. gambiae were used to transfect oocytes? More likely, it is because BBMVw were exposed to [Na(+)] in the micromolar range with the transporter(s) being surrounded by native lipids. By contrast, the oocyte measurements were made at [Na(+)] 100,000 times higher with AgNAT8 surrounded by foreign frog lipids. The results show that AaBBMVw are osmotically sealed; the time-course has a Na(+)-induced overshoot, the pH optimum is ∼7 and the K(0.5) values for Phe and Na(+) are very low. The transport is virtually unchanged when Na(+) is replaced by K(+) or Li(+) but decreased by Rb(+). This approach to resolving discrepancies between electrical data on solute transporters such as AgNAT8 that are over-expressed in oocytes and flux data on corresponding transporters that are highly expressed in native membrane vesicles, may serve as a model for similar studies that add membrane biochemistry to molecular biology in efforts to identify targets for the development of new methods to control disease-vector mosquitoes. PMID:22251673

  10. Villin and actin in the mouse kidney brush-border membrane bind to and are degraded by meprins, an interaction that contributes to injury in ischemia-reperfusion

    PubMed Central

    Ongeri, Elimelda Moige; Anyanwu, Odinaka; Reeves, W. Brian

    2011-01-01

    Meprins, metalloproteinases abundantly expressed in the brush-border membranes (BBMs) of rodent proximal kidney tubules, have been implicated in the pathology of renal injury induced by ischemia-reperfusion (IR). Disruption of the meprin β gene and actinonin, a meprin inhibitor, both decrease kidney injury resulting from IR. To date, the in vivo kidney substrates for meprins are unknown. The studies herein implicate villin and actin as meprin substrates. Villin and actin bind to the cytoplasmic tail of meprin β, and both meprin A and B are capable of degrading villin and actin present in kidney proteins as well as purified recombinant forms of these proteins. The products resulting from degradation of villin and actin were unique to each meprin isoform. The meprin B cleavage site in villin was Glu744-Val745. Recombinant forms of rat meprin B and homomeric mouse meprin A had Km values for villin and actin of ∼1 μM (0.6–1.2 μM). The kcat values varied substantially (0.6–128 s−1), resulting in different efficiencies for cleavage, with meprin B having the highest kcat/Km values (128 M−1·s−1 × 106). Following IR, meprins and villin redistributed from the BBM to the cytosol. A 37-kDa actin fragment was detected in protein fractions from wild-type, but not in comparable preparations from meprin knockout mice. The levels of the 37-kDa actin fragment were significantly higher in kidneys subjected to IR. The data establish that meprins interact with and cleave villin and actin, and these cytoskeletal proteins are substrates for meprins. PMID:21795642

  11. Functional Regulation of Sugar Assimilation by N-Glycan-specific Interaction of Pancreatic α-Amylase with Glycoproteins of Duodenal Brush Border Membrane*

    PubMed Central

    Asanuma-Date, Kimie; Hirano, Yuki; Le, Na; Sano, Kotone; Kawasaki, Nana; Hashii, Noritaka; Hiruta, Yoko; Nakayama, Ken-ichi; Umemura, Mariko; Ishikawa, Kazuhiko; Sakagami, Hiromi; Ogawa, Haruko

    2012-01-01

    Porcine pancreatic α-amylase (PPA) binds to N-linked glycans of glycoproteins (Matsushita, H., Takenaka, M., and Ogawa, H. (2002) J. Biol Chem., 277, 4680–4686). Immunostaining revealed that PPA is located at the brush-border membrane (BBM) of enterocytes in the duodenum and that the binding is inhibited by mannan but not galactan, indicating that PPA binds carbohydrate-specifically to BBM. The ligands for PPA in BBM were identified as glycoprotein N-glycans that are significantly involved in the assimilation of glucose, including sucrase-isomaltase (SI) and Na+/Glc cotransporter 1 (SGLT1). Binding of SI and SGLT1 in BBM to PPA was dose-dependent and inhibited by mannan. Using BBM vesicles, we found functional changes in PPA and its ligands in BBM due to the N-glycan-specific interaction. The starch-degrading activity of PPA and maltose-degrading activity of SI were enhanced to 240 and 175%, respectively, while Glc uptake by SGLT1 was markedly inhibited by PPA at high but physiologically possible concentrations, and the binding was attenuated by the addition of mannose-specific lectins, especially from Galanthus nivalis. Additionally, recombinant human pancreatic α-amylases expressed in yeast and purified by single-step affinity chromatography exhibited the same carbohydrate binding specificity as PPA in binding assays with sugar-biotinyl polymer probes. The results indicate that mammalian pancreatic α-amylases share a common carbohydrate binding activity and specifically bind to the intestinal BBM. Interaction with N-glycans in the BBM activated PPA and SI to produce much Glc on the one hand and to inhibit Glc absorption by enterocytes via SGLT1 in order to prevent a rapid increase in blood sugar on the other. PMID:22584580

  12. Chronic hypercapnia enhances V sub max of Na-H antiporter of renal brush-border membranes

    SciTech Connect

    Talor, Z.; Yang, Wuchang; Shuffield, J.; Sack, E.; Arruda, J.A.L. )

    1987-09-01

    Chronic hypercapnia is associated with increased proximal HCO{sub 3} reabsorption that is thought to be mediated by a Na-H antiporter. The authors hypothesized that chronic hypercapnia would be associated either with increased V{sub max} or with decreased K{sub m} of the Na-H antiporter. To test this hypothesis they made rabbits hypercapnic for 48 h by exposure to 10% CO{sub 2}. In both control and hypercapnic animals, cortical luminal membranes were enriched over the homogenate 16-fold in alkaline phosphatase and 10-fold in maltase activity. The kinetic activity of the Na-H antiporter was measured by the dissipation of the quenching of acridine orange by addition of different Na concentrations. Chronic hypercapnic rabbits had significantly higher V{sub max} of the Na-H antiporter of luminal membranes than controls. The K{sub m}, however, was not different between control and hypercapnic rabbits. {sup 22}Na uptake in presence of an outwardly directed pH gradient was significantly higher in vesicles from hypercapnic rabbits than controls. Amiloride inhibited the Na-H antiporter (as assessed by acridine orange quenching or {sup 22}Na uptake) to the same degree in membranes from both control and hypercapnic rabbits. The uptake of D-({sup 3}H)glucose by luminal membranes was not different between control and hypercapnic rabbits, indicating a specific enhancement of the Na-H antiporter. Thus chronic hypercapnia, but not acute hypercapnia, induces a selective and specific increase in the V{sub max} of Na-H antiporter, and this may mediate the adaptation to chronic hypercapnia.

  13. Age related changes in functions and physicochemical properties of rat jejunal brush border membrane after chronic ethanol administration.

    PubMed

    Lindi, C; Marciani, P; Omodeo-Sale, F

    1993-02-01

    1. We investigated the chronic effects of a 4 week treatment with ethanol on functions and physicochemical properties of BBM of young and adult rats (2 and 7 months old respectively). 2. In the ethanol treated groups the cholesterol/phospholipid and the protein/lipid ratios as well as the D-glucose uptake and lactase specific activity and Vmax were increased. In spite of a minor alcohol consumption the adult group was the more affected. 3. Membranes from the ethanol fed rats were less fluid and more tolerant to the in vitro addition of ethanol. PMID:8098680

  14. Structural state of the Na+/D-glucose cotransporter in calf kidney brush-border membranes. Target size analysis of Na+-dependent phlorizin binding and Na+-dependent D-glucose transport.

    PubMed

    Lin, J T; Szwarc, K; Kinne, R; Jung, C Y

    1984-11-01

    Target sizes of the renal sodium-D-glucose cotransport system in brush-border membranes of calf kidney cortex were estimated by radiation inactivation. In brush-border vesicles irradiated at -50 degrees C with 1.5 MeV electron beams, sodium-dependent phlorizin binding, and Na+-dependent D-glucose tracer exchange decreased exponentially with increasing doses of radiation (0.4-4.4 Mrad). Inactivation of phlorizin binding was due to a reduction in the number of high-affinity phlorizin binding sites but not in their affinity. The molecular weight of the Na+-dependent phlorizin binding unit was estimated to be 230 000 +/- 38 000. From the tracer exchange experiments a molecular weight of 345 000 +/- 24 500 was calculated for the D-glucose transport unit. The validity of these target size measurements was established by concomitant measurements of two brush-border enzymes, alkaline phosphatase and gamma-glutamyltransferase, whose target sizes were found to be 68 570 +/- 2670 and 73 500 +/- 2270, respectively. These findings provide further evidence for the assumption that the sodium-D-glucose cotransport system is a multimeric structure, in which distinct complexes are responsible for phlorizin binding and D-glucose translocation. PMID:6148966

  15. Plasticity of the brush border - the yin and yang of intestinal homeostasis.

    PubMed

    Delacour, Delphine; Salomon, Julie; Robine, Sylvie; Louvard, Daniel

    2016-03-01

    The brush border on the apical surface of enterocytes is a highly specialized structure well-adapted for efficient digestion and nutrient transport, whilst at the same time providing a protective barrier for the intestinal mucosa. The brush border is constituted of a densely ordered array of microvilli, protrusions of the plasma membrane, which are supported by actin-based microfilaments and interacting proteins and anchored in an apical network of actomyosin and intermediate filaments, the so-called terminal web. The highly dynamic, specialized apical domain is both an essential partner for the gut microbiota and an efficient signalling platform that enables adaptation to physiological stimuli from the external and internal milieu. Nevertheless, genetic alterations or various pathological stresses, such as infection, inflammation, and mechanical or nutritional alterations, can jeopardize this equilibrium and compromise intestinal functions. Long-time neglected, the intestinal brush-border shall be enlightening again as the central actor of the complex but essential intestinal homeostasis. Here, we review the processes and components involved in brush border organization and discuss pathological mechanisms that can induce brush border defects and their physiological consequences. PMID:26837713

  16. Differential Association of the Na+/H+ Exchanger Regulatory Factor (NHERF) Family of Adaptor Proteins with the Raft-and the Non-Raft Brush Border Membrane Fractions of NHE3

    PubMed Central

    Sultan, Ayesha; Luo, Min; Yu, Qin; Riederer, Brigitte; Xia, Weiliang; Chen, Mingmin; Lissner, Simone; Gessner, Johannes E.; Donowitz, Mark; Yun, C. Chris; deJonge, Hugo; Lamprecht, Georg; Seidler, Ursula

    2014-01-01

    Background/Aims Trafficking, brush border membrane (BBM) retention, and signal-specific regulation of the Na+/H+ exchanger NHE3 is regulated by the Na+/H+ Exchanger Regulatory Factor (NHERF) family of PDZ-adaptor proteins, which enable the formation of multiprotein complexes. It is unclear, however, what determines signal specificity of these NHERFs. Thus, we studied the association of NHE3, NHERF1 (EBP50), NHERF2 (E3KARP), and NHERF3 (PDZK1) with lipid rafts in murine small intestinal BBM. Methods Detergent resistant membranes (“lipid rafts”) were isolated by floatation of Triton X-incubated small intestinal BBM from a variety of knockout mouse strains in an Optiprep step gradient. Acid-activated NHE3 activity was measured fluorometrically in BCECF-loaded microdissected villi, or by assessment of CO2/HCO3− mediated increase in fluid absorption in perfused jejunal loops of anethetized mice. Results NHE3 was found to partially associate with lipid rafts in the native BBM, and NHE3 raft association had an impact on NHE3 transport activity and regulation in vivo. NHERF1, 2 and 3 were differentially distributed to rafts and non-rafts, with NHERF2 being most raft-associated and NHERF3 entirely non-raft associated. NHERF2 expression enhanced the localization of NHE3 to membrane rafts. The use of acid sphingomyelinase-deficient mice, which have altered membrane lipid as well as lipid raft composition, allowed us to test the validity of the lipid raft concept in vivo. Conclusions The differential association of the NHERFs with the raft-associated and the non-raft fraction of NHE3 in the brush border membrane is one component of the differential and signal-specific NHE3 regulation by the different NHERFs. PMID:24297041

  17. Identification of ABCC2 as a binding protein of Cry1Ac on brush border membrane vesicles from Helicoverpa armigera by an improved pull-down assay.

    PubMed

    Zhou, Zishan; Wang, Zeyu; Liu, Yuxiao; Liang, Gemei; Shu, Changlong; Song, Fuping; Zhou, Xueping; Bravo, Alejandra; Soberón, Mario; Zhang, Jie

    2016-08-01

    Cry1Ac toxin-binding proteins from Helicoverpa armigera brush border membrane vesicles were identified by an improved pull-down method that involves coupling Cry1Ac to CNBr agarose combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS). According to the LC-MS/MS results, Cry1Ac toxin could bind to six classes of aminopeptidase-N, alkaline phosphatase, cadherin-like protein, ATP-binding cassette transporter subfamily C protein (ABCC2), actin, ATPase, polycalin, and some other proteins not previously characterized as Cry toxin-binding molecules such as dipeptidyl peptidase or carboxyl/choline esterase and some serine proteases. This is the first report that suggests the direct binding of Cry1Ac toxin to ABCC2 in H. armigera. PMID:27037552

  18. Resistance in a laboratory population of Culex quinquefasciatus (Diptera: Culicidae) to Bacillus sphaericus binary toxin is due to a change in the receptor on midgut brush-border membranes.

    PubMed

    Nielsen-Leroux, C; Charles, J F; Thiéry, I; Georghiou, G P

    1995-02-15

    Direct binding experiments with isolated brush border membrane fractions (BBMF) from larvae of a susceptible laboratory strain of Culex quinquefasciatus Say, indicated the presence of a single class of Bacillus sphaericus binary toxin receptors. The dissociation constant (Kd) was approximately 11 nM and the maximum binding capacity (Bmax) approximately 8 pmol/mg BBMF protein. Similar binding experiments with a field population of C. quinquefasciatus that had been selected in the laboratory to more than 100,000-fold resistance to B. sphaericus binary toxin failed to reveal the presence of any specific binding. Thus this resistant strain had lost the functional receptor for B. sphaericus toxin. The binding characteristics of BBMF from the F1 larval progeny (susceptible females x resistant males) were very close to those of the parental susceptible strain, consistent with the resistance being recessive. PMID:7883005

  19. Deep-apical tubules: dynamic lipid-raft microdomains in the brush-border region of enterocytes.

    PubMed

    Hansen, Gert H; Pedersen, Jens; Niels-Christiansen, Lise-Lotte; Immerdal, Lissi; Danielsen, E Michael

    2003-07-01

    The brush border of small intestinal enterocytes is highly enriched in cholesterol- and glycosphingolipid-containing membrane microdomains, commonly termed as lipid 'rafts'. Functionally, transcytosis of IgA and exocytosis of newly made brush-border proteins in enterocytes occur through apical lipid raft-containing compartments, but little is otherwise known about these raft microdomains. We therefore studied in closer detail apical lipid-raft compartments in enterocytes by immunogold electron microscopy and biochemical analyses. Novel membrane structures, deep-apical tubules, were visualized by the non-permeable surface marker Ruthenium Red in the brush-border region of the cells. The surface-connected tubules were labelled by antibodies to caveolin-1 and the glycolipid asialo G(M1), and they were sensitive to cholesterol depletion by methyl-beta-cyclodextrin, indicating the presence of raft microdomains. Deep-apical tubules were positioned close to the actin rootlets of adjacent microvilli in the terminal web region, which had a diameter of 50-100 nm, and penetrated up to 1 microm into the cytoplasm. Markers for transcytosis, IgA and the polymeric immunoglobulin receptor, as well as the resident brush-border enzyme aminopeptidase N, were present in these deep-apical tubules. We propose that deep-apical tubules are a specialized lipid-raft microdomain in the brush-border region functioning as a hub in membrane trafficking at the brush border. In addition, the sensitivity to cholesterol depletion suggests that deep-apical tubules function as a cell-surface membrane reservoir for cholesterol and for rapid adaptive changes in the size of microvilli at the brush border. PMID:12689332

  20. Cordon bleu promotes the assembly of brush border microvilli

    PubMed Central

    Grega-Larson, Nathan E.; Crawley, Scott W.; Erwin, Amanda L.; Tyska, Matthew J.

    2015-01-01

    Microvilli are actin-based protrusions found on the surface of diverse cell types, where they amplify membrane area and mediate interactions with the external environment. In the intestinal tract, these protrusions play central roles in nutrient absorption and host defense and are therefore essential for maintaining homeostasis. However, the mechanisms controlling microvillar assembly remain poorly understood. Here we report that the multifunctional actin regulator cordon bleu (COBL) promotes the growth of brush border (BB) microvilli. COBL localizes to the base of BB microvilli via a mechanism that requires its proline-rich N-terminus. Knockdown and overexpression studies show that COBL is needed for BB assembly and sufficient to induce microvillar growth using a mechanism that requires functional WH2 domains. We also find that COBL acts downstream of the F-BAR protein syndapin-2, which drives COBL targeting to the apical domain. These results provide insight into a mechanism that regulates microvillar growth during epithelial differentiation and have significant implications for understanding the maintenance of intestinal homeostasis. PMID:26354418

  1. Brush border intestinal enzymes after multiple daily fractionation

    SciTech Connect

    Becciolini, A.; Giache, V.; Balzi, M.; Morrone, A.

    1987-03-01

    The modifications in brush border enzyme activity of the epithelial cell of the small intestine were studied after multiple daily fractionation (MDF) of 3 Gy X and 3 Gy X 2 X 2 (12 h split). Disaccharase and dipeptidase activities changed in the same way after irradiation. The results show that both total doses caused the three known phases of increase, decrease, and a return to normal. With MDF, activity at the end of irradiation was similar to or greater than that of controls and remained higher longer than a single dose of 8 Gy. However, the return to normal occurred sooner than after a single dose of 8 Gy. After 11 days, circadian oscillations of brush border enzyme activity appeared similar to those of controls in many segments of the intestine, reaching the highest activity during the night and the lowest in the afternoon.

  2. Probing endocytosis from the enterocyte brush border using fluorescent lipophilic dyes: lipid sorting at the apical cell surface.

    PubMed

    Danielsen, E Michael

    2015-05-01

    The small intestinal brush border is a specialized cell membrane that needs to withstand the solubilizing effect of bile salts during assimilation of dietary nutrients and to achieve detergent resistance; it is highly enriched in glycolipids organized in lipid raft microdomains. In the present work, the fluorescent lipophilic probes FM 1-43 (N-(3-triethylammoniumpropyl)-4-(4-(dibutylamino)styryl)pyridinium dibromide), FM 4-64 (N-(3-triethylammoniumpropyl)-4-(6-(4-(diethylamino) phenyl)hexatrienyl)pyridinium dibromide), TMA-DPH (1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene p-toluenesulfonate), and CellMask Orange plasma membrane stain were used to study endocytosis from the enterocyte brush border of organ-cultured porcine mucosal explants. All the dyes readily incorporated into the brush border but were not detectably endocytosed by 5 min, indicating a slow uptake compared with other cell types. At later time points, FM 1-43 clearly appeared in distinct punctae in the terminal web region, previously shown to represent early endosomes (TWEEs). In contrast, the other dyes were relatively "endocytosis resistant" to varying degrees for periods up to 2 h, indicating an active sorting of lipids in the brush border prior to internalization. For some of the dyes, a diphenylhexatriene motif in the lipophilic tail seemed to confer the relative endocytosis resistance. Lipid sorting by selective endocytosis therefore may be a process in the enterocytes aimed to generate and maintain a unique lipid composition in the brush border. PMID:25526697

  3. Binding Sites for Bacillus thuringiensis Cry2Ae Toxin on Heliothine Brush Border Membrane Vesicles Are Not Shared with Cry1A, Cry1F, or Vip3A Toxin ▿

    PubMed Central

    Gouffon, C.; Van Vliet, A.; Van Rie, J.; Jansens, S.; Jurat-Fuentes, J. L.

    2011-01-01

    The use of combinations of Bacillus thuringiensis (Bt) toxins with diverse modes of action for insect pest control has been proposed as the most efficient strategy to increase target range and delay the onset of insect resistance. Considering that most cases of cross-resistance to Bt toxins in laboratory-selected insect colonies are due to alteration of common toxin binding sites, independent modes of action can be defined as toxins sharing limited or no binding sites in brush border membrane vesicles (BBMV) prepared from the target insect larvae. In this paper, we report on the specific binding of Cry2Ae toxin to binding sites on BBMV from larvae of the three most commercially relevant heliothine species, Heliothis virescens, Helicoverpa zea, and Helicoverpa armigera. Using chromatographic purification under reducing conditions before labeling, we detected specific binding of radiolabeled Cry2Ae, which allowed us to perform competition assays using Cry1Ab, Cry1Ac, Cry1Fa, Vip3A, Cry2Ae, and Cry2Ab toxins as competitors. In these assays, Cry2Ae binding sites were shared with Cry2Ab but not with the tested Cry1 or Vip3A toxins. Our data support the use of Cry2Ae toxin in combination with Cry1 or Vip3A toxins in strategies to increase target range and delay the onset of heliothine resistance. PMID:21441333

  4. Taurine uptake across the human intestinal brush-border membrane is via two transporters: H+-coupled PAT1 (SLC36A1) and Na+- and Cl−-dependent TauT (SLC6A6)

    PubMed Central

    Anderson, Catriona M H; Howard, Alison; Walters, Julian R F; Ganapathy, Vadivel; Thwaites, David T

    2009-01-01

    Taurine is an essential amino acid in some mammals and is conditionally essential in humans. Taurine is an abundant component of meat and fish-based foods and has been used as an oral supplement in the treatment of disorders such as cystic fibrosis and hypertension. The purpose of this investigation was to identity the relative contributions of the solute transporters involved in taurine uptake across the luminal membrane of human enterocytes. Distinct transport characteristics were revealed following expression of the candidate solute transporters in Xenopus laevis oocytes: PAT1 (SLC36A1) is a H+-coupled, pH-dependent, Na+- and Cl−-independent, low-affinity, high-capacity transporter for taurine and β-alanine; TauT (SLC6A6) is a Na+- and Cl−-dependent, high-affinity, low-capacity transporter of taurine and β-alanine; ATB0,+ (SLC6A14) is a Na+- and Cl−-dependent, high-affinity, low-capacity transporter which accepts β-alanine but not taurine. Taurine uptake across the brush-border membrane of human intestinal Caco-2 cell monolayers showed characteristics of both PAT1- and TauT-mediated transport. Under physiological conditions, Cl−-dependent TauT-mediated uptake predominates at low taurine concentrations, whereas at higher concentrations typical of diet, Cl−-independent PAT1-mediated uptake is the major absorptive mechanism. Real-time PCR analysis of human duodenal and ileal biopsy samples demonstrates that PAT1, TauT and ATB0,+ mRNA are expressed in each tissue but to varying degrees. In conclusion, this study is the first to demonstrate both taurine uptake via PAT1 and functional coexpression of PAT1 and TauT at the apical membrane of the human intestinal epithelium. PAT1 may be responsible for bulk taurine uptake during a meal whereas TauT may be important for taurine supply to the intestinal epithelium and for taurine capture between meals. PMID:19074966

  5. Binding of diarrheagenic Escherichia coli to 32- to 33-kilodalton human intestinal brush border proteins.

    PubMed Central

    Manjarrez-Hernandez, A; Gavilanes-Parra, S; Chavez-Berrocal, M E; Molina-Lopez, J; Cravioto, A

    1997-01-01

    We have detected human intestinal brush border proteins to which Escherichia coli strains adhere by means of a blotting-nitrocellulose method in which the binding of radiolabeled bacteria to sodium dodecyl sulfate-polyacrylamide gel electrophoresis-separated intestinal cell membranes was evaluated. The brush border fraction contained several polypeptides that bound only adherent E. coli strains. The most prominent and consistent of these proteins had apparent molecular masses of 32 to 33 kDa. Additional polypeptides ranging from 50 to 70, from 105 to 130, and from 180 to 200 kDa were also recognized by adherent E. coli strains, although with less intensity (in accordance with the number of bound bacteria to these polypeptides). Independently of the pattern of adherence (localized [LA], diffuse [DA], or aggregative [AggA]) all HEp-2-adhering strains recognized, with different intensities, the 32- to 33-kDa brush border proteins, whereas nonadhesive strains did not. The relative avidity of an LA strain to bind to the 32- to 33-kDa proteins was approximately seven- and sixfold higher than the binding of strains with aggregative and diffuse adherence, respectively. Thus, it is reasonable to think that LA, DA, and AggA strains have a common adhesin that mediates binding to the 32- to 33-kDa bands. Inhibition experiments using HEp-2 cells demonstrated that isolated 32- to 33-kDa proteins or specific antiserum blocked preferentially bacterial adherence of the LA pattern. Delipidization and protein digestion of the human brush borders confirmed that E. coli bound to structures of a proteinaceous nature. Deglycosylation studies and sodium meta-periodate oxidation of the intestinal cell membranes decreased bacterial binding activity significantly, indicating that E. coli bound to carbohydrate moieties in the glycoproteins. These results suggest that binding of E. coli strains, mainly of the LA phenotype, to the 32- to 33-kDa proteins could play a role in colonization through

  6. Role of lysophosphatidylcholine in brush-border intestinal alkaline phosphatase release and restoration.

    PubMed

    Nakano, Takanari; Inoue, Ikuo; Alpers, David H; Akiba, Yasutada; Katayama, Shigehiro; Shinozaki, Rina; Kaunitz, Jonathan D; Ohshima, Susumu; Akita, Masumi; Takahashi, Seiichiro; Koyama, Iwao; Matsushita, Makoto; Komoda, Tsugikazu

    2009-07-01

    Intestinal alkaline phosphatase (IAP) is a brush-border membrane ectoenzyme (BBM-IAP) that is released into the lumen (L-IAP) after a high-fat diet. We examined the effects of oil feeding and the addition of mixed-lipid micelles on the formation of L-IAP in oil-fed rat intestine, Caco-2 cell monolayers, and mouse intestinal loops. We localized IAP in the duodenum of rats fed corn oil using fluorescence microscopy with enzyme-labeled fluorescence-97 as substrate. Four hours after oil feeding, L-IAP increased approximately 10-fold accompanied by the loss of BBM-IAP, consistent with BBM-IAP release. Rat IAP isozyme mRNAs progressively increased 4-6 h after oil feeding, followed by the increase of IAP activity in the subapical location at 6 h, consistent with the restoration of IAP protein. Postprandial lipid-micelle components, sodium taurocholate with or without oleic acid, mono-oleylglycerol, cholesterol, or lysophosphatidylcholine (lysoPC) were applied singly or as mixed-lipid micelles to the apical surface of polarized Caco-2 cell monolayers. LysoPC increased L-IAP >10-fold over basal release. LysoPC released IAP into the apical medium more than other intestinal brush-border enzymes, 5'-nucleotidase, sucrase, aminopeptidase N, and lactase, without comparable lactate dehydrogenase release or cell injury. LysoPC increased human IAP mRNA levels by 1.5-fold in Caco-2 cells. Luminally applied lysoPC also increased release of IAP preferentially in mouse intestinal loops. These data show that lysoPC accelerates the formation of L-IAP from BBM-IAP, followed by enhanced IAP synthesis, suggesting the role that lysoPC might play in the turnover of brush-border proteins. PMID:19407215

  7. Small intestinal brush border enzymes in cystic fibrosis.

    PubMed

    Van Biervliet, S; Eggermont, E; Carchon, H; Veereman, G; Deboeck, K

    1999-01-01

    The study concerns the maltase, saccharase, lactase and alkaline phosphatase activity in small intestinal biopsy specimens from 61 consecutively admitted, untreated, Caucasian cystic fibrosis patients. A group of 319 age matched controls admitted during the same time period for undefined gastrointestinal or nutritional disorders acted as the controls. In order to eliminate morphological damage as a confounding factor, the enzyme activities were studied in small intestinal biopsy specimens having both normal stereomicroscopic and histological features. It was shown that neither maltase nor saccharase activity was different in the two groups, in contrast to lactase and alkaline phophatase activity, that was significantly lower in cystic fibrosis patients. The differences could not be explained by the nutritional status as judged by the body mass index. Lactase activity is known to be easily affected by numerous enteropathies. As the information on alkaline phosphatase activity is limited, the low activity is discussed in more detail. Taking into account the literature data, the low alkaline phosphatase activity is tentatively attributed either to enhanced release from the brush border or to the faulty handling of alkaline phophatase protein in the post-golgi compartments secondary to the accumulation of incorrectly glycosylated CFTR in the same cell structures. PMID:10547891

  8. Location of a protein of the fodrin-spectrin-TW260/240 family in the mouse intestinal brush border.

    PubMed

    Hirokawa, N; Cheney, R E; Willard, M

    1983-03-01

    We have determined that a protein of the fodrin-spectrin-TW260/240 (FST) family is a component of the thin fibrils (approximately 5 nm wide, 100-200 nm long) that cross-link bundles of actin filaments to adjacent actin bundles and to the plasma membrane in the terminal web of the brush border of the intestinal epithelium. When isolated brush borders were incubated with anti-fodrin antibodies and prepared for electron microscopy by the quick-freeze, deep-etch technique, these approximately 5 nm fibrils were specifically decorated with the antibody. In addition, these cross-linking fibrils disappeared when the anti-fodrin-reactive proteins were extracted from the brush border. We conclude that FST is a component of a cross-linking system composed of approximately 5 nm fibrils that are morphologically distinct from the approximately 8 nm myosin-containing fibrils which were identified by anti-myosin decoration. In addition to linking actin bundles to adjacent actin bundles and to the plasma membrane, these FST fibrils may mediate actin-vesicle, actin-intermediate filament and vesicle-plasma membrane linkages. PMID:6831563

  9. Lymphocyte expression of a 90 kD brush border antigen.

    PubMed Central

    Van Leer, E H; Moullier, P; Ronco, P; Verroust, P

    1987-01-01

    This study analyses the expression by rat lymphocytes of six renal brush border (BB) antigens defined by a set of monoclonal antibodies and demonstrates that a 90 kD protein (gp 90), synthesized by BB and glomeruli is found on the surface of T and B cells. The other antigens, including the 330 kD protein involved in Heymann's nephritis--a model of epimembranous glomerulonephritis--are not detectable on the surface of lymphocytes. Immunochemical studies indicate that gp 90 and the related protein immunoprecipitated from thymocyte membranes co-migrate in SDS-PAGE. Quantitative binding analysis shows that the number of antigenic sites is in the same order of magnitude on thymocytes, spleen and bone-marrow lymphocytes as well as on two lines of pre-B and pre-T cells, but considerably lower on a highly differentiated helper T cell line. Images Fig. 4 PMID:3301100

  10. Small molecule pinocytosis and clathrin-dependent endocytosis at the intestinal brush border: Two separate pathways into the enterocyte.

    PubMed

    Michael Danielsen, E; Hansen, Gert H

    2016-02-01

    Pinocytosis at the small intestinal brush border was studied in postweaned porcine cultured mucosal explants, using the fluorescent polar probes Alexa hydrazide (AH, MW 570), Texas red dextran (TRD, MW ~ 3000), and Cascade blue dextran (CBD, MW ~ 10,000). Within 1 h, AH appeared in a string of subapical punctae in enterocytes, indicative of an ongoing constitutive pinocytosis. By comparison, TRD was taken up less efficiently into the same compartment, and no intracellular labeling of CBD was detectable, indicating that only small molecules are pinocytosed from the postweaned gut lumen. AH remained in the terminal web region in EEA-1-positive endosomes (“TWEEs”) for at least 2 h, implying that the pinocytic uptake does not proceed towards a transcytic pathway. Like AH, cholera toxin B subunit (CTB) was readily internalized, but the two probes appeared in completely non-overlapping subapical compartments, indicating the existence of two different uptake mechanisms operating simultaneously at the brush border. CTB is internalized by clathrin-dependent receptor mediated endocytosis, but surprisingly the toxin also caused a rapid disappearance from the apical cell surface of two major brush border enzymes, alkaline phosphatase and aminopeptidase N, demonstrating the disruptive effect of this pathway. By immunofluorescence, caveolin-1 was hardly detectable in enterocytes, arguing against a caveolae-mediated uptake of AH, whereas the pinocytosis/phagocytosis inhibitors dimethyl amiloride and cytochalasin D both arrested AH uptake. We propose that the constitutive pinocytic mechanism visualized by AH contributes to maintenance of membrane homeostasis and to enrich the contents of lipid raft constituents at the brush border. PMID:26615917

  11. Duodenal brush-border mucosal glucose transport and enzyme activities in aging man and effect of bacterial contamination of the small intestine.

    PubMed

    Wallis, J L; Lipski, P S; Mathers, J C; James, O F; Hirst, B H

    1993-03-01

    Duodenal biopsies were collected from 38 subjects (24 female and 14 male) ranging in age from 55 to 91 years. Evidence of bacterial contamination of the small bowel (BCSB) was sought at the same time by bacterial culture of duodenal aspirates and by hydrogen and [14C]glycocholic acid breath tests; subjects were considered to be positive for BCSB if any one of the three tests was abnormal. Biopsies were analyzed for six brush-border membrane enzyme activities: maltase, sucrase, lactase, alkaline phosphatase, leucine aminopeptidase, and alpha-glucosidase. Analysis of covariance with age as the covariate indicated no significant effect of age on the specific activities of these enzymes. Mucosal Na(+)-dependent glucose transport was quantified in brush-border membrane vesicles prepared from the biopsies. In all groups, glucose transport at 20-30 sec was greater (ranging from mean values of 2.45 to 3.66 times) than at 45 min, consistent with Na(+)-coupled glucose transport, and no significant effect of age was observed. BCSB had no significant effect on specific activities of any of the duodenal mucosal hydrolases but was associated with reduced (P = 0.05) brush-border glucose transport. None of the variables studied was significantly affected by the gender of subjects. In conclusion, these biochemical data do not support the contention that reduced capacity for carbohydrate absorption in the elderly is explained by reductions in duodenal brush-border mucosal disaccharidase activities or glucose transport. PMID:8444069

  12. Radiation inactivation studies of renal brush border water and urea transport

    SciTech Connect

    Verkman, A.S.; Dix, J.A.; Seifter, J.L.; Skorecki, K.L.; Jung, C.Y.; Ausiello, D.A.

    1985-12-01

    Radiation inactivation was used to determine the nature and molecular weight of water and urea transport pathways in brush border membrane vesicles (BBMV) isolated from rabbit renal cortex. BBMV were frozen to -50 degrees C, irradiated with 1.5 MeV electrons, thawed, and assayed for transport or enzyme activity. The freezing process had no effect on enzyme or transport kinetics. BBMV alkaline phosphatase activity gave linear ln(activity) vs. radiation dose plots with a target size of 68 +/- 3 kDa, similar to previously reported values. Water and solute transport were measured using the stopped-flow light-scattering technique. The rates of acetamide and osmotic water transport did not depend on radiation dose (0-7 Mrad), suggesting that transport of these substances does not require a protein carrier. In contrast, urea and thiourea transport gave linear ln(activity) vs. dose curves with a target size of 125-150 kDa; 400 mM urea inhibited thiourea flux by -50% at 0 and 4.7 Mrad, showing that radiation does not affect inhibitor binding to surviving transporters. These studies suggest that BBMV urea transport requires a membrane protein, whereas osmotic water transport does not.

  13. Renal brush-border Na/sup +/-H/sup +/ exchange activity in the aging rat

    SciTech Connect

    Kinsella, J.L.; Sacktor, B.

    1987-04-01

    Amiloride-sensitive Na/sup +/-H/sup +/ exchange activity in brush-border membrane vesicles isolated from male rat proximal tubules was decreased in the senescent rat (24 mo) compared with the young adult (6 mo). There was no significant loss in Na/sup +/-H/sup +/ exchange activity in the kidneys of animals between 6 and 18 mo of age. Amiloride-insensitive /sup 22/Na/sup +/ uptake and the rate of pH gradient dissipation were not altered during aging. The decrease in sodium-dependent (/sup 32/P) phosphate transport preceded the decline in Na/sup +/-H/sup +/ exchange activity by at least 6 mo. Sodium-dependent D-(/sup 3/H) glucose transport was not significantly altered during aging. Thus various renal plasma membrane transport functions were affected differently in the aging rat. The decrease in Na/sup +/-H/sup +/ exchange activity during aging contrasted with the increase in exchange activity reported previously in acute ablation models of chronic renal failure.

  14. Characterization of the fetal glucose transporter in rabbit kidney. Comparison with the adult brush border electrogenic Na+-glucose symporter.

    PubMed Central

    Beck, J C; Lipkowitz, M S; Abramson, R G

    1988-01-01

    Glucose transport was characterized in rabbit renal brush border membrane vesicles (BBMV) of the fetus late in gestation. Highly purified, osmotically reactive fetal BBMV contained a glucose transporter that was qualitatively indistinguishable from that in the adult: both are concentrative, Na+ dependent, electrogenic, stereospecific, and sensitive to phlorizin. Although the apparent Km for glucose is similar in the fetus and adult, the Vmax is significantly higher in the adult. When the membrane potential was clamped with a protonophore, this difference diminished; however, Vmax remained significantly higher in adult BBMV. This postnatal increase in Vmax was paralleled by a similar increase in the number of phlorizin binding sites. These findings indicate that the maturational increase in glucose transport is, in part, consequent to a more favorable electrical potential for Na+-dependent glucose transport and, in part, the result of the insertion of new transporters. The homogenate activity of several brush border enzymes also demonstrated significant maturational increases. The magnitude of these changes was variable and enzyme dependent. These combined observations suggest that mature expression of membrane proteins (transporters and enzymes) occurs at different stages of development of renal proximal tubule cells. PMID:3403709

  15. Disruption of the C. elegans Intestinal Brush Border by the Fungal Lectin CCL2 Phenocopies Dietary Lectin Toxicity in Mammals.

    PubMed

    Stutz, Katrin; Kaech, Andres; Aebi, Markus; Künzler, Markus; Hengartner, Michael O

    2015-01-01

    Lectins are non-immunoglobulin carbohydrate-binding proteins without enzymatic activity towards the bound carbohydrates. Many lectins of e.g. plants or fungi have been suggested to act as toxins to defend the host against predators and parasites. We have previously shown that the Coprinopsis cinerea lectin 2 (CCL2), which binds to α1,3-fucosylated N-glycan cores, is toxic to Caenorhabditis elegans and results in developmental delay and premature death. In this study, we investigated the underlying toxicity phenotype at the cellular level by electron and confocal microscopy. We found that CCL2 directly binds to the intestinal apical surface and leads to a highly damaged brush border with loss of microvilli, actin filament depolymerization, and invaginations of the intestinal apical plasma membrane through gaps in the terminal web. We excluded several possible toxicity mechanisms such as internalization and pore-formation, suggesting that CCL2 acts directly on intestinal apical plasma membrane or glycocalyx proteins. A genetic screen for C. elegans mutants resistant to CCL2 generated over a dozen new alleles in bre 1, ger 1, and fut 1, three genes required for the synthesis of the sugar moiety recognized by CCL2. CCL2-induced intestinal brush border defects in C. elegans are similar to the damage observed previously in rats after feeding the dietary lectins wheat germ agglutinin or concanavalin A. The evolutionary conserved reaction of the brush border between mammals and nematodes might allow C. elegans to be exploited as model organism for the study of dietary lectin-induced intestinal pathology in mammals. PMID:26057124

  16. Disruption of the C. elegans Intestinal Brush Border by the Fungal Lectin CCL2 Phenocopies Dietary Lectin Toxicity in Mammals

    PubMed Central

    Stutz, Katrin; Kaech, Andres; Aebi, Markus; Künzler, Markus; Hengartner, Michael O.

    2015-01-01

    Lectins are non-immunoglobulin carbohydrate-binding proteins without enzymatic activity towards the bound carbohydrates. Many lectins of e.g. plants or fungi have been suggested to act as toxins to defend the host against predators and parasites. We have previously shown that the Coprinopsis cinerea lectin 2 (CCL2), which binds to α1,3-fucosylated N-glycan cores, is toxic to Caenorhabditis elegans and results in developmental delay and premature death. In this study, we investigated the underlying toxicity phenotype at the cellular level by electron and confocal microscopy. We found that CCL2 directly binds to the intestinal apical surface and leads to a highly damaged brush border with loss of microvilli, actin filament depolymerization, and invaginations of the intestinal apical plasma membrane through gaps in the terminal web. We excluded several possible toxicity mechanisms such as internalization and pore-formation, suggesting that CCL2 acts directly on intestinal apical plasma membrane or glycocalyx proteins. A genetic screen for C. elegans mutants resistant to CCL2 generated over a dozen new alleles in bre 1, ger 1, and fut 1, three genes required for the synthesis of the sugar moiety recognized by CCL2. CCL2-induced intestinal brush border defects in C. elegans are similar to the damage observed previously in rats after feeding the dietary lectins wheat germ agglutinin or concanavalin A. The evolutionary conserved reaction of the brush border between mammals and nematodes might allow C. elegans to be exploited as model organism for the study of dietary lectin-induced intestinal pathology in mammals. PMID:26057124

  17. Sucrase-isomaltase and other brush border glycosidases in colorectal tumors.

    PubMed

    Lojda, Z; Fric, P

    1996-07-01

    Sucrase-isomaltase (SI), trehalase (T) and lactase-beta-glucosidase (LG) activities were assessed histochemically in samples of colorectal adenomas (11 tubular, 12 tubulovillous, 10 villous) and 30 adenocarcinomas obtained by biopsy during colonoscopy or from specimens removed by surgical intervention. Small samples of tumor tissue, tissue of the transitional zone and of macroscopically normal mucosa were quenched in heptan cooled in an acetone-dry ice mixture. Cryostat sections, transferred to non-precooled slides and in some cases to semipermeable membranes, were dried and subjected to the histochemical reactions for SI, T and LG. Sucrose, 2-naphthyl, 6-Br-2-naphthyl, and 5-Br-4-Cl-3-indoxyl alpha-D-glucosides, trehalose, and 5-Br-4-Cl-3-indoxyl-beta-D-fucoside were used as substrates. Sections of jejunal biopsies with normal activities of brush border glycosidases were used as controls. From samples of 5 adenomas, 5 adenocarcinomas and collected rests of jejunal biopsies with a normal finding 10% (w/vol) homogenates in 2% Triton X-100 were prepared. Homogenates were frozen and thawed 3 times and their supernatants subjected to isoelectric focusing on polyacrylamide gel plates. Zymograms were developed with the same methods as for the detection of alpha-glucosidases in sections. In no colorectal tumor LG was present. SI was found in 70% adenocarcinomas, 50% villous, 25% tubulovillous and 19% tubular adenomas when the method with sucrose, glucose oxidase-peroxidase and 3,3'-diaminobenzidine was used. Hardly discernible traces of activity were found in tumors with azo-coupling reactions applied at pH 5, 6 and 6.5. No reaction was detected with the indigogenic method applied at pH above 6.0. However, jejunal biopsies displayed very strong reactions confined to the brush border of enterocytes under the same conditions. A strongly positive reaction was seen in 7 of 12 tumors investigated recently when the indigogenic reaction was applied at pH below 6.0 (particularly

  18. Inhibition of Na -P/sub i/ cotransporter in small gut brush border by phosphonocarboxylic acids

    SciTech Connect

    Loghman-Adham, M.; Szczepsanska-Konkel, M.; Yusufi, A.N.K.; Van Scoy, M.; Dousa, T.P.

    1987-02-01

    The authors examined the effect of phosphonoformic acid (PFA) and phosphonoacetic acid (PAA) upon Na -P/sub i/ cotransport in brush-border membrane (BBM) from small gut of rat. Both PFA and PAA inhibited the Na gradient-dependent uptake of TSP/sub i/ by BBM vesicles (BBMV) prepared from intestinal mucosa but had no effect on Na -dependent uptakes of D-(TH)glucose, L-(TH)proline, or ( UC)succinate. The uptake in the absence of Na gradient, or uptake at equilibrium period (180 min), was not affected by PFA or by PAA. A chemical analogue of PFA and PAA, phosphonopropionic acid, had only a minor inhibitory effect and phenylphosphonic acid was inactive. Neither PFA nor PAA influenced the activity of rat intestinal BBM alkaline phosphatase. The BBMV from rat jejunum had a much higher capacity for Na gradient-dependent uptake of TSP/sub i/ than BBMV from duodenum or ileum. The inhibition of BBMV TSP/sub i/ transport across rat jejunum by PFA is competitive. They suggest that PFA and PAA are specific inhibitors of Na gradient-dependent uptake of P/sub i/ by BBMV from small intestinal mucosa and that they could serve as useful experimental tools for the studies of intestinal Na -P/sub i/ cotransport.

  19. Squalamine, a novel cationic steroid, specifically inhibits the brush-border Na+/H+ exchanger isoform NHE3.

    PubMed

    Akhter, S; Nath, S K; Tse, C M; Williams, J; Zasloff, M; Donowitz, M

    1999-01-01

    Squalamine, an endogenous molecule found in the liver and other tissues of Squalus acanthias, has antibiotic properties and causes changes in endothelial cell shape. The latter suggested that its potential targets might include transport proteins that control cell volume or cell shape. The effect of purified squalamine was examined on cloned Na+/H+ exchanger isoforms NHE1, NHE2, and NHE3 stably transfected in PS120 fibroblasts. Squalamine (1-h pretreatment) decreased the maximal velocity of rabbit NHE3 in a concentration-dependent manner (13, 47, and 57% inhibition with 3, 5, and 7 micrograms/ml, respectively) and also increased K'[H+]i. Squalamine did not affect rabbit NHE1 or NHE2 function. The inhibitory effect of squalamine was 1) time dependent, with no effect of immediate addition and maximum effect with 1 h of exposure, and 2) fully reversible. Squalamine pretreatment of the ileum for 60 min inhibited brush-border membrane vesicle Na+/H+ activity by 51%. Further investigation into the mechanism of squalamine's effects showed that squalamine required the COOH-terminal 76 amino acids of NHE3. Squalamine had no cytotoxic effect at the concentrations studied, as indicated by monitoring lactate dehydrogenase release. These results indicate that squalamine 1) is a specific inhibitor of the brush-border NHE isoform NHE3 and not NHE1 or NHE2, 2) acts in a nontoxic and fully reversible manner, and 3) has a delayed effect, indicating that it may influence brush-border Na+/H+ exchanger function indirectly, through an intracellular signaling pathway or by acting as an intracellular modulator. PMID:9886929

  20. Exposure to Engineered Nanomaterial Results in Disruption of Brush Borders in Epithelia Models in vitro

    NASA Astrophysics Data System (ADS)

    Faust, James J.

    Engineered nanoparticles (NP; 10-9 m) have found use in a variety of consumer goods and medical devices because of the unique changes in material properties that occur when synthesized on the nanoscale. Although many definitions for nanoparticle exist, from the perspective of size, nanoparticle is defined as particles with diameters less than 100 nm in any external dimension. Examples of their use include titanium dioxide added as a pigment in products intended to be ingested by humans, silicon dioxide NPs are used in foods as an anticaking agent, and gold or iron oxide NPs can be used as vectors for drug delivery or contrast agents for specialized medical imaging. Although the intended use of these NPs is often to improve human health, it has come to the attention of investigators that NPs can have unintended or even detrimental effects on the organism. This work describes one such unintended effect of NP exposure from the perspective of exposure via the oral route. First, this Dissertation will explain an event referred to as brush border disruption that occurred after nanoparticles interacted with an in vitro model of the human intestinal epithelium. Second, this Dissertation will identify and characterize several consumer goods that were shown to contain titanium dioxide that are intended to be ingested. Third, this Dissertation shows that sedimentation due to gravity does not artifactually result in disruption of brush borders as a consequence of exposure to food grade titanium dioxide in vitro. Finally, this Dissertation will demonstrate that iron oxide nanoparticles elicited similar effects after exposure to an in vitro brush border expressing model of the human placenta. Together, these data suggest that brush border disruption is not an artifact of the material/cell culture model, but instead represents a bona fide biological response as a result of exposure to nanomaterial.

  1. The cell adhesion molecule Fasciclin2 regulates brush border length and organization in Drosophila renal tubules.

    PubMed

    Halberg, Kenneth A; Rainey, Stephanie M; Veland, Iben R; Neuert, Helen; Dornan, Anthony J; Klämbt, Christian; Davies, Shireen-Anne; Dow, Julian A T

    2016-01-01

    Multicellular organisms rely on cell adhesion molecules to coordinate cell-cell interactions, and to provide navigational cues during tissue formation. In Drosophila, Fasciclin 2 (Fas2) has been intensively studied due to its role in nervous system development and maintenance; yet, Fas2 is most abundantly expressed in the adult renal (Malpighian) tubule rather than in neuronal tissues. The role Fas2 serves in this epithelium is unknown. Here we show that Fas2 is essential to brush border maintenance in renal tubules of Drosophila. Fas2 is dynamically expressed during tubule morphogenesis, localizing to the brush border whenever the tissue is transport competent. Genetic manipulations of Fas2 expression levels impact on both microvilli length and organization, which in turn dramatically affect stimulated rates of fluid secretion by the tissue. Consequently, we demonstrate a radically different role for this well-known cell adhesion molecule, and propose that Fas2-mediated intermicrovillar homophilic adhesion complexes help stabilize the brush border. PMID:27072072

  2. The cell adhesion molecule Fasciclin2 regulates brush border length and organization in Drosophila renal tubules

    PubMed Central

    Halberg, Kenneth A.; Rainey, Stephanie M.; Veland, Iben R.; Neuert, Helen; Dornan, Anthony J.; Klämbt, Christian; Davies, Shireen-Anne; Dow, Julian A. T.

    2016-01-01

    Multicellular organisms rely on cell adhesion molecules to coordinate cell–cell interactions, and to provide navigational cues during tissue formation. In Drosophila, Fasciclin 2 (Fas2) has been intensively studied due to its role in nervous system development and maintenance; yet, Fas2 is most abundantly expressed in the adult renal (Malpighian) tubule rather than in neuronal tissues. The role Fas2 serves in this epithelium is unknown. Here we show that Fas2 is essential to brush border maintenance in renal tubules of Drosophila. Fas2 is dynamically expressed during tubule morphogenesis, localizing to the brush border whenever the tissue is transport competent. Genetic manipulations of Fas2 expression levels impact on both microvilli length and organization, which in turn dramatically affect stimulated rates of fluid secretion by the tissue. Consequently, we demonstrate a radically different role for this well-known cell adhesion molecule, and propose that Fas2-mediated intermicrovillar homophilic adhesion complexes help stabilize the brush border. PMID:27072072

  3. Hormone induced expression of brush border lactase in suckling rat intestine.

    PubMed

    Chaudhry, Kamaljit Kaur; Mahmood, Safrun; Mahmood, Akhtar

    2008-05-01

    The postnatal development of intestine is associated with a decline in brush border lactase activity in rodents. This is similar to adulthood hypolactasia, a phenomenon prevalent in humans worldwide. In the present study, the effect of luminal proteases from adult rat intestine was studied in vitro on intestinal lactase activity in saline control, thyroxine, insulin and cortisone treated rat pups. Lactase levels were determined by enzyme analysis and Western blotting. mRNA levels encoding lactase were determined by Northern blotting. Administration of thyroxine for 4 days reduced (P<0.05) lactase activity, but insulin treatment had no effect in 8-day-old rat intestine. However, cortisone administration augmented (P<0.01) lactase activity, under these conditions. Western blot analysis showed decreased lactase signal corresponding to 220-kDa protein band in thyroxine treated animals. However, the intensity of lactase signal was high in cortisone treated animals compared to controls. mRNA levels encoding lactase showed a 6.8-kb mRNA transcript in saline and hormone treated rats. mRNA levels encoding lactase were increased in cortisone treated animals but were reduced in thyroxine injected pups compared to controls. Microvillus membranes from saline (P<0.01) and thyroxine (P<0.05) or insulin (P<0.01) treated rats upon incubation with luminal wash from adult rat intestine showed a significant decline in lactase activity. These findings suggest that thyroxine, insulin or cortisone induced changes in lactase expression in suckling rat intestine make it susceptible to luminal proteases, which may in part be responsible for observed maturational decline in lactase activity in adult rat intestine. PMID:18273561

  4. Effect of tannic acid on brush border disaccharidases in mammalian intestine.

    PubMed

    Chauhan, Ayesha; Gupta, Shiffalli; Mahmood, Akhtar

    2007-04-01

    Tannic acid is a glucoside (penta-m-digallolyl-glucose), which exhibits a wide variety of physiological functions. Around neutral pH, 0.4 mM tannic acid produced 84% inhibition of rat brush border sucrase activity, but 35-40% enzyme inhibition was observed in the rabbit intestine at 0.08 mM concentration. In the mice, 74-77% enzyme inhibition was observed at 0.05 mM concentration of tannic acid. The observed inhibition was reversible in rat intestine. Tannic acid (0.2 mM) also inhibited lactase (18% in adult and 71% in suckling animals), maltase (76%) and trehalase (88%) activities in rat intestine. pH versus activity curves showed that 0.2 mM tannic acid inhibited enzyme activity in rat by 91% at pH 5.5 which was reduced to 14% at pH 8.5 compared to the respective controls. In the rabbit 18-60% enzyme inhibition was noticed below pH 7.0, however at pH 8.5, it was of the order of 38%. Kinetic analysis revealed that tannic acid is a competitive inhibitor of rat brush border sucrase at pH 6.8. Effect of tannic acid together with various -SH group reacting reagents revealed that the enzyme inhibition is additive in nature, suggesting the distinct nature of binding sites on the enzyme for these compounds. The results suggest that tannic acid is a potent inhibitor of intestinal brush border disaccharidases, and could modulate the intestinal functions. PMID:17477307

  5. Mechanistic Basis of Organization of the Harmonin/USH1C-Mediated Brush Border Microvilli Tip-Link Complex.

    PubMed

    Li, Jianchao; He, Yunyun; Lu, Qing; Zhang, Mingjie

    2016-01-25

    Brush border microvilli are actin-based protrusions lining the apical surface of epithelial cells in intestines and proximal tubules of kidneys. While brush border microvilli resemble the relatively well-characterized stereocilia of hair cells, the mechanistic basis of tip-link complex organization in microvilli is poorly understood. Here, we have biochemically and structurally characterized the following pairs of interactions: protocadherin 24 and Harmonin (also known as USH1C or AIE-75), Harmonin and myosin VIIb (MYO7B), Harmonin and ANKS4B, and ANKS4B and MYO7B. We show that Harmonin, ANKS4B, and MYO7B form a stable ternary complex for anchoring microvilli tip-link cadherins. Despite having only Harmonin in common, the microvilli and the stereocilia tip-link complexes are formed via strikingly similar interaction modes. These results not only provide insight into the mechanistic bases of brush border microvilli formation and maintenance but may also be valuable for understanding some gut and/or kidney diseases caused by perturbations of brush border microvilli structures. PMID:26812017

  6. Freeze-thaw and high-voltage discharge allow macromolecule uptake into ileal brush-border vesicles

    SciTech Connect

    Donowitz, M.; Emmer, E.; McCullen, J.; Reinlib, L.; Cohen, M.E.; Rood, R.P.; Madara, J.; Sharp, G.W.G.; Murer, H.; Malmstrom, K.

    1987-06-01

    High-voltage discharge or one cycle of freeze-thawing are shown to transiently permeabilize rabbit ileal brush-border membrane vesicles to macromolecules. Uptake of the radiolabeled macromolecule dextran, mol wt 70,000, used as a marker for vesicle permeability, was determined by a rapid filtration technique, with uptake defined as substrate associated with the vesicle and releasable after incubation of vesicles with 0.1% saponin. Dextran added immediately after electric shock (2000 V) or at the beginning of one cycle of freeze-thawing was taken up approximately eightfold compared with control. ATP also was taken up into freeze-thawed vesicles, whereas there was no significant uptake into control vesicles. The increase in vesicle permeability was reversible, based on Na-dependent D-glucose uptake being decreased when studied 5 but not 15 min after electric shock, and was not significantly decreased after completion of one cycle of freeze-thawing. In addition, adenosine 3',5'-cyclic monophosphate and Ca/sup 2 +/-calmodulin-dependent protein kinase activity were similar in control vesicles and vesicles exposed to high-voltage discharge or freeze-thawing. Also, vesicles freeze-thawed with (/sup 32/P)ATP demonstrated increased phosphorylation compared with nonfrozen vesicles, while freeze-thawing did not alter vesicle protein as judged by Coomassie blue staining. These techniques should allow intestinal membrane vesicles to be used for studies of intracellular control of transport processes, for instance, studies of protein kinase regulation of transport.

  7. The prenatal development and glucocorticoid control of brush-border hydrolases in the pig small intestine.

    PubMed

    Sangild, P T; Sjöström, H; Norén, O; Fowden, A L; Silver, M

    1995-02-01

    The development of brush-border enzymes and the possible regulatory role of cortisol were investigated in the small intestine of the fetal and neonatal pig. With the sows under pentobarbitone anesthesia, osmotic minipumps containing either saline or cortisol were inserted s.c. into 25 fetuses from 10 pregnant sows (82-96 d gestation). Six d later, the infused fetuses were removed by cesarean section and samples of the proximal, middle, and distal intestine taken for analysis. Samples were also obtained from 48 piglets that did not undergo an operation (controls) and that were removed at intervals from 82 d gestation until term (114 +/- 2 d). In the proximal and middle intestine, the mean levels of lactase-phlorizin hydrolase (EC 3.2.1.23-62), maltaseglucoamylase (EC 3.2.1.20), aminopeptidase N (EC 3.4.11.2), and aminopeptidase A (EC 3.4.11.7) increased during the last 10-15 d before term, correlated positively with log10 plasma cortisol values, and were higher in cortisol-infused than in saline-infused fetuses (p < 0.05). Activity of sucrase-isomaltase (EC 3.2.1.48-10) was low in fetal pigs, and this enzyme and dipeptidyl peptidase IV (EC 3.4.14.5) were not significantly affected by fetal age or exogenous cortisol. Maltase (EC 3.2.1.48-10 and EC 3.2.1.20) activity was significantly decreased in the middle and distal intestine of cortisol-infused fetuses. The results suggest that the prepartum rise in endogenous cortisol secretion stimulates the prenatal expression of certain brush-border enzymes in the pig small intestine at this critical time. However, the effects of cortisol on the developing intestine were highly idiosyncratic for particular enzymes and intestinal regions. PMID:7731759

  8. Brush border enzyme activities in the small intestine after long-term gliadin feeding in animal models of human coeliac disease.

    PubMed

    Kozáková, H; Stĕpánková, R; Kolínská, J; Farré, M A; Funda, D P; Tucková, L; Tlaskalová-Hogenová, H

    1998-01-01

    Coeliac disease is a human, genetically linked, disorder which develops in gluten-sensitive persons. The aim of this study was to investigate the effect of prolonged feeding of gliadin, a major fraction of gluten, on enzyme activities of enterocyte brush border membrane enzymes in rats, mice and pigs. Brush-border membranes were isolated from mucosal scrapings of the small intestine of 21-d-old rat pups hand-fed with formula milk diet, two-month-old nu/nu and +/+ BALB/c mice and two-month-old piglets fed three times a week starting at birth with high doses of gliadin. Activities of lactase, sucrase and dipeptidyl peptidase IV (DPP IV) were determined. Individual animal models differed in their response to gliadin feeding. In comparison with albumin fed controls the activities of DPP IV and lactase were decreased in rat pups, nu/nu BALB/c mice and piglets. DPP IV activity was mostly affected in the ileum of rats and piglets fed with gliadin starting at birth. On the other hand, lactase and sucrase activities of nu/nu BALB/c mice and piglets decreased to the largest extent in jejunum. PMID:9821309

  9. Apical Invasion of Intestinal Epithelial Cells by Salmonella typhimurium Requires Villin to Remodel the Brush Border Actin Cytoskeleton

    PubMed Central

    Lhocine, Nouara; Arena, Ellen T.; Bomme, Perrine; Ubelmann, Florent; Prévost, Marie-Christine; Robine, Sylvie; Sansonetti, Philippe J.

    2015-01-01

    Summary Salmonella invasion of intestinal epithelial cells requires extensive, though transient, actin modifications at the site of bacterial entry. The actin-modifying protein villin is present in the brush border where it participates in the constitution of microvilli and in epithelial restitution after damage through its actin-severing activity. We investigated a possible role for villin in Salmonella invasion. The absence of villin, which is normally located at the bacterial entry site, leads to a decrease in Salmonella invasion. Villin is necessary for early membrane-associated processes and for optimal ruffle assembly by balancing the steady-state level of actin. The severing activity of villin is important for Salmonella invasion in vivo. The bacterial phosphatase SptP tightly regulates villin phosphorylation, while the actin-binding effector SipA protects F-actin and counterbalances villin-severing activity. Thus, villin plays an important role in establishing the balance between actin polymerization and actin severing to facilitate the initial steps of Salmonella entry. PMID:25600187

  10. Effects of acetaldehyde on brush border enzyme activities in human colon adenocarcinoma cell line Caco-2.

    PubMed

    Koivisto, T; Salaspuro, M

    1997-12-01

    The treatment of Caco-2 cells, a human colon adenocarcinoma cell line that closely resembles normal human small intestinal epithelial cells, with acetaldehyde resulted in significantly decreased activities of brush border enzymes sucrase, maltase, lactase, and gamma-glutamyltransferase; alkaline phosphatase activity was not affected. In the case of sucrase and maltase, the activities were also decreased by a combination of acetaldehyde and ethanol, although ethanol alone markedly increased them. The possibility that intraintestinal acetaldehyde, formed by intestinal microbes, might play a role in some small intestinal enzyme deficiencies observed earlier in alcoholics should therefore be considered. The mechanism by which acetaldehyde alters these enzyme activities remains unclear. The observation that acetaldehyde also disturbed cell polarization, an initial step in the process of differentiation in Caco-2 cells, indicates that acetaldehyde might decrease these enzyme activities by interfering with cell differentiation. Because ethanol and acetaldehyde metabolizing enzymes have not been previously studied from Caco-2 cells, alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) activities were also measured from these cells, and their ALDH isoenzyme pattern was characterized. Like many cancerous cell lines, Caco-2 cells were found to express no ADH. They, however, possessed ALDH activity that was comparable with normal colonic mucosal activity and also expressed the same ALDH classes (ALDHs 1 to 3) than normal human colonic mucosa. PMID:9438518

  11. Natural Selection at the Brush-Border: Adaptations to Carbohydrate Diets in Humans and Other Mammals

    PubMed Central

    Pontremoli, Chiara; Mozzi, Alessandra; Forni, Diego; Cagliani, Rachele; Pozzoli, Uberto; Menozzi, Giorgia; Vertemara, Jacopo; Bresolin, Nereo; Clerici, Mario; Sironi, Manuela

    2015-01-01

    Dietary shifts can drive molecular evolution in mammals and a major transition in human history, the agricultural revolution, favored carbohydrate consumption. We investigated the evolutionary history of nine genes encoding brush-border proteins involved in carbohydrate digestion/absorption. Results indicated widespread adaptive evolution in mammals, with several branches experiencing episodic selection, particularly strong in bats. Many positively selected sites map to functional protein regions (e.g., within glucosidase catalytic crevices), with parallel evolution at SI (sucrase-isomaltase) and MGAM (maltase-glucoamylase). In human populations, five genes were targeted by positive selection acting on noncoding variants within regulatory elements. Analysis of ancient DNA samples indicated that most derived alleles were already present in the Paleolithic. Positively selected variants at SLC2A5 (fructose transporter) were an exception and possibly spread following the domestication of specific fruit crops. We conclude that agriculture determined no major selective event at carbohydrate metabolism genes in humans, with implications for susceptibility to metabolic disorders. PMID:26319403

  12. Down-Regulation of Brush Border Efflux Transporter Expression in the Kidneys of Pregnant Mice

    PubMed Central

    Yacovino, Lindsay L.; Gibson, Christopher J.

    2013-01-01

    Pregnancy increases the urinary excretion of chemicals in women and rodents. It is unknown whether the enhanced clearance of drugs during pregnancy involves changes in the expression of transporters that mediate chemical secretion and reabsorption. The purpose of this study was to quantify the mRNA and protein expression of efflux transporters in kidneys from virgin and pregnant mice on gestational days 7, 11, 14, and 17 and postnatal days 1, 15, and 30 with use of quantitative polymerase chain reaction, Western blot, and immunofluorescence. Multidrug resistance protein (Mdr) 1b mRNA, multidrug resistance-associated protein (Mrp) 4 mRNA, and protein levels decreased significantly by 25–75% throughout pregnancy and lactation. Similarly, Mrp2 and multidrug and toxin extrusion transporter (Mate) 1 mRNA expression were down-regulated 20–40% during mid to late gestation but returned to control levels by postnatal day 15. In contrast, Mrp3 mRNA and protein increased 225% and 31%, respectively, at gestational day 14. Coordinated down-regulation of brush border transporters Mate1, Mrp2, and Mrp4 and up-regulation of the basolateral Mrp3 transporter would reduce chemical secretion into urine. PMID:22896729

  13. Natural Selection at the Brush-Border: Adaptations to Carbohydrate Diets in Humans and Other Mammals.

    PubMed

    Pontremoli, Chiara; Mozzi, Alessandra; Forni, Diego; Cagliani, Rachele; Pozzoli, Uberto; Menozzi, Giorgia; Vertemara, Jacopo; Bresolin, Nereo; Clerici, Mario; Sironi, Manuela

    2015-09-01

    Dietary shifts can drive molecular evolution in mammals and a major transition in human history, the agricultural revolution, favored carbohydrate consumption. We investigated the evolutionary history of nine genes encoding brush-border proteins involved in carbohydrate digestion/absorption. Results indicated widespread adaptive evolution in mammals, with several branches experiencing episodic selection, particularly strong in bats. Many positively selected sites map to functional protein regions (e.g., within glucosidase catalytic crevices), with parallel evolution at SI (sucrase-isomaltase) and MGAM (maltase-glucoamylase). In human populations, five genes were targeted by positive selection acting on noncoding variants within regulatory elements. Analysis of ancient DNA samples indicated that most derived alleles were already present in the Paleolithic. Positively selected variants at SLC2A5 (fructose transporter) were an exception and possibly spread following the domestication of specific fruit crops. We conclude that agriculture determined no major selective event at carbohydrate metabolism genes in humans, with implications for susceptibility to metabolic disorders. PMID:26319403

  14. Membrane-enhanced surface acoustic wave analysis of grafted polymer brushes

    NASA Astrophysics Data System (ADS)

    Brass, David A.; Shull, Kenneth R.

    2008-04-01

    An analysis is developed for the frequency response of a quartz crystal resonator (often referred to as a quartz crystal microbalance) that is modified with a grafted solvent-swollen polymer brush and placed in contact with a membrane capping layer. The shear wave generated at the resonator surface couples into the membrane layer with an efficiency that is strongly dependent on the thickness of the swollen brush layer. As a result, the resonant frequency changes by a maximum amount that is closely approximated by the Sauerbrey shift for the capping layer. The calculated shift substantially decreases for increases in the brush thickness of approximately 10nm, which gives a net frequency response that is extremely sensitive to the degree of swelling of the polymer brush. An optimum capping layer thickness is determined by balancing the Sauerbrey shift against dissipative effects that weaken the crystal resonance. This optimum membrane thickness depends only weakly on the properties of the membrane material and is in the micron range. Detailed multilayer calculations are presented for the specific case of a poly(ethylene glycol) brush swollen with water and brought into contact with an elastomeric water-permeable membrane. These calculations confirm that the method is sensitive to the properties of the brush layer in the experimentally relevant thickness regime. Connections are also made to conceptually simpler two and three layer models of the acoustic impedance of the material systems that are brought into contact with the resonator.

  15. The Responses of Rat Intestinal Brush Border and Cytosol Peptide Hydrolase Activities to Variation in Dietary Protein Content DIETARY REGULATION OF INTESTINAL PEPTIDE HYDROLASES

    PubMed Central

    Nicholson, J. Alex; McCarthy, Denis M.; Kim, Young S.

    1974-01-01

    The effects of variation in dietary protein content on small intestinal brush border and cytosol peptide hydrolase activities have been investigated. One group of rats was fed a high protein diet (55% casein) and another group was fed a low protein diet (10% casein). After 1 wk, brush border peptide hydrolase activity (L-leucyl-β-naphthylamide as substrate) and cytosol peptide hydrolase activity (L-prolyl-L-leucine as substrate) were determined in mucosae taken from the proximal, middle, and distal small intestine. As judged by several parameters, brush border peptide hydrolase activity was significantly greater in rats fed the high protein diet when data for corresponding segments were compared. In contrast, no significant difference was seen in cytosol peptide hydrolase activity. In a second study, brush border and cytosol peptide hydrolase activities were determined in the proximal intestine by utilizing an additional three peptide substrates: L-leucyl-L-alanine, L-phenylalanylglycine, and glycyl-L-phenylalanine. Sucrase, maltase, and alkaline phosphatase activities were also determined. As before, brush border peptide hydrolase activities were significantly greater in rats fed the high protein diet. However, activities of the nonproteolytic brush border enzymes did not vary significantly with diet. In contrast to the results obtained with L-prolyl-L-leucine as substrate for the cytosol enzymes, cytosol activity against the three additional peptide substrates was greater in rats fed the high protein diet. It is suggested that the brush border peptide hydrolase response to variation in dietary protein content represents a functional adaptation analogous to the regulation of intestinal disaccharidases by dietary carbohydrates. The implication of the differential responses of the cytosol peptide hydrolases is uncertain, since little is known of the functional role of these nonorgan-specific enzymes. PMID:4430719

  16. Borders as membranes :metaphors and models for improved policy in border regions.

    SciTech Connect

    Malczynski, Leonard A.; Passell, Howard David; Forster, Craig B.; Cockerill, Kristan

    2005-10-01

    Political borders are controversial and contested spaces. In an attempt to better understand movement along and through political borders, this project applied the metaphor of a membrane to look at how people, ideas, and things ''move'' through a border. More specifically, the research team employed this metaphor in a system dynamics framework to construct a computer model to assess legal and illegal migration on the US-Mexico border. Employing a metaphor can be helpful, as it was in this project, to gain different perspectives on a complex system. In addition to the metaphor, the multidisciplinary team utilized an array of methods to gather data including traditional literature searches, an experts workshop, a focus group, interviews, and culling expertise from the individuals on the research team. Results from the qualitative efforts revealed strong social as well as economic drivers that motivate individuals to cross the border legally. Based on the information gathered, the team concluded that legal migration dynamics were of a scope we did not want to consider hence, available demographic models sufficiently capture migration at the local level. Results from both the quantitative and qualitative data searches were used to modify a 1977 border model to demonstrate the dynamic nature of illegal migration. Model runs reveal that current US-policies based on neo-classic economic theory have proven ineffective in curbing illegal migration, and that proposed enforcement policies are also likely to be ineffective. We suggest, based on model results, that improvement in economic conditions within Mexico may have the biggest impact on illegal migration to the U.S. The modeling also supports the views expressed in the current literature suggesting that demographic and economic changes within Mexico are likely to slow illegal migration by 2060 with no special interventions made by either government.

  17. An all-aqueous route to polymer brush-modified membranes with remarkable permeabilites and protein capture rates

    PubMed Central

    Anuraj, Nishotha; Bhattacharjee, Somnath; Geiger, James H.; Baker, Gregory L.; Bruening, Merlin L.

    2011-01-01

    Microporous membranes are attractive for protein purification because convection rapidly brings proteins to binding sites. However, the low binding capacity of such membranes limits their applications. This work reports a rapid, aqueous procedure to create highly permeable, polymer brush-modified membranes that bind large amounts of protein. The synthetic method includes a 10-min adsorption of a macroinitiator in a hydroxylated nylon membrane and a subsequent 5-min aqueous atom transfer radical polymerization of 2-(methacryloyloxy)ethyl succinate from the immobilized initiator to form poly(acid) brushes. This procedure likely leads to more swollen, less dense brushes than polymerization from silane initiators, and thus requires less polymer to achieve the same binding capacity. The hydraulic permeability of the poly(acid) membranes is 4-fold higher than that of similar membranes prepared by growing brushes from immobilized silane initiators. These brush-containing nylon membranes bind 120 mg/cm3 of lysozyme using solution residence times as short as 35 ms, and when functionalized with nitrilotriacetate (NTA)-Ni2+ complexes, they capture 85 mg/cm3 of histidine6-tagged (His-tagged) Ubiquitin. Additionally the NTA-Ni2+-functionalized membranes isolate His-tagged myo-inositol-1-phosphate synthase directly from cell extracts and show >90% recovery of His-tagged proteins. PMID:22287817

  18. Ultrathin, freestanding, stimuli-responsive, porous membranes from polymer hydrogel-brushes

    NASA Astrophysics Data System (ADS)

    Kang, Chengjun; Ramakrishna, Shivaprakash N.; Nelson, Adrienne; Cremmel, Clement V. M.; Vom Stein, Helena; Spencer, Nicholas D.; Isa, Lucio; Benetti, Edmondo M.

    2015-07-01

    The fabrication of freestanding, sub-100 nm-thick, pH-responsive hydrogel membranes with controlled nano-morphology, based on modified poly(hydroxyethyl methacrylate) (PHEMA) is presented. Polymer hydrogel-brush films were first synthesized by surface-initiated atom transfer radical polymerization (SI-ATRP) and subsequently detached from silicon substrates by UV-induced photo-cleavage of a specially designed linker within the initiator groups. The detachment was also assisted by pH-induced osmotic forces generated within the films in the swollen state. The mechanical properties and morphology of the freestanding films were studied by atomic force microscopy (AFM). Inclusion of nanopores of controlled diameter was accomplished by performing SI-ATRP from initiator-coated surfaces that had previously been patterned with polystyrene nanoparticles. Assembly parameters and particle sizes could be varied, in order to fabricate nanoporous hydrogel-brush membranes with tunable pore coverage and characteristics. Additionally, due to the presence of weak polyacid functions within the hydrogel, the membranes exhibited pH-dependent thickness in water and reversible opening/closing of the pores.The fabrication of freestanding, sub-100 nm-thick, pH-responsive hydrogel membranes with controlled nano-morphology, based on modified poly(hydroxyethyl methacrylate) (PHEMA) is presented. Polymer hydrogel-brush films were first synthesized by surface-initiated atom transfer radical polymerization (SI-ATRP) and subsequently detached from silicon substrates by UV-induced photo-cleavage of a specially designed linker within the initiator groups. The detachment was also assisted by pH-induced osmotic forces generated within the films in the swollen state. The mechanical properties and morphology of the freestanding films were studied by atomic force microscopy (AFM). Inclusion of nanopores of controlled diameter was accomplished by performing SI-ATRP from initiator-coated surfaces that had

  19. Ultrathin, freestanding, stimuli-responsive, porous membranes from polymer hydrogel-brushes.

    PubMed

    Kang, Chengjun; Ramakrishna, Shivaprakash N; Nelson, Adrienne; Cremmel, Clement V M; vom Stein, Helena; Spencer, Nicholas D; Isa, Lucio; Benetti, Edmondo M

    2015-08-14

    The fabrication of freestanding, sub-100 nm-thick, pH-responsive hydrogel membranes with controlled nano-morphology, based on modified poly(hydroxyethyl methacrylate) (PHEMA) is presented. Polymer hydrogel-brush films were first synthesized by surface-initiated atom transfer radical polymerization (SI-ATRP) and subsequently detached from silicon substrates by UV-induced photo-cleavage of a specially designed linker within the initiator groups. The detachment was also assisted by pH-induced osmotic forces generated within the films in the swollen state. The mechanical properties and morphology of the freestanding films were studied by atomic force microscopy (AFM). Inclusion of nanopores of controlled diameter was accomplished by performing SI-ATRP from initiator-coated surfaces that had previously been patterned with polystyrene nanoparticles. Assembly parameters and particle sizes could be varied, in order to fabricate nanoporous hydrogel-brush membranes with tunable pore coverage and characteristics. Additionally, due to the presence of weak polyacid functions within the hydrogel, the membranes exhibited pH-dependent thickness in water and reversible opening/closing of the pores. PMID:26169114

  20. Molecular characterization of a family of metalloendopeptidases from the intestinal brush border of Haemonchus contortus.

    PubMed

    Newlands, G F J; Skuce, P J; Nisbet, A J; Redmond, D L; Smith, S K; Pettit, D; Smith, W D

    2006-09-01

    Substantial protection against the economically important parasitic nematode Haemonchus contortus has been achieved by immunizing sheep with a glycoprotein fraction isolated from the intestinal membranes of the worm (H-gal-GP). Previous studies showed that one of the major components of H-gal-GP is a family of at least 4 zinc metalloendopeptidases, designated MEPs 1-4. This paper describes aspects of the molecular architecture of this protease family, including the proteomic analysis of the MEP fraction of the H-gal-GP complex. These enzymes belong to the M13 zinc metalloendopeptidase family (EC 3.4.24.11), also known as neutral endopeptidases or neprilysins. The sequences of MEPs 1 and 3 suggested a typical Type II integral membrane protein structure, whilst MEPs 2 and 4 had putative cleavable signal peptides, typical of secreted proteins. Proteomic analysis of H-gal-GP indicated that the extracellular domain of all 4 MEPs had been cleaved close to the transmembrane region/signal peptide with additional cleavage sites mid-way along the polypeptide. MEP3 was present as a homo-dimer in H-gal-GP, whereas MEP1 or MEP2 formed hetero-dimers with MEP4. It was found that expression of MEP3 was confined to developing 4th-stage larvae and to adult worms, the stages of Haemonchus which feed on blood. MEP-like activity was detected in the H-gal-GP complex over a broad pH range (5-9). Since all 4 MEPs must share a similar microenvironment in the complex, this suggests that each might have a different substrate specificity. PMID:16740178

  1. Effect of nutritional rehabilitation on the development of intestinal brush border disaccharidases of postnatally malnourished weanling rats.

    PubMed

    Rossi, T M; Lee, P C; Young, C M; Lerner, A; Lebenthal, E

    1986-08-01

    The reversibility of the effects of postnatal malnutrition on the intestinal brush border enzymes and somatic and intestinal weights were examined using either ad libitum or restricted feedings. Malnutrition was induced in the immediate postnatal period by expanding newborn rat litters to 20 pups/dam. At 21 days of age, malnourished pups exhibited significantly decreased body and intestinal weights as compared to those from control litters. Malnourished pups also had significantly elevated lactase specific activities whereas sucrase and maltase activities were not affected in the proximal small intestine. With subsequent nutritional rehabilitation by an ad libitum (food available 24 h/day) or restricted feeding regimen (food available 2 h/day), body and intestinal weights remained significantly depressed by 56 days in malnourished as compared to control animals. Rats on 2-h feedings consumed approximately 35% of the food consumed by their ad libitum-fed counterparts. Comparison of the ratio of weight gained to the amount of food consumed did not demonstrate a greater food efficiency with any particular feeding pattern. With ad libitum or restricted feedings, lactase specific activity in the proximal segment attained control values by 14 days. Restricted feedings resulted in an apparent elevation of specific activity of sucrase and of maltase, when rats were sacrificed at one chosen time point. Multiple time studies in a 24-h cycle showed that maximal elevations in enzyme activities were associated with feeding time. There were no significant differences in mean specific daily enzyme activities between the two feeding regimens. Restricted feedings show no advantage in enzyme efficiency or in promoting the rate of recovery of the intestine after postnatal malnutrition. PMID:3090509

  2. Smart polymer brush nanostructures guide the self-assembly of pore-spanning lipid bilayers with integrated membrane proteins

    NASA Astrophysics Data System (ADS)

    Wilhelmina de Groot, G.; Demarche, Sophie; Santonicola, M. Gabriella; Tiefenauer, Louis; Vancso, G. Julius

    2014-01-01

    Nanopores in arrays on silicon chips are functionalized with pH-responsive poly(methacrylic acid) (PMAA) brushes and used as supports for pore-spanning lipid bilayers with integrated membrane proteins. Robust platforms are created by the covalent grafting of polymer brushes using surface-initiated atom transfer radical polymerization (ATRP), resulting in sensor chips that can be successfully reused over several assays. His-tagged proteins are selectively and reversibly bound to the nitrilotriacetic acid (NTA) functionalization of the PMAA brush, and consequently lipid bilayer membranes are formed. The enhanced membrane resistance as determined by electrochemical impedance spectroscopy and free diffusion of dyed lipids observed as fluorescence recovery after photobleaching confirmed the presence of lipid bilayers. Immobilization of the His-tagged membrane proteins on the NTA-modified PMAA brush near the pore edges is characterized by fluorescence microscopy. This system allows us to adjust the protein density in free-standing bilayers, which are stabilized by the polymer brush underneath. The potential application of the integrated platform for ion channel protein assays is demonstrated.

  3. Fabrication of cell outer membrane mimetic polymer brush on polysulfone surface via RAFT technique

    NASA Astrophysics Data System (ADS)

    Ma, Qian; Zhang, Hui; Zhao, Jiang; Gong, Yong-Kuan

    2012-10-01

    Cell membrane mimetic antifouling polymer brush was grown on polysulfone (PSF) membrane by surface-induced reversible addition-fragmentation chain transfer (RAFT) polymerization of 2-methacryloyloxyethyl phosphorylcholine (MPC). The RAFT agent immobilized PSF substrate was prepared by successive chloromethylation, amination with ethylenediamine (EDA) and amidation of the amine group of grafted EDA with the carboxylic group of 4-cyanopentanoic acid dithiobenzoate (CPAD). The surface RAFT polymerization of MPC was initiated in aqueous solution by 4,4‧-azobis-4-cyanopentanoic acid (ACPA). The formation of PMPC brush coating is evidenced by X-ray photoelectron spectroscopy and water contact angle measurements. The degree of polymerization of PMPC and the polymer grafting density were calculated from the high resolution XPS spectra. The platelet adhesion and protein adsorption results showed that the PMPC-grafted PSF surface has excellent antifouling ability to resist platelet adhesion completely and suppress protein adsorption significantly. This biomimetic and bio-friendly surface RAFT polymerization strategy could be promising for a variety of biomedical applications.

  4. Constructing proton-conductive highways within an ionomer membrane by embedding sulfonated polymer brush modified graphene oxide

    NASA Astrophysics Data System (ADS)

    Zhao, Liping; Li, Yifan; Zhang, Haoqin; Wu, Wenjia; Liu, Jindun; Wang, Jingtao

    2015-07-01

    Sulfonated polymer brush modified graphene oxide (SP-GO) fillers with controllable brush length are synthesized via the facile distillation-precipitation polymerization, and then incorporated into sulfonated poly(ether ether ketone) (SPEEK) matrix to fabricate composite membranes. The influences of SP-GO upon the microstructures, including thermal and mechanical properties, water uptake/swelling, proton conduction, H2 permeability and single PEMFC performances of composite membranes are intensively investigated. It is found that the SP-GO fillers are uniformly dispersed and tend to lie perpendicularly to the cross-section surface of the whole membrane, which allow SP-GO fillers creating inter-connected and broad ionic pathways through the sulfonic acid groups in polymer brushes. Meanwhile, the SP-GO fillers connect the ionic clusters in SPEEK matrix via interfacial interactions. In such a way, proton-transfer highways are constructed along the SPEEK/SP-GO interface, which lower the proton transfer activation energy and enhance the proton conductivities of the composite membranes under both hydrated and anhydrous conditions. Furthermore, elevating the brush length on SP-GO could further enhance the proton conductivity. Compared to SPEEK control membrane, a 95.5% increase in hydrated conductivity, an 178% increase in anhydrous conductivity and a 37% increase in maximum power density are obtained for the optimal composite membrane.

  5. Liquid Crystalline Block Copolymers with Brush Type Architecture: Toward Functional Membranes by Magnetic Field Alignment

    NASA Astrophysics Data System (ADS)

    Choo, Youngwoo; Gopinadhan, Manesh; Mahajan, Lalit; Kasi, Rajeswari; Osuji, Chinedum

    2015-03-01

    We introduce a novel liquid crystalline block copolymer with brush type architecture for membrane applications by magnetic field directed self-assembly. Ring-opening metathesis of n-alkyloxy cyanobiphenyl and polylactide (PLA) functionalized norbornene monomers provides efficient polymerization yielding low polydispersity block copolymers. The molecular weight of the PLA side chains, spacer length of the cyanobiphenyl mesogens are systematically varied to form well-ordered BCP morphologies at varying volume fractions. Interestingly, the system features morphology dependent anchoring condition where mesogens adopt planar anchoring on cylindrical interface while homeotropic anchoring was preferred on a planar block interface. The minority PLA domains from highly aligned materials can be readily degraded by hydrolysis to produce vertically aligned nanoporous polymer films which exhibit reversible thermal switching behavior. The polymers introduced here provide a versatile platform for scalable fabrication of aligned membranes and further functional materials based on such templates. This work was supported by NSF(CCMI-1246804).

  6. Short-term fasting induces intra-hepatic lipid accumulation and decreases intestinal mass without reduced brush-border enzyme activity in mink (Mustela vison) small intestine.

    PubMed

    Bjornvad, C R; Elnif, J; Sangild, P T

    2004-11-01

    For many mammalian species short-term fasting is associated with intestinal atrophy and decreased digestive capacity. Under natural conditions, strictly carnivorous animals often experience prey scarcity during winter, and they may therefore be particularly well adapted to short-term food deprivation. To examine how the carnivorous gastrointestinal tract is affected by fasting, small-intestinal structure, brush-border enzyme activities and hepatic structure and function were examined in fed mink (controls) and mink that had been fasted for 1-10 days. During the first 1-2 days of fasting, intestinal mass decreased more rapidly than total body mass and villus heights were reduced 25-40%. In contrast, tissue-specific activity of the brush-border enzymes sucrase, maltase, lactase, aminopeptidase A and dipeptidylpeptidase IV increased 0.5- to 1.5-fold at this time, but returned to prefasting levels after 6 days of fasting. After 6-10 days of fasting there was a marked increase in the activity of hepatic enzymes and accumulation of intra-hepatic lipid vacuoles. Thus, mink may be a useful model for studying fasting-induced intestinal atrophy and adaptation as well as mechanisms involved in accumulation of intra-hepatic lipids following food deprivation in strictly carnivorous domestic mammals, such as cats and ferrets. PMID:15503054

  7. Affinity Separation of Lectins Using Porous Membranes Immobilized with Glycopolymer Brushes Containing Mannose or N-Acetyl-d-Glucosamine

    PubMed Central

    Ogata, Yutaro; Seto, Hirokazu; Murakami, Tatsuya; Hoshino, Yu; Miura, Yoshiko

    2013-01-01

    Porous membranes with glycopolymer brushes were prepared as biomaterials for affinity separation. Glycopolymer brushes contained acrylic acid and D-mannose or N-acetyl-D-glucosamine, and were formed on substrates by surface-initiated atom transfer radical polymerization. The presence of glycopolymer brush was confirmed by X-ray photoelectron spectroscopy, contact angle, and ellipsometry measurements. The interaction between lectin and the glycopolymer immobilized on glass slides was confirmed using fluorescent-labeled proteins. Glycopolymer-immobilized surfaces exhibited specific adsorption of the corresponding lectin, compared with bovine serum albumin. Lectins were continuously rejected by the glycopolymer-immobilized membranes. When the protein solution was permeated through the glycopolymer-immobilized membrane, bovine serum albumin was not adsorbed on the membrane surface. In contrast, concanavalin A and wheat germ agglutinin were rejected by membranes incorporating D-mannose or N-acetyl-D-glucosamine, respectively. The amounts of adsorbed concanavalin A and wheat germ agglutinin was increased five- and two-fold that of adsorbed bovine serum albumin, respectively. PMID:24956944

  8. Differential effect of Bacillus firmus on immune response and enterocyte brush-border enzyme levels in BALB/c and B10.BR mice.

    PubMed

    Kozáková, H; Mlcková, P; Kolínská, J; Cechová, D; Stĕpánková, R; Reháková, Z; Prokesová, L

    2002-01-01

    A nonpathogenic bacterium of external environment possessing remarkable immunomodulatory activity, Bacillus firmus (BF) inactivated with formaldehyde, was given intragastrically to two genetically different mouse strains BALB/c (H-2d) and B10.BR/SnPh (B10.BR, H-2k) reared in conventional (CV) and B10.BR strain also in germ-free (GF) conditions. Repeated intragastric administration of BF (500 micrograms every other day over two weeks, starting at the age of 3 months) significantly enhanced intestinal IgA levels in CV BALB/c mice but did not affect intestinal IgA in CV B10.BR mice. In GF B10.BR mice, IgG levels in sera and intestinal washings increased after BF administration compared to CV B10.BR mice. In CV BALB/c mice, specific activity of enterocyte brush-border enzymes (lactase, gamma-glutamyltransferase, alkaline phosphatase) decreased after BF treatment; sucrase (sucrose alpha-glucosidase) activity was not affected. On the other hand, in B10.BR mice, specific activity of gamma-glutamyltransferase and dipeptidyl peptidase IV were higher after administration of BF in both CV and GF groups relative to untreated controls. The activities of lactase and glucoamylase (glucan 1,4-alpha-glucosidase) were significantly stimulated only in the group of GF B10.BR mice treated with formolized BF. The stimulation of immunoglobulin production after BF treatment was accompanied by changes in the levels of enterocyte brush-border enzymes; this responsiveness to BF treatment was genetically regulated. PMID:12630333

  9. Effect of bacterial monoassociation on brush-border enzyme activities in ex-germ-free piglets: comparison of commensal and pathogenic Escherichia coli strains.

    PubMed

    Kozakova, Hana; Kolinska, Jirina; Lojda, Zdenek; Rehakova, Zuzana; Sinkora, Jiri; Zakostelecka, Marie; Splichal, Igor; Tlaskalova-Hogenova, Helena

    2006-09-01

    This study was designed to investigate the effect of monoassociation of germ-free piglets with Escherichia coli strains on the development of intestinal brush-border enzyme activities. Piglets were delivered by hysterectomy, reared for seven days under germ-free conditions and fed milk formula diet. One group was maintained germ-free, the other four groups were monoassociated on day eight with one of four E. coli strains: non-pathogenic O86 or O83 and G58-1, or pathogenic 933D. The development of brush-border digestive enzyme functions in the small intestine was evaluated after 15 days. Germ-free controls exhibited slower developmental declines of lactase, gamma-glutamyltranspeptidase and alkaline phosphatase, and delayed increases of sucrase and glucoamylase compared to conventionally grown animals. Association of germ-free piglets with the non-pathogenic E. coli strains O86 and O83 resulted in increased enterocyte differentiation along the length of the small intestine, accompanied by declining activities of lactase, gamma-glutamyltranspeptidase and alkaline phosphatase, and elevated activities of maturational markers such as sucrase and glucoamylase. Maturational changes also occurred along the villus-crypt axis, as revealed by histochemical localization of aminopeptidase N on the villi tips in piglets colonized with E. coli O83. Interestingly, colonization with the pathogenic E. coli strain 933D stimulated changes in the main differentiation enzyme markers lactase, sucrase and glucoamylase to an extent comparable with those produced by the non-pathogenic and probiotic E. coli strains. In conclusion, germ-free piglets represent a valuable tool to study the consequences of colonization of the immature sterile gut with defined strains of bacteria. PMID:16949322

  10. FERRIREDUCTASE ACTIVITY IN LOBSTER HEPATOPANCREATIC BRUSH BORDER MEMBRANE VESICLES (BBMV). (R823068)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  11. ZINC UPTAKE BY LOBSTER HEPATOPANCREATIC BRUSH BORDER MEMBRANE VESICLES. (R823068)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  12. CALCIUM TRANSPORT PROCESSES OF LOBSTER HEPATOPANCREATIC BRUSH BORDER MEMBRANE VESICLES. (R823068)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  13. New poly(amino acid methacrylate) brush supports the formation of well-defined lipid membranes.

    PubMed

    Blakeston, Anita C; Alswieleh, Abdullah M; Heath, George R; Roth, Johannes S; Bao, Peng; Cheng, Nan; Armes, Steven P; Leggett, Graham J; Bushby, Richard J; Evans, Stephen D

    2015-03-31

    A novel poly(amino acid methacrylate) brush comprising zwitterionic cysteine groups (PCysMA) was utilized as a support for lipid bilayers. The polymer brush provides a 12-nm-thick cushion between the underlying hard support and the aqueous phase. At neutral pH, the zeta potential of the PCysMA brush was ∼-10 mV. Cationic vesicles containing >25% DOTAP were found to form a homogeneous lipid bilayer, as determined by a combination of surface analytical techniques. The lipid mobility as measured by FRAP (fluorescence recovery after photobleaching) gave diffusion coefficients of ∼1.5 μm(2) s(-1), which are comparable to those observed for lipid bilayers on glass substrates. PMID:25746444

  14. New Poly(amino acid methacrylate) Brush Supports the Formation of Well-Defined Lipid Membranes

    PubMed Central

    2015-01-01

    A novel poly(amino acid methacrylate) brush comprising zwitterionic cysteine groups (PCysMA) was utilized as a support for lipid bilayers. The polymer brush provides a 12-nm-thick cushion between the underlying hard support and the aqueous phase. At neutral pH, the zeta potential of the PCysMA brush was ∼−10 mV. Cationic vesicles containing >25% DOTAP were found to form a homogeneous lipid bilayer, as determined by a combination of surface analytical techniques. The lipid mobility as measured by FRAP (fluorescence recovery after photobleaching) gave diffusion coefficients of ∼1.5 μm2 s–1, which are comparable to those observed for lipid bilayers on glass substrates. PMID:25746444

  15. Grafting of carboxybetaine brush onto cellulose membranes via surface-initiated ARGET-ATRP for improving blood compatibility.

    PubMed

    Wang, Miao; Yuan, Jiang; Huang, Xiaobo; Cai, Xianmei; Li, Li; Shen, Jian

    2013-03-01

    Grafting-from has proven to be a very effective way to create high grafting densities and well-controlled polymer chains on different kinds of surfaces. In this work, we aim to graft zwitterionic brush from cellulose membrane (CM) via ARGET-ATRP (Activator Regenerated by Electron Transfer ATRP) method indirectly for blood compatibility improvement. Characterization of the CM substrates before and after modification was carried out by attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR), water contact angle measurements, X-ray photoelectron spectroscopy analysis, and atomic force microscopy, respectively. The results demonstrated zwitterionic brushes were successfully grafted on the CM surfaces, and the content of the grafted layer increased gradually with the polymerization time. The platelet adhesion, hemolytic test and plasma protein adsorption results indicated the cellulose membrane had significantly excellent blood compatibility featured on lower platelet adhesion and protein adsorption without causing hemolysis. The functionalized cellulose substrate could have a great potential usage for biomedical applications. PMID:23201719

  16. Sucrase-isomaltase deficiency in humans. Different mutations disrupt intracellular transport, processing, and function of an intestinal brush border enzyme.

    PubMed Central

    Naim, H Y; Roth, J; Sterchi, E E; Lentze, M; Milla, P; Schmitz, J; Hauri, H P

    1988-01-01

    Eight cases of congenital sucrase-isomaltase deficiency were studied at the subcellular and protein level with monoclonal antibodies against sucrase-isomaltase. At least three phenotypes were revealed: one in which sucrase-isomaltase protein accumulated intracellularly probably in the endoplasmic reticulum, as a membrane-associated high-mannose precursor, one in which the intracellular transport of the enzyme was apparently blocked in the Golgi apparatus, and one in which catalytically altered enzyme was transported to the cell surface. All patients expressed electrophoretically normal or near normal high-mannose sucrase-isomaltase. The results suggest that different, probably small, mutations in the sucrase-isomaltase gene lead to the synthesis of transport-incompetent or functionally altered enzyme which results in congenital sucrose intolerance. Images PMID:3403721

  17. Membrane distribution of sodium-hydrogen and chloride-bicarbonate exchangers in crypt and villus cell membranes from rabbit ileum.

    PubMed Central

    Knickelbein, R G; Aronson, P S; Dobbins, J W

    1988-01-01

    Present evidence suggests that in the small intestine, villus cells are primarily absorptive and crypt cells are primarily secretory. In order to further confirm that there are differences in transport properties between villus and crypt cells, we have separated villus from crypt cells, using calcium chelations techniques, and determined the distribution of Na:H and Cl:HCO3 exchange activity on brush border membrane and basolateral membrane preparations from these two cell populations. Separation of cells was determined utilizing alkaline phosphatase and maltase activity as a marker of villus cells and thymidine kinase activity as a marker of crypt cells. Utilizing these techniques, we were able to sequentially collect cells along the villus-crypt axis. Na-stimulated glucose and alanine uptake in brush border membrane vesicles diminished from the villus to the crypt region in the sequentially collected cells fractions, further suggesting separation of these cells. Brush border and basolateral membranes were then prepared from cells from the villus and crypt areas, utilizing a continuous sucrose gradient. In the villus cells, Na:H exchange activity was found associated with both the brush border and basolateral membrane, whereas, in crypt cells, Na:H exchange activity was only found on the basolateral membrane. Cl:HCO3 exchange activity was found only on the brush border membrane, in both villus and crypt cells. These studies suggest functional heterogeneity in ion transport between villus and crypt cells. PMID:2848868

  18. Humic acid removal and easy-cleanability using temperature-responsive ZrO2 tubular membranes grafted with poly(N-isopropylacrylamide) brush chains.

    PubMed

    Zhao, Yijiang; Zhou, Shouyong; Li, Meisheng; Xue, Ailian; Zhang, Yan; Wang, Jingang; Xing, Weihong

    2013-05-01

    New poly(N-isopropylacrylamide) brushes grafted with ZrO2 (PNIPAAm-g-ZrO2) composite membranes, which had been prepared in our laboratory, were used for humic acid (HA) removal. We found that the fluxes associated with such membranes, when compared to those obtained from unmodified ZrO2 membranes, declined slightly at both 25 °C and 35 °C. The PNIPAAm-g-ZrO2 membrane achieved a high rejection, of 98.0%, at a suitable steady flux of 111.9 L m(-2) h(-1) at 25 °C. This membrane exhibited good anti-fouling properties as well as improved membrane performance during filtration of HA. The important role of pH and Ca(2+) concentration in HA removal was also investigated. Lower adsorption fouling and a higher rejection were obtained at higher pH levels. The Ca(2+) ions reduced the electrostatic exclusion and played a cross-linking role between HA and the PNIPAAm-g-ZrO2 membrane surface. Fouling was severe in the presence of Ca(2+). These tests led to the development of an environment-friendly membrane cleaning method, by means of temperature-change water elution around LCST of PNIPAAm-brushes. After the alternate temperature-change (25 °C/35 °C) cleaning, a flux recovery of 98.2% was obtained for the PNIPAAm-g-ZrO2 membrane. Moreover, after four repeated experiments, the anti-fouling and easy-cleaning properties were still maintained. It is implied that PNIPAAm-brushes were firmly "stuck" to the membrane surface, and could not easily be removed by water cleaning or HA filtration. The PNIPAAm-g-ZrO2 membranes exhibited good stability and great potential for HA removal. PMID:23466218

  19. Proper Brushing

    MedlinePlus

    ... 3 teeth using a vibrating back & forth rolling motion. A rolling motion is when the brush makes contact with the ... gumline. Gently brush using back, forth, and rolling motion along all of the inner tooth surfaces. Tilt ...

  20. Endothelial Src kinase regulates membrane recycling from the lateral border recycling compartment during leukocyte transendothelial migration.

    PubMed

    Dasgupta, Bidisha; Muller, William A

    2008-12-01

    When leukocytes cross endothelial cells during the inflammatory response, membrane from the recently described lateral border recycling compartment (LBRC) is selectively targeted around diapedesing leukocytes. This "targeted recycling" is critical for leukocyte transendothelial migration. Blocking homophilic PECAM interactions between leukocytes and endothelial cells blocks targeted recycling from the LBRC and blocks diapedesis. However, the cellular signaling pathways that trigger targeted recycling are not known. We show that targeted recycling from the LBRC is dependent on Src kinase. The selective Src kinase inhibitor PP2 blocked targeted recycling and blocked diapedesis by over 70%. However, Src kinase inhibition did not affect the structure or normal constitutive recycling of membrane from the LBRC in the absence of leukocytes. PECAM, a Src kinase substrate, traffics between the LBRC and the endothelial surface at the cell border. However, virtually all of the PECAM in the cell that was phosphorylated on tyrosine residues was found in the LBRC. These findings demonstrate that Src kinase activity is critical for the targeted recycling of membrane from the LBRC to the site of transendothelial migration and that the PECAM in the LBRC is qualitatively different from the PECAM on the surface of endothelial cells. PMID:18991269

  1. The apical membrane of intestinal brush cells possesses a specialised, but species-specific, composition of glycoconjugates--on-section and in vivo lectin labelling in rats, guinea-pigs and mice.

    PubMed

    Gebert, A; al-Samir, K; Werner, K; Fassbender, S; Gebhard, A

    2000-05-01

    Brush cells are specialised epithelial cells that are assumed to represent chemoreceptors of the digestive tract. They comprise a small population of the epithelial cells lining the intestine, possess a unique ultrastructure and, in many aspects, resemble the receptor cells of taste buds. To characterise glycoconjugates possibly involved in a sensory function, we investigated brush cells in the small intestine of three species using lectin histochemistry in confocal light and thin-section electron microscopy. Brush cells of rats were selectively labelled by the sialic acid-specific lectin Maackia amurensis agglutinin, those of guinea-pigs by the D-galactose-specific lectin Bandeiraea simplicifolia agglutinin, isolectin B4 and those of mice by the L-fucose-specific lectin Ulex europaeus agglutinin lectin I. Lectin binding sites were consistently located in the glycocalyx of the apical membrane and in that of cytoplasmic vesicles. In vivo lectin labelling revealed that the glycoconjugates of the apical membrane are accessible under physiological conditions, that brush cells do not endocytose and that they probably possess a high membrane turnover rate. The results show that specialisations exist in the composition of glycoconjugates forming the glycocalyx of brush cells in all species investigated. The presence of brush cell-specific glycoconjugates would be in accordance with the current hypothesis of a receptive function of brush cells. Differences in the specific glycosylation patterns among rats, guinea-pigs and mice indicate that species-specific adaptations exist. PMID:10883398

  2. Inhibition of renal membrane binding and nephrotoxicity of aminoglycosides

    SciTech Connect

    Williams, P.D.; Hottendorf, G.H.; Bennett, D.B.

    1986-06-01

    The initial event in the renal tubular reabsorption of nephrotoxic aminoglycosides involves binding to brush border membranes. This primary event was measured in renal brush border membrane vesicles prepared from rat renal cortex utilizing (3H)gentamicin. In order to gain structure-activity information regarding this interaction the effect of substances having chemical similarities to aminoglycosides (sugars, polyamines and amino acids) on gentamicin binding to brush border membranes was determined. Polyamino acids were found to possess the greatest inhibitory potency. In addition to polymers of cationic amino acids (lysine, ornithine, arginine and histidine), polymers of neutral (asparagine) and acidic (aspartic and glutamic acid) amino acids also exhibited inhibition of the membrane binding of gentamicin. Inasmuch as inhibition of renal membrane binding has the potential to decrease aminoglycoside nephrotoxicity, several polyamino acids that inhibited membrane binding were tested in vivo for potential protective activity vs. gentamicin- and amikacin-induced nephrotoxicity. Polyasparagine90 and polyaspartic acid100 inhibited gentamicin and amikacin nephrotoxicity completely when coadministered to rats with the aminoglycosides. Polylysine20 provided complete and partial inhibition of gentamicin and amikacin nephrotoxicity, respectively. Whereas in vivo distribution studies revealed that cortical levels of (3H)amikacin were elevated slightly by the coadministration of polyaspartic acid, brush border and basolateral membranes contained significantly lower levels of the aminoglycoside (46 and 41% inhibition, respectively). These results question the role of charge per se in the binding of aminoglycosides to renal membranes and further confirm the importance of membrane binding in the pathogenesis of aminoglycoside nephrotoxicity.

  3. Hemocompatibility of poly(vinylidene fluoride) membrane grafted with network-like and brush-like antifouling layer controlled via plasma-induced surface PEGylation.

    PubMed

    Chang, Yung; Shih, Yu-Ju; Ko, Chao-Yin; Jhong, Jheng-Fong; Liu, Ying-Ling; Wei, Ta-Chin

    2011-05-01

    In this work, the hemocompatibility of PEGylated poly(vinylidene fluoride) (PVDF) microporous membranes with varying grafting coverage and structures via plasma-induced surface PEGylation was studied. Network-like and brush-like PEGylated layers on PVDF membrane surfaces were achieved by low-pressure and atmospheric plasma treatment. The chemical composition, physical morphology, grafting structure, surface hydrophilicity, and hydration capability of prepared membranes were determined to illustrate the correlations between grafting qualities and hemocompatibility of PEGylated PVDF membranes in contact with human blood. Plasma protein adsorption onto different PEGylated PVDF membranes from single-protein solutions and the complex medium of 100% human plasma were measured by enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies. Hemocompatibility of the PEGylated membranes was evaluated by the antifouling property of platelet adhesion observed by scanning electron microscopy (SEM) and the anticoagulant activity of the blood coagulant determined by testing plasma-clotting time. The control of grafting structures of PEGylated layers highly regulates the PVDF membrane to resist the adsorption of plasma proteins, the adhesion of platelets, and the coagulation of human plasma. It was found that PVDF membranes grafted with brush-like PEGylated layers presented higher hydration capability with binding water molecules than with network-like PEGylated layers to improve the hemocompatible character of plasma protein and blood platelet resistance in human blood. This work suggests that the hemocompatible nature of grafted PEGylated polymers by controlling grafting structures gives them great potential in the molecular design of antithrombogenic membranes for use in human blood. PMID:21449586

  4. CD2+ AND H+ STIMULATE UPTAKE OF 59FE2+ IN LOBSTER HEPATOPANCREATIC BRUSH BORDER MEMBRANE VESICLES. (R823068)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  5. ZINC STIMULATES L-PROLINE TRANSPORT BY THE IMINO SYSTEM IN LOBSTER HEPATOPANCREATIC BRUSH BORDER MEMBRANE VESICLES. (R823068)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  6. Ureteral retrograde brush biopsy

    MedlinePlus

    Biopsy - brush - urinary tract; Retrograde ureteral brush biopsy cytology; Cytology - ureteral retrograde brush biopsy ... to be biopsied is rubbed with the brush. Biopsy forceps may be used instead to collect a ...

  7. Secreted maltase-glucoamylase is upregulated in membrane maltase-glucoamylase null mice

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The known mammalian brush-border glucohydrolases are membrane-bound, small intestine lumenal enzymes that digest dietary starches and sugars into monosaccharides for absorption. The phenotype of maltase-glucoamylase deficiency is not understood. To address this, we ablated membrane-bound maltase-gl...

  8. Electrospun regenerated cellulose nanofibrous membranes surface-grafted with polymer chains/brushes via the atom transfer radical polymerization method for catalase immobilization.

    PubMed

    Feng, Quan; Hou, Dayin; Zhao, Yong; Xu, Tao; Menkhaus, Todd J; Fong, Hao

    2014-12-10

    In this study, an electrospun regenerated cellulose (RC) nanofibrous membrane with fiber diameters of ∼200-400 nm was prepared first; subsequently, 2-hydroxyethyl methacrylate (HEMA), 2-dimethylaminoethyl methacrylate (DMAEMA), and acrylic acid (AA) were selected as the monomers for surface grafting of polymer chains/brushes via the atom transfer radical polymerization (ATRP) method. Thereafter, four nanofibrous membranes (i.e., RC, RC-poly(HEMA), RC-poly(DMAEMA), and RC-poly(AA)) were explored as innovative supports for immobilization of an enzyme of bovine liver catalase (CAT). The amount/capacity, activity, stability, and reusability of immobilized catalase were evaluated, and the kinetic parameters (Vmax and Km) for immobilized and free catalase were determined. The results indicated that the respective amounts/capacities of immobilized catalase on RC-poly(HEMA) and RC-poly(DMAEMA) nanofibrous membranes reached 78 ± 3.5 and 67 ± 2.7 mg g(-1), which were considerably higher than the previously reported values. Meanwhile, compared to that of free CAT (i.e., 18 days), the half-life periods of RC-CAT, RC-poly(HEMA)-CAT, RC-poly(DMAEMA)-CAT, and RC-poly(AA)-CAT were 49, 58, 56, and 60 days, respectively, indicating that the storage stability of immobilized catalase was also significantly improved. Furthermore, the immobilized catalase exhibited substantially higher resistance to temperature variation (tested from 5 to 70 °C) and lower degree of sensitivity to pH value (tested from 4.0 and 10.0) than the free catalase. In particular, according to the kinetic parameters of Vmax and Km, the nanofibrous membranes of RC-poly(HEMA) (i.e., 5102 μmol mg(-1) min(-1) and 44.89 mM) and RC-poly(DMAEMA) (i.e., 4651 μmol mg(-1) min(-1) and 46.98 mM) had the most satisfactory biocompatibility with immobilized catalase. It was therefore concluded that the electrospun RC nanofibrous membranes surface-grafted with 3-dimensional nanolayers of polymer chains/brushes would be

  9. Modelling the mechanical behaviour of pit membranes in bordered pits with respect to cavitation resistance in angiosperms

    PubMed Central

    Tixier, Aude; Herbette, Stephane; Jansen, Steven; Capron, Marie; Tordjeman, Philippe; Cochard, Hervé; Badel, Eric

    2014-01-01

    Background and Aims Various correlations have been identified between anatomical features of bordered pits in angiosperm xylem and vulnerability to cavitation, suggesting that the mechanical behaviour of the pits may play a role. Theoretical modelling of the membrane behaviour has been undertaken, but it requires input of parameters at the nanoscale level. However, to date, no experimental data have indicated clearly that pit membranes experience strain at high levels during cavitation events. Methods Transmission electron microscopy (TEM) was used in order to quantify the pit micromorphology of four tree species that show contrasting differences in vulnerability to cavitation, namely Sorbus aria, Carpinus betulus, Fagus sylvatica and Populus tremula. This allowed anatomical characters to be included in a mechanical model that was based on the Kirchhoff–Love thin plate theory. A mechanistic model was developed that included the geometric features of the pits that could be measured, with the purpose of evaluating the pit membrane strain that results from a pressure difference being applied across the membrane. This approach allowed an assessment to be made of the impact of the geometry of a pit on its mechanical behaviour, and provided an estimate of the impact on air-seeding resistance. Key Results The TEM observations showed evidence of residual strains on the pit membranes, thus demonstrating that this membrane may experience a large degree of strain during cavitation. The mechanical modelling revealed the interspecific variability of the strains experienced by the pit membrane, which varied according to the pit geometry and the pressure experienced. The modelling output combined with the TEM observations suggests that cavitation occurs after the pit membrane has been deflected against the pit border. Interspecific variability of the strains experienced was correlated with vulnerability to cavitation. Assuming that air-seeding occurs at a given pit membrane

  10. Effects of Protease, Phytase and a Bacillus sp. Direct-Fed Microbial on Nutrient and Energy Digestibility, Ileal Brush Border Digestive Enzyme Activity and Cecal Short-Chain Fatty Acid Concentration in Broiler Chickens

    PubMed Central

    Murugesan, Ganapathi R.; Romero, Luis F.; Persia, Michael E.

    2014-01-01

    Two experiments were conducted to determine the effects of protease and phytase (PP) and a Bacillus sp. direct-fed microbial (DFM) on dietary energy and nutrient utilization in broiler chickens. In the first experiment, Ross 308 broiler chicks were fed diets supplemented with PP and DFM in a 2×2 factorial arrangement. The 4 diets (control (CON), CON + PP, CON + DFM, and CON + PP + DFM) were fed from 15–21 days of age. In Experiment 1, significant interaction (P≤0.01) between PP and DFM on the apparent ileal digestibility coefficient for starch, crude protein, and amino acid indicated that both additives increased the digestibility. Both additives increased the nitrogen retention coefficient with a significant interaction (P≤0.01). Although no interaction was observed, significant main effects (P≤0.01) for nitrogen-corrected apparent ME (AMEn) for PP or DFM indicated an additive response. In a follow-up experiment, Ross 308 broiler chicks were fed the same experimental diets from 1–21 days of age. Activities of ileal brush border maltase, sucrase, and L-alanine aminopeptidase were increased (P≤0.01) by PP addition, while a trend (P = 0.07) for increased sucrase activity was observed in chickens fed DFM, in Experiment 2. The proportion of cecal butyrate was increased (P≤0.01) by DFM addition. Increased nutrient utilization and nitrogen retention appear to involve separate but complementary mechanisms for PP and DFM, however AMEn responses appear to have separate and additive mechanisms. PMID:25013936

  11. Life at the border: Adaptation of proteins to anisotropic membrane environment

    PubMed Central

    Pogozheva, Irina D; Mosberg, Henry I; Lomize, Andrei L

    2014-01-01

    This review discusses main features of transmembrane (TM) proteins which distinguish them from water-soluble proteins and allow their adaptation to the anisotropic membrane environment. We overview the structural limitations on membrane protein architecture, spatial arrangement of proteins in membranes and their intrinsic hydrophobic thickness, co-translational and post-translational folding and insertion into lipid bilayers, topogenesis, high propensity to form oligomers, and large-scale conformational transitions during membrane insertion and transport function. Special attention is paid to the polarity of TM protein surfaces described by profiles of dipolarity/polarizability and hydrogen-bonding capacity parameters that match polarity of the lipid environment. Analysis of distributions of Trp resides on surfaces of TM proteins from different biological membranes indicates that interfacial membrane regions with preferential accumulation of Trp indole rings correspond to the outer part of the lipid acyl chain region—between double bonds and carbonyl groups of lipids. These “midpolar” regions are not always symmetric in proteins from natural membranes. We also examined the hydrophobic effect that drives insertion of proteins into lipid bilayer and different free energy contributions to TM protein stability, including attractive van der Waals forces and hydrogen bonds, side-chain conformational entropy, the hydrophobic mismatch, membrane deformations, and specific protein–lipid binding. PMID:24947665

  12. ANTIBODIES TO INTESTINAL MICROVILLOUS MEMBRANES

    PubMed Central

    Mackenzie, Iain L.; Donaldson, Robert M.; Kopp, William L.; Trier, Jerry S.

    1968-01-01

    Microvillous membranes isolated from the distal, but not proximal, half of hamster small bowel induced in rabbits the formation of antisera which inhibited intrinsic factor-mediated uptake of vitamin B12 by hamster brush borders. The extent of inhibition was directly proportional to the concentration of antiserum, and an excess of IF-bound vitamin B12 could overcome the inhibitory effect. The inhibitory factor was absorbed from antisera by brush borders isolated from the distal, but not proximal, half of the hamster intestine. Fractionation of antisera by gel filtration and DEAE-cellulose chromatography established that immunoglobulin G contained the inhibitory factor. Antisera capable of completely blocking uptake of IF-bound vitamin B12 did not react with hamster IF or with the IF-vitamin B12 complex, did not inhibit brush border disaccharidase activity and did not impair glucose transport by everted sacs of hamster intestine. These results demonstrate that an antibody to distal microvillous membranes competes with the IF-vitamin B12 complex for a specific binding site or receptor located on the surface of distal hamster intestine. PMID:19867301

  13. Brushing your infant's teeth

    MedlinePlus

    ... gov/ency/patientinstructions/000769.htm Brushing Your Child’s Teeth To use the sharing features on this page, ... how to brush their teeth on their own. Tooth and Gum Care for Young Children You should ...

  14. Impact of Electroviscosity on the Hydraulic Conductance of the Bordered Pit Membrane: A Theoretical Investigation1[C

    PubMed Central

    Santiago, Michael; Pagay, Vinay; Stroock, Abraham D.

    2013-01-01

    In perfusion experiments, the hydraulic conductance of stem segments () responds to changes in the properties of the perfusate, such as the ionic strength (), pH, and cationic identity. We review the experimental and theoretical work on this phenomenon. We then proceed to explore the hypothesis that electrokinetic effects in the bordered pit membrane (BPM) contribute to this response. In particular, we develop a model based on electroviscosity in which hydraulic conductance of an electrically charged porous membrane varies with the properties of the electrolyte. We use standard electrokinetic theory, coupled with measurements of electrokinetic properties of plant materials from the literature, to determine how the conductance of BPMs, and therefore , may change due to electroviscosity. We predict a nonmonotonic variation of with with a maximum reduction of 18%. We explore how this reduction depends on the characteristics of the sap and features of the BPM, such as pore size, density of chargeable sites, and their dissociation constant. Our predictions are consistent with changes in observed for physiological values of sap and pH. We conclude that electroviscosity is likely responsible, at least partially, for the electrolyte dependence of conductance through pits and that electroviscosity may be strong enough to play an important role in other transport processes in xylem. We conclude by proposing experiments to differentiate the impact of electroviscosity on from that of other proposed mechanisms. PMID:24014573

  15. Proline transport across the intestinal microvillus membrane may be regulated by membrane physical properties.

    PubMed

    Sadowski, D C; Gibbs, D J; Meddings, J B

    1992-03-23

    There is now abundant evidence that integral membrane protein function may be modulated by the physical properties of membrane lipids. The intestinal brush border membrane represents a membrane system highly specialized for nutrient absorption and, thus, provides an opportunity to study the interaction between integral membrane transport proteins and their lipid environment. We have previously demonstrated that alterations in this environment may modulate the function of the sodium-dependent glucose transporter in terms of its affinity for glucose. In this communication we report that membrane lipid-protein interactions are distinctly different for the proline transport proteins. Maximal transport rates for L-proline by either the neutral brush border or imino transport systems are reduced 10-fold when the surrounding membrane environment is made more fluid over the physiological range that exists along the crypt-villus axis. Furthermore, in microvillus membrane vesicles prepared from enterocytes isolated from along the crypt-villus axis a similar gradient exists in the functional activity of these transport systems. This would imply that either the functional activity of these transporters are regulated by membrane physical properties or that the synthesis and insertion of these proteins is coordinated in concert with membrane physical properties as the enterocyte migrates up the crypt-villus axis. PMID:1567897

  16. Applications of Polymer Brushes in Protein Analysis and Purification

    NASA Astrophysics Data System (ADS)

    Jain, Parul; Baker, Gregory L.; Bruening, Merlin L.

    2009-07-01

    This review examines the application of polymer brush-modified flat surfaces, membranes, and beads for protein immobilization and isolation. Modification of porous substrates with brushes yields membranes that selectively bind tagged proteins to give 99% pure protein at capacities as high as 100 mg of protein per cubic centimeter of membrane. Moreover, enrichment of phosphopeptides on brush-modified matrix-assisted laser desorption/ionization (MALDI) plates allows detection and characterization of femtomole levels of phosphopeptides by MALDI mass spectrometry. Because swollen hydrophilic brushes can resist nonspecific protein adsorption while immobilizing a high density of proteins, they are attractive as substrates for protein microarrays. This review highlights the advantages of polymer brush-modified surfaces over self-assembled monolayers and identifies some research needs in this area.

  17. Adsorption of Anionic or Cationic Surfactants in Polyanionic Brushes and Its Effect on Brush Swelling and Fouling Resistance during Emulsion Filtration.

    PubMed

    Yang, Zhefei; Tarabara, Volodymyr V; Bruening, Merlin L

    2015-11-01

    Atom transfer radical polymerization of ionic monomers from membrane surfaces yields polyelectrolyte brushes that swell in water and repel oil droplets to resist fouling during filtration of oil-in-water emulsions. However, surfactant adsorption to polyelectrolyte brushes may overcome this fouling resistance. This work examines adsorption of cationic and anionic surfactants in polyanionic brushes and the effect of these surfactants on emulsion filtration. In situ ellipsometry with films on flat surfaces shows that brushes composed of poly(3-sulfopropyl methacrylate salts) (pSPMK) swell 280% in water and do not adsorb sodium dodecyl sulfate (SDS). pSPMK-modified microfiltration membranes reject >99.9% of the oil from SDS-stabilized submicron emulsions, and the specific flux through these modified membranes is comparable to that through NF270 nanofiltration membranes. Moreover, the brush-modified membranes show no decline in flux over a 12 h filtration, whereas the flux through NF270 membranes decreases by 98.7%. In contrast, pSPMK brushes adsorb large quantities of cetyltrimethylammonium bromide (CTAB), and at low chain densities the brushes collapse in the presence of this cationic surfactant. Filtration of CTAB-stabilized emulsions through pSPMK-modified membranes gives minimal oil rejection, presumably due to the brush collapse. Thus, the fouling resistance of polyelectrolyte brush-modified membranes clearly depends on the surfactant composition in a particular emulsion. PMID:26442835

  18. Ceramic brush seals development

    NASA Technical Reports Server (NTRS)

    Howe, Harold

    1994-01-01

    The following topics are discussed in this viewgraph presentation: ceramic brush seals, research and development, manufacturing, brazed assembly development, controlling braze flow, fiber selection, and braze results.

  19. Rotating Brush Seal

    NASA Technical Reports Server (NTRS)

    Lattime, S. B.; Braun, M. J.; Choy. F. K.; Hendricks, R. C.; Steinetz, B. M.

    2006-01-01

    The proven technology of brush seals has been extended to the mitigation of problems arising from friction and wear at the bristle-rotor interface at high surface speeds. In prototype testing, the brush is mounted on, and free to rotate with the shaft, thus providing a complaint primary seal. A face seal positioned between the backing plate of the brush seal and the housing provides a secondary seal. The purpose of this paper is to demonstrate the interaction between the brush bristles and the shaft at high surface speeds as well as introduce a numerical model to simulate the bristle behavior. A test facility was constructed to study the effects of centrifugal forces on bristle deflection in a single rotating brush seal. The bristle-rotor interface was observed through a video camera, which utilized a high magnification borescope and a high frequency strobe light source. Rotational speeds of the rotor and the brush seal were measured by a magnetic and optical speed sensor, respectively. Preliminary results with speeds up to 11,000 rpm show no speed differential between the brush seal and rotor, or any instability problems associated with the brush seal. Bristle liftoff from the rotor is successfully captured on video.

  20. Genetic diversity of the Plasmodium falciparum apical membrane antigen I gene in parasite population from the China-Myanmar border area.

    PubMed

    Zhu, Xiaotong; Zhao, Zhenjun; Feng, Yonghui; Li, Peipei; Liu, Fei; Liu, Jun; Yang, Zhaoqing; Yan, Guiyun; Fan, Qi; Cao, Yaming; Cui, Liwang

    2016-04-01

    To investigate the genetic diversity of the Plasmodium falciparum apical membrane antigen 1 (PfAMA1) gene in Southeast Asia, we determined PfAMA1 sequences from 135 field isolates collected from the China-Myanmar border area and compared them with 956 publically available PfAMA1 sequences from seven global P. falciparum populations. This analysis revealed high genetic diversity of PfAMA1 in global P. falciparum populations with a total of 229 haplotypes identified. The genetic diversity of PfAMA1 gene from the China-Myanmar border is not evenly distributed in the different domains of this gene. Sequence diversity in PfAMA1 from the China-Myanmar border is lower than that observed in Thai, African and Oceanian populations, but higher than that in the South American population. This appeared to correlate well with the levels of endemicity of different malaria-endemic regions, where hyperendemic regions favor genetic cross of the parasite isolates and generation of higher genetic diversity. Neutrality tests show significant departure from neutrality in the entire ectodomain and Domain I of PfAMA1 in the China-Myanmar border parasite population. We found evidence supporting a substantial continent-wise genetic structure among P. falciparum populations, with the highest genetic differentiation detected between the China-Myanmar border and the South American populations. Whereas no alleles were unique to a specific region, there were considerable geographical differences in major alleles and their frequencies, highlighting further necessity to include more PfAMA1 alleles in vaccine designs. PMID:26825252

  1. Comb and brush acne.

    PubMed

    Petrozzi, J W

    1980-12-01

    Frequent combing or brushing of the hair may result in an exacerbation of existing acne or be the cause of localized acne. Because of today's hair styling habits and methods, this type of acne is commonly encountered but poorly appreciated by physicians. Patient education is the most important factor in the management of this condition. Comb and brush acne is another external factor to be added to the list of causes of "acne mechanica." PMID:6449356

  2. Almond brush module cutter

    SciTech Connect

    Zohns, M.A.; Jenkins, B.M.; Mehlschau, J.J.; Morrison, D.

    1983-06-01

    This paper addresses the design, construction, and evaluation of an almond brush module cutter. The module cutter is one link in a system which processes tree prunings for fuel and fiber. This system includes a modified cotton module builder, a module mover, the cutter, and a tub grinder. An economic analysis of the cutter is presented along with the problems involved in cutting brush modules.

  3. Lubrication by glycoprotein brushes.

    NASA Astrophysics Data System (ADS)

    Zappone, Bruno; Ruths, Marina; Greene, George W.; Israelachvili, Jacob

    2006-03-01

    Grafted polyelectrolyte brushes show excellent lubricating properties under water and have been proposed as a model to study boundary lubrication in biological system. Lubricin, a glycoprotein of the synovial fluid, is considered the major boundary lubricant of articular joints. Using the Surface Force Apparatus, we have measured normal and friction forces between model surfaces (negatively charged mica, positively charged poly-lysine and aminothiol, hydrophobic alkanethiol) bearing adsorbed layers of lubricin. Lubricin layers acts like a versatile anti-adhesive, adsorbing on all the surfaces considered and creating a repulsion similar to the force between end-grafted polymer brushes. Analogies with polymer brushes also appear from bridging experiment, where proteins molecules are end-adsorbed on two opposing surfaces at the same time. Lubricin `brushes' show good lubricating ability at low applied pressures (P<0.5MPa), especially on negatively charged surfaces like mica. At higher load, the adsorbed layers wears and fails lubricating the surfaces, while still protecting the underlying substrate from wearing. Lubricin might thus be a first example of biological polyelectrolytes providing `brush-like' lubrication and wear-protection.

  4. Bidirectional Brush Seals

    NASA Technical Reports Server (NTRS)

    Hendricks, Robert C.; Wilson, Jack; Wu, Tom; Flower, Ralph

    1997-01-01

    Presented is a study of the use of a set of I.D./O.D. bidirectional press seals to reduce the leakage losses in a wave rotor. Relative to the baseline configuration, data indicate the use of brush seals enhanced wave rotor efficiency from 36 to 45 percent at low leakages (small rotor endwall gap spacings) and from 15 to 33 percent at high leakages (larger endwall gap spacings). These brush seals are capable of sealing positive or negative pressure drops with respect to the axial direction. Surface tribology for these tests suggested little evidence of grooving although the bristles appeared polished.

  5. Temperature responsive behavior of polymer brush/polyelectrolyte multilayer composites.

    PubMed

    Micciulla, Samantha; Soltwedel, Olaf; Löhmann, Oliver; von Klitzing, Regine

    2016-01-28

    The complex interaction of polyelectrolyte multilayers (PEMs) physisorbed onto end-grafted polymer brushes with focus on the temperature-responsive behavior of the system is addressed in this work. The investigated brush/multilayer composite consists of a poly(styrene sulfonate)/poly(diallyldimethylammonium chloride) (PSS/PDADMAC) multilayer deposited onto the poly(N-isopropylacrylamide-b-dimethylaminoethyl methacrylate) P(NIPAM-b-DMAEMA) brush. Ellipsometry and neutron reflectometry were used to monitor the brush collapse with the thickness decrease as a function of temperature and the change in the monomer distribution perpendicular to the substrate at temperatures below, across and above the phase transition, respectively. It was found that the adsorption of PEMs onto polymer brushes had a hydrophobization effect on PDMAEMA, inducing the shift of its phase transition to lower temperatures, but without suppressing its temperature-responsiveness. Moreover, the diffusion of the free polyelectrolyte chains inside the charged brush was proved by comparing the neutron scattering length density profile of pure and the corresponding PEM-capped brushes, eased by the enhanced contrast between hydrogenated brushes and deuterated PSS chains. The results presented herein demonstrate the possibility of combining a temperature-responsive brush with polyelectrolyte multilayers without quenching the responsive behavior, even though significant interpolyelectrolyte interactions are present. This is of importance for the design of multicompartment coatings, where the brush can be used as a reservoir for the controlled release of substances and the multilayer on the top as a membrane to control the diffusion in/out by applying different stimuli. PMID:26612742

  6. A Brush-Creeper

    ERIC Educational Resources Information Center

    Mak, Se-yuen; Wong, Siu-ling

    2006-01-01

    In this note, we introduce a simple homemade toy called the brush-creeper, which can glide forward with no propellers, limbs, wheels, and seemingly no movement of any kind that can push forward against the ground. The toy arouses pupils' interest and their incentive to ask "Why?" in lessons related to friction.

  7. Brush potential curve tracer

    DOEpatents

    Finch, H.A.

    1985-06-21

    A device for analyzing commutating characteristics of a motor or generator includes a holder for supporting a plurality of probes adjacent a brush of the motor or generator. Measurements of electrical current characteristics of the probes provides information useful in analyzing operation of the machine. Methods for employing a device in accordance with the invention are also disclosed.

  8. Smart photonic carbon brush

    NASA Astrophysics Data System (ADS)

    Morozov, Oleg G.; Kuznetsov, Artem A.; Morozov, Gennady A.; Nureev, Ilnur I.; Sakhabutdinov, Airat Z.; Faskhutdinov, Lenar M.; Artemev, Vadim I.

    2016-03-01

    Aspects of the paper relate to a wear monitoring system for smart photonic carbon brush. There are many applications in which regular inspection is not feasible because of a number of factors including, for example, time, labor, cost and disruptions due to down time. Thus, there is a need for a system that can monitor the wear of a component while the component is in operation or without having to remove the component from its operational position. We propose a new smart photonic method for characterization of carbon brush wear. It is based on the usage of advantages of the multiplicative response of FBG and LPFG sensors and its double-frequency probing. Additional measuring parameters are the wear rate, the brush temperature, the engine rotation speed, the hangs control, and rotor speed. Sensor is embedded in brush. Firstly the change of sensor length is used to measure wear value and its central wavelength shift for temperature ones. The results of modeling and experiments are presented.

  9. Pericellular Brush and Mechanics of Guinea Pig Fibroblast Cells Studied with AFM.

    PubMed

    Dokukin, Maxim; Ablaeva, Yulija; Kalaparthi, Vivekanand; Seluanov, Andrei; Gorbunova, Vera; Sokolov, Igor

    2016-07-12

    The atomic force microscopy (AFM) indentation method combined with the brush model can be used to separate the mechanical response of the cell body from deformation of the pericellular layer surrounding biological cells. Although self-consistency of the brush model to derive the elastic modulus of the cell body has been demonstrated, the model ability to characterize the pericellular layer has not been explicitly verified. Here we demonstrate it by using enzymatic removal of hyaluronic content of the pericellular brush for guinea pig fibroblast cells. The effect of this removal is clearly seen in the AFM force-separation curves associated with the pericellular brush layer. We further extend the brush model for brushes larger than the height of the AFM probe, which seems to be the case for fibroblast cells. In addition, we demonstrate that an extension of the brush model (i.e., double-brush model) is capable of detecting the hierarchical structure of the pericellular brush, which, for example, may consist of the pericellular coat and the membrane corrugation (microridges and microvilli). It allows us to quantitatively segregate the large soft polysaccharide pericellular coat from a relatively rigid and dense membrane corrugation layer. This was verified by comparison of the parameters of the membrane corrugation layer derived from the force curves collected on untreated cells (when this corrugation membrane part is hidden inside the pericellular brush layer) and on treated cells after the enzymatic removal of the pericellular coat part (when the corrugations are exposed to the AFM probe). We conclude that the brush model is capable of not only measuring the mechanics of the cell body but also the parameters of the pericellular brush layer, including quantitative characterization of the pericellular layer structure. PMID:27410750

  10. Wire brush fastening device

    DOEpatents

    Meigs, Richard A.

    1995-01-01

    A fastening device is provided which is a variation on the conventional nut and bolt. The bolt has a longitudinal axis and threading helically affixed thereon along the longitudinal axis. A nut having a bore extending therethrough is provided. The bore of the nut has a greater diameter than the diameter of the bolt so the bolt can extend through the bore. An array of wire bristles are affixed within the bore so as to form a brush. The wire bristles extend inwardly from the bore and are constructed and arranged of the correct size, length and stiffness to guide the bolt within the bore and to restrain the bolt within the bore as required. A variety of applications of the wire brush nut are disclosed, including a bolt capture device and a test rig apparatus.

  11. Wire brush fastening device

    DOEpatents

    Meigs, R.A.

    1995-09-19

    A fastening device is provided which is a variation on the conventional nut and bolt. The bolt has a longitudinal axis and threading helically affixed thereon along the longitudinal axis. A nut having a bore extending therethrough is provided. The bore of the nut has a greater diameter than the diameter of the bolt so the bolt can extend through the bore. An array of wire bristles are affixed within the bore so as to form a brush. The wire bristles extend inwardly from the bore and are constructed and arranged of the correct size, length and stiffness to guide the bolt within the bore and to restrain the bolt within the bore as required. A variety of applications of the wire brush nut are disclosed, including a bolt capture device and a test rig apparatus. 13 figs.

  12. Wire brush fastening device

    SciTech Connect

    Meigs, R.A.

    1993-08-31

    A fastening device is provided which is a variation on the conventional nut and bolt. The bolt has a longitudinal axis and threading helically affixed thereon along the longitudinal axis. A nut having a bore extending therethrough is provided. The bore of the nut has a greater diameter than the diameter of the bolt so the bolt can extend through the bore. An array of wire bristles are affixed within the bore so as to form a brush. The wire bristles extend inwardly from the bore and are constructed and arranged of the correct size, length and stiffness to guide the bolt within the bore and to restrain the bolt within the bore as required. A variety of applications of the wire brush nut are disclosed, including a bolt capture device and a test rig apparatus.

  13. Interpenetrations in polymer brushes

    NASA Astrophysics Data System (ADS)

    Chakrabarti, Amitabha; Nelson, Peter; Toral, Raúl

    1994-01-01

    We carry out Monte Carlo simulations in both lattice and off-lattice geometry, to study the configurational properties of a system of two polymer-coated plates. We compare our results with numerical and analytical self-consistent-field theories. For the short chain-lengths considered here, we find both compression and interpenetration of the polymer brushes as the plate separation is decreased, instead of the ``classical-limit'' picture of compression with no interpenetration.

  14. Brush seals for cryogenic applications

    NASA Astrophysics Data System (ADS)

    Proctor, Margaret P.

    1994-07-01

    This viewgraph presentation presents test results of brush seals for cryogenic applications. Leakage for a single brush seal was two to three times less than for a 12-tooth labyrinth seal. The maximum temperature rise for a single brush seal was less than 50 R and occurred at 25 psid across the seal and 35,000 rpm. A static blowout test demonstrated sealing capability up to 550 psid. The seal limit was not obtained. The power loss for a single brush at 35,000 rpm and 175 psid was 2.45 hp. Two brushes far apart leak less than two brushes tight packed. Rotor wear was approximately 0.00075 mils and bristle wear was 1-3 mils after 4-1/2 hours.

  15. Brush seals for cryogenic applications

    NASA Technical Reports Server (NTRS)

    Proctor, Margaret P.

    1994-01-01

    This viewgraph presentation presents test results of brush seals for cryogenic applications. Leakage for a single brush seal was two to three times less than for a 12-tooth labyrinth seal. The maximum temperature rise for a single brush seal was less than 50 R and occurred at 25 psid across the seal and 35,000 rpm. A static blowout test demonstrated sealing capability up to 550 psid. The seal limit was not obtained. The power loss for a single brush at 35,000 rpm and 175 psid was 2.45 hp. Two brushes far apart leak less than two brushes tight packed. Rotor wear was approximately 0.00075 mils and bristle wear was 1-3 mils after 4-1/2 hours.

  16. Trans-Membrane Area Asymmetry Controls the Shape of Cellular Organelles

    PubMed Central

    Beznoussenko, Galina V.; Pilyugin, Sergei S.; Geerts, Willie J. C.; Kozlov, Michael M.; Burger, Koert N. J.; Luini, Alberto; Derganc, Jure; Mironov, Alexander A.

    2015-01-01

    Membrane organelles often have complicated shapes and differ in their volume, surface area and membrane curvature. The ratio between the surface area of the cytosolic and luminal leaflets (trans-membrane area asymmetry (TAA)) determines the membrane curvature within different sites of the organelle. Thus, the shape of the organelle could be critically dependent on TAA. Here, using mathematical modeling and stereological measurements of TAA during fast transformation of organelle shapes, we present evidence that suggests that when organelle volume and surface area are constant, TAA can regulate transformation of the shape of the Golgi apparatus, endosomal multivesicular bodies, and microvilli of brush borders of kidney epithelial cells. Extraction of membrane curvature by small spheres, such as COPI-dependent vesicles within the Golgi (extraction of positive curvature), or by intraluminal vesicles within endosomes (extraction of negative curvature) controls the shape of these organelles. For instance, Golgi tubulation is critically dependent on the fusion of COPI vesicles with Golgi cisternae, and vice versa, for the extraction of membrane curvature into 50–60 nm vesicles, to induce transformation of Golgi tubules into cisternae. Also, formation of intraluminal ultra-small vesicles after fusion of endosomes allows equilibration of their TAA, volume and surface area. Finally, when microvilli of the brush border are broken into vesicles and microvilli fragments, TAA of these membranes remains the same as TAA of the microvilli. Thus, TAA has a significant role in transformation of organelle shape when other factors remain constant. PMID:25761238

  17. An atrophic, telangiectatic patch at the distal border of the tongue: a mucous membrane manifestation of xeroderma pigmentosum.

    PubMed

    Bologna, Sheyla Batista; Harumi Nakajima Teshima, Tathyane; Lourenço, Silvia Vanessa; Nico, Marcello Menta Simonsen

    2014-01-01

    Xeroderma pigmentosum (XP) is a rare autosomal recessive disorder characterized by clinical and cellular sensitivity, pigmentary changes, and early development of malignancies in sun-exposed mucocutaneous and ocular structures due to a defective ability to repair intracellular DNA damage. Individuals with XP also have a greater frequency of oral cancer, particularly squamous cell carcinoma of the anterior third of the tongue. The current study reports four cases of XP that exhibited a characteristic crescent-shaped, atrophic, telangiectatic area on the distal border of the tongue and correlates this lesion with the development of tumors at this site during follow-up. The tongue lesion was photographed and biopsied in the four patients. During routine follow-up visits, new biopsies were performed if additional tongue lesions were observed. The studied lesions were similar in the four patients. During follow-up, squamous cell carcinoma developed in one patient and pyogenic granuloma developed in three patients and was relapsing in one. The lesion remained stable in one patient during the study. The atrophic and telangiectatic patches probably occur because of chronic sun damage to the exposed portion of the tongue, and this area has a high predisposition for the development of benign and malignant tumors. PMID:24456184

  18. Electrostatically anchored branched brush layers.

    PubMed

    Liu, Xiaoyan; Dedinaite, Andra; Rutland, Mark; Thormann, Esben; Visnevskij, Ceslav; Makuska, Ricardas; Claesson, Per M

    2012-11-01

    A novel type of block copolymer has been synthesized. It consists of a linear cationic block and an uncharged bottle-brush block. The nonionic bottle-brush block contains 45 units long poly(ethylene oxide) side chains. This polymer was synthesized with the intention of creating branched brush layers firmly physisorbed to negatively charged surfaces via the cationic block, mimicking the architecture (but not the chemistry) of bottle-brush molecules suggested to be present on the cartilage surface, and contributing to the efficient lubrication of synovial joints. The adsorption properties of the diblock copolymer as well as of the two blocks separately were studied on silica surfaces using quartz crystal microbalance with dissipation monitoring (QCM-D) and optical reflectometry. The adsorption kinetics data highlight that the diblock copolymers initially adsorb preferentially parallel to the surface with both the cationic block and the uncharged bottle-brush block in contact with the surface. However, as the adsorption proceeds, a structural change occurs within the layer, and the PEO bottle-brush block extends toward solution, forming a surface-anchored branched brush layer. As the adsorption plateau is reached, the diblock copolymer layer is 46-48 nm thick, and the water content in the layer is above 90 wt %. The combination of strong electrostatic anchoring and highly hydrated branched brush structures provide strong steric repulsion, low friction forces, and high load bearing capacity. The strong electrostatic anchoring also provides high stability of preadsorbed layers under different ionic strength conditions. PMID:23046176

  19. Spontaneous Curvature of Polymer Brushes

    NASA Astrophysics Data System (ADS)

    Sheiko, Sergei; da Silva, Marcelo; Shirvaniants, David; Rodrigues, Carlos; Beers, Kathryn; Matyjaszewski, Krzysztof; Potemkin, Igor; Moeller, Martin

    2003-03-01

    Experimental studies of cylindrical brushes on surfaces revealed peculiar shape of brush molecules with a curved backbone. According to scaling analysis, spontaneous bending of the backbone can be driven by entropic elasticity of the side chains: smaller extension of the chains is attained due to their asymmetric distribution with respect to the backbone. An equilibrium, i.e. spontaneous curvature results from the balance of the elasticity of the side chains and the elasticity of the interface. The curvature is predicted to increase with the side chain length. The system is of general interest because cylindrical brushes confined to a flat surface represent a two-dimensional cross-section of a planar brush grafted on both sides. Here we present systematic studies of cylindrical brushes with different length of the side chains. The curvature of cylindrical brushes confined to a flat substrate was investigated by atomic force microscopy. The method allowed visualization of individual molecules and quantitative analysis of their conformation. In agreement with theory, adsorbed brushes demonstrated spontaneous curvature, however the curvature was shown to decrease with the side chain length.

  20. Current developments in brush seals

    NASA Astrophysics Data System (ADS)

    Loewenthal, Bob

    1994-07-01

    The objective of the brush seal development program is to develop a comprehensive design methodology for brush seals using application requirements from engine manufacturers and experimental characterization of seal design and tribological pairs. The goals are to substantially lower leakage compared to labyrinth seals, seal life consistent with man-rated mission requirements, to investigate single and multiple staged brush seals, temperature up to 1200 F and surface speed up to 900 fps, and pressure drop across the seal of 50 psid. Test results are presented in viewgraph format.

  1. Transcription rates in DNA brushes.

    PubMed

    Yamamoto, Tetsuya; Safran, S A

    2015-04-21

    We theoretically predict the rate of transcription (TX) in DNA brushes by introducing the concept of TX dipoles that takes into account the unidirectional motion of enzymes (RNAP) along DNA during transcription as correlated pairs of sources and sinks in the relevant diffusion equation. Our theory predicts that the TX rates dramatically change upon the inversion of the orientation of the TX dipoles relative to the substrate because TX dipoles modulate the concentrations of RNAP in the solution. Comparing our theory with experiments suggests that, in some cases, DNA chain segments are relatively uniformly distributed in the brush, in contrast to the parabolic profile expected for flexible polymer brushes. PMID:25736601

  2. Lectin binding studies on a glycopolymer brush flow-through biosensor by localized surface plasmon resonance.

    PubMed

    Rosencrantz, Ruben R; Nguyen, Vu Hoa; Park, Hyunji; Schulte, Christine; Böker, Alexander; Schnakenberg, Uwe; Elling, Lothar

    2016-08-01

    A localized surface plasmon resonance biosensor in a flow-through configuration was applied for investigating kinetics of lectin binding to surface-grafted glycopolymer brushes. Polycarbonate filter membranes with pore sizes of 400 nm were coated with a 114-nm thick gold layer and used as substrate for surface-initiated atom-transfer radical polymerization of a glycomonomer. These grafted from glycopolymer brushes were further modified with two subsequent enzymatic reactions on the surface to yield an immobilized trisaccharide presenting brush. Specific binding of lectins including Clostridium difficile toxin A receptor domain to the glycopolymer brush surface could be investigated in a microfluidic setup with flow-through of the analytes and transmission surface plasmon resonance spectroscopy. Graphical abstract Glycopolymer brushes serve as high affinity ligands for lectin and toxin interactions in a sensitive, disposable flow-through LSPR biosensor. PMID:27277814

  3. Distribution of Chains in Polymer Brushes Produced by a “Grafting From” Mechanism

    DOE PAGESBeta

    Martinez, Andre P.; Carrillo, Jan-Michael Y.; Dobrynin, Andrey V.; Adamson, Douglas H.

    2016-01-11

    The molecular weight and polydispersity of the chains in a polymer brush are critical parameters determining the brush properties. However, the characterization of polymer brushes is hindered by the vanishingly small mass of polymer present in brush layers. In this study, in order to obtain sufficient quantities of polymer for analysis, polymer brushes were grown from high surface area fibrous nylon membranes by ATRP. We synthesized the brushes with varying surface initiator densities, polymerization times, and amounts of sacrificial initiator, then cleaved from the substrate, and analyzed by GPC and NMR. Characterization showed that the surface-grown polymer chains were moremore » polydisperse and had lower average molecular weight compared to solution-grown polymers synthesized concurrently. Furthermore, the molecular weight distribution of the polymer brushes was observed to be bimodal, with a low molecular weight population of chains representing a significant mass fraction of the polymer chains at high surface initiator densities. Moreover, the origin of this low MW polymer fraction is proposed to be the termination of growing chains by recombination during the early stages of polymerization, a mechanism confirmed by molecular dynamics simulations of brush polymerization.« less

  4. Osmotic and Salted Brush Phase of Polyelectrolyte Brushes

    NASA Astrophysics Data System (ADS)

    Helm, Christane A.; Ahrens, Heiko; Förster, Stephan

    2004-03-01

    Amphiphilic block copolymers consisting of a fluid hydrophobic Poly(ethyletylene) (PEE), and a Poly(styrenesulfonate) (PSS) part form monolayers at the air/water interface. With x-ray reflectivity it is shown that the hydrophobic blocks of PEE_114PSS_83 and PEE_144PSS_136 constitute a nm-thick melt, while the polyelectrolyte forms an osmotically swollen brush with counterion incorporation. A slight thickness increase on monolayer compression is found which can be explained by the strong stretching of the brushes. Only at high salt conditions (above 0.1 M), the brush shrinks and the thickness scales with the molecular area (exponent -1/3), and with the salt concentration (exponent ca. -1/5). With Grazing Incidence Diffraction, the lateral order of the polyelectrolyte chains can be detected.

  5. Cryogenic Brush Seal Test Results

    NASA Technical Reports Server (NTRS)

    Proctor, Margaret P.; Walker, James F.

    1996-01-01

    Brush seals are compliant, contact seals that have long-life, low-leakage characteristics desirable for use in rocket engine turbopumps. 50.8-mm (2.0 inch) diameter brush seals with a nominal initial radial interference of 0.127-mm (0.005 inch) were tested in liquid nitrogen at shaft speeds up to 35,000 rpm and differential pressure loads up to 1.21 MPa (175 psi) per brush. The measured leakage rate of a single brush was 2-3 times less than that measured for a 12-tooth, 0.127-mm (0.005 inch) radial clearance labyrinth seal used as a baseline. Stage effects were studied and it was found that two brush seals with a large separation distance leaked less than two brushes tightly packed together. The maximum measured groove depth on the Inconel 718 rotor was 25.4 (mu)m (0.001 inch) after 4.31 hours of shaft rotation. The Haynes-25 bristles wore approximately 25.4-76.2 (mu)m (0.001-0.003 inch) under the same conditions. Three seal runner coatings, chromium carbide, Teflon impregnated chromium, and zirconium oxide, were tested in liquid hydrogen at 35,000 and 65,000 rpm with separate 50.8 mm diameter brush seals made of Haynes-25 bristles and having a nominal initial radial interference of 129 rpm. Two bare Inconel-718 rotors were also tested as a baseline. The test results revealed significant differences between the wear characteristics of the uncoated and coated seal runners. At both speeds the brush seal with the bare Inconel-718 seal runner exhibited significant bristle wear with excessive material transferring to the runner surface. In contrast, the coated seal runners inhibited the transfer and deposit of bristle material. The chromium carbide coating showed only small quantities of bristle material transferring to its surface. The Teflon impregnated chromium coating also inhibited material transfer and provided some lubrication. This coating, however, is self-sacrificing. The Teflon remained present on the low speed runner, but it was completely removed from the

  6. Mesoscale modeling of polyelectrolyte brushes with salt.

    PubMed

    Ibergay, Cyrille; Malfreyt, Patrice; Tildesley, Dominic J

    2010-06-01

    We report dissipative particle dynamics (DPD) simulations of a polyelectrolyte brush under athermal solvent conditions. The electrostatic interactions are calculated using the particle-particle particle-mesh (PPPM) method with charges distributed over the particles. The polymer beads, counterions, co-ions, and solvent particles are modeled explicitly. The DPD simulations show a dependence of the brush height on the grafting density and the charge fraction that is typical of the nonlinear osmotic brush regime. We report the effect of the addition of salt on the structural properties of the brush. In the case of a polyelectrolyte brush with a high surface coverage, the simulations reproduce the transition between the nonlinear osmotic brush regime where the thickness of the brush is independent of the salt concentration and the salted regime where the brush height decreases weakly with the salt concentration. PMID:20455593

  7. Fabrication of protein-resistant blend based on PVDF-HFP and amphiphilic brush copolymer made from PMMA and PEGMA

    NASA Astrophysics Data System (ADS)

    Hwangbo, Kyung-Hee; Kim, Yu-Jeong; Cho, Kuk Young

    2012-12-01

    Polymeric blends provide a facile route to obtaining materials with various synergistic properties arising from the individual components. Poly(vinylidene fluoride-co-hexafluoropropylene) (PVDF-HFP), a hydrophobic polymer, is finding new applications in polymer electrolytes, membranes, and heat-resistant structural materials owing to its high thermal stability, mechanical strength, and weatherability. In this report, blends of PVDF-HFP and polymer brush were prepared with enhanced water uptake and protein resistance, which are important requirements for membranes used in food and biological applications. Polymer brush is composed of poly(methyl methacrylate) main chains, which are miscible with PVDF-HFP, and hydrophilic poly(ethylene glycol) (PEG) brush chains. Incorporation of PEG chains through the polymer brush structure not only enhanced water uptake and protein adsorption resistance but also produced a well-distributed morphology of the blending components through the matrix as evidenced by observation of the morphology after selective extraction of polymer brush from the matrix.

  8. Brush seals for cryogenic applications

    NASA Technical Reports Server (NTRS)

    Proctor, Margaret P.

    1993-01-01

    Brush seals are compliant, contacting seals and have significantly lower leakage than labyrinth seals in gas turbine applications. Their characteristics of long life and low leakage make them candidates for use in rocket engine turbopumps. Two-inch diameter brush seals with a nominal 0.005 inch radial interference were tested in liquid nitrogen at shaft speeds up to 35,000 rpm and pressure drops up to 175 psid per seal. A labyrinth seal was also tested to provide a baseline. Performance, staging effects, and wear results are presented.

  9. Brush seals for cryogenic applications

    NASA Astrophysics Data System (ADS)

    Proctor, Margaret P.

    1993-11-01

    Brush seals are compliant, contacting seals and have significantly lower leakage than labyrinth seals in gas turbine applications. Their characteristics of long life and low leakage make them candidates for use in rocket engine turbopumps. Two-inch diameter brush seals with a nominal 0.005 inch radial interference were tested in liquid nitrogen at shaft speeds up to 35,000 rpm and pressure drops up to 175 psid per seal. A labyrinth seal was also tested to provide a baseline. Performance, staging effects, and wear results are presented.

  10. Charge transferred in brush discharges

    NASA Astrophysics Data System (ADS)

    Talarek, M.; Kacprzyk, R.

    2015-10-01

    Electrostatic discharges from surfaces of plastic materials can be a source of ignition, when appear in explosive atmospheres. Incendivity of electrostatic discharges can be estimated using the transferred charge test. In the case of brush discharges not all the energy stored at the tested sample is released and the effective surface charge density (or surface potential) crater is observed after the discharge. Simplified model, enabling calculation of a charge transferred during electrostatic brush discharge, was presented. Comparison of the results obtained from the simplified model and from direct measurements of transferred charge are presented in the paper.

  11. Brush/Fin Thermal Interfaces

    NASA Technical Reports Server (NTRS)

    Knowles, Timothy R.; Seaman, Christopher L.; Ellman, Brett M.

    2004-01-01

    Brush/fin thermal interfaces are being developed to increase heat-transfer efficiency and thereby enhance the thermal management of orbital replaceable units (ORUs) of electronic and other equipment aboard the International Space Station. Brush/fin thermal interfaces could also be used to increase heat-transfer efficiency in terrestrial electronic and power systems. In a typical application according to conventional practice, a replaceable heat-generating unit includes a mounting surface with black-anodized metal fins that mesh with the matching fins of a heat sink or radiator on which the unit is mounted. The fins do not contact each other, but transfer heat via radiation exchange. A brush/fin interface also includes intermeshing fins, the difference being that the gaps between the fins are filled with brushes made of carbon or other fibers. The fibers span the gap between intermeshed fins, allowing heat transfer by conduction through the fibers. The fibers are attached to the metal surfaces as velvet-like coats in the manner of the carbon fiber brush heat exchangers described in the preceding article. The fiber brushes provide both mechanical compliance and thermal contact, thereby ensuring low contact thermal resistance. A certain amount of force is required to intermesh the fins due to sliding friction of the brush s fiber tips against the fins. This force increases linearly with penetration distance, reaching 1 psi (6.9 kPa) for full 2-in. (5.1 cm) penetration for the conventional radiant fin interface. Removal forces can be greater due to fiber buckling upon reversing the sliding direction. This buckling force can be greatly reduced by biasing the fibers at an angle perpendicularly to the sliding direction. Means of containing potentially harmful carbon fiber debris, which is electrically conductive, have been developed. Small prototype brush/fin thermal interfaces have been tested and found to exhibit temperature drops about onesixth of that of conventional

  12. Macroion Interaction at Polyelectrolyte Brush Interfaces

    NASA Astrophysics Data System (ADS)

    Qu, Chen

    2015-03-01

    The effect of macroions, including synthetic polyelectrolytes, DNA and proteins, on the structure and surface properties of charged polymer thin films remains inadequately understood partially due to the complexity involving the hydrophobic effect and the conformational change of polymeric macroions. In this work, we explore a group of inorganic nanocluster based macroions, hydrophilic polyoxometalates (POMs) of robust nanocluster structure and carrying high surface charges (~ 2-42 negative charges) to investigate their interaction with surface tethered poly-2-vinylpyridine (P2VP) brush-like thin films immersed in aqueous solution. We observe the collapse of swollen P2VP chains by adding POM macroions of increased concentration by AFM, QCM and contact goniometer measurements, in sharp contrast to the increased chain stretching by adding monovalent salts. A careful comparison is made between distinct POMs based on their charge, size and chemical nature. These findings serve as a good reference for theoretical model modification and design of new mesoporous composite membranes.

  13. Putative interaction of brush cells with bicarbonate secreting cells in the proximal corpus mucosa.

    PubMed

    Eberle, Julia Anna-Maria; Müller-Roth, Kai L; Widmayer, Patricia; Chubanov, Vladimir; Gudermann, Thomas; Breer, Heinz

    2013-01-01

    The gastric epithelium is protected from the highly acidic luminal content by alkaline mucus which is secreted from specialized epithelial cells. In the stomach of mice strong secretion of alkaline fluid was observed at the "gastric groove," the border between corpus and fundus mucosa. Since this region is characterized by numerous brush cells it was proposed that these cells might secrete alkaline solution as suggested for brush cells in the bile duct. In fact, it was found that in this region multiple cells express elements which are relevant for the secretion of bicarbonate, including carbonic anhydrase (CAII), the cystic fibrosis transmembrane conductance regulator (CFTR) and the Na(+)/H(+) exchanger (NHE1). However, this cell population was distinct from brush cells which express the TRP-channel TRPM5 and are considered as putative sensory cells. The location of both cell populations in close proximity implies the possibility for a paracrine interaction. This view was substantiated by the finding that brush cells express prostaglandin synthase-1 (COX-1) and the neighboring cells a specific receptor type for prostaglandins. The notion that brush cells may be able to sense a local acidification was supported by the observation that they express the channel PKD1L3 which contributes to the acid responsiveness of gustatory sensory cells. The results support the concept that brush cells may sense the luminal content and influence via prostaglandins the secretion of alkaline solution. PMID:23874305

  14. Putative interaction of brush cells with bicarbonate secreting cells in the proximal corpus mucosa

    PubMed Central

    Eberle, Julia Anna-Maria; Müller-Roth, Kai L.; Widmayer, Patricia; Chubanov, Vladimir; Gudermann, Thomas; Breer, Heinz

    2013-01-01

    The gastric epithelium is protected from the highly acidic luminal content by alkaline mucus which is secreted from specialized epithelial cells. In the stomach of mice strong secretion of alkaline fluid was observed at the “gastric groove,” the border between corpus and fundus mucosa. Since this region is characterized by numerous brush cells it was proposed that these cells might secrete alkaline solution as suggested for brush cells in the bile duct. In fact, it was found that in this region multiple cells express elements which are relevant for the secretion of bicarbonate, including carbonic anhydrase (CAII), the cystic fibrosis transmembrane conductance regulator (CFTR) and the Na+/H+ exchanger (NHE1). However, this cell population was distinct from brush cells which express the TRP-channel TRPM5 and are considered as putative sensory cells. The location of both cell populations in close proximity implies the possibility for a paracrine interaction. This view was substantiated by the finding that brush cells express prostaglandin synthase-1 (COX-1) and the neighboring cells a specific receptor type for prostaglandins. The notion that brush cells may be able to sense a local acidification was supported by the observation that they express the channel PKD1L3 which contributes to the acid responsiveness of gustatory sensory cells. The results support the concept that brush cells may sense the luminal content and influence via prostaglandins the secretion of alkaline solution. PMID:23874305

  15. Teach children to brush (image)

    MedlinePlus

    ... child's overall good health. Without proper dental care tooth decay and gum disease can lead to serious problems such as cavities and gingivitis, swollen and bleeding gums. Regular visits to the dentist, brushing twice each day, and flossing, are ways to ...

  16. Wear of metal fiber brushes

    NASA Astrophysics Data System (ADS)

    Brown, Lloyd Perryman, Jr.

    The goal of this dissertation was determining the wear mechanism of metal fiber brushes on commutators and slip rings with the goal of achieving the lowest possible wear rate. To this end, metal fiber brushes were operated, while conducting direct current, on gold-plated copper rotors with and without unfilled gaps to simulate slip rings and commutators, respectively. Wear rates on unfilled-groove commutators were found to be only modestly higher than on slip ring style rotors. Three possible causes for enhanced metal fiber brush wear on commutators were considered: (i) accelerated "adhesive" wear controlled by contact spots, (ii) fatigue induced wear and (iii) "fiber chopping". Similarly, SEM analysis of fiber tips and wear particles produced scant, if any, evidence of fiber chopping, which would occur as, again, fiber tips extend elastically into the commutator grooves and small slices of them would be "chopped" off by oncoming edges of commutator bars. Finally considered was "modified chopping", wherein fiber tips would be dragged over groove edges, resulting in tensile fracture and chopping. Only a single fiber fragment showed damage that might be compatible with that mechanism. Moreover, the fact that it was exemplified by a single tenuous case, rules it out as significant. The same conclusion also follows from comparing commutator wear rates with that on slip rings. These show good correlation in terms of effective brush pressure, which on commutators is increased because only bars conduct current and gaps do not support load. (Abstract shortened by UMI.)

  17. Polymer brushes for molecular transport

    NASA Astrophysics Data System (ADS)

    Tu, Huilin

    Polymer brushes and self-assembled monolayers (SAMs) are used as solid-state, 2-dimensional transport media to confine molecular diffusion. Microcontact printing and photolithography combined with surface-initiated atom transfer radical polymerization (ATRP) are the major techniques to construct patterned polymer brushes. Fluorescence recovery after photobleaching (FRAP) is used to determine diffusion coefficients of fluorescent dye, Prodan, in the polymer brushes and SAMs. Similar diffusion coefficients of Prodan are found for the SAMs formed by chlorotrimethyl silane (CTS) and silanated poly(ethylene glycol) (PEG). No fluorescence recovery is observed on the octadecyltrichlorosilane (OTS) SAM or on the clean silica surface. Patterned poly(N-isopropylacrylamide) (PNIPAAm) brushes were fabricated on silica substrates by surface-initiated ATRP of N-isopropylacrylamide from a micropatterned initiator. Variable temperature ellipsometry indicated that the lower critical solution temperature (LCST) of the hydrated PNIPAAm brush was broad, occurring over the range of 20-35°C. FRAP results of Prodan in PNIPAAm layers indicate that bulk translational diffusion is very slow relative to other diffusion mechanisms. Poly(oligoethylene glycol acrylate) (POEGA) chains are grafted onto silica substrates by surface-initiated ATRP. The diffusion of Prodan in dry POEGA is very fast, which is attributable to the low glass transition temperature of POEGA. The diffusion of Prodan in POEGA under variable humidity can be described reasonably well using WLF equation. Patterned POEGA brushes are not able to confine Prodan to diffuse exclusively inside the polymer regions, and possible explanations are rotational diffusion and a combination of fast surface diffusion and slow bulk diffusion. PNIPAAm chains are tethered onto silica particles via surface-initiated polymerization. Thermal annealing slightly improved the crystallinity of the colloidal assembly. Video microscopy of the two

  18. Turbine-Driven Pipe-Cleaning Brush

    NASA Technical Reports Server (NTRS)

    Werlink, Rudy J.; Rowell, David E.

    1994-01-01

    Simple pipe-cleaning device includes small turbine wheel axially connected, by standoff, to circular brush. Turbine wheel turns on hub bearing attached to end of upstream cable. Turbine-and-brush assembly inserted in pipe with cable trailing upstream and brush facing downstream. Water or cleaning solution pumped through pipe. Cable held at upstream end, so it holds turbine and brush in pipe at location to be cleaned. Flow in pipe turns turbine, which turns wheel, producing desired cleaning action. In addition to brushing action, device provides even mixing of cleaning solution in pipe.

  19. Comparative binding of Cry1Ab and Cry1F Bacillus thuringiensis toxins to brush border membrane proteins from Ostrinia nubilalis, Ostrinia furnacalis and Diatraea saccharalis (Lepidoptera: Crambidae) midgut tissue.

    PubMed

    Tan, Sek Yee; Cayabyab, Bonifacio F; Alcantara, Edwin P; Huang, Fangneng; He, Kanglai; Nickerson, Kenneth W; Siegfried, Blair D

    2013-11-01

    The European (Ostrinia nubilalis Hübner) and Asian corn borers (Ostrinia furnacalis Guenée) are closely related and display similar sensitivity to Cry1 toxins. In this study, we compared the binding patterns of Cry1Ab and Cry1F toxins between both Ostrinia spp., as well as the expression of putative cadherin- and aminopeptidase-N (APN)-like protein receptors. Additionally, cDNA sequences of these putative toxin receptors from both Ostrinia species were compared. Ligand blots for both species indicated a similar binding pattern for Cry1Ab with the strongest immunoreactive band at 260 kDa in both species. In addition, similar expression of the putative cadherin- and APN-like protein receptors were observed at 260 and 135 kDa, respectively. A high degree of similarity (98% amino acid sequence identity) of cDNA sequences for both putative receptor sequences was observed. The Cry1F ligand blot revealed that O. furnacalis and O. nubilalis BBMV exhibited slightly different binding patterns, with strong binding to putative proteins at 150 and 140 kDa, respectively. Both proteins appeared to also bind Cry1Ab, although the signal intensity was much reduced with Cry1Ab. O. furnacalis showed an additional but weaker band at 210 kDa relative to the 150 kDa band. Diatraea saccharalis (Fabricius), which was used as an outgroup species, exhibited different binding patterns than either Ostrinia species, with both Cry1Ab and Cry1F toxins binding to a 210 kDa protein. These results support the previous experiments indicating that O. nubilalis and O. furnacalis share similar patterns of susceptibility to Cry toxins. PMID:23999243

  20. Planar dipolar polymer brush: field theoretical investigations

    NASA Astrophysics Data System (ADS)

    Mahalik, Jyoti; Kumar, Rajeev; Sumpter, Bobby

    2015-03-01

    Physical properties of polymer brushes bearing monomers with permanent dipole moments and immersed in a polar solvent are investigated using self-consistent field theory (SCFT). It is found that mismatch between the permanent dipole moments of the monomer and the solvent plays a significant role in determining the height of the polymer brush. Sign as well as magnitude of the mismatch determines the extent of collapse of the polymer brush. The mismatch in the dipole moments also affects the force-distance relations and interpenetration of polymers in opposing planar brushes. In particular, an attractive force between the opposing dipolar brushes is predicted for stronger mismatch parameter. Furthermore, effects of added monovalent salt on the structure of dipolar brushes will also be presented. This investigation highlights the significance of dipolar interactions in affecting the physical properties of polymer brushes. Csmd division, Oak Ridge National Laboratory, 1 Bethel Valley Rd, Oak Ridge, TN 37831, USA.

  1. Brush Seals for Improved Steam Turbine Performance

    NASA Technical Reports Server (NTRS)

    Turnquist, Norman; Chupp, Ray; Baily, Fred; Burnett, Mark; Rivas, Flor; Bowsher, Aaron; Crudgington, Peter

    2006-01-01

    GE Energy has retrofitted brush seals into more than 19 operating steam turbines. Brush seals offer superior leakage control compared to labyrinth seals, owing to their compliant nature and ability to maintain very tight clearances to the rotating shaft. Seal designs have been established for steam turbines ranging in size from 12 MW to over 1200 MW, including fossil, nuclear, combined-cycle and industrial applications. Steam turbines present unique design challenges that must be addressed to ensure that the potential performance benefits of brush seals are realized. Brush seals can have important effects on the overall turbine system that must be taken into account to assure reliable operation. Subscale rig tests are instrumental to understanding seal behavior under simulated steam-turbine operating conditions, prior to installing brush seals in the field. This presentation discusses the technical challenges of designing brush seals for steam turbines; subscale testing; performance benefits of brush seals; overall system effects; and field applications.

  2. Loop polymer brushes from polymer single crystals

    NASA Astrophysics Data System (ADS)

    Zhou, Tian; Li, Christopher

    2014-03-01

    Loop polymer brushes represent a category of polymer brushes with both chain ends being tethered to a surface or interface with sufficiently high density. Due to this morphological difference, loop brushes exhibit distinct properties compared with traditional polymer brushes with single chain end being tethered. In our study, α, ω-functionalized polycaprolactone (PCL) single crystals were prepared as templates for polymer brush synthesis. By carefully controlling crystallization condition and immobilization, looped polymer brushes were successfully prepared. Comprehensive studies on the morphology and physical properties of these polymer brushes were carried out using Atomic Force Microscopy and FTIR. Advantages of using this method include exclusive loop morphology, high grafting density, controlled tethering sites and tunable loop size.

  3. Explicit Solvent Simulations of Friction between Brush Layers of Charged and Neutral Bottle-Brush Macromolecules

    SciTech Connect

    Carrillo, Jan-Michael; Brown, W Michael; Dobrynin, Andrey

    2012-01-01

    We study friction between charged and neutral brush layers of bottle-brush macromolecules using molecular dynamics simulations. In our simulations the solvent molecules were treated explicitly. The deformation of the bottle-brush macromolecules under the shear were studied as a function of the substrate separation and shear stress. For charged bottle-brush layers we study effect of the added salt on the brush lubricating properties to elucidate factors responsible for energy dissipation in charged and neutral brush systems. Our simulations have shown that for both charged and neutral brush systems the main deformation mode of the bottle-brush macromolecule is associated with the backbone deformation. This deformation mode manifests itself in the backbone deformation ratio, , and shear viscosity, , to be universal functions of the Weissenberg number W. The value of the friction coefficient, , and viscosity, , are larger for the charged bottle-brush coatings in comparison with those for neutral brushes at the same separation distance, D, between substrates. The additional energy dissipation generated by brush sliding in charged bottle-brush systems is due to electrostatic coupling between bottle-brush and counterion motion. This coupling weakens as salt concentration, cs, increases resulting in values of the viscosity, , and friction coefficient, , approaching corresponding values obtained for neutral brush systems.

  4. Polyelectrolyte brushes: theory, modelling, synthesis and applications.

    PubMed

    Das, Siddhartha; Banik, Meneka; Chen, Guang; Sinha, Shayandev; Mukherjee, Rabibrata

    2015-11-28

    Polyelectrolyte (PE) brushes are a special class of polymer brushes (PBs) containing charges. Polymer chains attain "brush"-like configuration when they are grafted or get localized at an interface (solid-fluid or liquid-fluid) with sufficiently close proximity between two-adjacent grafted polymer chains - such a proximity triggers a particular nature of interaction between the adjacent polymer molecules forcing them to stretch orthogonally to the grafting interface, instead of random-coil arrangement. In this review, we discuss the theory, synthesis, and applications of PE brushes. The theoretical discussion starts with the standard scaling concepts for polymer and PE brushes; following that, we shed light on the state of the art in continuum modelling approaches for polymer and PE brushes directed towards analysis beyond the scaling calculations. A special emphasis is laid in pinpointing the cases for which the PE electrostatic effects can be de-coupled from the PE entropic and excluded volume effects; such de-coupling is necessary to appropriately probe the complicated electrostatic effects arising from pH-dependent charging of the PE brushes and the use of these effects for driving liquid and ion transport at the interfaces covered with PE brushes. We also discuss the atomistic simulation approaches for polymer and PE brushes. Next we provide a detailed review of the existing approaches for the synthesis of polymer and PE brushes on interfaces, nanoparticles, and nanochannels, including mixed brushes and patterned brushes. Finally, we discuss some of the possible applications and future developments of polymer and PE brushes grafted on a variety of interfaces. PMID:26399305

  5. Nanotribology of charged polymer brushes

    NASA Astrophysics Data System (ADS)

    Klein, Jacob

    Polymers at surfaces, whose modern understanding may be traced back to early work by Sam Edwards1, have become a paradigm for modification of surface properties, both as steric stabilizers and as remarkable boundary lubricants2. Charged polymer brushes are of particular interest, with both technological implications and especially biological relevance where most macromolecules are charged. In the context of biolubrication, relevant in areas from dry eye syndrome to osteoarthritis, charged polymer surface phases and their complexes with other macromolecules may play a central role. The hydration lubrication paradigm, where tenaciously-held yet fluid hydration shells surrounding ions or zwitterions serve as highly-efficient friction-reducing elements, has been invoked to understand the excellent lubrication provided both by ionized3 and by zwitterionic4 brushes. In this talk we describe recent advances in our understanding of the nanotribology of such charged brush systems. We consider interactions between charged end-grafted polymers, and how one may disentangle the steric from the electrostatic surface forces5. We examine the limits of lubrication by ionized brushes, both synthetic and of biological origins, and how highly-hydrated zwitterionic chains may provide extremely effective boundary lubrication6. Finally we describe how the lubrication of articular cartilage in the major joints, a tribosystem presenting some of the greatest challenges and opportunities, may be understood in terms of a supramolecular synergy between charged surface-attached polymers and zwitterionic groups7. Work supported by European Research Council (HydrationLube), Israel Science Foundation (ISF), Petroleum Research Fund of the American Chemical Society, ISF-NSF China Joint Program.

  6. Does apical membrane GLUT2 have a role in intestinal glucose uptake?

    PubMed Central

    Naftalin, Richard J

    2014-01-01

    It has been proposed that the non-saturable component of intestinal glucose absorption, apparent following prolonged exposure to high intraluminal glucose concentrations, is mediated via the low affinity glucose and fructose transporter, GLUT2, upregulated within the small intestinal apical border. The evidence that the non-saturable transport component is mediated via an apical membrane sugar transporter is that it is inhibited by phloretin, after exposure to phloridzin. Since the other apical membrane sugar transporter, GLUT5, is insensitive to inhibition by either cytochalasin B, or phloretin, GLUT2 was deduced to be the low affinity sugar transport route. As in its uninhibited state, polarized intestinal glucose absorption depends both on coupled entry of glucose and sodium across the brush border membrane and on the enterocyte cytosolic glucose concentration exceeding that in both luminal and submucosal interstitial fluids, upregulation of GLUT2 within the intestinal brush border will usually stimulate downhill glucose reflux to the intestinal lumen from the enterocytes; thereby reducing, rather than enhancing net glucose absorption across the luminal surface. These states are simulated with a computer model generating solutions to the differential equations for glucose, Na and water flows between luminal, cell, interstitial and capillary compartments. The model demonstrates that uphill glucose transport via SGLT1 into enterocytes, when short-circuited by any passive glucose carrier in the apical membrane, such as GLUT2, will reduce transcellular glucose absorption and thereby lead to increased paracellular flow. The model also illustrates that apical GLUT2 may usefully act as an osmoregulator to prevent excessive enterocyte volume change with altered luminal glucose concentrations. PMID:25671087

  7. Myosin 1b Regulates Amino Acid Transport by Associating Transporters with the Apical Plasma Membrane of Kidney Cells

    PubMed Central

    Komaba, Shigeru; Coluccio, Lynne M.

    2015-01-01

    Amino acid transporters (AATers) in the brush border of the apical plasma membrane (APM) of renal proximal tubule (PT) cells mediate amino acid transport (AAT). We found that the membrane-associated class I myosin myosin 1b (Myo1b) localized at the apical brush border membrane of PTs. In opossum kidney (OK) 3B/2 epithelial cells, which are derived from PTs, expressed rat Myo1b-GFP colocalized in patched microvilli with expressed mouse V5-tagged SIT1 (SIT1-V5), which mediates neutral amino acid transport in OK cells. Lentivirus-mediated delivery of opossum Myo1b-specific shRNA resulted in knockdown (kd) of Myo1b expression, less SIT1-V5 at the APM as determined by localization studies, and a decrease in neutral AAT as determined by radioactive uptake assays. Myo1b kd had no effect on Pi transport or noticeable change in microvilli structure as determined by rhodamine phalloidin staining. The studies are the first to define a physiological role for Myo1b, that of regulating renal AAT by modulating the association of AATers with the APM. PMID:26361046

  8. Brush seals for turbine engine fuel conservation

    NASA Astrophysics Data System (ADS)

    Sousa, Mike

    1994-07-01

    The program objective is to demonstrate brush seals for replacing labyrinth seals in turboprop engines. The approach taken was to design and procure brush seals with assistance from Sealol, modify and instrument an existing T407 low pressure turbine test rig, replace inner balance piston and outer balance piston labyrinth seals with brush seals, conduct cyclic tests to evaluate seal leakage at operating pressures and temperatures, and evaluate effect of seal pack width and rotor eccentricity. Results are presented in viewgraph format and show that brush seals offer performance advantages over labyrinth seals.

  9. Brush seals for turbine engine fuel conservation

    NASA Technical Reports Server (NTRS)

    Sousa, Mike

    1994-01-01

    The program objective is to demonstrate brush seals for replacing labyrinth seals in turboprop engines. The approach taken was to design and procure brush seals with assistance from Sealol, modify and instrument an existing T407 low pressure turbine test rig, replace inner balance piston and outer balance piston labyrinth seals with brush seals, conduct cyclic tests to evaluate seal leakage at operating pressures and temperatures, and evaluate effect of seal pack width and rotor eccentricity. Results are presented in viewgraph format and show that brush seals offer performance advantages over labyrinth seals.

  10. Nanopatterned polymer brushes: conformation, fabrication and applications

    NASA Astrophysics Data System (ADS)

    Yu, Qian; Ista, Linnea K.; Gu, Renpeng; Zauscher, Stefan; López, Gabriel P.

    2015-12-01

    Surfaces with end-grafted, nanopatterned polymer brushes that exhibit well-defined feature dimensions and controlled chemical and physical properties provide versatile platforms not only for investigation of nanoscale phenomena at biointerfaces, but also for the development of advanced devices relevant to biotechnology and electronics applications. In this review, we first give a brief introduction of scaling behavior of nanopatterned polymer brushes and then summarize recent progress in fabrication and application of nanopatterned polymer brushes. Specifically, we highlight applications of nanopatterned stimuli-responsive polymer brushes in the areas of biomedicine and biotechnology.

  11. Nanopatterned polymer brushes: conformation, fabrication and applications.

    PubMed

    Yu, Qian; Ista, Linnea K; Gu, Renpeng; Zauscher, Stefan; López, Gabriel P

    2016-01-14

    Surfaces with end-grafted, nanopatterned polymer brushes that exhibit well-defined feature dimensions and controlled chemical and physical properties provide versatile platforms not only for investigation of nanoscale phenomena at biointerfaces, but also for the development of advanced devices relevant to biotechnology and electronics applications. In this review, we first give a brief introduction of scaling behavior of nanopatterned polymer brushes and then summarize recent progress in fabrication and application of nanopatterned polymer brushes. Specifically, we highlight applications of nanopatterned stimuli-responsive polymer brushes in the areas of biomedicine and biotechnology. PMID:26648412

  12. Development of a Brush Seals Program Leading to Ceramic Brush Seals

    NASA Technical Reports Server (NTRS)

    Hendricks, Robert C.; Flower, Ralph; Howe, Harold

    1994-01-01

    Some events of a U.S. Army/NASA Lewis Research Center brush seals program are reviewed, and the development of ceramic brush seals is described. Some preliminary room-temperature flow data are given, and the results of testing metallic brushes in cryogenic nitrogen are discussed.

  13. Brush seal numerical simulation: Concepts and advances

    NASA Astrophysics Data System (ADS)

    Braun, M. J.; Kudriavtsev, V. V.

    1994-07-01

    The development of the brush seal is considered to be most promising among the advanced type seals that are presently in use in the high speed turbomachinery. The brush is usually mounted on the stationary portions of the engine and has direct contact with the rotating element, in the process of limiting the 'unwanted' leakage flows between stages, or various engine cavities. This type of sealing technology is providing high (in comparison with conventional seals) pressure drops due mainly to the high packing density (around 100 bristles/sq mm), and brush compliance with the rotor motions. In the design of modern aerospace turbomachinery leakage flows between the stages must be minimal, thus contributing to the higher efficiency of the engine. Use of the brush seal instead of the labyrinth seal reduces the leakage flow by one order of magnitude. Brush seals also have been found to enhance dynamic performance, cost less, and are lighter than labyrinth seals. Even though industrial brush seals have been successfully developed through extensive experimentation, there is no comprehensive numerical methodology for the design or prediction of their performance. The existing analytical/numerical approaches are based on bulk flow models and do not allow the investigation of the effects of brush morphology (bristle arrangement), or brushes arrangement (number of brushes, spacing between them), on the pressure drops and flow leakage. An increase in the brush seal efficiency is clearly a complex problem that is closely related to the brush geometry and arrangement, and can be solved most likely only by means of a numerically distributed model.

  14. Brush seal numerical simulation: Concepts and advances

    NASA Technical Reports Server (NTRS)

    Braun, M. J.; Kudriavtsev, V. V.

    1994-01-01

    The development of the brush seal is considered to be most promising among the advanced type seals that are presently in use in the high speed turbomachinery. The brush is usually mounted on the stationary portions of the engine and has direct contact with the rotating element, in the process of limiting the 'unwanted' leakage flows between stages, or various engine cavities. This type of sealing technology is providing high (in comparison with conventional seals) pressure drops due mainly to the high packing density (around 100 bristles/sq mm), and brush compliance with the rotor motions. In the design of modern aerospace turbomachinery leakage flows between the stages must be minimal, thus contributing to the higher efficiency of the engine. Use of the brush seal instead of the labyrinth seal reduces the leakage flow by one order of magnitude. Brush seals also have been found to enhance dynamic performance, cost less, and are lighter than labyrinth seals. Even though industrial brush seals have been successfully developed through extensive experimentation, there is no comprehensive numerical methodology for the design or prediction of their performance. The existing analytical/numerical approaches are based on bulk flow models and do not allow the investigation of the effects of brush morphology (bristle arrangement), or brushes arrangement (number of brushes, spacing between them), on the pressure drops and flow leakage. An increase in the brush seal efficiency is clearly a complex problem that is closely related to the brush geometry and arrangement, and can be solved most likely only by means of a numerically distributed model.

  15. 21 CFR 884.1100 - Endometrial brush.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Endometrial brush. 884.1100 Section 884.1100 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES OBSTETRICAL AND GYNECOLOGICAL DEVICES Obstetrical and Gynecological Diagnostic Devices § 884.1100 Endometrial brush. (a) Identification....

  16. 21 CFR 884.1100 - Endometrial brush.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Endometrial brush. 884.1100 Section 884.1100 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES OBSTETRICAL AND GYNECOLOGICAL DEVICES Obstetrical and Gynecological Diagnostic Devices § 884.1100 Endometrial brush. (a) Identification....

  17. Brush-Wheel Samplers for Planetary Exploration

    NASA Technical Reports Server (NTRS)

    Rivellini, Tommaso

    2003-01-01

    A report proposes brush-wheel mechanisms for acquiring samples of soils from remote planets. In simplest terms, such a mechanism would contain brush wheels that would be counter-rotated at relatively high speed. The mechanism would be lowered to the ground from a spacecraft or other exploratory vehicle. Upon contact with the ground, the counter-rotating brush wheels would kick soil up into a collection chamber. Thus, in form and function, the mechanism would partly resemble traditional street and carpet sweepers. The main advantage of using of brush wheels (in contradistinction to cutting wheels or other, more complex mechanisms) is that upon encountering soil harder than expected, the brushes could simply deflect and the motor(s) could continue to turn. That is, sufficiently flexible brushes would afford resistance to jamming and to overloading of the motors used to rotate the brushes, and so the motors could be made correspondingly lighter and less power hungry. Of course, one could select the brush stiffnesses and motor torques and speeds for greatest effectiveness in sampling soil of a specific anticipated degree of hardness.

  18. Dual wound dc brush motor gearhead

    NASA Technical Reports Server (NTRS)

    Henson, Barrie W.

    1986-01-01

    The design requirements, the design, development tests and problems, the qualification and life test and the findings of the strip examination of a dual wound DC brushed motor gearhead are described. It is the only space qualified dual wound dc brushed motor gearhead in Europe.

  19. 21 CFR 886.1090 - Haidinger brush.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Haidinger brush. 886.1090 Section 886.1090 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES OPHTHALMIC DEVICES Diagnostic Devices § 886.1090 Haidinger brush. (a) Identification. A...

  20. A Brush Seals Program Modeling and Developments

    NASA Technical Reports Server (NTRS)

    Hendricks, Robert C.; Flower, Ralph; Howe, Harold

    1996-01-01

    Some events of a U.S. Army/NASA Lewis Research Center brush seals program are reviewed, and the development of ceramic brush seals is described. Some preliminary room-temperature flow data are modeled and compare favorably to the results of Ergun.

  1. Border Trade Facilitation System

    Energy Science and Technology Software Center (ESTSC)

    1999-06-07

    System for managing commercial trans-border shipments via the Internet. This application assists geographically distributed commercial and governmental users with the transshipment of goods across the U.S. - Mexico border.

  2. Resistance to the Bacillus thuringiensis bioinsecticide in a field population of Plutella xylostella is due to a change in a midgut membrane receptor.

    PubMed

    Ferré, J; Real, M D; Van Rie, J; Jansens, S; Peferoen, M

    1991-06-15

    The biochemical mechanism for resistance to Bacillus thuringiensis crystal proteins was studied in a field population of diamondback moths (Plutella xylostella) with a reduced susceptibility to the bioinsecticidal spray. The toxicity and binding characteristics of three crystal proteins [CryIA(b), CryIB, and CryIC] were compared between the field population and a laboratory strain. The field population proved resistant (greater than 200-fold compared with the laboratory strain) to CryIA(b), one of the crystal proteins in the insecticidal formulation. Binding studies showed that the two strains differ in a membrane receptor that recognizes CryIA(b). This crystal protein did not bind to the brush-border membrane of the midgut epithelial cells of the field population, either because of strongly reduced binding affinity or because of the complete absence of the receptor molecule. Both strains proved fully susceptible to the CryIB and CryIC crystal proteins, which were not present in the B. thuringiensis formulation used in the field. Characteristics of CryIB and CryIC binding to brush-border membranes of midgut epithelial cells were virtually identical in the laboratory and the field population. PMID:2052591

  3. Zwitterionic poly(amino acid methacrylate) brushes.

    PubMed

    Alswieleh, Abdullah M; Cheng, Nan; Canton, Irene; Ustbas, Burcin; Xue, Xuan; Ladmiral, Vincent; Xia, Sijing; Ducker, Robert E; El Zubir, Osama; Cartron, Michael L; Hunter, C Neil; Leggett, Graham J; Armes, Steven P

    2014-07-01

    A new cysteine-based methacrylic monomer (CysMA) was conveniently synthesized via selective thia-Michael addition of a commercially available methacrylate-acrylate precursor in aqueous solution without recourse to protecting group chemistry. Poly(cysteine methacrylate) (PCysMA) brushes were grown from the surface of silicon wafers by atom-transfer radical polymerization. Brush thicknesses of ca. 27 nm were achieved within 270 min at 20 °C. Each CysMA residue comprises a primary amine and a carboxylic acid. Surface zeta potential and atomic force microscopy (AFM) studies of the pH-responsive PCysMA brushes confirm that they are highly extended either below pH 2 or above pH 9.5, since they possess either cationic or anionic character, respectively. At intermediate pH, PCysMA brushes are zwitterionic. At physiological pH, they exhibit excellent resistance to biofouling and negligible cytotoxicity. PCysMA brushes undergo photodegradation: AFM topographical imaging indicates significant mass loss from the brush layer, while XPS studies confirm that exposure to UV radiation produces surface aldehyde sites that can be subsequently derivatized with amines. UV exposure using a photomask yielded sharp, well-defined micropatterned PCysMA brushes functionalized with aldehyde groups that enable conjugation to green fluorescent protein (GFP). Nanopatterned PCysMA brushes were obtained using interference lithography, and confocal microscopy again confirmed the selective conjugation of GFP. Finally, PCysMA undergoes complex base-catalyzed degradation in alkaline solution, leading to the elimination of several small molecules. However, good long-term chemical stability was observed when PCysMA brushes were immersed in aqueous solution at physiological pH. PMID:24884533

  4. Zwitterionic Poly(amino acid methacrylate) Brushes

    PubMed Central

    2014-01-01

    A new cysteine-based methacrylic monomer (CysMA) was conveniently synthesized via selective thia-Michael addition of a commercially available methacrylate-acrylate precursor in aqueous solution without recourse to protecting group chemistry. Poly(cysteine methacrylate) (PCysMA) brushes were grown from the surface of silicon wafers by atom-transfer radical polymerization. Brush thicknesses of ca. 27 nm were achieved within 270 min at 20 °C. Each CysMA residue comprises a primary amine and a carboxylic acid. Surface zeta potential and atomic force microscopy (AFM) studies of the pH-responsive PCysMA brushes confirm that they are highly extended either below pH 2 or above pH 9.5, since they possess either cationic or anionic character, respectively. At intermediate pH, PCysMA brushes are zwitterionic. At physiological pH, they exhibit excellent resistance to biofouling and negligible cytotoxicity. PCysMA brushes undergo photodegradation: AFM topographical imaging indicates significant mass loss from the brush layer, while XPS studies confirm that exposure to UV radiation produces surface aldehyde sites that can be subsequently derivatized with amines. UV exposure using a photomask yielded sharp, well-defined micropatterned PCysMA brushes functionalized with aldehyde groups that enable conjugation to green fluorescent protein (GFP). Nanopatterned PCysMA brushes were obtained using interference lithography, and confocal microscopy again confirmed the selective conjugation of GFP. Finally, PCysMA undergoes complex base-catalyzed degradation in alkaline solution, leading to the elimination of several small molecules. However, good long-term chemical stability was observed when PCysMA brushes were immersed in aqueous solution at physiological pH. PMID:24884533

  5. Effectiveness of oscillatory gutter brushes in removing street sweeping waste.

    PubMed

    Vanegas-Useche, Libardo V; Abdel-Wahab, Magd M; Parker, Graham A

    2015-09-01

    In this paper, the novel concept of oscillatory gutter brushes of road sweepers is studied experimentally. Their effectiveness in removing different debris types is studied by means of a brushing test rig. The debris types dealt with are medium-size gravel, small and fine particles, wet thin debris, and compacted debris. The performance of two types of brushes, cutting and F128, under diverse operating conditions is investigated. The purpose of the tests is to ascertain whether brush oscillations superimposed onto brush rotation improve sweeping effectiveness. According to the results, brush oscillations seem to be useful for increasing brushing effectiveness in the case of bonded particles and wet thin debris, especially for bonded debris, but they seem to be of no value for other loosed debris. Also, appropriate values of brush penetration, sweeper speed, brush angle of attack, rotational speed, and frequency of brush speed oscillations, for the debris types studied are provided. PMID:26026947

  6. Semiflexible polymer brushes and the brush-mushroom crossover.

    PubMed

    Egorov, Sergei A; Hsu, Hsiao-Ping; Milchev, Andrey; Binder, Kurt

    2015-04-01

    Semiflexible polymers end-grafted to a repulsive planar substrate under good solvent conditions are studied by scaling arguments, computer simulations, and self-consistent field theory. Varying the chain length N, persistence length lp, and grafting density σg, the chain linear dimensions and distribution functions of all monomers and of the free chain ends are studied. Particular attention is paid to the limit of very small σg, where the grafted chains behave as "mushrooms" no longer interacting with each other. Unlike a flexible mushroom, which has a self-similar structure from the size (a) of an effective monomer up to the mushroom height (h/a ∝ N(v), ν ≈ 3/5), a semiflexible mushroom (like a free semiflexible chain) exhibits three different scaling regimes, h/a ∝ N for contour length L = Na < lp, a Gaussian regime, h/a ∝ (Llp)(1/2)/a for lp ≪ L ≪ R* ∝ (lp(2)/a), and a regime controlled by excluded volume, h/a ∝ (lp/a)(1/5)N(ν). The semiflexible brush is predicted to scale as h/a ∝ (lpaσg)(1/3)N in the excluded volume regime, and h/a ∝ (lpa(3)σ(2))(1/4)N in the Gaussian regime. Since in the volume taken by a semiflexible mushroom excluded-volume interactions are much weaker in comparison to a flexible mushroom, there occurs an additional regime where semiflexible mushrooms overlap without significant chain stretching. Moreover, since the size of a semiflexible mushroom is much larger than the size of a flexible mushroom with the same N, the crossover from mushroom to brush behavior is predicted to take place at much smaller densities than for fully flexible chains. The numerical results, however, confirm the scaling predictions only qualitatively; for chain lengths that are relevant for experiments, often intermediate effective exponents are observed due to extended crossovers. PMID:25687784

  7. Tooth brushing inhibits oral bacteria in dogs.

    PubMed

    Watanabe, Kazuhiro; Hayashi, Kotaro; Kijima, Saku; Nonaka, Chie; Yamazoe, Kazuaki

    2015-10-01

    In this study, scaling, polishing and daily tooth brushing were performed in 20 beagle dogs, and the number of oral bacteria was determined using a bacterial counter. The dogs were randomized into the scaling (S), scaling + polishing (SP), scaling + tooth daily brushing (SB) and scaling + polishing + tooth daily brushing (SPB) groups. Samples were collected from the buccal surface of the maxillary fourth premolars of the dogs immediately after scaling and every week thereafter from weeks 1 to 8. Throughout the study, the number of bacteria was significantly lower in the SB and SPB groups compared with the S group. The findings suggest that daily tooth brushing inhibited oral bacterial growth in the dogs. PMID:25994486

  8. 3D-patterned polymer brush surfaces

    NASA Astrophysics Data System (ADS)

    Zhou, Xuechang; Liu, Xuqing; Xie, Zhuang; Zheng, Zijian

    2011-12-01

    Polymer brush-based three-dimensional (3D) structures are emerging as a powerful platform to engineer a surface by providing abundant spatially distributed chemical and physical properties. In this feature article, we aim to give a summary of the recent progress on the fabrication of 3D structures with polymer brushes, with a particular focus on the micro- and nanoscale. We start with a brief introduction on polymer brushes and the challenges to prepare their 3D structures. Then, we highlight the recent advances of the fabrication approaches on the basis of traditional polymerization time and grafting density strategies, and a recently developed feature density strategy. Finally, we provide some perspective outlooks on the future directions of engineering the 3D structures with polymer brushes.

  9. Brushing Your Child’s Teeth

    MedlinePlus

    ... the toothpaste rather than swallowing it. For children under age 3, use just a small amount of toothpaste ... floss and brush your teeth every day. Children under age 6 may be able to handle a toothbrush ...

  10. Three-Wheel Brush-Wheel Sampler

    NASA Technical Reports Server (NTRS)

    Duckworth, Geoffrey A.; Liu, Jun; Brown, Mark G.

    2010-01-01

    A new sampler is similar to a common snow blower, but is robust and effective in sample collection. The brush wheels are arranged in a triangle shape, each driven by a brushless DC motor and planetary gearhead embedded in the wheel shaft. Its speed can be varied from 800 - 2,000 rpm, depending on the surface regolith resistance. The sample-collecting flow path, and internal features, are designed based on flow dynamics, and the sample-collecting rates have consistently exceeded the requirement under various conditions that span the range of expected surface properties. The brush-wheel sampler (BWS) is designed so that the flow channel is the main body of the apparatus, and links the brush-wheel assembly to the sample canister. The combination of the three brush wheels, the sample flow path, and the canister location make sample collection, storage, and transfer an easier task.

  11. Flow visualization in a simulated brush seal

    NASA Technical Reports Server (NTRS)

    Braun, M. J.; Canacci, V.; Hendricks, R. C.

    1990-01-01

    A method to visualize and characterize the complex flow fields in simulated brush seals is presented. The brush seal configuration was tested in a water and then in an oil tunnel. The visualization procedure revealed typical regions that are rivering, jetting, vortical or lateral flows and exist upstream, downstream or within the seal. Such flows are engendered by variations in fiber void that are spatial and temporal and affect changes in seal leakage and stability. While the effects of interface motion for linear or cylindrical configurations have not been considered herein, it is believed that the observed flow fields characterize flow phenomenology in both circular and linear brush seals. The axial pressure profiles upstream, across and downstream of the brush in the oil tunnel have been measured under a variety of inlet pressure conditions and the ensuing pressure maps are presented and discussed.

  12. Water driven turbine/brush pipe cleaner

    NASA Astrophysics Data System (ADS)

    Werlink, Rudy J.

    1995-04-01

    Assemblies are disclosed for cleaning the inside walls of pipes and tubes. A first embodiment includes a small turbine with angled blades axially mounted on one end of a standoff support. An O-ring for stabilizing the assembly within the pipe is mounted in a groove within the outer ring. A replaceable circular brush is fixedly mounted on the opposite end of the standoff support and can be used for cleaning tubes and pipes of various diameters, lengths and configurations. The turbine, standoff support, and brush spin in unison relative to a hub bearing that is fixedly attached to a wire upstream of the assembly. The nonrotating wire is for retaining the assembly in tension and enabling return of the assembly to the pipe entrance. The assembly is initially placed in the pipe or tube to be cleaned. A pressurized water or solution source is provided at a required flow-rate to propel the assembly through the pipe or tube. The upstream water pressure propels and spins the turbine, standoff support and brush. The rotating brush combined with the solution cleans the inside of the pipe. The solution flows out of the other end of the pipe with the brush rotation controlled by the flow-rate. A second embodiment is similar to the first embodiment but instead includes a circular shaped brush with ring backing mounted in the groove of the exterior ring of the turbine, and also reduces the size of the standoff support or eliminates the standoff support.

  13. Water driven turbine/brush pipe cleaner

    NASA Technical Reports Server (NTRS)

    Werlink, Rudy J. (Inventor)

    1995-01-01

    Assemblies are disclosed for cleaning the inside walls of pipes and tubes. A first embodiment includes a small turbine with angled blades axially mounted on one end of a standoff support. An O-ring for stabilizing the assembly within the pipe is mounted in a groove within the outer ring. A replaceable circular brush is fixedly mounted on the opposite end of the standoff support and can be used for cleaning tubes and pipes of various diameters, lengths and configurations. The turbine, standoff support, and brush spin in unison relative to a hub bearing that is fixedly attached to a wire upstream of the assembly. The nonrotating wire is for retaining the assembly in tension and enabling return of the assembly to the pipe entrance. The assembly is initially placed in the pipe or tube to be cleaned. A pressurized water or solution source is provided at a required flow-rate to propel the assembly through the pipe or tube. The upstream water pressure propels and spins the turbine, standoff support and brush. The rotating brush combined with the solution cleans the inside of the pipe. The solution flows out of the other end of the pipe with the brush rotation controlled by the flow-rate. A second embodiment is similar to the first embodiment but instead includes a circular shaped brush with ring backing mounted in the groove of the exterior ring of the turbine, and also reduces the size of the standoff support or eliminates the standoff support.

  14. Rotating Intershaft Brush Seal Project

    NASA Technical Reports Server (NTRS)

    Krawiecki, Stephen; Mehta, Jayesh; Holloway, Gary

    2006-01-01

    The pursuit of high Mach number flight presents several challenges to the airframe and engine design engineers. Most obvious is the resulting high temperatures encountered as the aircraft approaches Mach 3 and above. The encountered high temperatures and shaft speeds of engines require rethinking in the areas of material selections, component design and component operating life. In the area of sump compartment sealing, one of the most difficult sealing applications is the sealing of an engine s rear sump. Normally this sump will need some method of sealing between two rotating shafts. This sealing operation is done with an intershaft seal. The aft sump region also presents an additional design requirement for the intershaft seal. This region has to absorb the engine s thermal growth, which means that in the seal area, axial movement, on the order of 0.30 in., between the rotating shafts must be tolerated. A new concept or new technology of sealing an intershaft sump configuration is being developed. This concept, called a rotating intershaft brush seal has key attributes that will allow this seal to perform better, in the demanding environment of sealing an aft sump with two rotating shafts, when compared to today s sealing technology of labyrinth and carbon sea

  15. Thermoresponsive PDMAEMA Brushes: Effect of Gold Nanoparticle Deposition.

    PubMed

    Yenice, Zuleyha; Schön, Sebastian; Bildirir, Hakan; Genzer, Jan; von Klitzing, Regine

    2015-08-13

    The paper addresses the effect of gold nanoparticle (Au-NP) deposition on the thermoresponsive volume phase transition of the weak polyelectrolyte poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) brushes. PDMAEMA brushes were synthesized via surface-initiated atom transfer radical polymerization (SI-ATRP). The PDMAEMA/Au-NP composite brushes were fabricated by immersing the brush modified wafer in the Au-NP suspension. Atomic force microscopy (AFM), ellipsometry, and scanning electron microscopy (SEM) have been employed to characterize the neat PDMAEMA brushes and PDMAEMA/Au-NP composite brushes. All neat PDMAEMA brushes swelled below the volume phase transition temperature and collapsed with increasing temperature over a broad temperature range independent of the initial brush thickness. Water uptake of the brushes is also independent of initial brush thickness. The adsorption of the charged Au-NPs significantly affects the degree of swelling and the thermoresponsive properties of the brushes. PDMAEMA/Au-NP composite brushes do not exhibit any noticeable phase transition at the experimental temperature range irrespective of the initial brush thickness. The reason for this behavior is attributed to a combination of the following: the decreased conformational entropy of the Au-NP adsorbed polymer chains, the increased hydrophilicity of the system due to the charged Au-NPs, and the ≈13 nm diameter Au-NPs causing steric hindrance. We have also shown that the AFM full-indentation method can be successfully applied to determine the polymer brush thicknesses. PMID:26132296

  16. Cold plasma brush generated at atmospheric pressure

    SciTech Connect

    Duan Yixiang; Huang, C.; Yu, Q. S.

    2007-01-15

    A cold plasma brush is generated at atmospheric pressure with low power consumption in the level of several watts (as low as 4 W) up to tens of watts (up to 45 W). The plasma can be ignited and sustained in both continuous and pulsed modes with different plasma gases such as argon or helium, but argon was selected as a primary gas for use in this work. The brush-shaped plasma is formed and extended outside of the discharge chamber with typical dimension of 10-15 mm in width and less than 1.0 mm in thickness, which are adjustable by changing the discharge chamber design and operating conditions. The brush-shaped plasma provides some unique features and distinct nonequilibrium plasma characteristics. Temperature measurements using a thermocouple thermometer showed that the gas phase temperatures of the plasma brush are close to room temperature (as low as 42 deg. C) when running with a relatively high gas flow rate of about 3500 ml/min. For an argon plasma brush, the operating voltage from less than 500 V to about 2500 V was tested, with an argon gas flow rate varied from less than 1000 to 3500 ml/min. The cold plasma brush can most efficiently use the discharge power as well as the plasma gas for material and surface treatment. The very low power consumption of such an atmospheric argon plasma brush provides many unique advantages in practical applications including battery-powered operation and use in large-scale applications. Several polymer film samples were tested for surface treatment with the newly developed device, and successful changes of the wettability property from hydrophobic to hydrophilic were achieved within a few seconds.

  17. The Border Pedagogy Revisited

    ERIC Educational Resources Information Center

    Kazanjian, Christopher John

    2011-01-01

    Border pedagogy is a multicultural educational approach utilized in multicultural settings to help students understand their histories and experiences and how it affects their identities and cultures. The approach seeks to produce intellectuals that transcend physical and metaphysical boundaries. The goal of border pedagogy is to remove cultural…

  18. Performance characteristics of brush seals for limited-life engines

    SciTech Connect

    Chupp, R.E. ); Dowler, C.A. )

    1993-04-01

    Brush seals are potential replacements for air-to-air labyrinth seals in gas turbine engines. An investigation has been conducted to determine the performance characteristics of brush seals for application in limited-life gas turbine engines. An elevated temperature, rotating test rig was designed and built to test labyrinth and brush seals in simulated subsonic and supersonic engine conditions. Results from initial tests for subsonic applications demonstrated that brush seals exhibit appreciably lower leakage compared to labyrinth seals, and thus offer significant engine performance improvements. Performance results have been obtained showing the effect of various brush seal parameters, including: initial interference, backplate gap, and multiple brush seals in series.

  19. Motor Brush Testing for Mars and Vacuum

    NASA Technical Reports Server (NTRS)

    Noon, Don E.

    1999-01-01

    Brush motors have been qualified and flown successfully on Mars missions, but upcoming missions require longer life and higher power. A test program was therefore undertaken to identify the best brush material for operation in the Mars atmosphere. Six different brush materials were used in 18 identical motors and operated under various load conditions for a period of four weeks in low-pressure CO2. All motors performed acceptably, with accumulated motor revolutions between 98 and 144 million revolutions, depending on load. A proprietary silver-graphite material from Superior Carbon (SG54-27) appears to be the best choice for long life, but even the stock copper-graphite brushes performed reliably with acceptable wear. The motors from the CO2 test were then cleaned and run in vacuum for 2 weeks. The difference in results was dramatic, with 5 motors failing catastrophically and wear rates increasing by orders of magnitude for the SG54-27 material. Three brush materials survived the test with no failures: SG54-27 with a proprietary Ball Aerospace impregnation, a silver-graphite-molybdenum disulfide material from Superior Carbon (SG59), and a copper sulfide-graphite material also from Superior Carbon (BG91).

  20. Bidirectional Brush Seals: Post-Test Analysis

    NASA Technical Reports Server (NTRS)

    Hendricks, Robert C.; Wilson, Jack; Wu, Tom Y.; Flower, Ralph; Mullen, Robert L.

    1997-01-01

    A post-test analysis of a set of inside-diameter/outside-diameter (ID/OD) bidirectional brush seals used in three-port wave rotor tests was undertaken to determine brush bristle and configuration wear, pullout, and rotor coating wear. The results suggest that sharp changes in the pressure profiles were not well reflected in bristle tip configuration patterns or wear. Also, positive-to-negative changes in axial pressure gradients appeared to have little effect on the backing plates. Although the brushes had similar porosities, they had very different unpacked arrays. This difference could explain the departure of experimental data from computational fluid dynamics flow predictions for well-packed arrays at higher pressure drops. The rotor wear led to "car-track" scars (upper and lower wear bands) with a whipped surface between the bands. Those bands may have resulted from bristle stiffening at the fence and gap plates during alternate portions of the rotor cycle. Within the bristle response range the wear surface reflected the pressure distribution effect on bristle motion. No sacrificial metallurgical data were taken. The bristles did wear, with correspondingly more wear on the ID brush configurations than on the OD configurations; the complexity in constructing the ID brush was a factor.

  1. Two-stage eutectic metal brushes

    DOEpatents

    Hsu, John S [Oak Ridge, TN

    2009-07-14

    A two-stage eutectic metal brush assembly having a slip ring rigidly coupled to a shaft, the slip ring being electrically coupled to first voltage polarity. At least one brush is rigidly coupled to a second ring and slidingly engaged to the slip ring. Eutectic metal at least partially fills an annulus between the second ring and a stationary ring. At least one conductor is rigidly coupled to the stationary ring and electrically coupled to a second voltage polarity. Electrical continuity is maintained between the first voltage polarity and the second voltage polarity. Periodic rotational motion is present between the stationary ring and the second ring. Periodic rotational motion is also present between the brush and the slip ring.

  2. Improved molybdenum disulfide-silver motor brushes have extended life

    NASA Technical Reports Server (NTRS)

    Horton, J. C.; King, H. M.

    1964-01-01

    Motor brushes of proper quantities of molybdenum disulfide and copper or silver are manufactured by sintering techniques. Graphite molds are used. These brushes operate satisfactorily for long periods in normal atmosphere or in a high-vacuum environment.

  3. Heterogeneous nucleation of calcium oxalate crystals in the presence of membrane vesicles

    NASA Astrophysics Data System (ADS)

    Khan, Saeed R.; Whalen, Patrick O.; Glenton, Patricia A.

    1993-12-01

    Membrane-assisted crystallization of calcium oxalate was studied in vitro, using constant composition methodology. Rat renal tubular brush border membrane vesicles were incubated in supersaturated solution of calcium oxalate. Calcium and oxalate depletion started much earlier in the presence of the vesicles than in their absence; within 8, 32, or 258 min of the incubation of vesicles in calcium oxalate solutions of relative supersaturation of 12, 10 or 6 respectively. Thin plate-like crystals with jagged edges formed in association with the membrane vesicles. Since crystal nucleation in the presence of membrane vesicles started within 8 min at a relative supersaturation as low as 12, it will start significantly earlier in the urine of stone formers which is known to have higher relative supersaturation with respect to calcium oxalate. These results demonstrate that cellular membranes can efficiently induce nucleation of calcium oxalate crystals from a metastable solution in an vitro system. Similar membrane induced heterogeneous nucleation of calcium oxalate in vivo within the renal tubules is a distinct possibility.

  4. Synthesis and antibacterial properties of ZnO brush pens

    NASA Astrophysics Data System (ADS)

    Wang, Dan; Zhang, Rong; Li, Yilin; Weng, Yuan; Liang, Weiquan; Zhang, Wenfeng; Zheng, Weitao; Hu, Haimei

    2015-12-01

    In this paper, ZnO with a novel hierarchical nanostructure has been synthesized by a new solution method. The novel hierarchical structure is named a ‘brush pen’. The biocompatibility and antibacterial properties of ZnO brush pens have been evaluated. The results demonstrate that ZnO brush pens show good antibacterial activity against Staphylococcus aureus.

  5. Bordering on environmental disaster.

    PubMed Central

    Schmidt, C W

    2000-01-01

    Millions of people live on the U.S.-Mexico border, drawn by employment opportunities from industry and agriculture, and booming growth is expected to continue in the coming decades. But the influx of people has long outstripped the capacity of border towns and cities to provide for their citizens, and a large percentage of the population lives in sprawling, underserviced shantytowns. Although investment in environmental infrastructure on both sides has led to improvements, shortages among necessities such as drinking water, wastewater treatment, and solid waste disposal are at crisis levels. In addition, a host of diseases including gastrointestinal infections, asthma, tuberculosis, multiple myeloma, systemic lupus erythematosus, hepatitis A, neural tube defects, and lead poisoning have been linked to environmental conditions on the border. Ongoing binational studies are attempting to define the breadth of the public and environmental health crisis and find ways to help make the border a healthier place. PMID:10903625

  6. 21 CFR 884.1100 - Endometrial brush.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... perforation, or a recent cesarean section, and (3) Design and testing: (i) The sampling component is covered... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Endometrial brush. 884.1100 Section 884.1100 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED)...

  7. 21 CFR 884.1100 - Endometrial brush.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... perforation, or a recent cesarean section, and (3) Design and testing: (i) The sampling component is covered... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Endometrial brush. 884.1100 Section 884.1100 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED)...

  8. 21 CFR 884.1100 - Endometrial brush.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... perforation, or a recent cesarean section, and (3) Design and testing: (i) The sampling component is covered... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Endometrial brush. 884.1100 Section 884.1100 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED)...

  9. Multifunctional brushes made from carbon nanotubes

    NASA Astrophysics Data System (ADS)

    Cao, Anyuan; Veedu, Vinod P.; Li, Xuesong; Yao, Zhaoling; Ghasemi-Nejhad, Mehrdad N.; Ajayan, Pulickel M.

    2005-07-01

    Brushes are common tools for use in industry and our daily life, performing a variety of tasks such as cleaning, scraping, applying and electrical contacts. Typical materials for constructing brush bristles include animal hairs, synthetic polymer fibres and metal wires (see, for example, ref. 1). The performance of these bristles has been limited by the oxidation and degradation of metal wires, poor strength of natural hairs, and low thermal stability of synthetic fibres. Carbon nanotubes, having a typical one-dimensional nanostructure, have excellent mechanical properties, such as high modulus and strength, high elasticity and resilience, thermal conductivity and large surface area (50-200 m2 g-1). Here we construct multifunctional, conductive brushes with carbon nanotube bristles grafted on fibre handles, and demonstrate their several unique tasks such as cleaning of nanoparticles from narrow spaces, coating of the inside of holes, selective chemical adsorption, and as movable electromechanical brush contacts and switches. The nanotube bristles can also be chemically functionalized for selective removal of heavy metal ions.

  10. Charge Transport Properties in Polymer Brushes

    NASA Astrophysics Data System (ADS)

    Moog, Mark; Tsui, Frank; Vonwald, Ian; You, Wei

    Electrical transport properties in poly(3-methyl)thiophene (P3MT) brushes have been studied. The P3MT brushes correspond to a new type of surface-tethered, vertically oriented conjugated molecular wires, sandwiched between two metallic electrodes to form the electrode-molecule-electrode (EME) devices. P3MT is a highly conjugated polymer, a ''workhorse'' material for organic electronics and photonics. The P3MT brushes were grown on ITO surfaces with controlled length (between 2 and 100 nm). The top electrodes were transfer-printed Au films with lateral dimensions between 200 nm and 50 μm. I-V and differential conductance measurements were performed using conductive AFM and 4-terminal techniques. Tunneling and field-emission measurements in EME devices with molecular lengths < 5 nm show HOMO mediated direct hole tunneling with energy barriers of 0.3 and 0.5 eV at the respective interfaces with ITO and Au. The transport properties in longer brushes are indicative of the two quasi-Ohmic interfaces with a characteristic offset in the conductance minimum of 0.12 V biased toward the ITO. Temperature dependent parameters have been examined at various molecular lengths. The drift mobility and the interplay between intra- and intermolecular transport have been investigated.