Intestinal alkaline phosphatase regulates protective surface microclimate pH in rat duodenum.

PubMed

Mizumori, Misa; Ham, Maggie; Guth, Paul H; Engel, Eli; Kaunitz, Jonathan D; Akiba, Yasutada

2009-07-15

Regulation of localized extracellular pH (pH(o)) maintains normal organ function. An alkaline microclimate overlying the duodenal enterocyte brush border protects the mucosa from luminal acid. We hypothesized that intestinal alkaline phosphatase (IAP) regulates pH(o) due to pH-sensitive ATP hydrolysis as part of an ecto-purinergic pH regulatory system, comprised of cell-surface P2Y receptors and ATP-stimulated duodenal bicarbonate secretion (DBS). To test this hypothesis, we measured DBS in a perfused rat duodenal loop, examining the effect of the competitive alkaline phosphatase inhibitor glycerol phosphate (GP), the ecto-nucleoside triphosphate diphosphohydrolase inhibitor ARL67156, and exogenous nucleotides or P2 receptor agonists on DBS. Furthermore, we measured perfusate ATP concentration with a luciferin-luciferase bioassay. IAP inhibition increased DBS and luminal ATP output. Increased luminal ATP output was partially CFTR dependent, but was not due to cellular injury. Immunofluorescence localized the P2Y(1) receptor to the brush border membrane of duodenal villi. The P2Y(1) agonist 2-methylthio-ADP increased DBS, whereas the P2Y(1) antagonist MRS2179 reduced ATP- or GP-induced DBS. Acid perfusion augmented DBS and ATP release, further enhanced by the IAP inhibitor l-cysteine, and reduced by the exogenous ATPase apyrase. Furthermore, MRS2179 or the highly selective P2Y(1) antagonist MRS2500 co-perfused with acid induced epithelial injury, suggesting that IAP/ATP/P2Y signalling protects the mucosa from acid injury. Increased DBS augments IAP activity presumably by raising pH(o), increasing the rate of ATP degradation, decreasing ATP-mediated DBS, forming a negative feedback loop. The duodenal epithelial brush border IAP-P2Y-HCO(3-) surface microclimate pH regulatory system effectively protects the mucosa from acid injury.

Intraluminal polyethylene glycol stabilizes tight junctions and improves intestinal preservation in the rat.

PubMed

Oltean, M; Joshi, M; Björkman, E; Oltean, S; Casselbrant, A; Herlenius, G; Olausson, M

2012-08-01

Rapidly progressing mucosal breakdown limits the intestinal preservation time below 10 h. Recent studies indicate that intraluminal solutions containing polyethylene glycol (PEG) alleviate preservation injury of intestines stored in UW-Viaspan. We investigated whether a low-sodium PEG solution is beneficial for intestines stored in histidine-tryptophane-ketoglutarate (HTK) preservation solution. Rat intestines used as control tissue (group 1) were perfused with HTK, groups 2 and 3 received either a customized PEG-3350 (group 2) or an electrolyte solution (group 3) intraluminally before cold storage. Tissue injury, brush-border maltase activity, zonula occludens-1 (ZO-1) and claudin-3 expression in the tight junctions (TJ) were analyzed after 8, 14 and 20 h. We measured epithelial resistance and permeability (Ussing chamber) after 8 and 14 h. Group 2 had superior morphology while maltase activity was similar in all groups. TJ proteins rapidly decreased and decolocalized in groups 1 3; these negative events were delayed in group 2, where colocalization persisted for about 14 h. Intestines in group 2 had higher epithelial resistance and lower permeability than the other groups. These results suggest that a customized PEG solution intraluminally reduces the intestinal preservation injury by improving several major epithelial characteristics without negatively affecting the brush-border enzymes or promoting edema. © Copyright 2012 The American Society of Transplantation and the American Society of Transplant Surgeons.

Localization of Bacillus thuringiensis Cry1A toxin-binding molecules in gypsy moth larval gut sections using fluorescence microscopy

Treesearch

Algimantas P. Valaitis

2011-01-01

The microbial insecticide Bacillus thuringiensis (Bt) produces Cry toxins, proteins that bind to the brush border membranes of gut epithelial cells of insects that ingest it, disrupting the integrity of the membranes, and leading to cell lysis and insect death. In gypsy moth, Lymantria dispar, two toxin-binding molecules for the...

Role of Sphingolipids in Infant Gut Health and Immunity.

PubMed

Nilsson, Åke

2016-06-01

Sphingomyelin (SM), glycosphingolipids, and gangliosides are important polar lipids in the milk fat globule membrane but are not found in standard milk replacement formulas. Because digestion and absorption of SM and glycosphingolipids generate the bioactive metabolites ceramide, sphingosine, and sphingosine-1-phosphate (S1P), and because intact gangliosides may have beneficial effects in the gut, this may be important for gut integrity and immune maturation in the neonate. The brush border enzymes that hydrolyze milk SM, alkaline sphingomyelinase (nucleotide phosphodiesterase pyrophosphatase 7), and neutral ceramidase are expressed at birth in both term and preterm infants. Released sphingosine is absorbed, phosphorylated to S1P, and converted to palmitic acid via S1P-lyase in the gut mucosa. Hypothetically, S1P also may be released from absorptive cells and exert important paracrine actions favoring epithelial integrity and renewal, as well as immune function, including secretory IgA production and migration of T lymphocyte subpopulations. Gluco-, galacto-, and lactosylceramide are hydrolyzed to ceramide by lactase-phlorizin hydrolase, which also hydrolyzes lactose. Gangliosides may adhere to the brush border and is internalized, modified, and possibly transported into blood, and may exert protective functions by their interactions with bacteria, bacterial toxins, and the brush border. Copyright © 2016 Elsevier Inc. All rights reserved.

Changes in intestinal absorption of nutrients and brush border glycoproteins after total parenteral nutrition in rats.

PubMed Central

Miura, S; Tanaka, S; Yoshioka, M; Serizawa, H; Tashiro, H; Shiozaki, H; Imaeda, H; Tsuchiya, M

1992-01-01

The effect of total parenteral nutrition on nutrients absorption and glycoprotein changes of brush border membrane was examined in rat small intestine. In total parenteral nutrition rats, a marked decrease in activity of brush border enzymes was observed mainly in the proximal and middle segments of the intestine. Galactose perfusion of jejunal segment showed that hexose absorption was significantly inhibited, while intestinal absorption of glycine or dipeptide, glycylglycine was not significantly affected by total parenteral nutrition treatment. When brush border membrane glycoprotein profile was examined by [3H]-glucosamine or [3H]-fucose incorporation into jejunal loops, significant changes were observed in the glycoprotein pattern of brush border membrane especially in the high molecular weight range over 120 kDa after total parenteral nutrition treatment, suggesting strong dependency of glycoprotein synthesis on luminal substances. Molecular weight of sucrase isomaltase in brush border membrane detected by specific antibody showed no significant difference, however, in total parenteral nutrition and control rats. Also, molecular weight of specific sodium glucose cotransporter of intestinal brush border membrane detected by selective photoaffinity labelling was not altered in total parenteral nutrition rats. It may be that prolonged absence of oral food intake may produce significant biochemical changes in brush border membrane glycoprotein and absorptive capacity of small intestine, but these changes were not observed in all brush border membrane glycoproteins. Images Figure 1 Figure 2 Figure 3 Figure 4 PMID:1582592

Dimerization and actin-bundling properties of villin and its role in the assembly of epithelial cell brush borders.

PubMed

George, Sudeep P; Wang, Yaohong; Mathew, Sijo; Srinivasan, Kamalakkannan; Khurana, Seema

2007-09-07

Villin is a major actin-bundling protein in the brush border of epithelial cells. In this study we demonstrate for the first time that villin can bundle actin filaments using a single F-actin binding site, because it has the ability to self-associate. Using fluorescence resonance energy transfer, we demonstrate villin self-association in living cells in microvilli and in growth factor-stimulated cells in membrane ruffles and lamellipodia. Using sucrose density gradient, size-exclusion chromatography, and matrix-assisted laser desorption ionization time-of-flight, the majority of villin was identified as a monomer or dimer. Villin dimers were also identified in Caco-2 cells, which endogenously express villin and Madin-Darby canine kidney cells that ectopically express villin. Using truncation mutants of villin, site-directed mutagenesis, and fluorescence resonance energy transfer, an amino-terminal dimerization site was identified that regulated villin self-association in parallel conformation as well as actin bundling by villin. This detailed analysis describes for the first time microvillus assembly by villin, redefines the actin-bundling function of villin, and provides a molecular mechanism for actin bundling by villin, which could have wider implications for other actin cross-linking proteins that share a villin-like headpiece domain. Our study also provides a molecular basis to separate the morphologically distinct actin-severing and actin-bundling properties of villin.

Characterization of brush borders purified in iso-osmotic medium and microvillar membranes subfractionated from mouse small intestine.

PubMed Central

Fujita, M; Ohta, H; Uezato, T

1981-01-01

Brush borders free of nuclei were isolated by repeated homogenization and centrifugation in iso-osmotic medium. They showed typical morphology under electron microscopy. The mean recovery and enrichment of alkaline phosphatase activity in the brush-border fraction were 50% and 17.5-fold respectively. gamma-Glutamyl transpeptidase showed a close parallelism with alkaline phosphatase and sucrase in subcellular distribution. Microvillar membranes were purified from isolated brush borders; they showed a further enrichment for alkaline phosphatase and were composed of homogeneous vesicles. Both brush-border and microvillar-membrane preparations were analysed for contamination by basolateral and endoplasmic-reticular membranes. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of the microvillar-membrane preparation in six different systems revealed approx. 40 components in the mol.wt. range 15 000-232 000. They were grouped into seven major classes on the basis of molecular weight and electrophoretic patterns. Images PLATE 1 PLATE 2 PMID:7317008

Structural specificity of mucosal-cell transport and metabolism of peptide drugs: implication for oral peptide drug delivery

NASA Technical Reports Server (NTRS)

Bai, J. P.; Amidon, G. L.

1992-01-01

The brush border membrane of intestinal mucosal cells contains a peptide carrier system with rather broad substrate specificity and various endo- and exopeptidase activities. Small peptide (di-/tripeptide)-type drugs with or without an N-terminal alpha-amino group, including beta-lactam antibiotics and angiotensin-converting enzyme (ACE) inhibitors, are transported by the peptide transporter. Polypeptide drugs are hydrolyzed by brush border membrane proteolytic enzymes to di-/tripeptides and amino acids. Therefore, while the intestinal brush border membrane has a carrier system facilitating the absorption of di-/tripeptide drugs, it is a major barrier limiting oral availability of polypeptide drugs. In this paper, the specificity of peptide transport and metabolism in the intestinal brush border membrane is reviewed.

Jejunal brush border microvillous alterations in Giardia muris-infected mice: role of T lymphocytes and interleukin-6.

PubMed

Scott, K G; Logan, M R; Klammer, G M; Teoh, D A; Buret, A G

2000-06-01

Intestinal colonization with the protozoan Giardia causes diffuse brush border microvillous alterations and disaccharidase deficiencies, which in turn are responsible for intestinal malabsorption and maldigestion. The role of T cells and/or cytokines in the pathogenesis of Giardia-induced microvillous injury remains unclear. The aim of this study was to assess the role of T cells and interleukin-6 (IL-6) in the brush border pathophysiology of acute murine giardiasis in vivo. Athymic nude (nu(-)/nu(-)) CD-1 mice and isogenic immunocompetent (nu(+)/nu(+)) CD-1 mice (4 weeks old) received an axenic Giardia muris trophozoite inoculum or vehicle (control) via orogastric gavage. Weight gain and food intake were assessed daily. On day 6, segments of jejunum were assessed for parasite load, brush border ultrastructure, IL-6 content, maltase and sucrase activities, villus-crypt architecture, and intraepithelial lymphocyte (IEL) infiltration. Despite similar parasitic loads on day 6, infected immunocompetent animals, but not infected nude mice, showed a diffuse loss of brush border microvillous surface area, which was correlated with a significant reduction in maltase and sucrase activities and a decrease in jejunal IL-6 concentration. In both athymic control and infected mice, jejunal brush border surface area and disaccharidases were high, but levels of tissue IL-6 were low and comparable to the concentration measured in immunocompetent infected animals. In both immunocompetent and nude mice, infection caused a small but significant increase in the numbers of IELs. These findings suggest that the enterocyte brush border injury and malfunction seen in giardiasis is, at least in part, mediated by thymus-derived T lymphocytes and that suppressed jejunal IL-6 does not necessarily accompany microvillous shortening.

Computer simulations of the mechanical response of brushes on the surface of cancerous epithelial cells

NASA Astrophysics Data System (ADS)

Goicochea, A. Gama; Guardado, S. J. Alas

2015-08-01

We report a model for atomic force microscopy by means of computer simulations of molecular brushes on surfaces of biological interest such as normal and cancerous cervical epithelial cells. Our model predicts that the force needed to produce a given indentation on brushes that can move on the surface of the cell (called “liquid” brushes) is the same as that required for brushes whose ends are fixed on the cell’s surface (called “solid” brushes), as long as the tip of the microscope covers the entire area of the brush. Additionally, we find that cancerous cells are softer than normal ones, in agreement with various experiments. Moreover, soft brushes are found to display larger resistance to compression than stiff ones. This phenomenon is the consequence of the larger equilibrium length of the soft brushes and the cooperative association of solvent molecules trapped within the brushes, which leads to an increase in the osmotic pressure. Our results show that a careful characterization of the brushes on epithelial cells is indispensable when determining the mechanical response of cancerous cells.

Diagnostic and Research Aspects of Small Intestinal Disaccharidases in Coeliac Disease

PubMed Central

Ciclitira, Paul J.

2017-01-01

Disaccharidases (DS) are brush border enzymes embedded in the microvillous membrane of small intestinal enterocytes. In untreated coeliac disease (CD), a general decrease of DS activities is seen. This manuscript reviews different aspects of DS activities in CD: their utility in the diagnosis and their application to in vitro toxicity testing. The latter has never been established in CD research. However, with the recent advances in small intestinal organoid techniques, DS might be employed as a biomarker for in vitro studies. This includes establishment of self-renewing epithelial cells raised from tissue, which express differentiation markers, including the brush border enzymes. Determining duodenal DS activities may provide additional information during the diagnostic workup of CD: (i) quantify the severity of the observed histological lesions, (ii) provide predictive values for the grade of mucosal villous atrophy, and (iii) aid diagnosing CD where minor histological changes are seen. DS can also provide additional information to assess the response to a gluten-free diet as marked increase of their activities occurs four weeks after commencing it. Various endogenous and exogenous factors affecting DS might also be relevant when considering investigating the role of DS in other conditions including noncoeliac gluten sensitivity and DS deficiencies. PMID:28512643

The effect of amino acids and dipeptides on sodium-ion transport in rat enterocytes.

PubMed

Cheeseman, C I; Devlin, D

1985-02-14

Sodium efflux from isolated intestinal epithelial cells was measured during incubation with several different free amino acids and dipeptides. L-Leucine, which is cotransported with sodium across the brush border membrane, significantly stimulated the total sodium efflux and almost all of this increase involved the ouabain-sensitive flux, i.e., the active component. In contrast, glycyl-L-leucine had little or no effect on active sodium efflux either in the presence or absence of 0.1 mM bestatin, a peptide hydrolase inhibitor. A second dipeptide L-carnosine (beta-alanyl-L-histidine) which is poorly hydrolysed by enterocytes also had no effect upon sodium efflux. However, glycylglycine, which has been shown to be cotransported with sodium, did stimulate the ionic efflux. In addition, measurement of sodium uptake by sheets of small intestine showed that glycyl-L-leucine, carnosine and glycyl-L-proline failed to increase the uptake of the ion, while glycylglycine did significantly stimulate sodium uptake. These data indicate that some dipeptides are not cotransported with sodium, while others are. This suggests that there may well be multiple peptide transporters with very different characteristics in the brush border membrane of enterocytes.

F4+ enterotoxigenic Escherichia coli (ETEC) adhesion mediated by the major fimbrial subunit FaeG.

PubMed

Xia, Pengpeng; Song, Yujie; Zou, Yajie; Yang, Ying; Zhu, Guoqiang

2015-09-01

The FaeG subunit is the major constituent of F4(+) fimbriae, associated with glycoprotein and/or glycolipid receptor recognition and majorly contributes to the pathogen attachment to the host cells. To investigate the key factor involved in the fimbrial binding of F4(+) Escherichia coli, both the recombinant E. coli SE5000 strains carrying the fae operon gene clusters that express the different types of fimbriae in vitro, named as rF4ab, rF4ac, and rF4ad, respectively, corresponding to the fimbrial types F4ab, F4ac, and F4ad, and the three isogenic in-frame faeG gene deletion mutants were constructed. The adhesion assays and adhesion inhibition assays showed that ΔfaeG mutants had a significant reduction in the binding to porcine brush border as well as the intestinal epithelial cell lines, while the complemented strain ΔfaeG/pfaeG restored the adhesion function. The recombinant bacterial strains rF4ab, rF4ac, and rF4ad have the same binding property as wild-type F4(+) E. coli strains do and improvement in terms of binding to porcine brush border and the intestinal epithelial cells, and the adherence was blocked by the monoclonal antibody anti-F4 fimbriae. These data demonstrate that the fimbrial binding of F4(+) E. coli is directly mediated by the major FaeG subunit. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Generation of stable lipid raft microdomains in the enterocyte brush border by selective endocytic removal of non-raft membrane.

PubMed

Danielsen, E Michael; Hansen, Gert H

2013-01-01

The small intestinal brush border has an unusually high proportion of glycolipids which promote the formation of lipid raft microdomains, stabilized by various cross-linking lectins. This unique membrane organization acts to provide physical and chemical stability to the membrane that faces multiple deleterious agents present in the gut lumen, such as bile salts, digestive enzymes of the pancreas, and a plethora of pathogens. In the present work, we studied the constitutive endocytosis from the brush border of cultured jejunal explants of the pig, and the results indicate that this process functions to enrich the contents of lipid raft components in the brush border. The lipophilic fluorescent marker FM, taken up into early endosomes in the terminal web region (TWEEs), was absent from detergent resistant membranes (DRMs), implying an association with non-raft membrane. Furthermore, neither major lipid raft-associated brush border enzymes nor glycolipids were detected by immunofluorescence microscopy in subapical punctae resembling TWEEs. Finally, two model raft lipids, BODIPY-lactosylceramide and BODIPY-GM1, were not endocytosed except when cholera toxin subunit B (CTB) was present. In conclusion, we propose that constitutive, selective endocytic removal of non-raft membrane acts as a sorting mechanism to enrich the brush border contents of lipid raft components, such as glycolipids and the major digestive enzymes. This sorting may be energetically driven by changes in membrane curvature when molecules move from a microvillar surface to an endocytic invagination.

The mRNA expression of amino acid and sugar transporters, aminopeptidase, as well as the di- and tri-peptide transporter PepT1 in the intestines of Eimeria infected broiler chickens.

PubMed

Miska, K B; Fetterer, R H

2017-02-01

Coccidiosis in chickens is caused by infection of gut epithelial cells with protozoan parasites of the genus Eimeria This disease causes losses to the poultry industry since infected birds fail to gain weight as rapidly as non-infected birds and efficiency of feed conversion is compromised. For the present study the effect of Eimeria on expression of components of amino acid and sugar uptake mechanisms was determined. Broiler chicks were infected with Eimeria maxima, which infects the jejunum; Eimeria acervulina, which infects the duodenum; or Eimeria tenella, which infects the ceca. Sections of the jejunum, duodenum, and ceca (depending on species of Eimeria) were taken at several time points between d zero and 14 post infection (PI) for mRNA expression analysis. Genes examined included one digestive enzyme, 7 peptide and amino acid transporters located on the brush border, 8 transporters located at the basolateral surface of the gut epithelium, and 5 sugar transporters. All 3 Eimeria species examined caused decrease in expression of brush border transporters particularly at d 5 to 7 PI, which corresponds to the time when pathology is greatest. The same pattern was seen in expression of sugar transporters. However, the expression of basolateral transporters differed among species. Eimeria tenella infection resulted in decreased expression of all basolateral transporters, while E. maxima infection caused increased expression of 2 genes and slight decrease in expression of the remaining 5 genes. Infection with E. acervulina resulted in increased expression at the height of infection of all but one basolateral transporter. In conclusion, Eimeria infection causes a general decrease in gene expression of sugar transporter and brush border AATs at the height of infection. However the expression of basolateral transporters is increased in E. maxima and E. acervulina infected birds. It is possible that decreased expression of brush border transporters in combination with increased expression of basolateral transporters leads to decrease of nutrients available for the parasite, thus limiting parasite reproduction. Published by Oxford University Press on behalf of Poultry Science Association 2016. This work is written by (a) US Government employee(s) and is in the public domain in the US.

Plastin 1 Binds to Keratin and Is Required for Terminal Web Assembly in the Intestinal Epithelium

PubMed Central

Grimm-Günter, Eva-Maria S.; Revenu, Céline; Ramos, Sonia; Hurbain, Ilse; Smyth, Neil; Ferrary, Evelyne; Louvard, Daniel; Robine, Sylvie

2009-01-01

Plastin 1 (I-plastin, fimbrin) along with villin and espin is a prominent actin-bundling protein of the intestinal brush border microvilli. We demonstrate here that plastin 1 accumulates in the terminal web and interacts with keratin 19, possibly contributing to anchoring the rootlets to the keratin network. This prompted us to investigate the importance of plastin 1 in brush border assembly. Although in vivo neither villin nor espin is required for brush border structure, plastin 1-deficient mice have conspicuous ultrastructural alterations: microvilli are shorter and constricted at their base, and, strikingly, their core actin bundles lack true rootlets. The composition of the microvilli themselves is apparently normal, whereas that of the terminal web is profoundly altered. Although the plastin 1 knockout mice do not show any overt gross phenotype and present a normal intestinal microanatomy, the alterations result in increased fragility of the epithelium. This is seen as an increased sensitivity of the brush border to biochemical manipulations, decreased transepithelial resistance, and increased sensitivity to dextran sodium sulfate-induced colitis. Plastin 1 thus emerges as an important regulator of brush border morphology and stability through a novel role in the organization of the terminal web, possibly by connecting actin filaments to the underlying intermediate filament network. PMID:19321664

The effect of essential fatty acid deficiency on the stimulation of intestinal calcium transport by 1,25-dihydroxyvitamin D3.

PubMed

Kreutter, D; Matsumoto, T; Peckham, R; Zawalich, K; Wen, W H; Zolock, D T; Rasmussen, H

1983-04-25

The effect of altering the lipid composition of the brush-border membrane on the ability of 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) to stimulate calcium transport across the intestinal mucosa was examined by raising chicks on a vitamin D, essential fatty acid-deficient diet (-DEFAD) and measuring calcium absorption from duodenal sacs in situ and calcium uptake into brush-border membrane vesicles in vitro. Administration of 1,25-(OH)2D3 to -DEFAD and to -D control chicks led to the same increase in calcium transport in situ, whereas calcium transport in isolated brush-border membrane vesicles was not stimulated in the EFAD group, but responded normally in the control group. When the incubation temperature was increased to 34 degrees C, brush-border membrane vesicles from 1,25-(OH)2D3-treated essential fatty acid-deficient (+DE-FAD) chicks accumulated calcium at a faster rate than did vesicles from -DEFAD chicks. There was a marked decrease in the linoleic acid content and an increase in the oleic acid content of both the total lipid extract of the brush-border membrane as well as the phosphatidylcholine and phosphatidylethanolamine fractions, which could explain the temperature sensitivity of the in vitro system. When the diet of the EFAD chicks was supplemented with linoleic acid, the rate of calcium uptake into subsequently isolated vesicles from +DE-FAD chicks correlated with the amount of linoleic acid in the brush-border membranes. These results support the concept that the action of 1,25-(OH)2D3 on membrane lipid turnover and structure plays a critically important role in the 1,25-(OH)2D3-mediated cellular transport responses.

Expression of nutrient transporters in duodenum, jejunum, and ileum of Eimeria maxima-infected broiler chickens.

PubMed

Fetterer, Raymond H; Miska, Katarzyna B; Jenkins, Mark C; Wong, Eric A

2014-10-01

The uptake of amino acids is mediated by active transporters located on the basolateral and brush border membranes of intestinal epithelial cells. The current study investigated the expression of amino acid transporters (AAT) and other genes in the intestine of chicks infected with Eimeria maxima. At 7-day postinfection (PI), tissue from each intestinal segment (duodenum, jejunum, and ileum) was taken from birds inoculated with 3 × 10(3) oocysts/bird and processed to recover RNA. Analysis of gene expression was performed using real-time reverse transcription polymerase chain reaction (qRT-PCR). Results were given as relative expression using β₂-microglobulin as an endogenous control. All the genes studied were expressed in three segments of the intestines, and expression of the genes was altered by infection with E. maxima. Even though the jejunum is considered the parasite's primary predilection site, there was no segment-related difference in expression of most of the genes studied. The antimicrobial peptide (LEAP2) was downregulated in all three segments of the intestine. The results also demonstrate that transporters associated with brush border membranes were downregulated while transporters associated with the basolateral membranes were upregulated and that E. maxima alters the expression of AAT and LEAP2 throughout the small intestine.

Control of apico-basal epithelial polarity by the microtubule minus-end-binding protein CAMSAP3 and spectraplakin ACF7.

PubMed

Noordstra, Ivar; Liu, Qingyang; Nijenhuis, Wilco; Hua, Shasha; Jiang, Kai; Baars, Matthijs; Remmelzwaal, Sanne; Martin, Maud; Kapitein, Lukas C; Akhmanova, Anna

2016-11-15

The microtubule cytoskeleton regulates cell polarity by spatially organizing membrane trafficking and signaling processes. In epithelial cells, microtubules form parallel arrays aligned along the apico-basal axis, and recent work has demonstrated that the members of CAMSAP/Patronin family control apical tethering of microtubule minus ends. Here, we show that in mammalian intestinal epithelial cells, the spectraplakin ACF7 (also known as MACF1) specifically binds to CAMSAP3 and is required for the apical localization of CAMSAP3-decorated microtubule minus ends. Loss of ACF7 but not of CAMSAP3 or its homolog CAMSAP2 affected the formation of polarized epithelial cysts in three-dimensional cultures. In short-term epithelial polarization assays, knockout of CAMSAP3, but not of CAMSAP2, caused microtubule re-organization into a more radial centrosomal array, redistribution of Rab11-positive (also known as Rab11A) endosomes from the apical cell surface to the pericentrosomal region and inhibition of actin brush border formation at the apical side of the cell. We conclude that ACF7 is an important regulator of apico-basal polarity in mammalian intestinal cells and that a radial centrosome-centered microtubule organization can act as an inhibitor of epithelial polarity. © 2016. Published by The Company of Biologists Ltd.

In Vivo and In Vitro Binding of Vip3Aa to Spodoptera frugiperda Midgut and Characterization of Binding Sites by 125I Radiolabeling

PubMed Central

Chakroun, Maissa

2014-01-01

Bacillus thuringiensis vegetative insecticidal proteins (Vip3A) have been recently introduced in important crops as a strategy to delay the emerging resistance to the existing Cry toxins. The mode of action of Vip3A proteins has been studied in Spodoptera frugiperda with the aim of characterizing their binding to the insect midgut. Immunofluorescence histological localization of Vip3Aa in the midgut of intoxicated larvae showed that Vip3Aa bound to the brush border membrane along the entire apical surface. The presence of fluorescence in the cytoplasm of epithelial cells seems to suggest internalization of Vip3Aa or a fragment of it. Successful radiolabeling and optimization of the binding protocol for the 125I-Vip3Aa to S. frugiperda brush border membrane vesicles (BBMV) allowed the determination of binding parameters of Vip3A proteins for the first time. Heterologous competition using Vip3Ad, Vip3Ae, and Vip3Af as competitor proteins showed that they share the same binding site with Vip3Aa. In contrast, when using Cry1Ab and Cry1Ac as competitors, no competitive binding was observed, which makes them appropriate candidates to be used in combination with Vip3A proteins in transgenic crops. PMID:25002420

Direct reprogramming of human bone marrow stromal cells into functional renal cells using cell-free extracts.

PubMed

Papadimou, Evangelia; Morigi, Marina; Iatropoulos, Paraskevas; Xinaris, Christodoulos; Tomasoni, Susanna; Benedetti, Valentina; Longaretti, Lorena; Rota, Cinzia; Todeschini, Marta; Rizzo, Paola; Introna, Martino; Grazia de Simoni, Maria; Remuzzi, Giuseppe; Goligorsky, Michael S; Benigni, Ariela

2015-04-14

The application of cell-based therapies in regenerative medicine is gaining recognition. Here, we show that human bone marrow stromal cells (BMSCs), also known as bone-marrow-derived mesenchymal cells, can be reprogrammed into renal proximal tubular-like epithelial cells using cell-free extracts. Streptolysin-O-permeabilized BMSCs exposed to HK2-cell extracts underwent morphological changes-formation of "domes" and tubule-like structures-and acquired epithelial functional properties such as transepithelial-resistance, albumin-binding, and uptake and specific markers E-cadherin and aquaporin-1. Transmission electron microscopy revealed the presence of brush border microvilli and tight intercellular contacts. RNA sequencing showed tubular epithelial transcript abundance and revealed the upregulation of components of the EGFR pathway. Reprogrammed BMSCs integrated into self-forming kidney tissue and formed tubular structures. Reprogrammed BMSCs infused in immunodeficient mice with cisplatin-induced acute kidney injury engrafted into proximal tubuli, reduced renal injury and improved function. Thus, reprogrammed BMSCs are a promising cell resource for future cell therapy. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

Local pH domains regulate NHE3-mediated Na+ reabsorption in the renal proximal tubule

PubMed Central

Burford, James L.; McDonough, Alicia A.; Holstein-Rathlou, Niels-Henrik; Peti-Peterdi, Janos

2014-01-01

The proximal tubule Na+/H+ exchanger 3 (NHE3), located in the apical dense microvilli (brush border), plays a major role in the reabsorption of NaCl and water in the renal proximal tubule. In response to a rise in blood pressure NHE3 redistributes in the plane of the plasma membrane to the base of the brush border, where NHE3 activity is reduced. This NHE3 redistribution is assumed to provoke pressure natriuresis; however, it is unclear how NHE3 redistribution per se reduces NHE3 activity. To investigate if the distribution of NHE3 in the brush border can change the reabsorption rate, we constructed a spatiotemporal mathematical model of NHE3-mediated Na+ reabsorption across a proximal tubule cell and compared the model results with in vivo experiments in rats. The model predicts that when NHE3 is localized exclusively at the base of the brush border, it creates local pH microdomains that reduce NHE3 activity by >30%. We tested the model's prediction experimentally: the rat kidney cortex was loaded with the pH-sensitive fluorescent dye BCECF, and cells of the proximal tubule were imaged in vivo using confocal fluorescence microscopy before and after an increase of blood pressure by ∼50 mmHg. The experimental results supported the model by demonstrating that a rise of blood pressure induces the development of pH microdomains near the bottom of the brush border. These local changes in pH reduce NHE3 activity, which may explain the pressure natriuresis response to NHE3 redistribution. PMID:25298526

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

USDA-ARS?s Scientific Manuscript database

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...

Cation-dependent nutrient transport in shrimp digestive tract.

PubMed

Simmons, Tamla; Mozo, Julie; Wilson, Jennifer; Ahearn, Gregory A

2012-02-01

Purified epithelial brush border membrane vesicles (BBMV) were produced from the hepatopancreas of the Atlantic White shrimp, Litopeneaus setiferus, using standard methods originally developed for mammalian tissues and previously applied to other crustacean and echinoderm epithelia. These vesicles were used to study the cation dependency of sugar and amino acid transport across luminal membranes of hepatopancreatic epithelial cells. (3)H-D: -glucose uptake by BBMV against transient sugar concentration gradients occurred when either transmembrane sodium or potassium gradients were the only driving forces for sugar accumulation, suggesting the presence of a possible coupled transport system capable of using either cation. (3)H-L: -histidine transport was only stimulated by a transmembrane potassium gradient, while (3)H-L: -leucine uptake was enhanced by either a sodium or potassium gradient. These responses suggest the possible presence of a potassium-dependent transporter that accommodates either amino acid and a sodium-dependent system restricted only to L: -leucine. Uptake of (3)H-L: -leucine was significantly stimulated (P < 0.05) by several metallic cations (e.g., Zn(2+), Cu(2+), Mn(2+), Cd(2+), or Co(2+)) at external pH values of 7.0 or 5.0 (internal pH 7.0), suggesting a potential synergistic role of the cations in the transmembrane transfer of amino acids. (3)H-L: -histidine influxes (15 suptakes) were hyperbolic functions of external [zinc] or [manganese], following Michaelis-Menten kinetics. The apparent affinity constant (e.g., K (m)) for manganese was an order of magnitude smaller (K (m) = 0.22 μM Mn) than that for zinc (K (m) = 1.80 μM Zn), while no significant difference (P > 0.05) occurred between their maximal transport velocities (e.g., J (max)). These results suggest that a number of cation-dependent nutrient transport systems occur on the shrimp brush border membrane and aid in the absorption of these important dietary elements.

Colon luminal content and epithelial cell morphology are markedly modified in rats fed with a high-protein diet.

PubMed

Andriamihaja, Mireille; Davila, Anne-Marie; Eklou-Lawson, Mamy; Petit, Nathalie; Delpal, Serge; Allek, Fadhila; Blais, Anne; Delteil, Corine; Tomé, Daniel; Blachier, François

2010-11-01

Hyperproteic diets are used in human nutrition to obtain body weight reduction. Although increased protein ingestion results in an increased transfer of proteins from the small to the large intestine, there is little information on the consequences of the use of such diets on the composition of large intestine content and on epithelial cell morphology and metabolism. Rats were fed for 15 days with either a normoproteic (NP, 14% protein) or a hyperproteic isocaloric diet (HP, 53% protein), and absorptive colonocytes were observed by electron microscopy or isolated for enzyme activity studies. The colonic luminal content was recovered for biochemical analysis. Absorbing colonocytes were characterized by a 1.7-fold reduction in the height of the brush-border membranes (P = 0.0001) after HP diet consumption when compared with NP. This coincided in the whole colon content of HP animals with a 1.8-fold higher mass content (P = 0.0020), a 2.2-fold higher water content (P = 0.0240), a 5.2-fold higher protease activity (P = 0.0104), a 5.5-fold higher ammonia content (P = 0.0008), and a more than twofold higher propionate, valerate, isobutyrate, and isovalerate content (P < 0.05). The basal oxygen consumption of colonocytes was similar in the NP and HP groups, but ammonia was found to provoke a dose-dependent decrease of oxygen consumption in the isolated absorbing colonocytes. The activity of glutamine synthetase (which condenses ammonia and glutamate) was found to be much higher in colonocytes than in small intestine enterocytes and was 1.6-fold higher (P = 0.0304) in colonocytes isolated from HP animals than NP. Glutaminase activity remained unchanged. Thus hyperproteic diet ingestion causes marked changes both in the luminal environment of colonocytes and in the characteristics of these cells, demonstrating that hyperproteic diet interferes with colonocyte metabolism and morphology. Possible causal relationships between energy metabolism, reduced height of colonocyte brush-border membranes, and reduced water absorption are discussed.

Is there a tripeptidyl peptidase in the renal brush-border membrane?

PubMed Central

Kenny, A J; Ingram, J

1988-01-01

A recent claim that the renal brush border contains a tripeptidyl peptidase [Andersen & McDonald (1987) Am. J. Physiol. 253, F649-F655] was examined. In a fluorescent assay, the hydrolysis of Gly-Pro-Met-2-naphthylamide (-NH-Nap) and Gly-Pro-Leu-NH-Nap by pig kidney microvilli was strongly inhibited by amastatin or di-isopropyl phosphorofluoridate (inhibitors of aminopeptidases and dipeptidyl peptidase IV). The products formed were shown to be Gly-Pro and Met-NH-Nap (or Leu-NH-Nap) and free 2-naphthylamine. Specific antibodies to pig and rat aminopeptidase N abolished the apparent tripeptidyl peptidase activity. We conclude that these substrates are hydrolysed by the sequential attack of dipeptidyl peptidase IV and aminopeptidase N and that pig and rat brush borders lack a detectable tripeptidyl peptidase. Images Fig. 1. PMID:3058122

Isolation and partial characterization of gypsy moth BTR-270, an anionic brush border membrane glycoconjugate that binds Bacillus thuringiensis Cry1A toxins with high affinity

Treesearch

Algimantas P. Valaitis; Jeremy L. Jenkins; Mi Kyong Lee; Donald H. Dean; Karen J. Garner

2001-01-01

BTR-270, a gypsy moth (Lymantria dispar) brush border membrane molecule that binds Bacillus thuringiensis (Bt) Cry1A toxins with high affinity, was purified by preparative gel electrophoresis. Rabbit antibodies specific for the Bt toxin-binding molecule were raised. Attempts to label BTR-270 by protein-directed techniques were...

Shiga toxin 1 interaction with enterocytes causes apical protein mistargeting through the depletion of intracellular galectin-3

DOE Office of Scientific and Technical Information (OSTI.GOV)

Laiko, Marina; Murtazina, Rakhilya; Malyukova, Irina

Shiga toxins (Stx) 1 and 2 are responsible for intestinal and systemic sequelae of infection by enterohemorrhagic Escherichia coli (EHEC). However, the mechanisms through which enterocytes are damaged remain unclear. While secondary damage from ischemia and inflammation are postulated mechanisms for all intestinal effects, little evidence excludes roles for more primary toxin effects on intestinal epithelial cells. We now document direct pathologic effects of Stx on intestinal epithelial cells. We study a well-characterized rabbit model of EHEC infection, intestinal tissue and stool samples from EHEC-infected patients, and T84 intestinal epithelial cells treated with Stx1. Toxin uptake by intestinal epithelial cellsmore » in vitro and in vivo causes galectin-3 depletion from enterocytes by increasing the apical galectin-3 secretion. This Shiga toxin-mediated galectin-3 depletion impairs trafficking of several brush border structural proteins and transporters, including villin, dipeptidyl peptidase IV, and the sodium-proton exchanger 2, a major colonic sodium absorptive protein. The mistargeting of proteins responsible for the absorptive function might be a key event in Stx1-induced diarrhea. These observations provide new evidence that human enterocytes are directly damaged by Stx1. Conceivably, depletion of galectin-3 from enterocytes and subsequent apical protein mistargeting might even provide a means whereby other pathogens might alter intestinal epithelial absorption and produce diarrhea.« less

Water pumps.

PubMed

Loo, Donald D F; Wright, Ernest M; Zeuthen, Thomas

2002-07-01

The transport of water across epithelia has remained an enigma ever since it was discovered over 100 years ago that water was transported across the isolated small intestine in the absence of osmotic and hydrostatic pressure gradients. While it is accepted that water transport is linked to solute transport, the actual mechanisms are not well understood. Current dogma holds that active ion transport sets up local osmotic gradients in the spaces between epithelial cells, the lateral intercellular spaces, and this in turn drives water transport by local osmosis. In the case of the small intestine, which in humans absorbs about 8 l of water a day, there is no direct evidence for either local osmosis or aquaporin gene expression in enterocytes. Intestinal water absorption is greatly enhanced by glucose, and this is the basis for oral rehydration therapy in patients with secretory diarrhoea. In our studies of the intestinal brush border Na+-glucose cotransporter we have obtained evidence that there is a direct link between the transport of Na+, glucose and water transport, i.e. there is cotransport of water along with Na+ and sugar, that will account for about 50 % of the total water transport across the human intestinal brush border membrane. In this short review we summarize the evidence for water cotransport and propose how this occurs during the enzymatic turnover of the transporter. This is a general property of cotransporters and so we expect that this may have wider implications in the transport of water and other small polar molecules across cell membranes in animals and plants.

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

Human fetal enterocytes in vitro: modulation of the phenotype by extracellular matrix.

PubMed Central

Sanderson, I R; Ezzell, R M; Kedinger, M; Erlanger, M; Xu, Z X; Pringault, E; Leon-Robine, S; Louvard, D; Walker, W A

1996-01-01

The differentiation of small intestinal epithelial cells may require stimulation by microenvironmental factors in vivo. In this study, the effects of mesenchymal and luminal elements in nonmalignant epithelia] cells isolated from the human fetus were studied in vitro. Enterocytes from the human fetus were cultured and microenvironmental factors were added in stages, each stage more closely approximating the microenvironment in vivo. Four stages were examined: epithelial cells derived on plastic from intestinal culture and grown as a cell clone, the same cells grown on connective tissue support, primary epithelial explants grown on fibroblasts with a laminin base, and primary epithelial explants grown on fibroblasts and laminin with n-butyrate added to the incubation medium. The epithelial cell clone dedifferentiated when grown on plastic; however, the cells expressed cytokeratins and villin as evidence of their epithelial cell origin. Human connective tissue matrix from Engelbreth-Holm-Swarm sarcoma cells (Matrigel) modulated their phenotype: alkaline phosphatase activity increased, microvilli developed on their apical surface, and the profile of insulin-like growth factor binding proteins resembled that secreted by differentiated enterocytes. Epithelial cells taken directly from the human fetus as primary cultures and grown as explants on fibroblasts and laminin expressed greater specific enzyme activities in brush border membrane fractions than the cell clone. These activities were enhanced by the luminal molecule sodium butyrate. Thus the sequential addition of connective tissue and luminal molecules to nonmalignant epithelia] cells in vitro induces a spectrum of changes in the epithelial cell phenotype toward full differentiation. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 PMID:8755542

Influence of exocrine and endocrine pancreatic function on intestinal brush border enaymatic activities.

PubMed Central

Caspary, W F; Winckler, K; Lankisch, P G; Creutzfeldt, W

1975-01-01

Digestive enzymatic activities (disaccharidases, alkaline phosphatase, peptide hydrolases) have been determined in the mucosa of 14 patients with chronic pancreatitis. All had an abnormal secretin-pancreozymin test. Four patients had insulin-dependent diabetes mellitus, four a pathological glucose tolerance test. Nine patients had steatorrhoea. Maltase, sucrase, and alkaline phosphatase activity was significantly elevated in patients with exocrine pancreatic insufficiency, whereas those of lactase, trehalase, and peptide hydrolase were normal. Patients with steatorrhoea had higher maltase and sucrase activity than those without steatorrhoea, whereas decreased glucose tolerance had no effect on brush border enzymatic activity. It is suggested thatdecreased exocrine rather than decreased endocrine pancreatic function is responsible for the increase in intestinal disaccharidase and alkaline phosphatase activity, possible by the influence of pacreatic enzymes on the turnover of brush border enzymes from the luminal side of the mucosal membranes or by direct hormonal stimulation though cholecystokinin. PMID:1092602

Influence of time, storage temperature and freeze/thaw cycles on the activity of digestive enzymes from gilthead sea bream (Sparus aurata).

PubMed

Solovyev, Mikhail; Gisbert, Enric

2016-10-01

In this study, we tested the effects of long-term storage (2 years) at -20 °C and short-term storage (several hours) in ice and freeze/thaw cycles on the activities of pancreatic, gastric and intestinal (brush border and cytosolic) digestive enzymes in a teleost fish species. The results revealed a significant lose in activity of pancreatic (trypsin, chymotrypsin, total alkaline proteases and α-amylase) and intestinal cytosolic (leucine-alanine peptidase) enzymes between 140 and 270 days of storage at -20 °C, whereas in contrast, the activity of all the assayed brush border enzymes remained constant during the first 2 years of storage at -20 °C. During short-term storage conditions, the most stable enzymes assayed were those of the enterocytes of the brush border, which did not show any change in activity after being held for 5 h in ice. Five freezing and thawing cycles did not affect the activity of the intestinal brush border enzymes and the cytosolic ones, whereas the activity of trypsin, α-amylase and bile-salt-activated lipase was significantly affected by the number of freezing and thawing cycles. No changes in pepsin activity were found in samples exposed to 1 and 2 freezing and thawing cycles.

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.

Bacillus thuringiensis Cry1A toxin-binding glycoconjugates present on the brush border membrane and in the peritrophic membrane of the Douglas-fir tussock moth are peritrophins

Treesearch

Algimantas P. Valaitis; John D. Podgwaite

2013-01-01

Bacillus thuringiensis (Bt) Cry1A toxin-binding sites in the Douglas fir tussock moth (DFTM) larval gut were localized using immunofluorescence microscopy. Cry1Aa, Cry1Ab and Cry1Ac all bound strongly to the DFTM peritrophic membrane (PM); weaker binding of the Cry1A toxins was observed along the apical brush border of the midgut epithelium....

Effect of chronic hypo and hypervitaminosis C on the brush border enzymes and the intestinal uptake of glucose and alanine.

PubMed

Mahmood, A; Chauhan, V P; Lyall, V; Sarkar, A K

1979-08-15

Brush border sucrase and alkaline phosphatase activities are considerably enhanced in the intestine of ascorbic acid deficient guinea-pigs. Similar increase in the uptake of D-glucose and L-alanine also occurs in chronic vitamin C deficiency. However the permeability of D-glucose and L-alanine in the intestine of animals fed with large doses of vitamin C is severely depressed, with a reduction in the levels of sucrase and alkaline phosphatase activities.

Effect of cephalosporins on organic ion transport in renal membrane vesicles from rat and rabbit kidney cortex.

PubMed

Williams, P D; Hitchcock, M J; Hottendorf, G H

1985-03-01

The effects of cephaloridine and cephalothin on prototypical organic anion (p-aminohippurate, PAH) and cation (N-methylnicotinamide, NMN) transport were observed in brush border and basolateral membrane vesicles prepared from rat and rabbit renal cortex. The cephalosporins interacted with both the cationic and anionic transport systems. Cephalothin inhibited PAH transport in basolateral and brush border membrane in both rats and rabbits. Cephaloridine on the other hand inhibited PAH and NMN transport across rabbit basolateral membranes while it showed a lack of interaction with transport systems in rat basolateral membranes. Conversely, cephaloridine inhibited brush border transport of PAH and NMN in the rat but not in the rabbit. These results provide indirect evidence that cephalothin may be secreted across the renal tubule cell in rats and rabbits while cephaloridine may not accumulate in the rat kidney and becomes trapped in rabbit renal tubule cells. The differences in transport effects observed may explain intra- and interspecies differences in susceptibility to cephalosporin nephrotoxicity.

Effect of cisplatin on organic ion transport in membrane vesicles from rat kidney cortex.

PubMed

Williams, P D; Hottendorf, G H

1985-01-01

Purified renal membrane vesicles were utilized to gain indirect information regarding the renal handling of cisplatin. The effects of cisplatin on prototypical organic anion (p-amino-hippurate, PAH) and cation (N1-methylnicotinamide; tetraethylammonium, TEA) transport in brush border and basolateral membrane vesicles prepared from rat kidney cortex were observed. While cisplatin inhibited organic cation transport (N1-methylnicotinamide; TEA) in brush border and basolateral membranes, no interaction with the organic anion (p-amino-hippurate) system was observed. Kinetic analyses revealed that cisplatin is a competitive inhibitor of TEA transport in brush border membranes with a ki of 0.12 mM. While the relationship between organic cation transport inhibition and cisplatin nephrotoxicity is unknown, it may suggest that the cisplatin complex itself is transported into the kidney by the organic cation system. The reported effect of the organic anion, probenecid, on the renal handling of cisplatin is discussed in light of these results.

The mechanisms of brush border Na+/H+ exchanger activation by corticosteroids.

PubMed

Zallocchi, Marisa; Igarreta, Pilar; Calvo, Juan Carlos; Reboucas, Nancy Amaral; Damasco, María Christina

2003-02-01

Previously we showed that corticosterone and aldosterone increased proton fluxes in proximal tubule, by micropuncture and stationary microperfusion. Since the Na+/H+ exchanger is responsible for the main proximal proton secretion, we have now evaluated the effects aldosterone on Na+/H+ exchange activity in brush border vesicles. In order to evaluate the mechanism of action of glucocorticoids and mineralocorticoids, we studied the comparative effects of corticosterone and aldosterone on the abundance of NHE3 and NHE2 isoforms. We isolated renal brush border vesicles from rats by differential centrifugation in sham-operated, adrenalectomized, and adrenalectomized-aldosterone treated (ADX + aldosterone) animals. We measured the kinetics of H+ transport in response to increasing concentrations of Sodium Gluconate by fluorimetry using acridine orange. For Na+/H+ exchanger abundance we used Western blot analysis of brush border proteins in the above groups and in adrenalectomized-corticosterone treated rats. The Vmax in adrenalectomized animals was 22,162+/-1828 fluorescence units/min; in sham animals, 37,020+/-2722; and in ADX + aldosterone, 42,344+/-3044 (p<0.01 adrenalectomized vs others). No differences in Km were observed. Adrenalectomy decreased NHE3 abundance over Sham by 32% without modifying NHE2. Corticosterone-replacement enhanced NHE3 abundance by 76% and failed to increase NHE2. Aldosterone enhanced NHE2 abundance by 75% and did not increase NHE3. Mineralocorticoids enhance Na+/H+ exchange activity by increasing NHE2 abundance; glucocorticoids, by increasing NHE3 abundance.

Transmissible Gastroenteritis in Feeder Pigs: Observations on the Jejunal Epithelium of Normal Feeder Pigs and Feeder Pigs Infected with TGE Virus

PubMed Central

Morin, M.; Morehouse, L. G.

1974-01-01

Light and electron microscopy findings in the jejunal mucosa of the normal feeder pig and feeder pigs infected with transmissible gastroenteritis (TGE) virus are reported. Villi in the mid jejunum of the normal feeder pig were elongated, finger shaped and covered with a layer of columnar absorptive cells with a well developed and regular brush border. Severe lesions of villous atrophy were present in all jejunal segments of feeder swine killed 96 hours post infection with TGE virus. Atrophic villi were covered by flat to cuboidal cells with a poorly developed brush border in some areas. In other segments, cells varied in appearance from sub-columnar to columnar type of near normal appearance. The ultrastructure of the jejunal absorptive cells in the normal feeder pig was found to be similar to that described for the jejunal cells of other adult mammals. There were no significant indications of high pinocytotic activity. The epithelial cells covering the atrophic villi of TGE infected pigs had a fine structure similar to that described for the crypt cells, ranging in appearance from very immature to moderately differentiated cells. Microvilli were very short, decreased markedly in number and irregular in arrangement. The terminal web was poorly developed. Strands of rough endoplasmic reticulum were markedly diminished and an increase in free ribosomes was noted. The significance of these observations in explaining pathogenesis of TGE in feeder pigs is discussed. ImagesFig. 1.Fig. 2.Fig. 3.Fig. 4.Fig. 5.Fig. 6.Fig. 7.Fig. 8. PMID:4277743

Water pumps

PubMed Central

Loo, Donald D F; Wright, Ernest M; Zeuthen, Thomas

2002-01-01

The transport of water across epithelia has remained an enigma ever since it was discovered over 100 years ago that water was transported across the isolated small intestine in the absence of osmotic and hydrostatic pressure gradients. While it is accepted that water transport is linked to solute transport, the actual mechanisms are not well understood. Current dogma holds that active ion transport sets up local osmotic gradients in the spaces between epithelial cells, the lateral intercellular spaces, and this in turn drives water transport by local osmosis. In the case of the small intestine, which in humans absorbs about 8 l of water a day, there is no direct evidence for either local osmosis or aquaporin gene expression in enterocytes. Intestinal water absorption is greatly enhanced by glucose, and this is the basis for oral rehydration therapy in patients with secretory diarrhoea. In our studies of the intestinal brush border Na+-glucose cotransporter we have obtained evidence that there is a direct link between the transport of Na+, glucose and water transport, i.e. there is cotransport of water along with Na+ and sugar, that will account for about 50 % of the total water transport across the human intestinal brush border membrane. In this short review we summarize the evidence for water cotransport and propose how this occurs during the enzymatic turnover of the transporter. This is a general property of cotransporters and so we expect that this may have wider implications in the transport of water and other small polar molecules across cell membranes in animals and plants. PMID:12096049

Expression of Mucin-Type Glycoprotein K88 Receptors Strongly Correlates with Piglet Susceptibility to K88+ Enterotoxigenic Escherichia coli, but Adhesion of This Bacterium to Brush Borders Does Not

PubMed Central

Francis, David H.; Grange, Philippe A.; Zeman, David H.; Baker, Diane R.; Sun, Ronggai; Erickson, Alan K.

1998-01-01

Three antigenic variants of the K88 fimbrial adhesin exist in nature, K88ab, K88ac, and K88ad. Enterotoxigenic Escherichia coli (ETEC) strains that produce these fimbriae cause life-threatening diarrhea in some but not all young pigs. The susceptibility of pigs to these organisms has been correlated with the adherence of bacteria to isolated enterocyte brush borders. Whether that correlation holds for multiple K88 variants and over a broad genetic base of pigs is unknown and was the impetus for this study. We also desired to examine the correlation of the expression of a porcine intestinal brush border mucin-type glycoprotein (IMTGP) which binds K88ab and K88ac with the susceptibility of piglets to K88+ ETEC. Of 31 neonatal gnotobiotic pigs inoculated with K88ab+ or K88ac+ ETEC, 13 developed severe diarrhea, became dehydrated, and died or became moribund. Another pig became severely lethargic but not dehydrated. In vitro brush border adherence analysis was not possible for 10 of the severely ill pigs due to colonization by challenge strains. However, of the 17 pigs that did not become severely ill, 8 (47%) had brush borders that supported the adherence of K88ab+ and K88ac+ bacteria in vitro, suggesting a poor correlation between in vitro brush border adherence and piglet susceptibility to K88+ ETEC. By contrast, the expression of IMTGP was highly correlated with susceptibility to K88+ ETEC. Of the 12 pigs that produced IMTGP, 11 developed severe diarrhea. The other pig that produced IMTGP became lethargic but not severely diarrheic. Only 2 of 18 pigs that did not produce IMTGP became severely diarrheic. Colonizing bacteria were observed in histologic sections of intestines from all pigs that expressed IMTGP except for the one that did not develop severe diarrhea. However, colonizing bacteria were observed in histologic sections from only one pig that did not produce IMTGP. The bacterial concentration in the jejuna and ilea of pigs expressing IMTGP was significantly greater (P < 0.005) than that in pigs not expressing IMTGP. These observations suggest the IMTGP is a biologically relevant receptor for K88ab+ and K88ac+ E. coli or a correlate for expression for such a receptor. PMID:9712746

Impaired nutrient signaling and body weight control in a Na+ neutral amino acid cotransporter (Slc6a19)-deficient mouse.

PubMed

Bröer, Angelika; Juelich, Torsten; Vanslambrouck, Jessica M; Tietze, Nadine; Solomon, Peter S; Holst, Jeff; Bailey, Charles G; Rasko, John E J; Bröer, Stefan

2011-07-29

Amino acid uptake in the intestine and kidney is mediated by a variety of amino acid transporters. To understand the role of epithelial neutral amino acid uptake in whole body homeostasis, we analyzed mice lacking the apical broad-spectrum neutral (0) amino acid transporter B(0)AT1 (Slc6a19). A general neutral aminoaciduria was observed similar to human Hartnup disorder which is caused by mutations in SLC6A19. Na(+)-dependent uptake of neutral amino acids into the intestine and renal brush-border membrane vesicles was abolished. No compensatory increase of peptide transport or other neutral amino acid transporters was detected. Mice lacking B(0)AT1 showed a reduced body weight. When adapted to a standard 20% protein diet, B(0)AT1-deficient mice lost body weight rapidly on diets containing 6 or 40% protein. Secretion of insulin in response to food ingestion after fasting was blunted. In the intestine, amino acid signaling to the mammalian target of rapamycin (mTOR) pathway was reduced, whereas the GCN2/ATF4 stress response pathway was activated, indicating amino acid deprivation in epithelial cells. The results demonstrate that epithelial amino acid uptake is essential for optimal growth and body weight regulation.

Proton-coupled oligopeptide transport by rat renal cortical brush border membrane vesicles: a functional analysis using ACE inhibitors to determine the isoform of the transporter.

PubMed

Temple, C S; Boyd, C A

1998-08-14

We demonstrate that the angiotensin-converting enzyme inhibitors enalapril and captopril inhibit the transport of D-Phe-L-Gln into PepT1-expressing Xenopus oocytes and into rat renal cortical brush border membrane vesicles (BBMV). The kinetics of inhibition are competitive. Enalapril and captopril are not substrates for PepT2 (Boll et al., Proc. Natl. Acad. Sci. 93 (1996) 284-289). Therefore we conclude that in rat renal cortical BBMV this neutral dipeptide is transported via PepT1.

Vigabatrin transport across the human intestinal epithelial (Caco-2) brush-border membrane is via the H+-coupled amino-acid transporter hPAT1

PubMed Central

Abbot, Emily L; Grenade, Danielle S; Kennedy, David J; Gatfield, Kelly M; Thwaites, David T

2005-01-01

The aim of this investigation was to determine if the human proton-coupled amino-acid transporter 1 (hPAT1 or SLC36A1) is responsible for the intestinal uptake of the orally-administered antiepileptic agent 4-amino-5-hexanoic acid (vigabatrin). The Caco-2 cell line was used as a model of the human small intestinal epithelium. Competition experiments demonstrate that [3H]GABA uptake across the apical membrane was inhibited by vigabatrin and the GABA analogues trans-4-aminocrotonic acid (TACA) and guvacine, whereas 1-(aminomethyl)cyclohexaneacetic acid (gabapentin) had no affect. Experiments with 2′,7′-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF)-loaded Caco-2 cells demonstrate that apical exposure to vigabatrin and TACA induce comparable levels of intracellular acidification (due to H+/amino-acid symport) to that generated by GABA, suggesting that they are substrates for a H+-coupled absorptive transporter such as hPAT1. In hPAT1 and mPAT1-expressing Xenopus laevis oocytes [3H]GABA uptake was inhibited by vigabatrin, TACA and guvacine, whereas gabapentin failed to inhibit [3H]GABA uptake. In Na+-free conditions, vigabatrin and TACA evoked similar current responses (due to H+/amino-acid symport) in hPAT1-expressing oocytes under voltage-clamp conditions to that induced by GABA (whereas no current was observed in water-injected oocytes) consistent with the ability of these GABA analogues to inhibit [3H]GABA uptake. This study demonstrates that hPAT1 is the carrier responsible for the uptake of vigabatrin across the brush-border membrane of the small intestine and emphasises the therapeutic potential of hPAT1 as a delivery route for orally administered, clinically significant GABA-related compounds. PMID:16331283

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

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 ratemore » 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.« less

Polarizing intestinal epithelial cells electrically through Ror2

PubMed Central

Cao, Lin; McCaig, Colin D.; Scott, Roderick H.; Zhao, Siwei; Milne, Gillian; Clevers, Hans; Zhao, Min; Pu, Jin

2014-01-01

ABSTRACT The apicobasal polarity of enterocytes determines where the brush border membrane (apical membrane) will form, but how this apical membrane faces the lumen is not well understood. The electrical signal across the epithelium could serve as a coordinating cue, orienting and polarizing enterocytes. Here, we show that applying a physiological electric field to intestinal epithelial cells, to mimic the natural electric field created by the transepithelial potential difference, polarized phosphorylation of the actin-binding protein ezrin, increased expression of intestinal alkaline phosphatase (ALPI, a differentiation marker) and remodeled the actin cytoskeleton selectively on the cathode side. In addition, an applied electric field also activated ERK1/2 and LKB1 (also known as STK11), key molecules in apical membrane formation. Disruption of the tyrosine protein kinase transmembrane receptor Ror2 suppressed activation of ERK1/2 and LKB1 significantly, and subsequently inhibited apical membrane formation in enterocytes. Our findings indicate that the endogenous electric field created by the transepithelial potential difference might act as an essential coordinating signal for apical membrane formation at a tissue level, through activation of LKB1 mediated by Ror2–ERK signaling. PMID:24928904

Na/sup +/-independent, phloretin-sensitive monosaccharide transport system in isolated intestinal epithelial cells. [Chickens

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kimmich, G.A.; Randles, J.

1975-01-01

A monosaccharide transport system in addition to the active Na/sup +/-dependent system characteristic of the brush border surface of vertebrate intestinal tissue has been identified in isolated chick intestinal epithelial cells. The newly described system differs in several characteristics from the Na/sup +/-dependent process, including function in the absence of Na/sup +/; a high sensitivity to phloretin, relative insensitivity to phlorizin; different substrate specificity; and a very high K/sub T/ and V/sub max/. The system apparently functions only in a facilitated diffusion manner so that it serves to move monosaccharide across the cell membrane down its chemical gradient. An appreciablemore » fraction of total sugar efflux occurs via the Na/sup +/-independent carrier from cells which have accumulated sugar to a steady state. Phloretin selectively blocks this efflux so that a normal steady-state sugar gradient of seven- to eightfold is transformed to a new steady-state gradient which is greater than 14-fold. Locus of the new system is tentatively ascribed to the serosal cell surface where it would serve for monosaccharide transfer between enterocyte and lamina propria of the villus. (auth)« less

[Effect of natural or synthetic detergents on the transport of D-glucose in the membranes of vesicles of the brush border of the intestine of the rabbit].

PubMed

Favilli, F; Iantomasi, T; Stio, M; Treves, C; Vanni, P; Vincenzini, M T

1988-01-01

We describe here the effects of natural and synthetic detergents on the D-glucose transport into brush-border membranes of vesicles of rabbit's intestine. Two synthetic detergents: Triton X-100 and dodecyltrimethylammonium bromide have been found very strong inhibitors (more than 50 p. 100 of inhibition of maximal D-glucose uptake). Kinetic studies showed that these detergents behaved as mixed type inhibitors. The Na+-dependent transport of amino acids (aspartic acid, lysine, phenylalanine) is only poorly affected by dodecyltrimethylammonium bromide, while Triton X-100 inhibits unspecifically all the transport studied.

The Secretion and Action of Brush Border Enzymes in the Mammalian Small Intestine.

PubMed

Hooton, Diane; Lentle, Roger; Monro, John; Wickham, Martin; Simpson, Robert

2015-01-01

Microvilli are conventionally regarded as an extension of the small intestinal absorptive surface, but they are also, as latterly discovered, a launching pad for brush border digestive enzymes. Recent work has demonstrated that motor elements of the microvillus cytoskeleton operate to displace the apical membrane toward the apex of the microvillus, where it vesiculates and is shed into the periapical space. Catalytically active brush border digestive enzymes remain incorporated within the membranes of these vesicles, which shifts the site of BB digestion from the surface of the enterocyte to the periapical space. This process enables nutrient hydrolysis to occur adjacent to the membrane in a pre-absorptive step. The characterization of BB digestive enzymes is influenced by the way in which these enzymes are anchored to the apical membranes of microvilli, their subsequent shedding in membrane vesicles, and their differing susceptibilities to cleavage from the component membranes. In addition, the presence of active intracellular components of these enzymes complicates their quantitative assay and the elucidation of their dynamics. This review summarizes the ontogeny and regulation of BB digestive enzymes and what is known of their kinetics and their action in the peripheral and axial regions of the small intestinal lumen.

A bioartificial environment for kidney epithelial cells based on a supramolecular polymer basement membrane mimic and an organotypical culture system.

PubMed

Mollet, Björne B; Bogaerts, Iven L J; van Almen, Geert C; Dankers, Patricia Y W

2017-06-01

Renal applications in healthcare, such as renal replacement therapies and nephrotoxicity tests, could potentially benefit from bioartificial kidney membranes with fully differentiated and functional human tubular epithelial cells. A replacement of the natural environment of these cells is required to maintain and study cell functionality cell differentiation in vitro. Our approach was based on synthetic supramolecular biomaterials to mimic the natural basement membrane (BM) on which these cells grow and a bioreactor to provide the desired organotypical culture parameters. The BM mimics were constructed from ureidopyrimidinone (UPy)-functionalized polymer and bioactive peptides by electrospinning. The resultant membranes were shown to have a hierarchical fibrous BM-like structure consisting of self-assembled nanofibres within the electrospun microfibres. Human kidney-2 (HK-2) epithelial cells were cultured on the BM mimics under organotypical conditions in a custom-built bioreactor. The bioreactor facilitated in situ monitoring and functionality testing of the cultures. Cell viability and the integrity of the epithelial cell barrier were demonstrated inside the bioreactor by microscopy and transmembrane leakage of fluorescently labelled inulin, respectively. Furthermore, HK-2 cells maintained a polarized cell layer and showed modulation of both gene expression of membrane transporter proteins and metabolic activity of brush border enzymes when subjected to a continuous flow of culture medium inside the new bioreactor for 21 days. These results demonstrated that both the culture and study of renal epithelial cells was facilitated by the bioartificial in vitro environment that is formed by synthetic supramolecular BM mimics in our custom-built bioreactor. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

Two stages of enteropathogenic Escherichia coli intestinal pathogenicity are up and down-regulated by the epithelial cell differentiation.

PubMed

Gabastou, J M; Kernéis, S; Bernet-Camard, M F; Barbat, A; Coconnier, M H; Kaper, J B; Servin, A L

1995-09-01

Pathogens and eucaryotic cells are active partners during the process of pathogenicity. To gain access to enterocytes and to cross the epithelial membrane, many enterovirulent microorganisms interact with the brush border membrane-associated components as receptors. Recent reports provide evidence that intestinal cell differentiation plays a role in microbial pathogenesis. Human enteropathogenic Escherichia coli (EPEC) develop their pathogenicity upon infecting enterocytes. To determine if intestinal epithelial cell differentiation influences EPEC pathogenicity, we examined the infection of human intestinal epithelial cells by JPN 15 (pMAR7) [EAF+ eae+] EPEC strain as a function of the cell differentiation. The human embryonic intestinal INT407 cells, the human colonic T84 cells, the human undifferentiated HT-29 cells (HT-29 Std) and two enterocytic cell lines, HT-29 glc-/+ and Caco-2 cells, were used as cellular models. Cells were infected apically with the EPEC strain and the cell-association and cell-entry were examined by quantitative determination using metabolically radiolabeled bacteria, as well as by light, scanning and transmission electron microscopy. [EAF+ eae+] EPEC bacteria efficiently colonized the cultured human intestinal cells. Diffuse bacterial adhesion occurred to undifferentiated HT-29 Std and INT407 cells, whereas characteristic EPEC cell clusters were observed on fully differentiated enterocytic HT-29 glc-/+ cells and on colonic crypt T84 cells. As shown using the Caco-2 cell line, which spontaneously differentiates in culture, the formation of EPEC clusters increased as a function of the epithelial cell differentiation. In contrast, efficient cell-entry of [EAF+ eae+] EPEC bacteria occurred in recently differentiated Caco-2 cells and decreased when the cells were fully differentiated.(ABSTRACT TRUNCATED AT 250 WORDS)

Pathogenesis of Human Enterovirulent Bacteria: Lessons from Cultured, Fully Differentiated Human Colon Cancer Cell Lines

PubMed Central

Liévin-Le Moal, Vanessa

2013-01-01

SUMMARY Hosts are protected from attack by potentially harmful enteric microorganisms, viruses, and parasites by the polarized fully differentiated epithelial cells that make up the epithelium, providing a physical and functional barrier. Enterovirulent bacteria interact with the epithelial polarized cells lining the intestinal barrier, and some invade the cells. A better understanding of the cross talk between enterovirulent bacteria and the polarized intestinal cells has resulted in the identification of essential enterovirulent bacterial structures and virulence gene products playing pivotal roles in pathogenesis. Cultured animal cell lines and cultured human nonintestinal, undifferentiated epithelial cells have been extensively used for understanding the mechanisms by which some human enterovirulent bacteria induce intestinal disorders. Human colon carcinoma cell lines which are able to express in culture the functional and structural characteristics of mature enterocytes and goblet cells have been established, mimicking structurally and functionally an intestinal epithelial barrier. Moreover, Caco-2-derived M-like cells have been established, mimicking the bacterial capture property of M cells of Peyer's patches. This review intends to analyze the cellular and molecular mechanisms of pathogenesis of human enterovirulent bacteria observed in infected cultured human colon carcinoma enterocyte-like HT-29 subpopulations, enterocyte-like Caco-2 and clone cells, the colonic T84 cell line, HT-29 mucus-secreting cell subpopulations, and Caco-2-derived M-like cells, including cell association, cell entry, intracellular lifestyle, structural lesions at the brush border, functional lesions in enterocytes and goblet cells, functional and structural lesions at the junctional domain, and host cellular defense responses. PMID:24006470

Alcohol versus brush PRK: visual outcomes and adverse effects.

PubMed

Sia, Rose K; Ryan, Denise S; Stutzman, Richard D; Psolka, Maximilian; Mines, Michael J; Wagner, Melvin E; Weber, Eric D; Wroblewski, Keith J; Bower, Kraig S

2012-08-01

A smooth corneal surface prior to laser ablation is important in order to achieve a favorable refractive outcome. In this study, we compare PRK outcomes following two commonly used methods of epithelial debridement: Amoils epithelial scrubber (brush) versus 20% ethanol (alcohol). We reviewed records of patients who underwent wavefront-optimized PRK for myopia or myopic astigmatism between January 2008 and June 2010. Two treatment groups (brush vs. alcohol) were compared in terms of uncorrected distance visual acuity (UDVA), manifest refraction spherical equivalent (MRSE), corrected distance visual acuity (CDVA), and complications at postoperative months 1, 3, 6, and 12. One thousand five hundred ninety-three eyes of 804 patients underwent PRK during the study period: 828 brush-treated eyes and 765 alcohol-treated eyes. At 6 months postoperatively UDVA was ≥20/20 in 94.7% of brush-treated eyes versus 94.4% of alcohol-treated eyes (P = 0.907). At 1 month a higher percentage of brush-treated eyes maintained or gained one or more lines CDVA compared to alcohol-treated eyes (P = 0.007), but there were no other differences in UDVA, MRSE, or CDVA at any point postoperatively. At 1 month 75.4% of brush-treated eyes versus 70.4% of alcohol-treated eyes were free of complications (P = 0.032), and there were fewer brush-treated eyes with corneal haze (4.0% vs. 6.9%, P = 0.012) and dry eye (8.9% vs. 14.4%, P = 0.001). Although corneal haze was slightly more frequent in the alcohol group, most was trace and not significant. Although alcohol-assisted PRK had more minor complications in the early postoperative period, including corneal haze and dry eye, results for both groups beyond 1 month were comparable. Copyright © 2012 Wiley Periodicals, Inc.

Binding of glycated ovocystatin to rat renal brush border membranes.

PubMed

Golab, Krzysztof; Gburek, Jakub; Konopska, Bogusława; Krotkiewski, Hubert; Warwas, Maria

2013-10-01

Glycated proteins are considered as one of the factors involved in the pathogenesis of diabetic complications, including nephropathy. These proteins are formed endogenously under conditions of hyperglycemia, as well as being provided with food containing sugars, which was subjected to high temperature. Examples are egg products. One of the proteins found in eggs in a relatively high concentration is chicken cystatin (ovocystatin). It is now believed that some proteins can passage the intestinal epithelium by transcytosis directly into the bloodstream. Thus, glycated protein present in food can be an additional source of glycotoxins. The aim of this study was to compare the affinity of native and glycated cystatin to the brush border membranes of rat kidney. Kinetic analysis was performed with surface plasmon resonance technique using sensor chip L1. Dissociation constants for native and glycated cystatin (Kd ) were 2.76 μmol/L and 3.82 μmol/L, respectively. The results of our study indicate that glycation only slightly affects binding of cystatin to brush border membranes. This suggests that glycated cystatin and other glycated proteins may also be efficiently taken up in the kidney proximal tubule. The observation may be important for understanding the mechanisms involved in the development of diabetic nephropathy. © 2013 Japanese Society of Animal Science.

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.

The In Ovo Feeding Administration (Gallus Gallus)—An Emerging In Vivo Approach to Assess Bioactive Compounds with Potential Nutritional Benefits

PubMed Central

2018-01-01

In recent years, the in ovo feeding in fertilized broiler (Gallus gallus) eggs approach was further developed and currently is widely applied in the evaluation process of the effects of functional foods (primarily plant origin compounds) on the functionality of the intestinal brush border membrane, as well as potential prebiotic properties and interactions with the intestinal microbial populations. This review collates the information of potential nutrients and their effects on the mineral absorption, gut development, brush border membrane functionality, and immune system. In addition, the advantages and limitations of the in ovo feeding method in the assessment of potential prebiotic effects of plant origin compounds is discussed. PMID:29597266

Proteomic changes during intestinal cell maturation in vivo

PubMed Central

Chang, Jinsook; Chance, Mark R.; Nicholas, Courtney; Ahmed, Naseem; Guilmeau, Sandra; Flandez, Marta; Wang, Donghai; Byun, Do-Sun; Nasser, Shannon; Albanese, Joseph M.; Corner, Georgia A.; Heerdt, Barbara G.; Wilson, Andrew J.; Augenlicht, Leonard H.; Mariadason, John M.

2008-01-01

Intestinal epithelial cells undergo progressive cell maturation as they migrate along the crypt-villus axis. To determine molecular signatures that define this process, proteins differentially expressed between the crypt and villus were identified by 2D-DIGE and MALDI-MS. Forty-six differentially expressed proteins were identified, several of which were validated by immunohistochemistry. Proteins upregulated in the villus were enriched for those involved in brush border assembly and lipid uptake, established features of differentiated intestinal epithelial cells. Multiple proteins involved in glycolysis were also upregulated in the villus, suggesting increased glycolysis is a feature of intestinal cell differentiation. Conversely, proteins involved in nucleotide metabolism, and protein processing and folding were increased in the crypt, consistent with functions associated with cell proliferation. Three novel paneth cell markers, AGR2, HSPA5 and RRBP1 were also identified. Notably, significant correlation was observed between overall proteomic changes and corresponding gene expression changes along the crypt-villus axis, indicating intestinal cell maturation is primarily regulated at the transcriptional level. This proteomic profiling analysis identified several novel proteins and functional processes differentially induced during intestinal cell maturation in vivo. Integration of proteomic, immunohistochemical, and parallel gene expression datasets demonstrate the coordinated manner in which intestinal cell maturation is regulated. PMID:18824147

Nephropathy associated with sickle cell anemia: an autologous immune complex nephritis. I. Studies on nature of glomerular-bound antibody and antigen identification in a patient with sickle cell disease and immune deposit glomerulonephritis.

PubMed

Strauss, J; Pardo, V; Koss, M N; Griswold, W; McIntosh, R M

1975-03-01

The nature of the glomerular-bound antibody and the putative antigen was investigated in one of the patients with sickle cell disease and immune deposit membranoproliferative glomerulonephritis by immunohistologic and glomerular antibody elution. Renal proximal tubular epithelial antigen was localized in association with immunoglobulins G (IgG), M (IgM), Clq fraction of the first component of complement (Clq) and the third component of complement (C3) in a granular pattern along the glomerular basement membrane of the patient's kidney. IgG and IgM were eluted from glomeruli. These immunoglobulins fixed to the proximal tubules of normal human kidney by direct immunofluorescence. This localization was abolished by absorption of the eluted immunoglobulins with renal tubular epithelial (RTE) antigen. The IgG eluted from the glomeruli blocked the fixation of rabbit anti-RTE antigen to normal proximal tubular brush border. These studies suggest that the nephritis in this patient was due to deposition of complexes or RTE antigen and specific antibody. An autologous immune complex nephritis may develop in some patients with sickle cell anemia secondary to RTE antigen released possibly after renal ischemia or some other phenomenon causing renal tubular damage.

Transcriptional Modulation of Genes Encoding Structural Characteristics of Differentiating Enterocytes During Development of a Polarized Epithelium In Vitro

PubMed Central

Halbleib, Jennifer M.; Sääf, Annika M.

2007-01-01

Although there is considerable evidence implicating posttranslational mechanisms in the development of epithelial cell polarity, little is known about the patterns of gene expression and transcriptional regulation during this process. We characterized the temporal program of gene expression during cell–cell adhesion–initiated polarization of human Caco-2 cells in tissue culture, which develop structural and functional polarity similar to that of enterocytes in vivo. A distinctive switch in gene expression patterns occurred upon formation of cell–cell contacts between neighboring cells. Expression of genes involved in cell proliferation was down-regulated concomitant with induction of genes necessary for functional specialization of polarized epithelial cells. Transcriptional up-regulation of these latter genes correlated with formation of important structural and functional features in enterocyte differentiation and establishment of structural and functional cell polarity; components of the apical microvilli were induced as the brush border formed during polarization; as barrier function was established, expression of tight junction transmembrane proteins peaked; transcripts encoding components of the apical, but not the basal-lateral trafficking machinery were increased during polarization. Coordinated expression of genes encoding components of functional cell structures were often observed indicating temporal control of expression and assembly of multiprotein complexes. PMID:17699590

Pancreatic α-Amylase Controls Glucose Assimilation by Duodenal Retrieval through N-Glycan-specific Binding, Endocytosis, and Degradation*

PubMed Central

Date, Kimie; Satoh, Ayano; Iida, Kaoruko; Ogawa, Haruko

2015-01-01

α-Amylase, a major pancreatic protein and starch hydrolase, is essential for energy acquisition. Mammalian pancreatic α-amylase binds specifically to glycoprotein N-glycans in the brush-border membrane to activate starch digestion, whereas it significantly inhibits glucose uptake by Na+/glucose cotransporter 1 (SGLT1) at high concentrations (Asanuma-Date, K., Hirano, Y., Le, N., Sano, K., Kawasaki, N., Hashii, N., Hiruta, Y., Nakayama, K., Umemura, M., Ishikawa, K., Sakagami, H., and Ogawa, H. (2012) Functional regulation of sugar assimilation by N-glycan-specific interaction of pancreatic α-amylase with glycoproteins of duodenal brush border membrane. J. Biol. Chem. 287, 23104–23118). However, how the inhibition is stopped was unknown. Here, we show a new mechanism for the regulation of intestinal glucose absorption. Immunohistochemistry revealed that α-amylase in the duodena of non-fasted, but not fasted, pigs was internalized from the pancreatic fluid and immunostained. We demonstrated that after N-glycan binding, pancreatic α-amylase underwent internalization into lysosomes in a process that was inhibited by α-mannoside. The internalized α-amylase was degraded, showing low enzymatic activity and molecular weight at the basolateral membrane. In a human intestinal Caco-2 cell line, Alexa Fluor 488-labeled pancreatic α-amylase bound to the cytomembrane was transported to lysosomes through the endocytic pathway and then disappeared, suggesting degradation. Our findings indicate that N-glycan recognition by α-amylase protects enterocytes against a sudden increase in glucose concentration and restores glucose uptake by gradual internalization, which homeostatically controls the postprandial blood glucose level. The internalization of α-amylase may also enhance the supply of amino acids required for the high turnover of small intestine epithelial cells. This study provides novel and significant insights into the control of blood sugar during the absorption stage in the intestine. PMID:26023238

Mixed organic solvents induce renal injury in rats.

PubMed

Qin, Weisong; Xu, Zhongxiu; Lu, Yizhou; Zeng, Caihong; Zheng, Chunxia; Wang, Shengyu; Liu, Zhihong

2012-01-01

To investigate the injury effects of organic solvents on kidney, an animal model of Sprague-Dawley (SD) rats treated with mixed organic solvents via inhalation was generated and characterized. The mixed organic solvents consisted of gasoline, dimethylbenzene and formaldehyde (GDF) in the ratio of 2:2:1, and were used at 12,000 PPM to treat the rats twice a day, each for 3 hours. Proteinuria appeared in the rats after exposure for 5-6 weeks. The incidences of proteinuria in male and female rats after exposure for 12 weeks were 43.8% (7/16) and 25% (4/16), respectively. Urinary N-Acetyl-β-(D)-Glucosaminidase (NAG) activity was increased significantly after exposure for 4 weeks. Histological examination revealed remarkable injuries in the proximal renal tubules, including tubular epithelial cell detachment, cloud swelling and vacuole formation in the proximal tubular cells, as well as proliferation of parietal epithelium and tubular reflux in glomeruli. Ultrastructural examination found that brush border and cytoplasm of tubular epithelial cell were dropped, that tubular epithelial cells were partially disintegrated, and that the mitochondria of tubular epithelial cells were degenerated and lost. In addition to tubular lesions, glomerular damages were also observed, including segmental foot process fusion and loss of foot process covering on glomerular basement membrane (GBM). Immunofluorescence staining indicated that the expression of nephrin and podocin were both decreased after exposure of GDF. In contrast, increased expression of desmin, a marker of podocyte injury, was found in some areas of a glomerulus. TUNEL staining showed that GDF induced apoptosis in tubular cells and glomerular cells. These studies demonstrate that GDF can induce both severe proximal tubular damage and podocyte injury in rats, and the tubular lesions appear earlier than that of glomeruli.

Mixed Organic Solvents Induce Renal Injury in Rats

PubMed Central

Qin, Weisong; Xu, Zhongxiu; Lu, Yizhou; Zeng, Caihong; Zheng, Chunxia; Wang, Shengyu; Liu, Zhihong

2012-01-01

To investigate the injury effects of organic solvents on kidney, an animal model of Sprague-Dawley (SD) rats treated with mixed organic solvents via inhalation was generated and characterized. The mixed organic solvents consisted of gasoline, dimethylbenzene and formaldehyde (GDF) in the ratio of 2∶2:1, and were used at 12,000 PPM to treat the rats twice a day, each for 3 hours. Proteinuria appeared in the rats after exposure for 5–6 weeks. The incidences of proteinuria in male and female rats after exposure for 12 weeks were 43.8% (7/16) and 25% (4/16), respectively. Urinary N-Acetyl-β-(D)-Glucosaminidase (NAG) activity was increased significantly after exposure for 4 weeks. Histological examination revealed remarkable injuries in the proximal renal tubules, including tubular epithelial cell detachment, cloud swelling and vacuole formation in the proximal tubular cells, as well as proliferation of parietal epithelium and tubular reflux in glomeruli. Ultrastructural examination found that brush border and cytoplasm of tubular epithelial cell were dropped, that tubular epithelial cells were partially disintegrated, and that the mitochondria of tubular epithelial cells were degenerated and lost. In addition to tubular lesions, glomerular damages were also observed, including segmental foot process fusion and loss of foot process covering on glomerular basement membrane (GBM). Immunofluorescence staining indicated that the expression of nephrin and podocin were both decreased after exposure of GDF. In contrast, increased expression of desmin, a marker of podocyte injury, was found in some areas of a glomerulus. TUNEL staining showed that GDF induced apoptosis in tubular cells and glomerular cells. These studies demonstrate that GDF can induce both severe proximal tubular damage and podocyte injury in rats, and the tubular lesions appear earlier than that of glomeruli. PMID:23029287

An Apical MRCK-driven Morphogenetic Pathway Controls Epithelial Polarity

PubMed Central

Zihni, Ceniz; Vlassaks, Evi; Terry, Stephen; Carlton, Jeremy; Leung, Thomas King Chor; Olson, Michael; Pichaud, Franck; Balda, Maria Susana; Matter, Karl

2017-01-01

Polarized epithelia develop distinct cell surface domains, with the apical membrane acquiring characteristic morphological features such as microvilli. Cell polarization is driven by polarity determinants including the evolutionarily conserved partitioning defective (PAR) proteins that are separated into distinct cortical domains. PAR protein segregation is thought to be a consequence of asymmetric actomyosin contractions. The mechanism of activation of apically polarized actomyosin contractility is unknown. Here we show that the Cdc42 effector MRCK activates Myosin-II at the apical pole to segregate aPKC-Par6 from junctional Par3, defining the apical domain. Apically polarized MRCK-activated actomyosin contractility is reinforced by cooperation with aPKC-Par6 downregulating antagonistic RhoA-driven junctional actomyosin contractility, and drives polarization of cytosolic brush border determinants and apical morphogenesis. MRCK-activated polarized actomyosin contractility is required for apical differentiation and morphogenesis in vertebrate epithelia and Drosophila photoreceptors. Our results identify an apical origin of actomyosin-driven morphogenesis that couples cytoskeletal reorganization to PAR polarity signalling. PMID:28825699

Phosphorous digestibility and activity of intestinal phytase in hybrid tilapia, Oreochromis niloticus X O. aureus

USGS Publications Warehouse

La Vorgna, M.W.; Hafez, Y.; Hughes, S.G.; Handwerker, T.

2003-01-01

Experiments were conducted to determine the degree to which phytate-bound phosphorus from plant protein sources could be used by hybrid tilapia (Oreochromis niloticus X O. aureus). Utilizing an inert marker technique with chromic oxide, hybrid tilapia in our study were effective at utilizing both inorganic and phytate phosphorus as evidenced by average apparent digestibility values of 93.2% and 90.0% for total and phytate phosphorus, respectively. Analysis of the intestinal brush border membrane of the tilapia revealed enzyme activity that was capable of hydrolyzing phytic acid. The presence of phytic acid hydrolyzing enzyme activity in the intestinal brush border provides a probable mechanism by which these hybrid tilapia are able to utilize phytate phosphorus effectively. ?? 2003 by The Haworth Press, Inc. All rights reserved.

Class B type I scavenger receptor is responsible for the high affinity cholesterol binding activity of intestinal brush border membrane vesicles

PubMed Central

Labonté, Eric D.; Howles, Philip N.; Granholm, Norman A.; Rojas, Juan C.; Davies, Joanna P.; Ioannou, Yiannis A.; Hui, David Y.

2007-01-01

Recent studies have documented the importance of Niemann Pick C1-like 1 protein (NPC1L1), a putative physiological target of the drug ezetimibe, in mediating intestinal cholesterol absorption. However, whether NPC1L1 is the high affinity cholesterol binding protein on intestinal brush border membranes is still controversial. In this study, brush border membrane vesicles (BBMV) from wild type and NPC1L1−/− mice were isolated and assayed for micellar cholesterol binding in the presence or absence of ezetimibe. Results confirmed the loss of the high affinity component of cholesterol binding when wild type BBMV preparations were incubated with antiserum against the class B type 1 scavenger receptor (SR-BI) in the reaction mixture similar to previous studies. Subsequently, second order binding of cholesterol was observed with BBMV from wild type and NPC1L1−/− mice. The inclusion of ezetimibe in these in vitro reaction assays resulted in the loss of the high affinity component of cholesterol interaction. Surprisingly, BBMVs from NPC1L1−/− mice maintained active binding of cholesterol. These results documented that SR-BI, not NPC1L1, is the major protein responsible for the initial high affinity cholesterol ligand binding process in the cholesterol absorption pathway. Additionally, ezetimibe may inhibit BBM cholesterol binding through targets such as SR-BI in addition to its inhibition of NPC1L1. PMID:17442616

Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes*

PubMed Central

Park, Dayoung; Brune, Kristin A.; Mitra, Anupam; Marusina, Alina I.; Maverakis, Emanual; Lebrilla, Carlito B.

2015-01-01

Changes in cell surface glycosylation occur during the development and differentiation of cells and have been widely correlated with the progression of several diseases. Because of their structural diversity and sensitivity to intra- and extracellular conditions, glycans are an indispensable tool for analyzing cellular transformations. Glycans present on the surface of intestinal epithelial cells (IEC) mediate interactions with billions of native microorganisms, which continuously populate the mammalian gut. A distinct feature of IECs is that they differentiate as they migrate upwards from the crypt base to the villus tip. In this study, nano-LC/ESI QTOF MS profiling was used to characterize the changes in glycosylation that correspond to Caco-2 cell differentiation. As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. At day 21, when cells appear to be completely differentiated, remodeling of the cell surface glycome ceases. Differential expression of glycans during IEC maturation appears to play a key functional role in regulating the membrane-associated hydrolases and contributes to the mucosal surface innate defense mechanisms. Developing methodologies to rapidly identify changes in IEC surface glycans may lead to a rapid screening approach for a variety of disease states affecting the GI tract. PMID:26355101

Protein tyrosine phosphatase SAP-1 protects against colitis through regulation of CEACAM20 in the intestinal epithelium.

PubMed

Murata, Yoji; Kotani, Takenori; Supriatna, Yana; Kitamura, Yasuaki; Imada, Shinya; Kawahara, Kohichi; Nishio, Miki; Daniwijaya, Edwin Widyanto; Sadakata, Hisanobu; Kusakari, Shinya; Mori, Munemasa; Kanazawa, Yoshitake; Saito, Yasuyuki; Okawa, Katsuya; Takeda-Morishita, Mariko; Okazawa, Hideki; Ohnishi, Hiroshi; Azuma, Takeshi; Suzuki, Akira; Matozaki, Takashi

2015-08-04

Intestinal epithelial cells contribute to regulation of intestinal immunity in mammals, but the detailed molecular mechanisms of such regulation have remained largely unknown. Stomach-cancer-associated protein tyrosine phosphatase 1 (SAP-1, also known as PTPRH) is a receptor-type protein tyrosine phosphatase that is localized specifically at microvilli of the brush border in gastrointestinal epithelial cells. Here we show that SAP-1 ablation in interleukin (IL)-10-deficient mice, a model of inflammatory bowel disease, resulted in a marked increase in the severity of colitis in association with up-regulation of mRNAs for various cytokines and chemokines in the colon. Tyrosine phosphorylation of carcinoembryonic antigen-related cell adhesion molecule (CEACAM) 20, an intestinal microvillus-specific transmembrane protein of the Ig superfamily, was greatly increased in the intestinal epithelium of the SAP-1-deficient animals, suggesting that this protein is a substrate for SAP-1. Tyrosine phosphorylation of CEACAM20 by the protein tyrosine kinase c-Src and the consequent association of CEACAM20 with spleen tyrosine kinase (Syk) promoted the production of IL-8 in cultured cells through the activation of nuclear factor-κB (NF-κB). In addition, SAP-1 and CEACAM20 were found to form a complex through interaction of their ectodomains. SAP-1 and CEACAM20 thus constitute a regulatory system through which the intestinal epithelium contributes to intestinal immunity.

Cholinergic urethral brush cells are widespread throughout placental mammals.

PubMed

Deckmann, Klaus; Krasteva-Christ, Gabriela; Rafiq, Amir; Herden, Christine; Wichmann, Judy; Knauf, Sascha; Nassenstein, Christina; Grevelding, Christoph G; Dorresteijn, Adriaan; Chubanov, Vladimir; Gudermann, Thomas; Bschleipfer, Thomas; Kummer, Wolfgang

2015-11-01

We previously identified a population of cholinergic epithelial cells in murine, human and rat urethrae that exhibits a structural marker of brush cells (villin) and expresses components of the canonical taste transduction signaling cascade (α-gustducin, phospholipase Cβ2 (PLCβ2), transient receptor potential cation channel melanostatin 5 (TRPM5)). These cells serve as sentinels, monitoring the chemical composition of the luminal content for potentially hazardous compounds such as bacteria, and initiate protective reflexes counteracting further ingression. In order to elucidate cross-species conservation of the urethral chemosensory pathway we investigated the occurrence and molecular make-up of urethral brush cells in placental mammals. We screened 11 additional species, at least one in each of the five mammalian taxonomic units primates, carnivora, perissodactyla, artiodactyla and rodentia, for immunohistochemical labeling of the acetylcholine synthesizing enzyme, choline acetyltransferase (ChAT), villin, and taste cascade components (α-gustducin, PLCβ2, TRPM5). Corresponding to findings in previously investigated species, urethral epithelial cells with brush cell shape were immunolabeled in all 11 mammals. In 8 species, immunoreactivities against all marker proteins and ChAT were observed, and double-labeling immunofluorescence confirmed the cholinergic nature of villin-positive and chemosensory (TRPM5-positive) cells. In cat and horse, these cells were not labeled by the ChAT antiserum used in this study, and unspecific reactions of the secondary antiserum precluded conclusions about ChAT-expression in the bovine epithelium. These data indicate that urethral brush cells are widespread throughout the mammalian kingdom and evolved not later than about 64.5millionyears ago. Copyright © 2015 Elsevier B.V. All rights reserved.

Identification of Epithelial Phospholipase A2 Receptor 1 as a Potential Target in Asthma

PubMed Central

Nolin, James D.; Ogden, H. Luke; Lai, Ying; Altemeier, William A.; Frevert, Charles W.; Bollinger, James G.; Naika, Gajendra S.; Kicic, Anthony; Stick, Stephen M.; Lambeau, Gerard; Henderson, William R.; Gelb, Michael H.

2016-01-01

Secreted phospholipase A2s (sPLA2s) regulate eicosanoid formation and have been implicated in asthma. Although sPLA2s function as enzymes, some of the sPLA2s bind with high affinity to a C-type lectin receptor, called PLA2R1, which has functions in both cellular signaling and clearance of sPLA2s. We sought to examine the expression of PLA2R1 in the airway epithelium of human subjects with asthma and the function of the murine Pla2r1 gene in a model of asthma. Expression of PLA2R1 in epithelial brushings was assessed in two distinct cohorts of children with asthma by microarray and quantitative PCR, and immunostaining for PLA2R1 was conducted on endobronchial tissue and epithelial brushings from adults with asthma. C57BL/129 mice deficient in Pla2r1 (Pla2r1−/−) were characterized in an ovalbumin (OVA) model of allergic asthma. PLA2R1 was differentially overexpressed in epithelial brushings of children with atopic asthma in both cohorts. Immunostaining for PLA2R1 in endobronchial tissue localized to submucosal glandular epithelium and columnar epithelial cells. After OVA sensitization and challenge, Pla2r1−/− mice had increased airway hyperresponsiveness, as well as an increase in cellular trafficking of eosinophils to the peribronchial space and bronchoalveolar lavage fluid, and an increase in airway permeability. In addition, Pla2r1−/− mice had more dendritic cells in the lung, higher levels of OVA-specific IgG, and increased production of both type-1 and type-2 cytokines by lung leukocytes. PLA2R1 is increased in the airway epithelium in asthma, and serves as a regulator of airway hyperresponsiveness, airway permeability, antigen sensitization, and airway inflammation. PMID:27448109

Identification of Epithelial Phospholipase A2 Receptor 1 as a Potential Target in Asthma.

PubMed

Nolin, James D; Ogden, H Luke; Lai, Ying; Altemeier, William A; Frevert, Charles W; Bollinger, James G; Naika, Gajendra S; Kicic, Anthony; Stick, Stephen M; Lambeau, Gerard; Henderson, William R; Gelb, Michael H; Hallstrand, Teal S

2016-12-01

Secreted phospholipase A 2 s (sPLA 2 s) regulate eicosanoid formation and have been implicated in asthma. Although sPLA 2 s function as enzymes, some of the sPLA 2 s bind with high affinity to a C-type lectin receptor, called PLA2R1, which has functions in both cellular signaling and clearance of sPLA 2 s. We sought to examine the expression of PLA2R1 in the airway epithelium of human subjects with asthma and the function of the murine Pla2r1 gene in a model of asthma. Expression of PLA2R1 in epithelial brushings was assessed in two distinct cohorts of children with asthma by microarray and quantitative PCR, and immunostaining for PLA2R1 was conducted on endobronchial tissue and epithelial brushings from adults with asthma. C57BL/129 mice deficient in Pla2r1 (Pla2r1 -/- ) were characterized in an ovalbumin (OVA) model of allergic asthma. PLA2R1 was differentially overexpressed in epithelial brushings of children with atopic asthma in both cohorts. Immunostaining for PLA2R1 in endobronchial tissue localized to submucosal glandular epithelium and columnar epithelial cells. After OVA sensitization and challenge, Pla2r1 -/- mice had increased airway hyperresponsiveness, as well as an increase in cellular trafficking of eosinophils to the peribronchial space and bronchoalveolar lavage fluid, and an increase in airway permeability. In addition, Pla2r1 -/- mice had more dendritic cells in the lung, higher levels of OVA-specific IgG, and increased production of both type-1 and type-2 cytokines by lung leukocytes. PLA2R1 is increased in the airway epithelium in asthma, and serves as a regulator of airway hyperresponsiveness, airway permeability, antigen sensitization, and airway inflammation.

Phosphatidylethanolamine N-methyltransferase activity is increased in rat intestinal brush-border membrane by chronic ethanol ingestion.

PubMed

Furtado, Valéria Cristina Soares; Takiya, Christina Maeda; Braulio, Valeria Bender

2002-01-01

Phosphatidylethanolamine N-methyltransferase (PEMT) catalyses the synthesis of phosphatidylcholine from phosphatidylethanolamine. The aim of this study was to evaluate the effect of chronic ethanol ingestion on PEMT activity in the jejunal brush-border membrane (BBM) of adequately nourished rats. For this purpose, rats were fed a liquid diet containing ethanol [ethanol-fed group (EFG)] or an isocaloric liquid diet without ethanol [pair-fed group (PFG)] for 4 weeks. Diet ingestion, body weight, nitrogen balance and urinary creatinine excretion were monitored during the experimental period, and serum transferrin levels were determined at the end. BBM was isolated for the determination of PEMT activity. PEMT activity was significantly increased in the jejunal BBM of the EFG. Nutritional parameters, however, did not differ between groups. The increase in PEMT activity may be attributed exclusively to chronic ethanol ingestion, since a major nutritional deficit was excluded.

Expression analysis of extracellular matrix components in brush biopsies of oral lesions.

PubMed

Driemel, Oliver; Kosmehl, Hartwig; Rosenhahn, Julia; Berndt, Alexander; Reichert, Torsten E; Zardi, Luciano; Dahse, Regine

2007-01-01

Oral brush biopsies have proved to be a promising new non-invasive methodology in the diagnosis of oral lesions. The extracellular matrix (ECM) molecules gamma2 chain of laminin-5 (L5gamma2), tenascin-c (Tn-C) and the fibronectin isoform containing EDB (EDB-fn) are involved in matrix remodeling during malignant transformation in oral carcinoma. Expression of L5gamma2, Tn-C and EDB-fn was analysed in brush biopsy-obtained cells of benign inflammatory or hyperproliferative lesions and primary oral squamous cell carcinoma (OSCC) using the Roche LightCycler 2.0 System. Oral carcinoma are detectable with mRNA resynthesis of the ECM molecules L5gamma2 and Tn-C in oral brush biopsies. EDB-fn mRNA was not detected--the stroma myofibroblasts are apparently a preferential source of EDB-fn and sampling by oral brush biopsy harvests epithelial cells and does not reach the cells which do express EDB-fn. The performance of gene expression analysis in brush biopsies is limited by a high RNase activity in the oral cavity.

Differential interaction of Escherichia coli heat-labile toxin and cholera toxin with pig intestinal brush border glycoproteins depending on their ABH and related blood group antigenic determinants.

PubMed

Balanzino, L E; Barra, J L; Monferran, C G; Cumar, F A

1994-04-01

The ability of glycoproteins from pig intestinal brush border membranes (BBM) to bind cholera toxin (CT) or heat-labile toxins from strains of Escherichia coli isolated from human (LTh) or pig (LTp) intestines was studied. Glycoproteins capable of binding the toxins are also recognized by antibodies or lectins specific for ABO(H) blood group and related antigens. Pigs expressing A, H, or I antigenic determinants were used for comparison. The toxin-binding capacity of a glycoprotein depends on the toxin type and the blood group epitope borne by the glycoprotein. LTh and LTp preferably bound to several blood group A-active glycoproteins rather than H-active glycoproteins. By contrast, CT practically did not recognize either blood group A- or blood group H-active glycoproteins, while glycoproteins from pigs expressing I antigenic determinants were able to interact with LTh, LTp, and CT. LTh, LTp, or CT glycoprotein binding was selectively inhibited by specific lectins or monosaccharides. Affinity purification of the toxin binding brush border glycoproteins on the basis of their blood group reactivity suggests that such glycoproteins are hydrolytic enzymes. BBM from A+ pigs contain about 27 times more LTh binding sites, in addition to those recognized by CT, than an equivalent membrane preparation from H+ pigs. The present findings may help clarify some previous unclear results on LTh binding to intestinal BBM glycoproteins obtained by use of animals not typed by their ABO(H) blood group phenotype.

Saccharomyces boulardii upgrades cellular adaptation after proximal enterectomy in rats

PubMed Central

Buts, J; De Keyser, N; Marandi, S; Hermans, D; Sokal, E; Chae, Y; Lambotte, L; Chanteux, H; Tulkens, P

1999-01-01

BACKGROUND—Saccharomyces boulardii is a non-pathogenic yeast which exerts trophic effects on human and rat small intestinal mucosa.
AIMS—To examine the effects of S boulardii on ileal adaptation after proximal enterectomy in rats.
METHODS—Wistar rats, aged eight weeks, underwent 60% proximal resection or transection and received by orogastric intubation either 1 mg/g body wt per day lyophilised S boulardii or the vehicle for seven days. The effects on ileal mucosal adaptation were assessed eight days after surgery.
RESULTS—Compared with transection, resection resulted in mucosal hyperplasia with significant decreases in the specific and total activities of sucrase, lactase, and maltase. Treatment of resected animals with S boulardii had no effect on mucosal hyperplasia but did upgrade disaccharidase activities to the levels of the transected group. Enzyme stimulation by S boulardii was associated with significant increases in diamine oxidase activity and mucosal polyamine concentrations. Likewise, sodium dependent D-glucose uptake by brush border membrane vesicles, measured as a function of time and glucose concentration in the incubation medium, was significantly (p<0.05) increased by 81% and three times respectively in the resected group treated with S boulardii. In agreement with this, expression of the sodium/glucose cotransporter-1 in brush border membranes of resected rats treated with S boulardii was enhanced twofold compared with resected controls.
CONCLUSION—Oral administration of S boulardii soon after proximal enterectomy improves functional adaptation of the remnant ileum.


Keywords: Saccharomyces boulardii; intestine; adaptation; polyamines; brush border membrane enzymes; glucose uptake PMID:10369710

Role of specific activators of intestinal amino acid transport in Bombyx mori larval growth and nutrition.

PubMed

Leonardi, M G; Casartelli, M; Fiandra, L; Parenti, P; Giordana, B

2001-12-01

Nutrient absorption and its modulation are critical for animal growth. In this paper, we demonstrate that leucine methyl ester (Leu-OMe) can greatly increase the activity of the transport system responsible for the absorption of most essential amino acids in the larval midgut of the silkworm Bombyx mori. We investigated leucine uptake activation by Leu-OMe in brush border membrane vesicles and in the apical membrane of epithelial cells in the midgut incubated in vitro. Moreover, the addition of this strong activator of amino acid absorption to diet significantly affected larval growth. Silkworms fed on artificial diet supplemented with Leu-OMe reached maximum body weight 12-18 h before control larvae, and produced cocoon shells up to 20% heavier than those of controls. The activation of amino acid absorption plays an essential role in larval development so that larval growth and cocoon production similar to controls reared on an artificial diet with 25% of dry mulberry leaf powder were observed in silkworms fed on an artificial diet with only 5% of mulberry powder. Arch. Copyright 2001 Wiley-Liss, Inc.

Midgut GPI-anchored proteins with alkaline phosphatase activity from the cotton boll weevil (Anthonomus grandis) are putative receptors for the Cry1B protein of Bacillus thuringiensis.

PubMed

Martins, Erica Soares; Monnerat, Rose Gomes; Queiroz, Paulo Roberto; Dumas, Vinicius Fiuza; Braz, Shélida Vasconcelos; de Souza Aguiar, Raimundo Wagner; Gomes, Ana Cristina Menezes Mendes; Sánchez, Jorge; Bravo, Alejandra; Ribeiro, Bergmann Morais

2010-02-01

Cry toxins from Bacillus thuringiensis (Bt) are used for insect control. They interact with specific receptors located on the host cell surface and are activated by host proteases following receptor binding resulting in midgut epithelial cells lysis. In this work we had cloned, sequenced and expressed a cry1Ba toxin gene from the B thuringiensis S601 strain which was previously shown to be toxic to Anthonomus grandis, a cotton pest. The Cry1Ba6 protein expressed in an acrystaliferous B. thuringiensis strain was toxic to A. grandis in bioassays. The binding of Cry1Ba6 toxin to proteins located in the midgut brush border membrane of A. grandis was analyzed and we found that Cry1Ba6 binds to two proteins (62 and 65kDa) that showed alkaline phosphatase (ALP) activity. This work is the first report that shows the localization of Cry toxin receptors in the midgut cells of A. grandis. 2009. Published by Elsevier Ltd.

Recovery of Na-glucose cotransport activity after renal ischemia is impaired in mice lacking vimentin.

PubMed

Runembert, Isabelle; Couette, Sylviane; Federici, Pierre; Colucci-Guyon, Emma; Babinet, Charles; Briand, Pascale; Friedlander, Gérard; Terzi, Fabiola

2004-11-01

Vimentin, an intermediate filament protein mainly expressed in mesenchyma-derived cells, is reexpressed in renal tubular epithelial cells under many pathological conditions, characterized by intense cell proliferation. Whether vimentin reexpression is only a marker of cell dedifferentiation or is instrumental in the maintenance of cell structure and/or function is still unknown. Here, we used vimentin knockout mice (Vim(-/-)) and an experimental model of acute renal injury (30-min bilateral renal ischemia) to explore the role of vimentin. Bilateral renal ischemia induced an initial phase of acute tubular necrosis that did not require vimentin and was similar, in terms of morphological and functional changes, in Vim(+/+) and Vim(-/-) mice. However, vimentin was essential to favor Na-glucose cotransporter 1 localization to brush-border membranes and to restore Na-glucose cotransport activity in regenerating tubular cells. We show that the effect of vimentin inactivation is specific and results in persistent glucosuria. We propose that vimentin is part of a structural network that favors carrier localization to plasma membranes to restore transport activity in injured kidneys.

Subcellular localization and distribution of the reduced folate carrier in normal rat tissues.

PubMed

Hinken, M; Halwachs, S; Kneuer, C; Honscha, W

2011-01-27

The reduced folate carrier (Rfc1; Slc19a1) mediated transport of reduced folates and antifolate drugs such as methotrexate (MTX) play an essential role in physiological folate homeostasis and MTX cancer chemotherapy. As no systematic reports are as yet available correlating Rfc1 gene expression and protein levels in all tissues crucial for folate and antifolate uptake, storage or elimination, we investigated gene and protein expression of rat Rfc1 (rRfc1) in selected tissues. This included the generation of a specific anti-rRfc1 antibody. Rabbits were immunised with isolated rRfc1 peptides producing specific anti-rRfc1 antiserum targeted to the intracellular C-terminus of the carrier. Using RT-PCR analysis, high rRfc1 transcript levels were detected in colon, kidney, brain, thymus, and spleen. Moderate rRfc1 gene expression was observed in small intestine, liver, bone marrow, lung, and testes whereas transcript levels were negligible in heart, skeletal muscle or leukocytes. Immunohistochemical analyses revealed strong carrier expression in the apical membrane of tunica mucosa epithelial cells of small intestine and colon, in the brush-border membrane of choroid plexus epithelial cells or in endothelial cells of small vessels in brain and heart. Additionally, high rRfc1 protein levels were localized in the basolateral membrane of renal tubular epithelial cells, in the plasma membrane of periportal hepatocytes, and sertoli cells of the testes. Taken together, our results demonstrated that rRfc1 is expressed almost ubiquitously but to very different levels. The predominant tissue distribution supports the essential role of Rfc1 in physiological folate homeostasis. Moreover, our results may contribute to understand antifolate pharmacokinetics and selected organ toxicity associated with MTX chemotherapy.

Uptake mechanism of valproic acid in human placental choriocarcinoma cell line (BeWo).

PubMed

Ushigome, F; Takanaga, H; Matsuo, H; Tsukimori, K; Nakano, H; Ohtani, H; Sawada, Y

2001-04-13

Valproic acid is an anticonvulsant widely used for the treatment of epilepsy. However, valproic acid is known to show fetal toxicity, including teratogenicity. In the present study, to elucidate the mechanisms of valproic acid transport across the blood-placental barrier, we carried out transcellular transport and uptake experiments with human placental choriocarcinoma epithelial cells (BeWo cells) in culture. The permeability coefficient of [3H]valproic acid in BeWo cells for the apical-to-basolateral flux was greater than that for the opposite flux, suggesting a higher unidirectional transport in the fetal direction. The uptake of [3H]valproic acid from the apical side was temperature-dependent and enhanced under acidic pH. In the presence of 50 microM carbonyl cyanide p-trifluoromethoxylhydrazone, the uptake of [3H]valproic acid was significantly reduced. A metabolic inhibitor, 10 mM sodium azide, also significantly reduced the uptake of [3H]valproic acid. Therefore, valproic acid is actively transported in a pH-dependent manner on the brush-border membrane of BeWo cells. Kinetic analysis of valproic acid uptake revealed the involvement of a non-saturable component and a saturable component. The Michaelis constant for the saturable transport (K(t)) was smaller under acidic pH, suggesting a proton-linked active transport mechanism for valproic acid in BeWo cells. In the inhibitory experiments, some short-chain fatty acids, such as acetic acid, lactic acid, propanoic acid and butyric acid, and medium-chain fatty acids, such as hexanoic acid and octanoic acid, inhibited the uptake of [3H]valproic acid. The uptake of [3H]valproic acid was also significantly decreased in the presence of 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid, salicylic acid and furosemide, which are well-known inhibitors of the anion exchange system. Moreover, p-aminohippuric acid significantly reduced the uptake of [3H]valproic acid. These results suggest that an active transport mechanism for valproic acid exists on the brush-border membrane of placental trophoblast cells and operates in a proton-linked manner.

Differential expression of pro-inflammatory cytokines in intra-epithelial T cells between trachea and bronchi distinguishes severity of COPD.

PubMed

Hodge, Greg; Reynolds, Paul N; Holmes, Mark; Hodge, Sandra

2012-12-01

Measuring T-cell production of intracellular cytokines by flow cytometry enables specific monitoring of airway inflammation and response to therapies in chronic lung diseases including chronic obstructive pulmonary disease (COPD). We have previously shown that T cells in the airways of ex- and current- smoker COPD patients and healthy smokers produce increased T-cell pro-inflammatory cytokines IFNγ and TNFα versus healthy controls. However, we could not differentiate between COPD groups and smokers due to a high degree of inter-patient variability. To address this limitation, we hypothesized that intraepithelial T cells obtained from brushings of trachea may serve as an ideal intra-patient control compared with cells obtained from left and right bronchi. Production of intracellular cytokines by intraepithelial T-cells obtained from trachea and right and left bronchi from 26 individuals with COPD (16 with GOLD I and 10 with GOLD II-III disease), 11 healthy controls and 8 smokers was measured by flow cytometry. There was a significant increase in intraepithelial T-cell IFNγ and TNFα in both right and left bronchi of GOLD II-III COPD patients compared to cells obtained from the trachea. There were no changes in T cell pro-inflammatory cytokines between the bronchi and trachea from control subjects, GOLD I COPD patients or healthy smokers. There was a significant negative correlation between increased intraepithelial IFNγ and TNFα in bronchial brushing T-cells compared with tracheal T-cells, and compared with FEV1. Monitoring intracellular intra-epithelial T-cell cytokine production in bronchial brushings using autologous tracheal brushings as controls provides improves the sensitivity of the technique. Therapeutic targeting of these pro-inflammatory cytokines and assessing the effects of drugs on immune reactivity has the potential to reduce lung inflammation caused by intra-epithelial T cells in COPD. Copyright © 2012 Elsevier Ltd. All rights reserved.

Patterns of gene expression in different histotypes of epithelial ovarian cancer correlate with those in normal fallopian tube, endometrium, and colon.

PubMed

Marquez, Rebecca T; Baggerly, Keith A; Patterson, Andrea P; Liu, Jinsong; Broaddus, Russell; Frumovitz, Michael; Atkinson, Edward N; Smith, David I; Hartmann, Lynn; Fishman, David; Berchuck, Andrew; Whitaker, Regina; Gershenson, David M; Mills, Gordon B; Bast, Robert C; Lu, Karen H

2005-09-01

Epithelial ovarian cancers are thought to arise from flattened epithelial cells that cover the ovarian surface or that line inclusion cysts. During malignant transformation, different histotypes arise that resemble epithelial cells from normal fallopian tube, endometrium, and intestine. This study compares gene expression in serous, endometrioid, clear cell, and mucinous ovarian cancers with that in the normal tissues that they resemble. Expression of 63,000 probe sets was measured in 50 ovarian cancers, in 5 pools of normal ovarian epithelial brushings, and in mucosal scrapings from 4 normal fallopian tube, 5 endometrium, and 4 colon specimens. Using rank-sum analysis, genes whose expressions best differentiated the ovarian cancer histotypes and normal ovarian epithelium were used to determine whether a correlation based on gene expression existed between ovarian cancer histotypes and the normal tissues they resemble. When compared with normal ovarian epithelial brushings, alterations in serous tumors correlated with those in normal fallopian tube (P = 0.0042) but not in other normal tissues. Similarly, mucinous cancers correlated with those in normal colonic mucosa (P = 0.0003), and both endometrioid and clear cell histotypes correlated with changes in normal endometrium (P = 0.0172 and 0.0002, respectively). Mucinous cancers displayed the greatest number of alterations in gene expression when compared with normal ovarian epithelial cells. Studies at a molecular level show distinct expression profiles of different histologies of ovarian cancer and support the long-held belief that histotypes of ovarian cancers come to resemble normal fallopian tube, endometrial, and colonic epithelium. Several potential molecular markers for mucinous ovarian cancers have been identified.

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...

Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes.

PubMed

Park, Dayoung; Brune, Kristin A; Mitra, Anupam; Marusina, Alina I; Maverakis, Emanual; Lebrilla, Carlito B

2015-11-01

Changes in cell surface glycosylation occur during the development and differentiation of cells and have been widely correlated with the progression of several diseases. Because of their structural diversity and sensitivity to intra- and extracellular conditions, glycans are an indispensable tool for analyzing cellular transformations. Glycans present on the surface of intestinal epithelial cells (IEC) mediate interactions with billions of native microorganisms, which continuously populate the mammalian gut. A distinct feature of IECs is that they differentiate as they migrate upwards from the crypt base to the villus tip. In this study, nano-LC/ESI QTOF MS profiling was used to characterize the changes in glycosylation that correspond to Caco-2 cell differentiation. As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. At day 21, when cells appear to be completely differentiated, remodeling of the cell surface glycome ceases. Differential expression of glycans during IEC maturation appears to play a key functional role in regulating the membrane-associated hydrolases and contributes to the mucosal surface innate defense mechanisms. Developing methodologies to rapidly identify changes in IEC surface glycans may lead to a rapid screening approach for a variety of disease states affecting the GI tract. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Apolipoprotein A-1 (apoA-1) deposition in, and release from, the enterocyte brush border: a possible role in transintestinal cholesterol efflux (TICE)?

PubMed

Danielsen, E Michael; Hansen, Gert H; Rasmussen, Karina; Niels-Christiansen, Lise-Lotte; Frenzel, Franz

2012-03-01

Transintestinal cholesterol efflux (TICE) has been proposed to represent a non-hepatobiliary route of cholesterol secretion directly "from blood to gut" and to play a physiologically significant role in excretion of neutral sterols, but so far little is known about the proteins involved in the process. We have previously observed that apolipoprotein A-1 (apoA-1) synthesized by enterocytes of the small intestine is mainly secreted apically into the gut lumen during fasting where its assembly into chylomicrons and basolateral discharge is at a minimal level. In the present work we showed, both by immunomicroscopy and subcellular fractionation, that a fraction of the apically secreted apoA-1 in porcine small intestine was not released from the cell surface but instead deposited in the brush border. Cholesterol was detected in immunoisolated microvillar apoA-1, and it was partially associated with detergent resistant membranes (DRMs), indicative of localization in lipid raft microdomains. The apolipoprotein was not readily released from microvillar vesicles by high salt or by incubation with phosphatidylcholine-specific phospholipase C or trypsin, indicating a relatively firm attachment to the membrane bilayer. However, whole bile or taurocholate efficiently released apoA-1 at low concentrations that did not solubilize the transmembrane microvillar protein aminopeptidase N. Based on these findings and the well known role played by apoA-1 in extrahepatic cellular cholesterol removal and reverse cholesterol transport (RCT), we propose that brush border-deposited apoA-1 in the small intestine acts in TICE by mediating cholesterol efflux into the gut lumen. Copyright © 2011 Elsevier B.V. All rights reserved.

Evidence of native starch degradation with human small intestinal maltase-glucoamylase (recombinant)

USDA-ARS?s Scientific Manuscript database

Action of human small intestinal brush border carbohydrate digesting enzymes is thought to involve only final hydrolysis reactions of oligosaccharides to monosaccharides. In vitro starch digestibility assays use fungal amyloglucosidase to provide this function. In this study, recombinant N-terminal ...

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

[Kinetic properties of the fructose influx across the brush border of the rat jejunum. Effects of a diet rich in fructose].

PubMed

Crouzoulon, G

1978-10-01

The unidirectional influx (i.e. initial rate of uptake) of D-fructose across the brush border of rat jejunum is a saturable function of concentration, with a Kt of 125 mM, which implicates a carrier mechanism. This mechanism appears to be very specific for fructose in view of the lack of influx inhibition observed in the presence of large concentrations of the sugars or polyols, D-glucose, D-galactose, D-mannose, D-xylose, L-sorbose, D-tagatose, sorbitol or mannitol. D-Fructose uptake is inhibited by incubation, preceded by a 30-min preincubation in the same inhibitory conditions, in the absence of Na, or in the presence of metabolic poisons, NaF, 2,4-dinitrophenol, monoiodoacetate. Phloridzin (10-3 M), with or without preincubation, has no effect on uptake. D-Fructose influx is stimulated by fructose feeding, mainly because the augmentation of the number of active sites of transfer: Jmax is increased two-fold, Kt is more weakly affected.

A Tenebrio molitor GPI-anchored alkaline phosphatase is involved in binding of Bacillus thuringiensis Cry3Aa to brush border membrane vesicles.

PubMed

Zúñiga-Navarrete, Fernando; Gómez, Isabel; Peña, Guadalupe; Bravo, Alejandra; Soberón, Mario

2013-03-01

Bacillus thuringiensis Cry toxins recognizes their target cells in part by the binding to glycosyl-phosphatidyl-inositol (GPI) anchored proteins such as aminopeptidase-N (APN) or alkaline phosphatases (ALP). Treatment of Tenebrio molitor brush border membrane vesicles (BBMV) with phospholipase C that cleaves out GPI-anchored proteins from the membranes, showed that GPI-anchored proteins are involved in binding of Cry3Aa toxin to BBMV. A 68 kDa GPI-anchored ALP was shown to bind Cry3Aa by toxin overlay assays. The 68 kDa GPI-anchored ALP was preferentially expressed in early instar larvae in comparison to late instar larvae. Our work shows for the first time that GPI-anchored ALP is important for Cry3Aa binding to T. molitor BBMV suggesting that the mode of action of Cry toxins is conserved in different insect orders. Copyright © 2012 Elsevier Inc. All rights reserved.

Alterations in gut microflora populations and brush border functionality following intra-amniotic daidzein administration

USDA-ARS?s Scientific Manuscript database

Daidzein is an isoflavone found primarily in soybean and various soy-based products such as tofu. In the intestines, daidzein is reductively transformed to its constituent metabolites equol and O-desmethylangolensin. Although the ingestion of daidzein has been associated with marked physiological im...

Adaptations to the air breathing in the posterior intestine of the catfish (Corydoras aeneus, Callichthyidae). A histological and ultrastructural study.

PubMed

Podkowa, Dagmara; Goniakowska-Witalińska, Lucyna

2002-01-01

A light and transmission electron microscopic study of the intestine of catfish C. aeneus shows that the anterior part of the intestine is a site of digestion and absorption and its structure is typical of that of other teleostean fishes. However, in this species the thin-walled posterior intestine is adapted to air breathing. In this region mucosa is smooth and lined with respiratory epithelium with capillary network. Several types of cells are observed in the epithelium: flattened respiratory epithelial cells with short microvili, goblet cells, scarce epithelial cells with numerous longer microvilli, and two types of endocrine cells (EC). The solitary brush cells with several long and thick microvilli described here are the first observation of such cells in the gastrointestinal tract of fishes. Bodies of respiratory epithelial cells lie between capillaries. Their cytoplasm, apart from typical organelles contains dense and lamellar bodies, which are a site of accumulation of surfactant. In regions where capillaries are covered by thin cytoplasmic sheets of respiratory epithelial cells, a thin (0.24-3.00 microm) air-blood barrier is formed, thus enabling gas exchange. Epithelial cells with longer microvilli do not participate in the formation of the air-blood barrier and are probably responsible for absorbtion. EC of the closed type are dispersed within the epithelium. Their cytoplasm contains characteristic round or oval dense core vesicles 69 to 230 nm in diameter. The role of EC and brush cells in the regulation of processes related to absorbtion, and to respiration, is disscused.

Cholinergic epithelial cell with chemosensory traits in murine thymic medulla.

PubMed

Panneck, Alexandra Regina; Rafiq, Amir; Schütz, Burkhard; Soultanova, Aichurek; Deckmann, Klaus; Chubanov, Vladimir; Gudermann, Thomas; Weihe, Eberhard; Krasteva-Christ, Gabriela; Grau, Veronika; del Rey, Adriana; Kummer, Wolfgang

2014-12-01

Specialized epithelial cells with a tuft of apical microvilli ("brush cells") sense luminal content and initiate protective reflexes in response to potentially harmful substances. They utilize the canonical taste transduction cascade to detect "bitter" substances such as bacterial quorum-sensing molecules. In the respiratory tract, most of these cells are cholinergic and are approached by cholinoceptive sensory nerve fibers. Utilizing two different reporter mouse strains for the expression of choline acetyltransferase (ChAT), we observed intense labeling of a subset of thymic medullary cells. ChAT expression was confirmed by in situ hybridization. These cells showed expression of villin, a brush cell marker protein, and ultrastructurally exhibited lateral microvilli. They did not express neuroendocrine (chromogranin A, PGP9.5) or thymocyte (CD3) markers but rather thymic epithelial (CK8, CK18) markers and were immunoreactive for components of the taste transduction cascade such as Gα-gustducin, transient receptor potential melastatin-like subtype 5 channel (TRPM5), and phospholipase Cβ2. Reverse transcription and polymerase chain reaction confirmed the expression of Gα-gustducin, TRPM5, and phospholipase Cβ2. Thymic "cholinergic chemosensory cells" were often in direct contact with medullary epithelial cells expressing the nicotinic acetylcholine receptor subunit α3. These cells have recently been identified as terminally differentiated epithelial cells (Hassall's corpuscle-like structures in mice). Contacts with nerve fibers (identified by PGP9.5 and CGRP antibodies), however, were not observed. Our data identify, in the thymus, a previously unrecognized presumptive chemosensitive cell that probably utilizes acetylcholine for paracrine signaling. This cell might participate in intrathymic infection-sensing mechanisms.

Computer-assisted brush-biopsy analysis for the detection of dysplasia in a high-risk Barrett's esophagus surveillance population.

PubMed

Anandasabapathy, Sharmila; Sontag, Stephen; Graham, David Y; Frist, Stephen; Bratton, Joan; Harpaz, Noam; Waye, Jerome D

2011-03-01

Barrett's epithelial dysplasia, the direct precursor to esophageal adenocarcinoma, is often unapparent and frequently missed during surveillance of Barrett's esophagus with four-quadrant forceps biopsy protocol. To determine whether the detection of dysplasia is improved by adding computer-assisted brush biopsy (EndoCDx©) to four-quadrant biopsy protocol. Patients with a history of Barrett's esophagus with dysplasia scheduled for endoscopic surveillance were recruited from four academic medical centers. Patients underwent brush biopsy followed by four-quadrant biopsy every 1-2 cm. The results from brush and forceps biopsy were reviewed independently by pathologists blinded to the other's results. Among 151 patients enrolled (124 men, 27 women; mean age: 65), 117 (77.5%) had forceps and brush-biopsy specimens adequate for interpretation. The mean number of forceps biopsies was 11.9 (median 10, range 2-40) and brush biopsies was 2.0 (median 2, range 1-4). The overall yield of forceps alone was 25.2% (n = 38). Brush biopsy added an additional 16 positive cases increasing the yield of dysplasia detection by 42% (95% CI: 20.7-72.7). The number needed to test (NNT) to detect one additional case of dysplasia was 9.4 (95% CI: 6.4-17.7). There were no significant differences in results among different centers, between standard versus jumbo forceps, or between forceps biopsies taken every 1 cm versus every 2 cm. These data suggest that computer-assisted brush biopsy is a useful adjunct to standard endoscopic surveillance regimens for the identification of dysplasia in Barrett's esophagus.

Morphology and histology of the alimentary canal of Lygus hesperus (Heteroptera: Cimicomoropha: Miridae)

USGS Publications Warehouse

Habibi, J.; Coudron, T.A.; Backus, E.A.; Brandt, S.L.; Wagner, R.M.; Wright, M.K.; Huesing, J.E.

2008-01-01

Microdissection and transverse semithin sections were used to perform a light microscopy survey of the gross morphology and cellular anatomy of the alimentary canal, respectively, of Lygus hesperus Knight, a key pest of cotton (Gossypium hirsutum L.), alfalfa (Medicago sativa L.), and other crops. The gross morphology of the alimentary canal showed a relatively unadorned tube compared with other hemipterans, with variably shaped compartments and one small diverticulum. However, the epithelial cell anatomy of the gut was relatively complex, with the midgut having the most diverse structure and cell types. The midgut was typical of the "Lygus-type gut" seen in the older literature, i.e., it consisted of three major regions, the first (descending), second (ascending), and third (descending) ventriculi, with different variants of three major epithelial cell types in each region. Our light microscopy (LM) study suggests that the three cell types are nondifferentiated regenerative cells (which sparsely occurred throughout the midgut but were abundant in the anterior region of the first ventriculus), endocrine cells, and columnar cells. Although the Lygus gut cells strongly resemble those cell types seen in other insects, their identification should be confirmed via transmission electron microscopy to be considered definitive. These cell types differed in the size and opacity of vesicles, geometry of cell surface in the gut lumen, and size, shape, and concentration of brush-border microvilli and location within the gut. Comparison of gut structure in L. hesperus with that of other hemipterans, especially in relation to hemipteran phylogeny and feeding strategies, is discussed.

Titanium Dioxide Nanoparticle Ingestion Alters Nutrient Absorption in an In Vitro Model of the Small Intestine

PubMed Central

Guo, Zhongyuan; Martucci, Nicole J.; Moreno-Olivas, Fabiola; Tako, Elad; Mahler, Gretchen J.

2017-01-01

Ingestion of titanium dioxide (TiO2) nanoparticles from products such as agricultural chemicals, processed food, and nutritional supplements is nearly unavoidable. The gastrointestinal tract serves as a critical interface between the body and the external environment, and is the site of essential nutrient absorption. The goal of this study was to examine the effects of ingesting the 30 nm TiO2 nanoparticles with an in vitro cell culture model of the small intestinal epithelium, and to determine how acute or chronic exposure to nano-TiO2 influences intestinal barrier function, reactive oxygen species generation, proinflammatory signaling, nutrient absorption (iron, zinc, fatty acids), and brush border membrane enzyme function (intestinal alkaline phosphatase). A Caco-2/HT29-MTX cell culture model was exposed to physiologically relevant doses of TiO2 nanoparticles for acute (four hours) or chronic (five days) time periods. Exposure to TiO2 nanoparticles significantly decreased intestinal barrier function following chronic exposure. Reactive oxygen species (ROS) generation, proinflammatory signaling, and intestinal alkaline phosphatase activity all showed increases in response to nano-TiO2. Iron, zinc, and fatty acid transport were significantly decreased following exposure to TiO2 nanoparticles. This is because nanoparticle exposure induced a decrease in absorptive microvilli in the intestinal epithelial cells. Nutrient transporter protein gene expression was also altered, suggesting that cells are working to regulate the transport mechanisms disturbed by nanoparticle ingestion. Overall, these results show that intestinal epithelial cells are affected at a functional level by physiologically relevant exposure to nanoparticles commonly ingested from food. PMID:28944308

An Integrative Review of Mechanotransduction in Endothelial, Epithelial (Renal) and Dendritic Cells (Osteocytes)

PubMed Central

Weinbaum, Sheldon; Duan, Yi; Thi, Mia M.; You, Lidan

2013-01-01

In this review we will examine from a biomechanical and ultrastructural viewpoint how the cytoskeletal specialization of three basic cell types, endothelial cells (ECs), epithelial cells (renal tubule) and dendritic cells (osteocytes), enables the mechano-sensing of fluid flow in both their native in vivo environment and in culture, and the downstream signaling that is initiated at the molecular level in response to fluid flow. These cellular responses will be discussed in terms of basic mysteries and paradoxes encountered by each cell type. In ECs fluid shear stress (FSS) is nearly entirely attenuated by the endothelial glycocalyx that covers their apical membrane and yet FSS is communicated to both intracellular and junctional molecular components in activating a wide variety of signaling pathways. The same is true in proximal tubule (PT) cells where a dense brush border of microvilli covers the apical surface and the flow at the apical membrane is negligible. A four decade old unexplained mystery is the ability of PT epithelia to reliably reabsorb 60% of the flow entering the tubule regardless of the glomerular filtration rate. In the cortical collecting duct (CCD) the flow rates are so low that a special sensing apparatus, a primary cilia is needed to detect very small variations in tubular flow. In bone it has been a century old mystery as to how osteocytes embedded in a stiff mineralized tissue are able to sense miniscule whole tissue strains that are far smaller than the cellular level strains required to activate osteocytes in vitro. PMID:23976901

Tracking the fate of pasta (T. Durum semolina) immunogenic proteins by in vitro simulated digestion.

PubMed

Mamone, Gianfranco; Nitride, Chiara; Picariello, Gianluca; Addeo, Francesco; Ferranti, Pasquale; Mackie, Alan

2015-03-18

The aim of the present study was to identify and characterize the celiacogenic/immunogenic proteins and peptides released during digestion of pasta (Triticum durum semolina). Cooked pasta was digested using a harmonized in vitro static model of oral-gastro-duodenal digestion. The course of pasta protein digestion was monitored by SDS-PAGE, and gluten proteins were specifically analyzed by Western blot using sera of celiac patients. Among the allergens, nonspecific lipid-transfer protein was highly resistant to gastro-duodenal hydrolysis, while other digestion-stable allergens such as α-amylase/trypsin inhibitors were not detected being totally released in the pasta cooking water. To simulate the final stage of intestinal degradation, the gastro-duodenal digesta were incubated with porcine jejunal brush-border membrane hydrolases. Sixty-one peptides surviving the brush-border membrane peptidases were identified by liquid chromatography-mass spectrometry, including several gluten-derived sequences encrypting different motifs responsible for the induction of celiac disease. These results provide new insights into the persistence of wheat-derived peptides during digestion of cooked pasta samples.

TRPM5, a taste-signaling transient receptor potential ion-channel, is a ubiquitous signaling component in chemosensory cells.

PubMed

Kaske, Silke; Krasteva, Gabriele; König, Peter; Kummer, Wolfgang; Hofmann, Thomas; Gudermann, Thomas; Chubanov, Vladimir

2007-07-04

A growing number of TRP channels have been identified as key players in the sensation of smell, temperature, mechanical forces and taste. TRPM5 is known to be abundantly expressed in taste receptor cells where it participates in sweet, amino acid and bitter perception. A role of TRPM5 in other sensory systems, however, has not been studied so far. Here, we systematically investigated the expression of TRPM5 in rat and mouse tissues. Apart from taste buds, where we found TRPM5 to be predominantly localized on the basolateral surface of taste receptor cells, TRPM5 immunoreactivity was seen in other chemosensory organs - the main olfactory epithelium and the vomeronasal organ. Most strikingly, we found solitary TRPM5-enriched epithelial cells in all parts of the respiratory and gastrointestinal tract. Based on their tissue distribution, the low cell density, morphological features and co-immunostaining with different epithelial markers, we identified these cells as brush cells (also known as tuft, fibrillovesicular, multivesicular or caveolated cells). In terms of morphological characteristics, brush cells resemble taste receptor cells, while their origin and biological role are still under intensive debate. We consider TRPM5 to be an intrinsic signaling component of mammalian chemosensory organs, and provide evidence for brush cells being an important cellular correlate in the periphery.

Regulation of protein expression and function of octn2 in forskolin-induced syncytialization in BeWo Cells.

PubMed

Huang, F-D; Kung, F-L; Tseng, Y-C; Chen, M-R; Chan, H-S; Lin, C-J

2009-02-01

Placental OCTN2 is a high-affinity carnitine transporter that can interact with a number of therapeutic agents. The process of syncytialization is associated with the expression of a variety of genes. However, the association between syncytialization and OCTN2 expression is not yet clear. Given that forskolin induces BeWo cells to undergo biochemical and morphological differentiation, the purpose of the present study was to investigate whether the function and expression of OCTN2 are influenced by forskolin treatment during syncytialization. The forskolin-induced differentiation of BeWo cells was validated by secretion of beta-human chorionic gonadotropin (beta-hCG) and syncytin expression. Cellular localization of OCTN2 was analyzed by confocal microscopy. Expression of OCTN2 and the modular proteins PDZK1, PDZK2, NHERF1 and NHERF2 was analyzed by Western blotting and carnitine uptake by BeWo cells was estimated and the kinetic properties of uptake measured. The results showed that forskolin treatment increased beta-hCG secretion and syncytin expression, suggesting induction of syncytialization. Confocal images of BeWo cells showed the localization of OCTN2 in the brush-border membrane. OCTN2 protein expression was upregulated in isolated brush-border membranes by long-term forskolin treatment, but the V(m) for carnitine uptake was unchanged, although the K(m) increased. PDZK1, NHERF1 and NHERF2 protein expression in the brush-border membrane was downregulated by forskolin treatment, whereas PDZK2 levels remained unchanged. In conclusion, protein expression and function of OCTN2 in BeWo cells can be regulated by forskolin treatment. While the presence of forskolin results in an increase in OCTN2 protein expression, the increase in uptake capacity may be compensated by the decreased expression of PDZK1, NHERF1 or NHERF2.

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

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

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

2013-01-01

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. 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. 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. 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. © 2013 S. Karger AG, Basel.

A Novel Role for Integrin-linked Kinase in Epithelial Sheet Morphogenesis

PubMed Central

Vespa, Alisa; D'Souza, Sudhir J.A.; Dagnino, Lina

2005-01-01

Integrin-linked kinase (ILK) is a multidomain protein involved in cell motility and cell-extracellular matrix interactions. ILK is found in integrin-containing focal adhesions in undifferentiated primary epidermal keratinocytes. Induction of keratinocyte differentiation by treatment with Ca2+ triggers formation of cell–cell junctions, loss of focal adhesions, and ILK distribution to cell borders. We now show that Ca2+ treatment of keratinocytes induces rapid (≤1 h) translocation to the cell membrane of the adherens junction (AJ) proteins E-cadherin and β-catenin. This is followed by slower (>6 h) localization of tight junction (TJ) proteins. The kinetics of ILK movement toward the cell periphery mimics that of AJ components, suggesting that ILK plays a role in the early formation of cell–cell contacts. Whereas the N terminus in ILK mediates localization to cell borders, expression of an ILK deletion mutant incapable of localizing to the cell membrane (ILK 191-452) interferes with translocation of E-cadherin/β-catenin to cell borders, precluding Ca2+-induced AJ formation. Cells expressing ILK 191-452 also fail to form TJ and sealed cell–cell borders and do not form epithelial sheets. Thus, we have uncovered a novel role for ILK in epithelial cell–cell adhesion, independent of its well-established role in integrin-mediated adhesion and migration. PMID:15975904

A novel role for integrin-linked kinase in epithelial sheet morphogenesis.

PubMed

Vespa, Alisa; D'Souza, Sudhir J A; Dagnino, Lina

2005-09-01

Integrin-linked kinase (ILK) is a multidomain protein involved in cell motility and cell-extracellular matrix interactions. ILK is found in integrin-containing focal adhesions in undifferentiated primary epidermal keratinocytes. Induction of keratinocyte differentiation by treatment with Ca(2+) triggers formation of cell-cell junctions, loss of focal adhesions, and ILK distribution to cell borders. We now show that Ca(2+) treatment of keratinocytes induces rapid (6 h) localization of tight junction (TJ) proteins. The kinetics of ILK movement toward the cell periphery mimics that of AJ components, suggesting that ILK plays a role in the early formation of cell-cell contacts. Whereas the N terminus in ILK mediates localization to cell borders, expression of an ILK deletion mutant incapable of localizing to the cell membrane (ILK 191-452) interferes with translocation of E-cadherin/beta-catenin to cell borders, precluding Ca(2+)-induced AJ formation. Cells expressing ILK 191-452 also fail to form TJ and sealed cell-cell borders and do not form epithelial sheets. Thus, we have uncovered a novel role for ILK in epithelial cell-cell adhesion, independent of its well-established role in integrin-mediated adhesion and migration.

Comprehensive fire prevention legislation enacted by the California legislature in 1992 after the East Bay firestorm

Treesearch

Rachel Richman

1995-01-01

Legislation was enacted by the California Legislature in 1992 after the East Bay firestorm on the Oakland/Berkeley border; it included roofing standards, brush clearance, and other safety requirements. Fire safety personnel and local government officials all participated in developing these measures and presenting them to the California Legislature.

Intra amniotic administration of raffinose and stachyose affects the intestinal brush border functionality and alters gut microflora populations

USDA-ARS?s Scientific Manuscript database

This study investigates the effectiveness of two types of prebiotics, stachyose and raffinose, which are present in staple food crops that are widely consumed in regions where dietary Fe deficiency is a health concern. The hypothesis is that these prebiotics will improve Fe status, intestinal functi...

Biological Activity of Bacillus thuringiensis and Associated Toxins against the Asian Longhorned Beetle (Coleoptera: Cerambycidae)

Treesearch

Vincent D' amico; John D. Podgwaite; Sara Duke; Sara Duke

2004-01-01

Bacillus thuringiensis Berliner var. tenebrionis and B. thuringiensis toxins were assayed against larval and adult Asian longhorned beetles, Anoplophora glabripennis (A. glabripennis). Preliminary in vitro assays showed some toxins to be active on whole midgut preparations in voltage clamp assays and in assays on brush border membrane vesicles formed from midgut...

The in ovo feeding administration (gallus gallus)-an emerging in vivo approach to assess bioactive compounds with potential nutritional benefits

USDA-ARS?s Scientific Manuscript database

In recent years, the in ovo feeding in fertilized broiler (Gallus gallus) eggs approach was further developed and currently is widely applied in the evaluation process of the effects of functional foods (primarily plant origin compounds) on the functionality of the intestinal brush border membrane, ...

DE-Cadherin Is Required for Intercellular Motility during Drosophila Oogenesis

PubMed Central

Niewiadomska, Paulina; Godt, Dorothea; Tepass, Ulrich

1999-01-01

Cadherins are involved in a variety of morphogenetic movements during animal development. However, it has been difficult to pinpoint the precise function of cadherins in morphogenetic processes due to the multifunctional nature of cadherin requirement. The data presented here indicate that homophilic adhesion promoted by Drosophila E-cadherin (DE-cadherin) mediates two cell migration events during Drosophila oogenesis. In Drosophila follicles, two groups of follicle cells, the border cells and the centripetal cells migrate on the surface of germline cells. We show that the border cells migrate as an epithelial patch in which two centrally located cells retain epithelial polarity and peripheral cells are partially depolarized. Both follicle cells and germline cells express DE-cadherin, and border cells and centripetal cells strongly upregulate the expression of DE-cadherin shortly before and during their migration. Removing DE-cadherin from either the follicle cells or the germline cells blocks migration of border cells and centripetal cells on the surface of germline cells. The function of DE-cadherin in border cells appears to be specific for migration as the formation of the border cell cluster and the adhesion between border cells are not disrupted in the absence of DE-cadherin. The speed of migration depends on the level of DE-cadherin expression, as border cells migrate more slowly when DE-cadherin activity is reduced. Finally, we show that the upregulation of DE-cadherin expression in border cells depends on the activity of the Drosophila C/EBP transcription factor that is essential for border cell migration. PMID:9971747

Chemosensory brush cells of the trachea. A stable population in a dynamic epithelium.

PubMed

Saunders, Cecil J; Reynolds, Susan D; Finger, Thomas E

2013-08-01

Tracheal brush cells (BCs) are specialized epithelial chemosensors that use the canonical taste transduction cascade to detect irritants. To test whether BCs are replaced at the same rate as other cells in the surrounding epithelium of adult mice, we used 5-bromo-2'-deoxyuridine (BrdU) to label dividing cells. Although scattered BrdU-labeled epithelial cells are present 5-20 days after BrdU, no BCs are labeled. These data indicate that BCs comprise a relatively static population. To determine how BCs are generated during development, we injected 5-day-old mice with BrdU and found labeled BCs and non-BC epithelial cells 5 days after BrdU. During the next 60 days, the percentage of labeled BCs increased, whereas the percentage of other labeled cell types decreased. These data suggest that BCs are generated from non-BC progenitor cells during postnatal tracheal growth. To test whether the adult epithelium retains the capacity to generate BCs, tracheal epithelial cells were recovered from adult mice and grown in an air-liquid interface (ALI) culture. After transition to differentiation conditions, BCs are detected, and comprise 1% of the total cell population by Day 14. BrdU added to cultures before the differentiation of BCs was chased into BCs, indicating that the increase in BC density is attributable to the proliferation of a non-BC progenitor. We conclude that: (1) BCs are normally a static population in adult mice; (2) BC progenitors proliferate and differentiate during neonatal development; and (3) BCs can be regenerated from a proliferative population resident in adult epithelium.

Polymers as directing agents for motions of chemical and biological species

NASA Astrophysics Data System (ADS)

Tanyeri, Nihan Yonet

This thesis involves descriptions of solid surface modifications with various polymeric materials which were used as a guiding agent for motion of chemical and biological species. Quasi-two dimensional poly(oligoethylene glycol) acrylate polymer brush based molecular conduits have been designed with the goal of regulating and controlling the diffusive transport of molecular, e.g. organic dyes, and ionic species, e.g. AuCl4-, and Cu2+ ions, along predefined 2-D pathways. The transport of these chemical species has been examined by both fluorescence and dark field microscopy. The polymer brushes were formed through microcontact printing of an initiator, followed by surface-initiated Atom Transfer Radical Polymerization (SI-ATRP). SI-ATRP enables both 2-D patterning with a resolution of about 1 micrometer, and control over the resultant polymer brush thickness (which was varied from 10-100 nm). A hydrophilic poly(oligoethylene glycol) acrylate brushe was selected because of its potential to dissolve a wide range of hydrophilic species. The transport of fluorescent species can be directly followed. A non-lithographic fabrication method was developed for mufluidic devices used in the diffusion studies. Singular channel mufluidic device was utilized to study the directed organic dye diffusion. The AuCl4-, and Cu 2+ ion transport was studied by designing molecular devices with two mufluidic channels. We have demonstrated that the various species of interest diffuse much more rapidly along the predefined pathway than along the bare (polymer brush free) regions of the substrate, demonstrating that diffusive conduits for molecular transport can indeed be formed. The protein resistance of poly(N-isopropylacrylamide) (PNIPAM) brushes grafted from silicon wafers was investigated as a function of the chain molecular weight, grafting density, and temperature. Above the lower critical solution temperature (LCST) of 32°C, the collapse of the water swollen chains, determined by ellipsometry, depends on the grafting density and molecular weight. Ellipsometry, radio assay, and fluorescence imaging demonstrated that, below the LCST, the brushes repel protein as effectively as oligoethylene oxide terminated monolayers. Above 32°C, very low levels of protein adsorb on densely grafted brushes, and the amounts of adsorbed protein increase with decreasing brush grafting densities. Brushes that do not exhibit a collapse transition also bind protein, even though the chains remain extended above the LCST. These findings suggest possible mechanisms underlying protein interactions with end-grafted PNIPAM brushes. 3D porous materials on solid surfaces were built to mimic the corneal basement membrane so that we can monitor direction of the corneal epithelia cells behaviors as the surface topography changes. We have used colloidal crystal templating approach to build the 3D porous structures. Polystyrene (PS) colloids were crystallized in a flow cell. The crystals were filled with acrylamide precursor (including photoinitiator, crosslinker) in the oxygen free aqueous solution. After polymerization of acrylamide under UV exposure, PS colloids were dissolved in chloroform. Thus, 3D porous polyacrylamide hydrogels have been fabricated. The various pore sizes at the 3D porous surface have been obtained by using PS colloids with the colloid diameters ranging from 450 nm to 4 mum. Human corneal epithelial cell growth, morphology change and adhesion studies have been conducted on the porous polyacrylamide scaffolds. The effect of pore size on human corneal epithelial cell function has been investigated.

Border Forces and Friction Control Epithelial Closure Dynamics

PubMed Central

Cochet-Escartin, Olivier; Ranft, Jonas; Silberzan, Pascal; Marcq, Philippe

2014-01-01

We study the closure dynamics of a large number of well-controlled circular apertures within an epithelial monolayer, where the collective cell migration responsible for epithelization is triggered by the removal of a spatial constraint rather than by scratching. Based on experimental observations, we propose a physical model that takes into account border forces, friction with the substrate, and tissue rheology. Border protrusive activity drives epithelization despite the presence of a contractile actomyosin cable at the periphery of the wound. The closure dynamics is quantified by an epithelization coefficient, defined as the ratio of protrusive stress to tissue-substrate friction, that allows classification of different phenotypes. The same analysis demonstrates a distinct signature for human cells bearing the oncogenic RasV12 mutation, demonstrating the potential of the approach to quantitatively characterize metastatic transformations. PMID:24411238

Kidney and Phosphate Metabolism

PubMed Central

2008-01-01

The serum phosphorus level is maintained through a complex interplay between intestinal absorption, exchange intracellular and bone storage pools, and renal tubular reabsorption. The kidney plays a major role in regulation of phosphorus homeostasis by renal tubular reabsorption. Type IIa and type IIc Na+/Pi transporters are important renal Na+-dependent inorganic phosphate (Pi) transporters, which are expressed in the brush border membrane of proximal tubular cells. Both are regulated by dietary Pi intake, vitamin D, fibroblast growth factor 23 (FGF23) and parathyroid hormone. The expression of type IIa Na+/Pi transporter result from hypophosphatemia quickly. However, type IIc appears to act more slowly. Physiological and pathophysiological alteration in renal Pi reabsorption are related to altered brush border membrane expression/content of the type II Na+/Pi cotransporter. Many studies of genetic and acquired renal phosphate wasting disorders have led to the identification of novel genes. Two novel Pi regulating genes, PHEX and FGF23, play a role in the pathophysiology of genetic and acquired renal phosphate wasting disorders and studies are underway to define their mechanism on renal Pi regulation. In recent studies, sodium-hydrogen exchanger regulatory factor 1 (NHERF1) is reported as another new regulator for Pi reabsorption mechanism. PMID:24459526

Saccharomyces cerevisiae CNCM I-3856 prevents colitis induced by AIEC bacteria in the transgenic mouse model mimicking Crohn's disease.

PubMed

Sivignon, Adeline; de Vallée, Amélie; Barnich, Nicolas; Denizot, Jérémy; Darcha, Claude; Pignède, Georges; Vandekerckove, Pascal; Darfeuille-Michaud, Arlette

2015-02-01

Adherent-invasive Escherichia coli (AIEC), which colonize the ileal mucosa of patients with Crohn's disease (CD), are able to adhere to and invade intestinal epithelial cells. Overexpression of the glycoprotein CEACAM6 on host cells favors AIEC attachment and inflammation. We investigated the ability of Saccharomyces cerevisiae CNCM I-3856 to inhibit AIEC adhesion and to reduce colitis. Adhesion experiments were performed on T84 cells and on enterocytes from patients with CD with AIEC LF82 in the presence of S. cerevisiae. Colonization and symptoms of colitis were assessed in LF82-infected transgenic CEABAC10 mice treated with live S. cerevisiae or S. cerevisiae derivatives. Proinflammatory cytokines were quantified by enzyme linked immunosorbent assay. Intestinal permeability was assessed by measuring the 4 kDa dextran-FITC flux in the serum. S. cerevisiae strongly inhibited LF82 adhesion to T84 cells and to the brush border of CD enterocytes. Yeasts decreased LF82 colonization and colitis in CEABAC10 mice and restored barrier function through prevention of the LF82-induced expression of pore-forming tight junction claudin-2 at the plasma membrane of intestinal epithelial cells. These effects were accompanied by a decrease in proinflammatory cytokines IL-6, IL-1β, and KC release by the gut mucosa. Yeast derivatives exerted similar effects on LF82 colonization and colitis demonstrating that yeast viability was not essential to exert beneficial effects. S. cerevisiae yeasts reduce colitis induced by AIEC bacteria in CEACAM6-expressing mice. Such a probiotic strategy could be envisaged in a subgroup of patients with CD abnormally expressing CEACAM6 at the ileal mucosa and therefore susceptible to being colonized by AIEC bacteria.

Insecticidal spectrum and mode of action of the Bacillus thuringiensis Vip3Ca insecticidal protein.

PubMed

Gomis-Cebolla, Joaquín; Ruiz de Escudero, Iñigo; Vera-Velasco, Natalia Mara; Hernández-Martínez, Patricia; Hernández-Rodríguez, Carmen Sara; Ceballos, Tomás; Palma, Leopoldo; Escriche, Baltasar; Caballero, Primitivo; Ferré, Juan

2017-01-01

The Vip3Ca protein, discovered in a screening of Spanish collections of Bacillus thuringiensis, was known to be toxic to Chrysodeixis chalcites, Mamestra brassicae and Trichoplusia ni. In the present study, its activity has been tested with additional insect species and we found that Cydia pomonella is moderately susceptible to this protein. Vip3Ca (of approximately 90kDa) was processed to an approximately 70kDa protein when incubated with midgut juice in all tested species. The kinetics of proteolysis correlated with the susceptibility of the insect species to Vip3Ca. The activation was faster to slower in the following order: M. brassicae (susceptible), Spodoptera littoralis (moderately susceptible), Agrotis ipsilon and Ostrinia nubilalis (slightly susceptible). Processing Vip3Ca by O. nubilalis or M. brassicae midgut juice did not significantly changed its toxicity to either insect species, indicating that the low susceptibility of O. nubilalis is not due to a problem in the midgut processing of the toxin. M. brassicae larvae fed with Vip3Ca showed binding of this toxin to the apical membrane of the midgut epithelial cells. Histopathological inspection showed sloughing of the epithelial cells with further disruption, which suggests that the mode of action of Vip3Ca is similar to that described for Vip3Aa. Biotin-labeled Vip3Ca and Vip3Aa bound specifically to M. brassicae brush border membrane vesicles and both toxins competed for binding sites. This result suggests that insects resistant to Vip3A may also be cross-resistant to Vip3C, which has implications for Insect Resistance Management (IRM). Copyright © 2016 Elsevier Inc. All rights reserved.

Cystic fibrosis epithelial cells are primed for apoptosis as a result of increased Fas (CD95).

PubMed

Chen, Qiwei; Pandi, Sudha Priya Soundara; Kerrigan, Lauren; McElvaney, Noel G; Greene, Catherine M; Elborn, J Stuart; Taggart, Clifford C; Weldon, Sinéad

2018-02-24

Previous work suggests that apoptosis is dysfunctional in cystic fibrosis (CF) airways with conflicting results. We evaluated the relationship between dysfunctional cystic fibrosis transmembrane conductance regulator (CFTR) and apoptosis in CF airway epithelial cells. Apoptosis and associated caspase activity were analysed in non-CF and CF tracheal and bronchial epithelial cell lines. Basal levels of apoptosis and activity of caspase-3 and caspase-8 were significantly increased in CF epithelial cells compared to controls, suggesting involvement of extrinsic apoptosis signalling, which is mediated by the activation of death receptors, such as Fas (CD95). Increased levels of Fas were observed in CF epithelial cells and bronchial brushings from CF patients compared to non-CF controls. Neutralisation of Fas significantly inhibited caspase-3 activity in CF epithelial cells compared to untreated cells. In addition, activation of Fas significantly increased caspase-3 activity and apoptosis in CF epithelial cells compared to control cells. Overall, these results suggest that CF airway epithelial cells are more sensitive to apoptosis via increased levels of Fas and subsequent activation of the Fas death receptor pathway, which may be associated with dysfunctional CFTR. Copyright © 2018 European Cystic Fibrosis Society. All rights reserved.

Efficient entrapment of large and small compounds during vesiculation of intestinal microvilli.

PubMed Central

van Dommelen, F S; Hamer, C M; De Jonge, H R

1986-01-01

An efficient method is described permitting the encapsulation of membrane-impermeable compounds at the interior of intestinal microvilli during vesicle formation. Rat intestinal epithelial cells were isolated by high-frequency vibration and exposed transiently to iso-osmotic medium containing 5 mM-EDTA. Vesiculation of microvilli was effected by freeze-thawing instead of mechanical fragmentation or hypo-osmotic lysis. Solutes to be entrapped were mixed with the extracellular medium before freezing in liquid N2. Microvillous vesicles were isolated from thawed cell suspensions by Ca2+- or Mg2+-aggregation of contaminants and differential centrifugation. The yield, purity, orientation and transport properties of the vesicles were similar, or superior, to preparations described in the literature. A high loading efficiency was demonstrated for small impermeants (cyclic GMP, ATP, Arsenazo III) as well as proteins (albumin); in contrast, loading of isolated vesicles by hypo-osmotic shock was only partially effective (cyclic GMP, ATP) or ineffective (albumin). Entrapment of an ATP-regenerating system could partially block a Mg2+-dependent conversion of intravesicular ATP into ADP. No evidence was obtained for the contribution of a proton pump to the intrinsic Mg2+-ATPase of the vesicle. Potential applications of the vesicle-loading technique in studies of brush-border transport regulation by intramicrovillar factors are discussed. Images Fig. 1. Fig. 2. PMID:3024625

Rapid diagnosis of primary ciliary dyskinesia: cell culture and soft computing analysis.

PubMed

Pifferi, Massimo; Bush, Andrew; Montemurro, Francesca; Pioggia, Giovanni; Piras, Martina; Tartarisco, Gennaro; Di Cicco, Maria; Chinellato, Iolanda; Cangiotti, Angela M; Boner, Attilio L

2013-04-01

Diagnosis of primary ciliary dyskinesia (PCD) sometimes requires repeated nasal brushing to exclude secondary ciliary alterations. Our aim was to evaluate whether the use of a new method of nasal epithelial cell culture can speed PCD diagnosis in doubtful cases and to identify which are the most informative parameters by means of a multilayer artificial neural network (ANN). A cross-sectional study was performed in patients with suspected PCD. All patients underwent nasal brushing for ciliary motion analysis, ultrastructural assessment and evaluation of ciliary function after ciliogenesis in culture by ANN. 151 subjects were studied. A diagnostic suspension cell culture was obtained in 117 nasal brushings. A diagnosis of PCD was made in 36 subjects (29 of whom were children). In nine out of the 36 patients the diagnosis was made only after a second brushing, because of equivocal results of both tests at first examination. In each of these subjects diagnosis of PCD was confirmed by cell culture results. Cell culture in suspension evaluated by means of ANN allows the separation of PCD from secondary ciliary dyskinesia patients after only 5 days of culture and allows diagnosis to be reached in doubtful cases, thus avoiding the necessity of a second sample.

[Primary culture of human normal epithelial cells].

PubMed

Tang, Yu; Xu, Wenji; Guo, Wanbei; Xie, Ming; Fang, Huilong; Chen, Chen; Zhou, Jun

2017-11-28

The traditional primary culture methods of human normal epithelial cells have disadvantages of low activity of cultured cells, the low cultivated rate and complicated operation. To solve these problems, researchers made many studies on culture process of human normal primary epithelial cell. In this paper, we mainly introduce some methods used in separation and purification of human normal epithelial cells, such as tissue separation method, enzyme digestion separation method, mechanical brushing method, red blood cell lysis method, percoll layered medium density gradient separation method. We also review some methods used in the culture and subculture, including serum-free medium combined with low mass fraction serum culture method, mouse tail collagen coating method, and glass culture bottle combined with plastic culture dish culture method. The biological characteristics of human normal epithelial cells, the methods of immunocytochemical staining, trypan blue exclusion are described. Moreover, the factors affecting the aseptic operation, the conditions of the extracellular environment, the conditions of the extracellular environment during culture, the number of differential adhesion, and the selection and dosage of additives are summarized.

Neuroglian stabilizes epithelial structure during Drosophila oogenesis.

PubMed

Wei, Jun; Hortsch, Michael; Goode, Scott

2004-08-01

The vertebrate L1 family of cell adhesion molecules (CAMs) and their fly homolog, Neuroglian, are members of the immunoglobulin (Ig) superfamily of CAMs. In general, Ig CAMs have been found to play critical roles in mediating axon guidance. One Ig CAM, NCAM, has also been implicated in maintaining epithelial integrity and suppressing metastatic dissemination of tumor cells. Other Ig CAMs, such as Nrg, are also expressed in epithelia. We thus tested the hypothesis that, like NCAM, Nrg might also be required for maintaining epithelial integrity and for inhibiting tumor invasion. We used the Drosophila follicular epithelium to determine the function of Nrg in vivo in maintaining epithelial structure, and in regulating the motility of migrating border cells and invasive tumorous follicle cells. Nrg(167) is expressed on the lateral membrane of follicle cells. Loss of Nrg(167) causes border cells to delay delamination and causes other follicle cells to delaminate inappropriately. The delaminated cells have aberrant epithelial polarity manifested as severe mislocalization of apical and basal membrane proteins, and uniform localization of lateral membrane proteins. Furthermore, loss of Nrg(167) dramatically enhances the invasive phenotype associated with loss of Discs Large, a neoplastic tumor suppressor. These results indicate that Nrg(167) stabilizes epithelial polarity by regulating junctional adhesion and function in normal and tumorous epithelia. Our data also suggest that Ig superfamily members have significant functional redundancy in maintaining epithelial polarity, with individual members playing subtle, unique roles during epithelial morphogenesis. Copyright 2004 Wiley-Liss, Inc.

Proximal tubule proteins are significantly elevated in bladder urine of patients with ureteropelvic junction obstruction and may represent novel biomarkers: A pilot study.

PubMed

Gerber, Claire; Harel, Miriam; Lynch, Miranda L; Herbst, Katherine W; Ferrer, Fernando A; Shapiro, Linda H

2016-04-01

Ureteropelvic junction obstruction (UPJO) is the major cause of hydronephrosis in children and may lead to renal injury and early renal dysfunction. However, diagnosis of the degree of obstruction and severity of renal injury relies on invasive and often inconclusive renal scans. Biomarkers from voided urine that detect early renal injury are highly desirable because of their noninvasive collection and their potential to assist in earlier and more reliable diagnosis of the severity of obstruction. Early in response to UPJO, increased intrarenal pressure directly impacts the proximal tubule brush border. We hypothesize that single-pass, apically expressed proximal tubule brush border proteins will be shed into the urine early and rapidly and will be reliable noninvasive urinary biomarkers, providing the tools for a more reliable stratification of UPJO patients. We performed a prospective cohort study at Connecticut Children's Medical Center. Bladder urine samples from 12 UPJO patients were obtained prior to surgical intervention. Control urine samples were collected from healthy pediatric patients presenting with primary nocturnal enuresis. We determined levels of NGAL, KIM-1 (previously identified biomarkers), CD10, CD13, and CD26 (potentially novel biomarkers) by ELISA in control and experimental urine samples. Urinary creatinine levels were used to normalize the urinary protein levels measured by ELISA. Each of the proximal tubule proteins outperformed the previously published biomarkers. No differences in urinary NGAL and KIM-1 levels were observed between control and obstructed patients (p = 0.932 and p = 0.799, respectively). However, levels of CD10, CD13, and CD26 were significantly higher in the voided urine of obstructed individuals when compared with controls (p = 0.002, p = 0.024, and p = 0.007, respectively) (Figure). Targeted identification of reliable, noninvasive biomarkers of renal injury is critical to aid in diagnosing patients at risk, guiding therapeutic decisions and monitoring treatment efficacy. Proximal tubule brush border proteins are reliably detected in the urine of obstructed patients and may be more effective at predicting UPJO. Copyright © 2015 Journal of Pediatric Urology Company. Published by Elsevier Ltd. All rights reserved.

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

PubMed

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.

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

Elasticity of biomembranes studied by dynamic light scattering

NASA Astrophysics Data System (ADS)

Fujime, Satoru; Miyamoto, Shigeaki

1991-05-01

Combination of osmotic swelling and dynamic light scattering makes it possible to measure the elastic modulus of biomembranes. By this technique we have observed a drastic increase in membrane flexibility on activation of Na/glucose cotransporters in membrane vesicles prepared from brush-borders of rat small intestine and on activation by micromolar [Ca2] of exocytosis in secretory granules isolated from rat pancreatic acinar cells and bovine adrenal chromaffin cells. 1 .

Human neutral brush border endopeptidase EC 3.4.24.11 in urine, its isolation, characterisation and activity in renal diseases.

PubMed

Vlaskou, D; Hofmann, W; Guder, W G; Siskos, P A; Dionyssiou-Asteriou, A

2000-07-01

Human neutral brush border endopeptidase (NEP) was purified from the urine of patients suffering from acute toxic tubulointerstitial nephropathy. An enzyme preparation with specific activity of 102 Ug(-1) protein was obtained. The urinary activities of neutral endopeptidase and alanine aminopeptidase were measured in patients with renal disease and in 30 control patients, resulting in a reference range from 0.1 to 0.7 Ug(-1) creatinine and 1.4-14.1 Ug(-1) creatinine, respectively. Urine enzyme activities were highest in patients with acute tubulotoxic renal diseases. Neutral endopeptidase and alanine aminopeptidase activities were found to be 6.5- and 10-fold higher than the upper value of the reference range, respectively. Smaller increases in the rate of excretion of these enzymes (2.5- and 3.5-fold), respectively, were observed in patients suffering from acute tubular insufficiency and even lower increases, 2- and 1.5-fold, respectively, were observed in patients with chronic renal diseases. In diabetics and kidney transplant patients the enzyme excretion rates were within the reference range. Assay of both transmembrane metalloproteinases in urine may prove valuable in serving as markers for renal toxicity. Together with beta-NAG these enzymes could be employed as differentiation markers between acute and chronic tubular insufficiency.

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

Digestive system dysfunction in cystic fibrosis: challenges for nutrition therapy.

PubMed

Li, Li; Somerset, Shawn

2014-10-01

Cystic fibrosis can affect food digestion and nutrient absorption. The underlying mutation of the cystic fibrosis trans-membrane regulator gene depletes functional cystic fibrosis trans-membrane regulator on the surface of epithelial cells lining the digestive tract and associated organs, where Cl(-) secretion and subsequently secretion of water and other ions are impaired. This alters pH and dehydrates secretions that precipitate and obstruct the lumen, causing inflammation and the eventual degradation of the pancreas, liver, gallbladder and intestine. Associated conditions include exocrine pancreatic insufficiency, impaired bicarbonate and bile acid secretion and aberrant mucus formation, commonly leading to maldigestion and malabsorption, particularly of fat and fat-soluble vitamins. Pancreatic enzyme replacement therapy is used to address this insufficiency. The susceptibility of pancreatic lipase to acidic and enzymatic inactivation and decreased bile availability often impedes its efficacy. Brush border digestive enzyme activity and intestinal uptake of certain disaccharides and amino acids await clarification. Other complications that may contribute to maldigestion/malabsorption include small intestine bacterial overgrowth, enteric circular muscle dysfunction, abnormal intestinal mucus, and intestinal inflammation. However, there is some evidence that gastric digestive enzymes, colonic microflora, correction of fatty acid abnormalities using dietary n-3 polyunsaturated fatty acid supplementation and emerging intestinal biomarkers can complement nutrition management in cystic fibrosis. Copyright © 2014 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved.

Diphenyl ditelluride intoxication triggers histological changes in liver, kidney, and lung of mice.

PubMed

da Luz, Sônia Cristina Almeida; Daubermann, Melissa Falster; Thomé, Gustavo Roberto; Dos Santos, Matheus Mülling; Ramos, Angelica; Torres Salazar, Gerson; da Rocha, João Batista Teixeira; Barbosa, Nilda Vargas

2015-01-01

Tellurium compounds may be cytotoxic to different cells types. Thus, this work evaluated the effect of diphenyl ditelluride ((PhTe)2), an organotellurium commonly used in organic synthesis, on the morphology of liver, kidney, and lung. Adult mice were acutely (a subcutaneous single dose: 250 μmol/kg) or subchronically (one daily subcutaneous dose: 10 or 50 μmol/kg for 7 and 14 days) exposed to (PhTe)2. Afterwards, the histological analyses of liver, kidney, and lungs were performed. Liver histology revealed that the hepatocytes of mice subchronically exposed to (PhTe)2 presented cytoplasmic vacuolization, hydropic degeneration, and hyperchromatic nuclei. Subchronic exposure to 50 μmol/kg (PhTe)2 also caused hepatic necrosis. Microvesicular and macrovesicular steatosis were identified in liver of mice acutely exposed to (PhTe)2. Acute and subchronic intoxication with (PhTe)2 induced changes on epithelial cells of renal tubules, namely, loss of brush border and cytoplasmatic vacuolization. Atrophy and hypertrophy, cast proteinaceous formation, and acute tubular necrosis were also identified in renal tissue. Mice subchronically exposed to 50 μmol/kg (PhTe)2 developed intra-alveolar edema and alveolar wall congestion in some areas of lungs. Acute exposure to (PhTe)2 did not cause histological changes in lungs. Our data show that (PhTe)2 may be considered a histotoxic agent for liver, kidney, and lung.

Evaluation of lipid oxidative stress status and inflammation in atopic ocular surface disease

PubMed Central

Wakamatsu, Tais H.; Ayako, Igarashi; Takano, Yoji; Matsumoto, Yukihiro; Ibrahim, Osama M.A.; Okada, Naoko; Satake, Yoshiyuki; Fukagawa, Kazumi; Shimazaki, Jun; Tsubota, Kazuo; Fujishima, Hiroshi

2010-01-01

Background Although the oxidative stress status in atopic skin disease has been reported to be elevated, there are still no studies related to the status of oxidative stress in atopic ocular surface disease. The purpose of this study was to evaluate the ocular surface lipid oxidative stress status and inflammation in atopic keratoconjunctivitis (AKC) patients and normal subjects. Methods Twenty eight eyes of 14 patients (9 males, 5 females) with AKC and 18 eyes of 9 age and sex matched (4 males and 5 females) normal healthy controls were examined in this prospective study. The severity of atopic dermatitis (AD) was scored by the SCORing Atopic Dermatitis (SCORAD) index. All subjects underwent Schirmer test, tear film break up time (BUT), fluorescein/Rose Bengal stainings, tear collection, and brush cytology from the upper palpebral conjunctiva. The brush cytology samples were stained with Diff-Quik for differentiation of inflammatory cells and immunohistochemistry (IHC) staining with HEL (hexanoyl-lysine) and 4-HNE (4-hydroxy-2-nonenal) to study lipid oxidation. HEL and cytokine (interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-10 (IL-10), tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ)) levels were measured by enzyme-linked immunosorbent assay (ELISA) from tear samples of AKC patients and control subjects. Toluidine Blue and IHC staining with HEL, 4-HNE and cluster of differentiation 45 (CD45) were performed on papillary samples of AKC patients. This study was conducted in compliance with the “Declaration of Helsinki.” Results The tear stability and vital staining scores were significantly worse in eyes of AKC patients (p<0.05) compared to the controls. Inflammatory cells and positively stained conjunctival epithelial cells for HEL and 4-HNE showed a significant elevation in brush cytology samples of AKC patients. Significantly higher levels of HEL and cytokines were detected in tears of AKC patients compared to controls. Papillary specimens also revealed many CD45 inflammatory cells as well as many cells positively stained with HEL and 4-HNE in IHC. A strong significant linear positive correlation between conjunctival inflammation and epithelial lipid oxidative stress status was observed. Conjunctival lipid oxidative stress also correlated strongly with tear HEL levels and epithelial damage scores. Conclusions The ocular surface disease in AKC was characterized by marked tear instability, ocular surface epithelial damage, increase in inflammatory infiltrates and presence of increased lipid oxidation. PMID:21139696

Fabrication of nonfouling, bactericidal, and bacteria corpse release multifunctional surface through surface-initiated RAFT polymerization.

PubMed

Wang, Bailiang; Ye, Zi; Tang, Yihong; Han, Yuemei; Lin, Quankui; Liu, Huihua; Chen, Hao; Nan, Kaihui

Infections after surgery or endophthalmitis are potentially blinding complications caused by bacterial adhesion and subsequent biofilm formation on the intraocular lens. Neither single-function anti-adhesion surface nor contacting killing surface can exhibit ideal antibacterial function. In this work, a novel (2-(dimethylamino)-ethyl methacrylate- co -2-methacryloyloxyethyl phosphorylcholine) (p (DMAEMA- co -MPC)) brush was synthesized by "grafting from" method through reversible-addition fragmentation chain transfer polymerization. 1-Bromoheptane was used to quaternize the p (DMAEMA- co -MPC) brush coating and to endow the surface with bactericidal function. The success of the surface functionalization was confirmed by atomic force microscopy, water contact angle, and spectroscopic ellipsometry. The quaternary ammonium salt units were employed as efficient disinfection that can eliminate bacteria through contact killing, whereas the 2-methacryloyloxyethyl phosphorylcholine units were introduced to suppress unwanted nonspecific adsorption. The functionalized poly(dimethyl siloxane) surfaces showed efficiency in reducing bovine serum albumin adsorption and in inhibiting bacteria adhesion and biofilm formation. The copolymer brushes also demonstrated excellent bactericidal function against gram-positive ( Staphylococcus aureus ) bacteria measured by bacteria live/dead staining and shake-flask culture methods. The surface biocompatibility was evaluated by morphology and activity measurement with human lens epithelial cells in vitro. The achievement of the p (DMAEMA + - co -MPC) copolymer brush coating with nonfouling, bactericidal, and bacteria corpse release properties can be used to modify intraocular lenses.

Fabrication of nonfouling, bactericidal, and bacteria corpse release multifunctional surface through surface-initiated RAFT polymerization

PubMed Central

Wang, Bailiang; Ye, Zi; Tang, Yihong; Han, Yuemei; Lin, Quankui; Liu, Huihua; Chen, Hao; Nan, Kaihui

2017-01-01

Infections after surgery or endophthalmitis are potentially blinding complications caused by bacterial adhesion and subsequent biofilm formation on the intraocular lens. Neither single-function anti-adhesion surface nor contacting killing surface can exhibit ideal antibacterial function. In this work, a novel (2-(dimethylamino)-ethyl methacrylate-co-2-methacryloyloxyethyl phosphorylcholine) (p (DMAEMA-co-MPC)) brush was synthesized by “grafting from” method through reversible–addition fragmentation chain transfer polymerization. 1-Bromoheptane was used to quaternize the p (DMAEMA-co-MPC) brush coating and to endow the surface with bactericidal function. The success of the surface functionalization was confirmed by atomic force microscopy, water contact angle, and spectroscopic ellipsometry. The quaternary ammonium salt units were employed as efficient disinfection that can eliminate bacteria through contact killing, whereas the 2-methacryloyloxyethyl phosphorylcholine units were introduced to suppress unwanted nonspecific adsorption. The functionalized poly(dimethyl siloxane) surfaces showed efficiency in reducing bovine serum albumin adsorption and in inhibiting bacteria adhesion and biofilm formation. The copolymer brushes also demonstrated excellent bactericidal function against gram-positive (Staphylococcus aureus) bacteria measured by bacteria live/dead staining and shake-flask culture methods. The surface biocompatibility was evaluated by morphology and activity measurement with human lens epithelial cells in vitro. The achievement of the p (DMAEMA+-co-MPC) copolymer brush coating with nonfouling, bactericidal, and bacteria corpse release properties can be used to modify intraocular lenses. PMID:28053527

A Bioinformatics Approach for Integrated Transcriptomic and Proteomic Comparative Analyses of Model and Non-sequenced Anopheline Vectors of Human Malaria Parasites*

PubMed Central

Mohien, Ceereena Ubaida; Colquhoun, David R.; Mathias, Derrick K.; Gibbons, John G.; Armistead, Jennifer S.; Rodriguez, Maria C.; Rodriguez, Mario Henry; Edwards, Nathan J.; Hartler, Jürgen; Thallinger, Gerhard G.; Graham, David R.; Martinez-Barnetche, Jesus; Rokas, Antonis; Dinglasan, Rhoel R.

2013-01-01

Malaria morbidity and mortality caused by both Plasmodium falciparum and Plasmodium vivax extend well beyond the African continent, and although P. vivax causes between 80 and 300 million severe cases each year, vivax transmission remains poorly understood. Plasmodium parasites are transmitted by Anopheles mosquitoes, and the critical site of interaction between parasite and host is at the mosquito's luminal midgut brush border. Although the genome of the “model” African P. falciparum vector, Anopheles gambiae, has been sequenced, evolutionary divergence limits its utility as a reference across anophelines, especially non-sequenced P. vivax vectors such as Anopheles albimanus. Clearly, technologies and platforms that bridge this substantial scientific gap are required in order to provide public health scientists with key transcriptomic and proteomic information that could spur the development of novel interventions to combat this disease. To our knowledge, no approaches have been published that address this issue. To bolster our understanding of P. vivax–An. albimanus midgut interactions, we developed an integrated bioinformatic-hybrid RNA-Seq-LC-MS/MS approach involving An. albimanus transcriptome (15,764 contigs) and luminal midgut subproteome (9,445 proteins) assembly, which, when used with our custom Diptera protein database (685,078 sequences), facilitated a comparative proteomic analysis of the midgut brush borders of two important malaria vectors, An. gambiae and An. albimanus. PMID:23082028

A bioinformatics approach for integrated transcriptomic and proteomic comparative analyses of model and non-sequenced anopheline vectors of human malaria parasites.

PubMed

Ubaida Mohien, Ceereena; Colquhoun, David R; Mathias, Derrick K; Gibbons, John G; Armistead, Jennifer S; Rodriguez, Maria C; Rodriguez, Mario Henry; Edwards, Nathan J; Hartler, Jürgen; Thallinger, Gerhard G; Graham, David R; Martinez-Barnetche, Jesus; Rokas, Antonis; Dinglasan, Rhoel R

2013-01-01

Malaria morbidity and mortality caused by both Plasmodium falciparum and Plasmodium vivax extend well beyond the African continent, and although P. vivax causes between 80 and 300 million severe cases each year, vivax transmission remains poorly understood. Plasmodium parasites are transmitted by Anopheles mosquitoes, and the critical site of interaction between parasite and host is at the mosquito's luminal midgut brush border. Although the genome of the "model" African P. falciparum vector, Anopheles gambiae, has been sequenced, evolutionary divergence limits its utility as a reference across anophelines, especially non-sequenced P. vivax vectors such as Anopheles albimanus. Clearly, technologies and platforms that bridge this substantial scientific gap are required in order to provide public health scientists with key transcriptomic and proteomic information that could spur the development of novel interventions to combat this disease. To our knowledge, no approaches have been published that address this issue. To bolster our understanding of P. vivax-An. albimanus midgut interactions, we developed an integrated bioinformatic-hybrid RNA-Seq-LC-MS/MS approach involving An. albimanus transcriptome (15,764 contigs) and luminal midgut subproteome (9,445 proteins) assembly, which, when used with our custom Diptera protein database (685,078 sequences), facilitated a comparative proteomic analysis of the midgut brush borders of two important malaria vectors, An. gambiae and An. albimanus.

EPA blocks TNF-α-induced inhibition of sugar uptake in Caco-2 cells via GPR120 and AMPK.

PubMed

Castilla-Madrigal, Rosa; Barrenetxe, Jaione; Moreno-Aliaga, María J; Lostao, María Pilar

2018-03-01

The aim of the present work was to investigate in Caco-2 cells whether eicosapentaenoic acid (EPA), an omega-3 polyunsaturated fatty acid, could block the inhibitory effect of tumor necrosis factor-α (TNF-α) on sugar transport, and identify the intracellular signaling pathways involved. After pre-incubation of the Caco-2 cells with TNF-α and EPA for 1 hr, EPA prevented the inhibitory effect of the cytokine on α-methyl-d-glucose (αMG) uptake (15 min) and on SGLT1 expression at the brush border membrane, measured by Western blot. The ERK1/2 inhibitor PD98059 and the AMPK activator AICAR also prevented the inhibitory effect of TNF-α on both αMG uptake and SGLT1 expression. Interestingly, the AMPK inhibitor, Compound C, abolished the ability of EPA to prevent TNF-α-induced reduction of sugar uptake and transporter expression. The GPR120 antagonist, AH7614, also blocked the preventive effect of EPA on TNF-α-induced decrease of αMG uptake and AMPK phosphorylation. In summary, TNF-α inhibits αMG uptake by decreasing SGLT1 expression in the brush border membrane through the activation of ERK1/2 pathway. EPA prevents the inhibitory effect of TNF-α through the involvement of GPR120 and AMPK activation. © 2017 Wiley Periodicals, Inc.

The epidermal growth factor receptor (EGF-R) is present on the basolateral, but not the apical, surface of enterocytes in the human gastrointestinal tract.

PubMed Central

Playford, R J; Hanby, A M; Gschmeissner, S; Peiffer, L P; Wright, N A; McGarrity, T

1996-01-01

BACKGROUND: While it is clear that luminal epidermal growth factor (EGF) stimulates repair of the damaged bowel, its significance in maintaining normal gut growth remains uncertain. If EGF is important in maintaining normal gut growth, the EGF receptor (EGF-R) should be present on the apical (luminal) surface in addition to the basolateral surface. AIMS/SUBJECTS/METHODS: This study examined the distribution of the EGF-R in the epithelium throughout the human gastro-intestinal tract using immunohistochemistry, electron microscopy, and western blotting of brush border preparations. RESULTS: Immunostaining of the oesophagus showed circumferential EGF-R positivity in the cells of the basal portions of the stratified squamous epithelium but surface cells were EGF-R negative. In the normal stomach, small intestine, and colon, immunostaining localised the receptor to the basolateral surface with the apical membranes being consistently negative. EGF-R positivity within the small intestine appeared to be almost entirely restricted to the proliferative (crypt) region. Western blotting demonstrated a 170 kDa protein in whole tissue homogenates but not in the brush border vesicle preparations. CONCLUSIONS: As the EGF-R is located only on the basolateral surfaces in the normal adult gastrointestinal tract, the major role of luminal EGF is probably to stimulate repair rather than to maintain normal gut growth. Images Figure 1 Figure 2 Figure 3 PMID:8977341

Morphology and innervation pattern of the feline urogenital junction.

PubMed

Wrobel, K H; Gürtler, A

2004-12-01

The feline urogenital junction is situated between the extratesticular rete and the spacious initial segments of the efferent ductules. The rete epithelium is cuboidal to low columnar. The rete cells forming the junction rest on a wavy basal lamina, display deep mutual invaginations, possess central nuclei with several infoldings and form a distinct border with the columnar epithelial cells of the initial segments of the ductuli efferentes. The epithelium of the initial segments is composed of ciliated cells and non-ciliated principal cells. The latter are the dominating type and characterized by an apical brush-border and a supranuclear endocytotic apparatus. The stroma of the extratesticular rete contains an abundance of collagen whereas contractile cells are here generally absent. In contrast, the initial segments of the efferent ductules are surrounded by elastic fibres and a layer of contractile cells. All nerves for the feline urogenital junction come from the nervus spermaticus superior. In the epididymal head, small nerve bundles deviate into the septa between the ductules. Single fibres establish a dense network within the muscular coat of the ductuli. At the transition to the extratesticular rete, this network ends abruptly. Nerve fibres in the confines of the rete are associated with blood vessels or proceed to the testicular interior, but establish no relationships with the rete epithelium or the myofibroblasts of the mediastinum. The nervous network in the walls of the efferent ductules and their initial segments is not only composed of sympathetic but also parasympathetic, non-myelinated fibres. Particularly noteworthy is the abundance of calcitonin gene-related peptide (CGRP)- and substance P (SP)-containing axons around the initial segments. Both neuroproteins are consistent markers for sensory neurones. Taken together, it can be assumed that the entry of seminal fluid and spermatozoa into the efferent ductules is controlled by a regulatory nervous chain provided with afferent and efferent components.

1-alpha,25-Dihydroxyvitamin D3 up-regulates the expression of 2 types of human intestinal alkaline phosphatase alternative splicing variants in Caco-2 cells and may be an important regulator of their expression in gut homeostasis.

PubMed

Noda, Seiko; Yamada, Asako; Nakaoka, Kanae; Goseki-Sone, Masae

2017-10-01

Vitamin D insufficiency is associated with a greater risk of osteoporosis and also influences skeletal muscle functions, differentiation, and development. The principal function of vitamin D in calcium homeostasis is to increase the absorption of calcium from the intestine, and the level of alkaline phosphatase (ALP) activity, a differentiation marker for intestinal epithelial cells, is regulated by vitamin D. Intestinal-type ALP is expressed at a high concentration in the brush border membrane of intestinal epithelial cells, and is known to be affected by several kinds of nutrients. Recent reviews have highlighted the importance of intestinal-type ALP in gut homeostasis. Intestinal-type ALP controls bacterial endotoxin-induced inflammation by dephosphorylating lipopolysaccharide and is a gut mucosal defense factor. In this study, we investigated the influence of vitamin D on the expression of 2 types of alternative mRNA variants encoding the human alkaline phosphatase, intestinal (ALPI) gene in human Caco-2 cells as an in vitro model of the small intestinal epithelium. After treatment with 1-alpha,25-dihydroxyvitamin D 3 , the biologically active form of vitamin D 3 , there were significant increases in the ALP activities of Caco-2 cells. Inhibitor and thermal inactivation experiments showed that the increased ALP had properties of intestinal-type ALP. Reverse transcription-polymerase chain reaction analysis revealed that expression of the 2 types of alternative mRNA variants from the ALPI gene was markedly enhanced by vitamin D in Caco-2 cells. In conclusion, these findings agree with the hypothesis: vitamin D up-regulated the expression of 2 types of human intestinal alkaline phosphatase alternative splicing variants in Caco-2 cells; vitamin D may be an important regulator of ALPI gene expression in gut homeostasis. Copyright © 2017 Elsevier Inc. All rights reserved.

Impaired airway epithelial cell responses from children with asthma to rhinoviral infection.

PubMed

Kicic, A; Stevens, P T; Sutanto, E N; Kicic-Starcevich, E; Ling, K-M; Looi, K; Martinovich, K M; Garratt, L W; Iosifidis, T; Shaw, N C; Buckley, A G; Rigby, P J; Lannigan, F J; Knight, D A; Stick, S M

2016-11-01

The airway epithelium forms an effective immune and physical barrier that is essential for protecting the lung from potentially harmful inhaled stimuli including viruses. Human rhinovirus (HRV) infection is a known trigger of asthma exacerbations, although the mechanism by which this occurs is not fully understood. To explore the relationship between apoptotic, innate immune and inflammatory responses to HRV infection in airway epithelial cells (AECs) obtained from children with asthma and non-asthmatic controls. In addition, to test the hypothesis that aberrant repair of epithelium from asthmatics is further dysregulated by HRV infection. Airway epithelial brushings were obtained from 39 asthmatic and 36 non-asthmatic children. Primary cultures were established and exposed to HRV1b and HRV14. Virus receptor number, virus replication and progeny release were determined. Epithelial cell apoptosis, IFN-β production, inflammatory cytokine release and epithelial wound repair and proliferation were also measured. Virus proliferation and release was greater in airway epithelial cells from asthmatics but this was not related to the number of virus receptors. In epithelial cells from asthmatic children, virus infection dampened apoptosis, reduced IFN-β production and increased inflammatory cytokine production. HRV1b infection also inhibited wound repair capacity of epithelial cells isolated from non-asthmatic children and exaggerated the defective repair response seen in epithelial cells from asthmatics. Addition of IFN-β restored apoptosis, suppressed virus replication and improved repair of airway epithelial cells from asthmatics but did not reduce inflammatory cytokine production. Collectively, HRV infection delays repair and inhibits apoptotic processes in epithelial cells from non-asthmatic and asthmatic children. The delayed repair is further exaggerated in cells from asthmatic children and is only partially reversed by exogenous IFN-β. © 2016 John Wiley & Sons Ltd.

A Noninvasive Test for MicroRNA Expression in Oral Squamous Cell Carcinoma.

PubMed

Gissi, Davide B; Morandi, Luca; Gabusi, Andrea; Tarsitano, Achille; Marchetti, Claudio; Cura, Francesca; Palmieri, Annalisa; Montebugnoli, Lucio; Asioli, Sofia; Foschini, Maria P; Scapoli, Luca

2018-06-16

MicroRNAs have recently been proposed as non-invasive biomarkers in Oral Squamous Cell Carcinoma (OSCC). The aim of this study was to analyze the expression of a panel of miRNAs in epithelial cells collected by oral brushing from OSCCs from regenerative areas after OSCC surgical resection and from their respective normal distant mucosa. Oral brushing specimens were collected from 24 healthy donors, 14 OSCC patients with specimens from tumour and normal distant mucosa, and from 13 patients who had OSCC resection, with samples from regenerative areas after OSCC resection and normal distant mucosa. Expression levels of eight targets (miR-21, miR-375, miR-345, miR-181b, miR-146a, miR-649, miR-518b, and miR-191) were evaluated by real-time Polymerase Chain Reaction (PCR). A highly significant between-group difference was found for miR-21 (F = 6.58, p < 0.001), miR-146a (F = 6.974, p < 0.001), and miR-191 (F = 17.07, p < 0.001). The major difference was observed between samples from healthy donors and from OSCC brushing, whereas no significant differences were observed between areas infiltrated by OSCC and their respective normal distant mucosa. Furthermore, altered expression of miR-146a and miR-191 was also observed in regenerative areas after OSCC resection. Oral brushing could be proposed as a noninvasive method to study microRNA expression in oral mucosa in OSCC patients.

Structural and functional changes associated with normal and abnormal fundus autofluorescence in patients with retinitis pigmentosa.

PubMed

Greenstein, Vivienne C; Duncker, Tobias; Holopigian, Karen; Carr, Ronald E; Greenberg, Jonathan P; Tsang, Stephen H; Hood, Donald C

2012-02-01

To analyze the structure and visual function of regions bordering the hyperautofluorescent ring/arcs in retinitis pigmentosa. Twenty-one retinitis pigmentosa patients (21 eyes) with rings/arcs and 21 normal individuals (21 eyes) were studied. Visual sensitivity in the central 10° was measured with microperimetry. Retinal structure was evaluated with spectral-domain optical coherence tomography. The distance from the fovea to disruption/loss of the inner outer segment (IS/OS) junction and thicknesses of the total receptor plus retinal pigment epithelial complex and outer segment plus retinal pigment epithelial complex layers were measured. Results were compared with measurements of the distance from the fovea to the inner and outer borders of the ring/arc seen on fundus autofluorescence. Disruption/loss of the inner outer segment junction occurred closer to the inner border of the ring/arc and it was closer to the fovea in eight eyes. For 19 eyes, outer segment plus and receptor plus RPE complex thicknesses were significantly decreased at locations closer to the fovea than the appearance of the inner border of hyperautofluorescence. Mean visual sensitivity was decreased inside, across, and outside the ring/arc by 3.5 ± 3.8, 8.9 ± 4.8, and 17.0 ± 2.4 dB, respectively. Structural and functional changes can occur inside the hyperfluorescent ring/arc in retinitis pigmentosa.

Nasal Epithelial Cells as Surrogates for Bronchial Epithelial Cells in Airway Inflammation Studies

PubMed Central

McDougall, Catherine M.; Blaylock, Morgan G.; Douglas, J. Graham; Brooker, Richard J.; Helms, Peter J.; Walsh, Garry M.

2008-01-01

The nose is an attractive source of airway epithelial cells, particularly in populations in which bronchoscopy may not be possible. However, substituting nasal cells for bronchial epithelial cells in the study of airway inflammation depends upon comparability of responses, and evidence for this is lacking. Our objective was to determine whether nasal epithelial cell inflammatory mediator release and receptor expression reflect those of bronchial epithelial cells. Paired cultures of undifferentiated nasal and bronchial epithelial cells were obtained from brushings from 35 subjects, including 5 children. Cells were subject to morphologic and immunocytochemical assessment. Mediator release from resting and cytokine-stimulated cell monolayers was determined, as was cell surface receptor expression. Nasal and bronchial cells had identical epithelial morphology and uniform expression of cytokeratin 19. There were no differences in constitutive expression of CD44, intercellular adhesion molecule-1, αvβ3, and αvβ5. Despite significantly higher constitutive release of IL-8, IL-6, RANTES (regulated on activation, normal T cell expressed and secreted), and matrix metalloproteinase (MMP)-9 from nasal compared with bronchial cells, the increments in release of all studied mediators in response to stimulation with IL-1β and TNF-α were similar, and there were significant positive correlations between nasal and bronchial cell secretion of IL-6, RANTES, vascular endothelial growth factor, monocyte chemoattractant protein-1, MMP-9, and tissue inhibitor of metalloproteinase-1. Despite differences in absolute mediator levels, the responses of nasal and bronchial epithelial cells to cytokine stimulation were similar, expression of relevant surface receptors was comparable, and there were significant correlations between nasal and bronchial cell mediator release. Therefore, nasal epithelial cultures constitute an accessible surrogate for studying lower airway inflammation. PMID:18483420

Nasal epithelial cells as surrogates for bronchial epithelial cells in airway inflammation studies.

PubMed

McDougall, Catherine M; Blaylock, Morgan G; Douglas, J Graham; Brooker, Richard J; Helms, Peter J; Walsh, Garry M

2008-11-01

The nose is an attractive source of airway epithelial cells, particularly in populations in which bronchoscopy may not be possible. However, substituting nasal cells for bronchial epithelial cells in the study of airway inflammation depends upon comparability of responses, and evidence for this is lacking. Our objective was to determine whether nasal epithelial cell inflammatory mediator release and receptor expression reflect those of bronchial epithelial cells. Paired cultures of undifferentiated nasal and bronchial epithelial cells were obtained from brushings from 35 subjects, including 5 children. Cells were subject to morphologic and immunocytochemical assessment. Mediator release from resting and cytokine-stimulated cell monolayers was determined, as was cell surface receptor expression. Nasal and bronchial cells had identical epithelial morphology and uniform expression of cytokeratin 19. There were no differences in constitutive expression of CD44, intercellular adhesion molecule-1, alphavbeta3, and alphavbeta5. Despite significantly higher constitutive release of IL-8, IL-6, RANTES (regulated on activation, normal T cell expressed and secreted), and matrix metalloproteinase (MMP)-9 from nasal compared with bronchial cells, the increments in release of all studied mediators in response to stimulation with IL-1beta and TNF-alpha were similar, and there were significant positive correlations between nasal and bronchial cell secretion of IL-6, RANTES, vascular endothelial growth factor, monocyte chemoattractant protein-1, MMP-9, and tissue inhibitor of metalloproteinase-1. Despite differences in absolute mediator levels, the responses of nasal and bronchial epithelial cells to cytokine stimulation were similar, expression of relevant surface receptors was comparable, and there were significant correlations between nasal and bronchial cell mediator release. Therefore, nasal epithelial cultures constitute an accessible surrogate for studying lower airway inflammation.

Aging Selectively Modulates Vitamin C Transporter Expression Patterns in the Kidney.

PubMed

Forman, Katherine; Martínez, Fernando; Cifuentes, Manuel; Bertinat, Romina; Salazar, Katterine; Nualart, Francisco

2017-09-01

In the kidney, vitamin C is reabsorbed from the glomerular ultrafiltrate by sodium-vitamin C cotransporter isoform 1 (SVCT1) located in the brush border membrane of the proximal tubules. Although we know that vitamin C levels decrease with age, the adaptive physiological mechanisms used by the kidney for vitamin C reabsorption during aging remain unknown. In this study, we used an animal model of accelerated senescence (SAMP8 mice) to define the morphological alterations and aging-induced changes in the expression of vitamin C transporters in renal tissue. Aging induced significant morphological changes, such as periglomerular lymphocytic infiltrate and glomerular congestion, in the kidneys of SAMP8 mice, although no increase in collagen deposits was observed using 2-photon microscopy analysis and second harmonic generation. The most characteristic histological alteration was the dilation of intracellular spaces in the basolateral region of proximal tubule epithelial cells. Furthermore, a combination of laser microdissection, qRT-PCR, and immunohistochemical analyses allowed us to determine that SVCT1 expression specifically increased in the proximal tubules from the outer strip of the outer medulla (segment S3) and cortex (segment S2) during aging and that these tubules also express GLUT1. We conclude that aging modulates vitamin C transporter expression and that renal over-expression of SVCT1 enhances vitamin C reabsorption in aged animals that may synthesize less vitamin C. J. Cell. Physiol. 232: 2418-2426, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Vitamin D-restricted high-fat diet down-regulates expression of intestinal alkaline phosphatase isozymes in ovariectomized rats.

PubMed

Nakaoka, Kanae; Yamada, Asako; Noda, Seiko; Goseki-Sone, Masae

2018-05-01

Intestinal alkaline phosphatase (IAP) is expressed at a high concentration in the brush border membrane of intestinal epithelial cells. Intestinal alkaline phosphatase controls bacterial endotoxin-induced inflammation by dephosphorylating lipopolysaccharide and is a gut mucosal defense factor. Previously, we reported that IAP activity in the duodenum was significantly decreased in male rats receiving a high-fat diet with vitamin D restriction. Here, we tested the hypothesis that IAP is also regulated by a vitamin D-restricted high-fat diet in an animal model of menopause. Twenty-four female rats were ovariectomized (OVX), and another 6 female rats were sham operated. The OVX rats were divided into 4 groups and fed experimental diets: a basic control diet, a basic control diet with vitamin D restriction, a high-fat diet, and a high-fat diet with vitamin D restriction. After 28days of the experimental diets, the vitamin D-restricted high-fat diet decreased alkaline phosphatase activity in the duodenum of the OVX groups. The vitamin D-restricted high-fat diet down-regulated mRNA expressions of IAP isozymes in the duodenum of the OVX groups. These findings support the hypothesis that the expression of IAP is suppressed by a vitamin D-restricted high-fat diet in OVX rats. An adequate vitamin D intake and prevention of low vitamin D levels may be important for IAP expression in gut homeostasis. Copyright © 2018 Elsevier Inc. All rights reserved.

Characterisation and cloning of a Na(+)-dependent broad-specificity neutral amino acid transporter from NBL-1 cells: a novel member of the ASC/B(0) transporter family.

PubMed

Pollard, Matthew; Meredith, David; McGivan, John D

2002-04-12

Na(+)-dependent neutral amino acid transport into the bovine renal epithelial cell line NBL-1 is catalysed by a broad-specificity transporter originally termed System B(0). This transporter is shown to differ in specificity from the B(0) transporter cloned from JAR cells [J. Biol. Chem. 271 (1996) 18657] in that it interacts much more strongly with phenylalanine. Using probes designed to conserved transmembrane regions of the ASC/B(0) transporter family we have isolated a cDNA encoding the NBL-1 cell System B(0) transporter. When expressed in Xenopus oocytes the clone catalysed Na(+)-dependent alanine uptake which was inhibited by glutamine, leucine and phenylalanine. However, the clone did not catalyse Na(+)-dependent phenylalanine transport, again as in NBL-1 cells. The clone encoded a protein of 539 amino acids; the predicted transmembrane domains were almost identical in sequence to those of the other members of the B(0)/ASC transporter family. Comparison of the sequences of NBL-1 and JAR cell transporters showed some differences near the N-terminus, C-terminus and in the loop between helices 3 and 4. The NBL-1 B(0) transporter is not the same as the renal brush border membrane transporter since it does not transport phenylalanine. Differences in specificity in this protein family arise from relatively small differences in amino acid sequence.

Modulation of Starch Digestion for Slow Glucose Release through “Toggling” of Activities of Mucosal α-Glucosidases*

PubMed Central

Lee, Byung-Hoo; Eskandari, Razieh; Jones, Kyra; Reddy, Kongara Ravinder; Quezada-Calvillo, Roberto; Nichols, Buford L.; Rose, David R.; Hamaker, Bruce R.; Pinto, B. Mario

2012-01-01

Starch digestion involves the breakdown by α-amylase to small linear and branched malto-oligosaccharides, which are in turn hydrolyzed to glucose by the mucosal α-glucosidases, maltase-glucoamylase (MGAM) and sucrase-isomaltase (SI). MGAM and SI are anchored to the small intestinal brush-border epithelial cells, and each contains a catalytic N- and C-terminal subunit. All four subunits have α-1,4-exohydrolytic glucosidase activity, and the SI N-terminal subunit has an additional exo-debranching activity on the α-1,6-linkage. Inhibition of α-amylase and/or α-glucosidases is a strategy for treatment of type 2 diabetes. We illustrate here the concept of “toggling”: differential inhibition of subunits to examine more refined control of glucogenesis of the α-amylolyzed starch malto-oligosaccharides with the aim of slow glucose delivery. Recombinant MGAM and SI subunits were individually assayed with α-amylolyzed waxy corn starch, consisting mainly of maltose, maltotriose, and branched α-limit dextrins, as substrate in the presence of four different inhibitors: acarbose and three sulfonium ion compounds. The IC50 values show that the four α-glucosidase subunits could be differentially inhibited. The results support the prospect of controlling starch digestion rates to induce slow glucose release through the toggling of activities of the mucosal α-glucosidases by selective enzyme inhibition. This approach could also be used to probe associated metabolic diseases. PMID:22851177

NOX1 loss-of-function genetic variants in patients with inflammatory bowel disease

PubMed Central

Schwerd, T.; Bryant, R. V.; Pandey, S.; Capitani, M.; Meran, L.; Cazier, J.-B.; Jung, J.; Mondal, K.; Parkes, M.; Mathew, CG; Fiedler, K.; McCarthy, D. J.; Sullivan, PB; Rodrigues, A.; Travis, SPL; Moore, C.; Sambrook, J.; Ouwehand, W. H.; Roberts, D. J.; Danesh, J.; Russell, R. K.; Wilson, D. C.; Kelsen, J. R.; Cornall, R.; Denson, L. A.; Kugathasan, S.; Knaus, U. G.; Goncalves Serra, E.; Anderson, C. A.; Duerr, R. H.; McGovern, D. P. B.; Cho, J.; Powrie, F.; Li, V. S. W.; Muise, A. M.; Uhlig, H. H.

2017-01-01

Genetic defects that affect intestinal epithelial barrier function can present with very early onset inflammatory bowel disease (VEOIBD). Using whole genome sequencing, a novel hemizygous defect in NOX1 encoding NAPDH oxidase 1 was identified in a patient with ulcerative colitis-like VEOIBD. Exome screening of 1,878 paediatric patients identified further seven male IBD patients with rare NOX1 mutations. Loss-of-function was validated in p.N122H and p.T497A, and to a lesser degree in p.Y470H, p.R287Q, p.I67M, p.Q293R as well as the previously described p.P330S and the common NOX1 SNP p.D360N (rs34688635) variant. The missense mutation p.N122H abrogated reactive oxygen species (ROS) production in cell lines, ex-vivo colonic explants and patient-derived colonic organoid cultures. Within colonic crypts, NOX1 constitutively generates a high level of ROS in the crypt lumen. Analysis of 9,513 controls and 11,140 IBD patients of non-Jewish European ancestry did not reveal an association between p.D360N and IBD. Our data suggest that loss-of-function variants in NOX1 do not cause a Mendelian disorder of high penetrance but are a context specific modifier. Our results implicate that variants in NOX1 change brush border ROS within colonic crypts at the interface between the epithelium and luminal microbes. PMID:29091079

Intestinal surfactant permeation enhancers and their interaction with enterocyte cell membranes in a mucosal explant system.

PubMed

Danielsen, E Michael; Hansen, Gert H

2017-07-03

Intestinal permeation enhancers (PEs) are agents aimed to improve oral delivery of therapeutic drugs with poor bioavailability. The main permeability barrier for oral delivery is the intestinal epithelium, and PEs act to increase the paracellular and/or transcellular passage of drugs. Transcellular passage can be achieved by cell membrane permeabilization and/or by endocytic uptake and subsequent transcytosis. One broad class of PEs is surfactants which act by inserting into the cell membrane, thereby perturbing its integrity, but little is known about how the dynamics of the membrane are affected. In the present work, the interaction of the surfactants lauroyl-L-carnitine, 1-decanoyl-rac-glycerol, and nonaethylene glycol monododecyl ether with the intestinal epithelium was studied in organ cultured pig jejunal mucosal explants. As expected, at 2 mM, these agents rapidly permeabilized the enterocytes for the fluorescent polar tracer lucifer yellow, but surprisingly, they all also blocked both constitutive -and receptor-mediated pathways of endocytosis from the brush border, indicating a complete arrest of apical membrane trafficking. At the ultrastructural level, the PEs caused longitudinal fusion of brush border microvilli. Such a membrane fusogenic activity could also explain the observed formation of vesicle-like structures and large vacuoles along the lateral cell membranes of the enterocytes induced by the PEs. We conclude that the surfactant action of the PEs selected in this study not only permeabilized the enterocytes, but profoundly changed the dynamic properties of their constituent cell membranes.

Studies on the Electrical Potential Profile across Rabbit Ileum

PubMed Central

Rose, Richard C.; Schultz, Stanley G.

1971-01-01

When isolated strips of mucosal rabbit ileum are bathed by physiological electrolyte solution the electrical potential difference (PD) across the brush border (ψmc) averages 36 mv, cell interior negative. Rapid replacement of Na in the mucosal solution with less permeant cations, Tris or choline, results in an immediate hyperpolarization of ψmc. Conversely, replacement of choline in the mucosal solution with Na results in an abrupt depolarization of ψmc. These findings indicate that Na contributes to the conductance across the brush border. The presence of actively transported sugars or amino acids in the mucosal solution brings about a marked depolarization of ψmc and a smaller increase in the transmural PD (Δψms). It appears that the Na influx that is coupled to the influxes of amino acids and sugars is electrogenic and responsible for the depolarization of ψmc. Under control conditions Δψms can be attributed to the depolarization of ψmc together with the presence of a low resistance transepithelial shunt, possibly the lateral intercellular spaces. However, quantitatively similar effects of amino acids on ψmc are also seen in tissues poisoned with metabolic inhibitors or ouabain. Under these conditions Δψmc is much smaller than under control conditions. Thus, the depolarization of ψmc might not account for the entire Δψms, observed in nonpoisoned tissue. An additional electromotive force which is directly coupled to metabolic processes might contribute to the normal Δψms. PMID:5576764

Disrupted apical exocytosis of cargo vesicles causes enteropathy in FHL5 patients with Munc18-2 mutations.

PubMed

Vogel, Georg F; van Rijn, Jorik M; Krainer, Iris M; Janecke, Andreas R; Posovszky, Carsten; Cohen, Marta; Searle, Claire; Jantchou, Prevost; Escher, Johanna C; Patey, Natalie; Cutz, Ernest; Müller, Thomas; Middendorp, Sabine; Hess, Michael W; Huber, Lukas A

2017-07-20

Familial hemophagocytic lymphohistiocytosis 5 (FHL5) is an autosomal recessive disease caused by mutations in STXBP2, coding for Munc18-2, which is required for SNARE-mediated membrane fusion. FHL5 causes hematologic and gastrointestinal symptoms characterized by chronic enteropathy that is reminiscent of microvillus inclusion disease (MVID). However, the molecular pathophysiology of FHL5-associated diarrhea is poorly understood. Five FHL5 patients, including four previously unreported patients, were studied. Morphology of duodenal sections was analyzed by electron and fluorescence microscopy. Small intestinal enterocytes and organoid-derived monolayers displayed the subcellular characteristics of MVID. For the analyses of Munc18-2-dependent SNARE-protein interactions, a Munc18-2 CaCo2-KO model cell line was generated by applying CRISPR/Cas9 technology. Munc18-2 is required for Slp4a/Stx3 interaction in fusion of cargo vesicles with the apical plasma membrane. Cargo trafficking was investigated in patient biopsies, patient-derived organoids, and the genome-edited model cell line. Loss of Munc18-2 selectively disrupts trafficking of certain apical brush-border proteins (NHE3 and GLUT5), while transport of DPPIV remained unaffected. Here, we describe the molecular mechanism how the loss of function of Munc18-2 leads to cargo-selective mislocalization of brush-border components and a subapical accumulation of cargo vesicles, as it is known from the loss of polarity phenotype in MVID.

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Donowitz, M.; Emmer, E.; McCullen, J.

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 intomore » 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.« less

Lipids in the proximal convoluted tubule of the cat kidney and the reabsorption of cholesterol.

PubMed

Bargmann, W; Krisch, B; Leonhardt, H

1977-02-14

Lipid deposits in the cat kidney are mainly located in the epithelium of the proximal tubuli contorti, particularly in the pars contorta. As the amount of fatty acids in the blood of renal arteries is higher than in renal veins, the lipid inclusions are likely to be formed in the proximal convoluted tubule. Whether fat occurring in the urine has been released from the nephron epithelium and the mode of this release remains obscure. The structural equivalent of lipid extrusion into the tubules has not been observed. Components of the tubular lipids include triglycerides, phosphoglycerides and cholesterol. The results of the digitonin-cholesterol reaction favour the assumption that cholesterol is eliminated in the glomeruli and pinocytotically reabsorbed by the brush border cells, this process possibly serving recycling of this compound. The dilated basal labyrinth and intercellular space contain perpendicularly oriented lipid accumulations that reach the basal lamina. The ultrastructure of the lipid storing cells of pars contorta reacting positively for phosphoglyceride and cholesterol is characterised mainly by bodies with marginal plates. As far as can be judged from their morphology, these bodies are interpreted as large peroxisomes. A special feature of the pars recta are dumbbell shaped bodies and elongated or cup-like mitochondria concentrically surrounding cytoplasmic areas, as well as a well-developed smooth ER. In what way the organelles of the brush border cells are involved in catabolic and anabolic processes as far as renal lipid metabolism is concerned remains to be answered.

Protein-Linked Glycan Degradation in Infants Fed Human Milk

PubMed Central

Dallas, David C.; Sela, David; Underwood, Mark A.; German, J. Bruce; Lebrilla, Carlito

2014-01-01

Many human milk proteins are glycosylated. Glycosylation is important in protecting bioactive proteins and peptide fragments from digestion. Protein-linked glycans have a variety of functions; however, there is a paucity of information on protein-linked glycan degradation in either the infant or the adult digestive system. Human digestive enzymes can break down dietary disaccharides and starches, but most of the digestive enzymes required for complex protein-linked glycan degradation are absent from both human digestive secretions and the external brush border membrane of the intestinal lining. Indeed, complex carbohydrates remain intact throughout their transit through the stomach and small intestine, and are undegraded by in vitro incubation with either adult pancreatic secretions or intact intestinal brush border membranes. Human gastrointestinal bacteria, however, produce a wide variety of glycosidases with regio- and anomeric specificities matching those of protein-linked glycan structures. These bacteria degrade a wide array of complex carbohydrates including various protein-linked glycans. That bacteria possess glycan degradation capabilities, whereas the human digestive system, perse, does not, suggests that most dietary protein-linked glycan breakdown will be of bacterial origin. In addition to providing a food source for specific bacteria in the colon, protein-linked glycans from human milk may act as decoys for pathogenic bacteria to prevent invasion and infection of the host. The composition of the intestinal microbiome may be particularly important in the most vulnerable humans-the elderly, the immunocompromised, and infants (particularly premature infants). PMID:24533224

Glucocorticoid receptors in bronchial epithelial cells in asthma.

PubMed

Vachier, I; Chiappara, G; Vignola, A M; Gagliardo, R; Altieri, E; Térouanne, B; Vic, P; Bousquet, J; Godard, P; Chanez, P

1998-09-01

The expression of the glucocorticoid receptor (GR) in untreated or in steroid-dependent asthmatic patients is poorly understood. We therefore studied GR mRNA and protein levels in bronchial biopsies obtained from seven untreated asthmatic patients, seven control volunteers, and seven patients with chronic bronchitis. We also studied in bronchial epithelial cells obtained by brushing from 13 untreated asthmatics, 18 steroid-dependent asthmatics, 11 control volunteers, and 12 patients with chronic bronchitis, GR and heat shock protein 90 kD (hsp90) mRNA as well as the immunoreactivity of GR, intercellular adhesion molecule (ICAM-1), and granulocyte macrophage-colony-stimulating factor (GM-CSF). GR mRNA and protein level was similar in all subject groups in both biopsies and bronchial epithelial cells. Hsp90 mRNA level was also similar in all subject groups. ICAM-1 expression was significantly increased in bronchial epithelial cells from untreated asthmatics, but ICAM-1 was not expressed in those from steroid-dependent asthmatic patients. GM-CSF expression was significantly increased in bronchial epithelial cells from untreated and steroid-dependent asthmatic patients. GR expression within the airways is unaltered by oral long-term steroid treatment in asthma, but the expression of some but not all specific markers for asthma is modified by oral steroid.

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-βI. However, the junk-food diet also increased SGLT1 and SGLT2 levels at the proximal tubule BBM. Glucose reabsorption across the proximal tubule BBM, via GLUT2, SGLT1 and SGLT2, is not solely dependent on glycaemic status, but is also influenced by diet-induced changes in glucose metabolism. We conclude that different metabolic disturbances result in complex adaptations in renal glucose transporter protein levels and function. © 2016 The Authors. Experimental Physiology © 2016 The Physiological Society.

Iran-Iraq Border Quake Region Imaged by NASA Satellite

NASA Image and Video Library

2017-11-15

On Sunday, Nov. 12, 2017, a magnitude 7.3 earthquake struck along the Iran-Iraq border near Halabjah, Iraq. The earthquake was felt as far away as Kuwait, Qatar, Turkey, Lebanon and Israel. Extensive damage and numerous casualties were reported in the area near the epicenter (yellow star on image). The earthquake occurred along the boundary between the Arabian and Eurasian tectonic plates. This is an earthquake-prone area, and has experienced many deadly earthquakes in the past. In this perspective-view image, bright red areas are crops in fields, pale red on mountain ridges are shrubs and trees, dark gray areas are traces of earlier brush fires, and gray and tan colors are different rock types. The image was acquired Sept. 8, 2017, and the star marks the earthquake epicenter at 34.9 degrees north, 45.9 degrees east. https://photojournal.jpl.nasa.gov/catalog/PIA22112

Analysis of endoscopic brush samples identified mucosa-associated dysbiosis in inflammatory bowel disease.

PubMed

Nishino, Kyohei; Nishida, Atsushi; Inoue, Ryo; Kawada, Yuki; Ohno, Masashi; Sakai, Shigeki; Inatomi, Osamu; Bamba, Shigeki; Sugimoto, Mitsushige; Kawahara, Masahiro; Naito, Yuji; Andoh, Akira

2018-01-01

The mucosa-associated gut microbiota directly modulates epithelial and mucosal function. In this study, we investigated the mucosa-associated microbial community in patients with inflammatory bowel disease (IBD), using endoscopic brush samples. A total of 174 mucus samples from 43 patients with ulcerative colitis (UC), 26 with Crohn's disease (CD) and 14 non-IBD controls were obtained by gentle brushing of mucosal surfaces using endoscopic cytology brushes. The gut microbiome was analyzed using 16S rRNA gene sequencing. There were no significant differences in microbial structure among different anatomical sites (the ileum, cecum and sigmoid colon) within individuals. There was, however, a significant difference in microbial structure between CD, UC and non-IBD controls. The difference between CD and non-IBD controls was more marked than that between UC patients and non-IBD controls. α-Diversity was significantly lower in UC and CD patients than non-IBD controls. When comparing CD patients with non-IBD controls, the phylum Proteobacteria was significantly increased and the phyla Firmicutes and Bacteroidetes were significantly reduced. These included a significant increase in the genera Escherichia, Ruminococcus (R. gnavus), Cetobacterium, Actinobacillus and Enterococcus, and a significant decrease in the genera Faecalibacterium, Coprococcus, Prevotella and Roseburia. Comparisons between CD and UC patients revealed a greater abundance of the genera Escherichia, Ruminococcus (R. gnavus), Clostridium, Cetobacterium, Peptostreptococcus in CD patients, and the genera Faecalibacterium, Blautia, Bifidobacterium, Roseburia and Citrobacter in UC patients. Mucosa-associated dysbiosis was identified in IBD patients. CD and UC may be distinguishable from the mucosa-associated microbial community structure.

Cell tracing reveals a dorsoventral lineage restriction plane in the mouse limb bud mesenchyme.

PubMed

Arques, Carlos G; Doohan, Roisin; Sharpe, James; Torres, Miguel

2007-10-01

Regionalization of embryonic fields into independent units of growth and patterning is a widespread strategy during metazoan development. Compartments represent a particular instance of this regionalization, in which unit coherence is maintained by cell lineage restriction between adjacent regions. Lineage compartments have been described during insect and vertebrate development. Two common characteristics of the compartments described so far are their occurrence in epithelial structures and the presence of signaling regions at compartment borders. Whereas Drosophila compartmental organization represents a background subdivision of embryonic fields that is not necessarily related to anatomical structures, vertebrate compartment borders described thus far coincide with, or anticipate, anatomical or cell-type discontinuities. Here, we describe a general method for clonal analysis in the mouse and use it to determine the topology of clone distribution along the three limb axes. We identify a lineage restriction boundary at the limb mesenchyme dorsoventral border that is unrelated to any anatomical discontinuity, and whose lineage restriction border is not obviously associated with any signaling center. This restriction is the first example in vertebrates of a mechanism of primordium subdivision unrelated to anatomical boundaries. Furthermore, this is the first lineage compartment described within a mesenchymal structure in any organism, suggesting that lineage restrictions are fundamental not only for epithelial structures, but also for mesenchymal field patterning. No lineage compartmentalization was found along the proximodistal or anteroposterior axes, indicating that patterning along these axes does not involve restriction of cell dispersion at specific axial positions.

[Current concepts of digestion and absorption of carbohydrates].

PubMed

Luz, S dos S; de Campos, P L; Ribeiro, S M; Tirapegui, J

1997-01-01

The aim of this paper is to review recent aspects of digestion and absorption of carbohydrates that are the main source of energy in human diets. Recent researches have found that starch is not largely hydrolysed and absorbed in the small bowel but one part of it is resistant to digestion. Several food factors may be responsible for digestion and absorption velocity and totality of carbohydrates. Therefore, carbohydrate classification must be based not only on molecular size to express the real carbohydrates utilization as an energy source by humans. In agreement with molecular size of carbohydrate, its classification can be: a) monosaccharides; b) disaccharides; c) oligosaccharides; d) polysaccharides. In agreement with carbohydrate digestibility or availability, its classification can be: a) digestible carbohydrates; b) undigestable carbohydrates (NSP). Carbohydrate digestibility can be altered by several factors like: Intrinsic factors: a) physical structure; b) molecular physical distribution; c) physical state of food; d) food antinutrients. Extrinsics factors: a) chewing; b) transit time of food; c) amount of starch present; d) diet antinutrients. Under influence of this factors, process of digestion happen by enzymatic activity a long the gastrointestinal tract. Salivary and pancreatic amylase; glycosidases of the duodenal enterocyte brush border (lactase, sacarase and maltase), whose activity happen by close interaction of digestive breakdown with transport. The summarized pathways of the absorptive process: 1. movement from the bulk phase of the lumenal or mucosal fluid to enterocyte surface; 2. movement across the brush border membrane through specific transporters: a) SGLT1; b) GLUT 5; c) passive diffusion. 3. movement across the basolateral membrane by the GLUT 2.

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

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.

Hypoglycemic property of soy isoflavones from hypocotyl in Goto-Kakizaki diabetic rats.

PubMed

Jin, Ming; Shen, Ming-Hua; Jin, Mei-Hua; Jin, Ai-Hua; Yin, Xue-Zhe; Quan, Ji-Shu

2018-03-01

The present study was carried out to investigate the hypoglycemic effect of soy isoflavones from hypocotyl in GK diabetic rats. A single administration and long-term administration tests were conducted in GK diabetic rats to test the hypoglycemic effect of soy isoflavones. At the end of long-term administration trial, blood protein, cholesterol, triglyceride, glycosylated serum protein, C-reactive protein, insulin, aminotransferase, lipid peroxide, interleukin-6, tumor necrosis factor-α were estimated. Inhibition of soy isoflavones against α-amylase and α-glucosidase, as well as on glucose uptake into brush border membrane vesicles or Caco-2 cells were determined in vitro . In single administration experiment, soy isoflavones reduced postprandial blood glucose levels in GK rats. In long-term administration, hypoglycemic effect of soy isoflavones was first observed at week 12 and maintained till week 16. A significant reduction in fasting blood glucose, C-reactive protein, and lipid peroxide was noted at week 16. However, there was no significant treatment effect on blood insulin. Furthermore, soy isoflavone administration resulted in significant decreases in glycosylated serum protein, tumor necrosis factor-α, and interleukin-6. Other biochemical parameters, such as protein, cholesterol, triglyceride and aminotransferases were not modified, however. The results in vitro showed that soy isoflavones showed a potent inhibitory effect on intestinal α-glucosidase, but not on pancreatic α-amylase. Soy isoflavones also decreased glucose transport potency into brush border membrane vesicles or Caco-2 cells. It is concluded that soy isoflavones from hypocotyl, performs hypoglycemic function in GK rats with type 2 diabetes, maybe via suppression of carbohydrate digestion and glucose uptake in small intestine.

Impaired organic ion transport in proximal tubules of rats with Heymann nephritis.

PubMed

Park, E K; Hong, S K; Goldinger, J; Andres, G; Noble, B

1985-10-01

Organic ion transport across the basolateral membrane of proximal tubules was measured by means of the tissue slice technique in each of the four different stages of Heymann nephritis. Impairment of both organic anion and cation transport was detected early in Stage 2, and became more severe in Stage 3 of Heymann nephritis. The decreased transport function was associated with extensive damage to proximal tubule cells, including loss of brush border microvilli and basal infoldings. Despite these abnormalities of structure and function, oxygen consumption of proximal tubule cells remained essentially normal. Partial recovery of organic cation transport was noted late in Heymann nephritis (Stage 4). Recovery of the cation transport function was associated with a partial restoration of brush border microvilli and basal infoldings to proximal tubule cells. However, organic anion transport remained depressed throughout the entire course of disease. Impairment of organic ion transport in rats with Heymann nephritis appeared to result from damage to basolateral membrane transport elements rather than general deterioration of the metabolic machinery of proximal tubule cells. Decreased organic cation transport appeared to be the consequence of a reduction in the number of carrier sites, a phenomenon that could have resulted from decreased membrane surface area. However, the depression of organic anion transport was associated with decreased substrate affinity of the anion carrier, indicating that qualitative, rather than quantitative changes, were primarily responsible for that defect. Specific antibody-mediated damage to the anion transport elements in basolateral membranes of proximal tubules is postulated to occur in Heymann nephritis.

Inhibition of gentamicin binding to rat renal brush-border membrane by megalin ligands and basic peptides.

PubMed

Nagai, Junya; Saito, Masaki; Adachi, Yoshinori; Yumoto, Ryoko; Takano, Mikihisa

2006-05-01

Our previous studies showed that coadministration of cytochrome c and a 20-residue basic peptide, N-WASP181-200 (NISHTKEKKKGKAKKKRLTK, pI=10.87) inhibits renal accumulation of gentamicin. In this study, we examined effects of ligands of megalin, an endocytic receptor involved in renal uptake of gentamicin, and basic peptides including N-WASP180-200 and its mutant peptides on gentamicin binding to isolated rat renal brush-border membrane (BBM). Gentamicin binding to BBM was inhibited by megalin ligands, basic peptide fragments of cytochrome c, and N-WASP181-200 in a concentration-dependent manner. Klotz plot analysis showed that N-WASP181-200 inhibited the binding of gentamicin in a competitive manner. By substituting glycines for lysines in N-WASP181-200 at positions 9 and 15, the inhibitory effect on gentamicin binding to BBM was reduced, which may be related to a decrease in the alpha-helix content in the peptide. Gentamicin binding to BBM treated with trypsin, in which megalin completely disappeared, was significantly but not completely decreased compared with the native BBM. In addition, treatment of BBM with trypsin led to a decrease in the inhibitory effect of N-WASP181-200 on gentamicin binding. These observations support that megalin ligands and basic peptides including N-WASP181-200 decrease renal accumulation of gentamicin by inhibiting its binding to BBM of proximal tubule cells, partly interacting with megalin. In addition, the alpha-helix conformation may play an important role in the inhibitory effect of N-WASP181-200 on the binding of gentamicin to BBM.

Na+-independent D-glucose transport in rabbit renal basolateral membranes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cheung, P.T.; Hammerman, M.R.

1988-05-01

To define the mechanism by which glucose is transported across the basolateral membrane of the renal proximal tubular cell, we measured D-(14C)glucose uptake in basolateral membrane vesicles from rabbit kidney. Na+-dependent D-glucose transport, demonstrable in brush-border vesicles, could not be demonstrated in basolateral membrane vesicles. In the absence of Na+, the uptake of D-(14C)glucose in basolateral vesicles was more rapid than that of L-(3H)glucose over a concentration range of 1-50 mM. Subtraction of the latter from the former uptakes revealed a saturable process with apparent Km of 9.9 mM and Vmax of 0.80 nmol.mg protein-1.s-1. To characterize the transport componentmore » of D-glucose uptake in basolateral vesicles, we measured trans stimulation of 2 mM D-(14C)glucose entry in the absence of Na+. Trans stimulation could be effected by preloading basolateral vesicles with D-glucose, 2-deoxy-D-glucose, or 3-O-methyl-D-glucose, but not with L-glucose or alpha-methyl-D-glucoside. Trans-stimulated D-(14C)glucose uptake was inhibited by 0.1 mM phloretin or cytochalasin B but not phlorizin. In contrast, Na+-dependent D-(14C)glucose transport in brush-border vesicles was inhibited by phlorizin but not phloretin or cytochalasin B. Our findings are consistent with the presence of a Na+-independent D-glucose transporter in the proximal tubular basolateral membrane with characteristics similar to those of transporters present in nonepithelial cells.« less

Cleistanthus collinus induces type I distal renal tubular acidosis and type II respiratory failure in rats.

PubMed

Maneksh, Delinda; Sidharthan, Anita; Kettimuthu, Kavithapriya; Kanthakumar, Praghalathan; Lourthuraj, Amala A; Ramachandran, Anup; Subramani, Sathya

2010-06-01

A water decoction of the poisonous shrub Cleistanthus collinus is used for suicidal purposes. The mortality rate is 28%. The clinical profile includes distal renal tubular acidosis (DRTA) and respiratory failure. The mechanism of toxicity is unclear. To demonstrate features of C. collinus toxicity in a rat model and to identify its mechanism(s) of action. Rats were anesthetized and the carotid artery was cannulated. Electrocardiogram and respiratory movements were recorded. Either aqueous extract of C. collinus or control solution was administered intraperitoneally. Serial measurements of blood gases, electrolytes and urinary pH were made. Isolated brush border and basolateral membranes from rat kidney were incubated with C. collinus extract and reduction in ATPase activity was assessed. Venous blood samples from human volunteers and rats were incubated with an acetone extract of C. collinus and plasma potassium was estimated as an assay for sodium-potassium pump activity. The mortality was 100% in tests and 17% in controls. Terminal event in test animals was respiratory arrest. Controls had metabolic acidosis, respiratory compensation acidic urine and hyperkalemia. Test animals showed respiratory acidosis, alkaline urine and low blood potassium as compared to controls. C. collinus extract inhibited ATPase activity in rat kidney. Plasma K(+) did not increase in human blood incubated with C. collinus extract. Active principles of C. collinus inhibit proton pumps in the renal brush border, resulting in type I DRTA in rats. There is no inhibition of sodium-potassium pump activity. Test animals develop respiratory acidosis, and the immediate cause of death is respiratory arrest.

Design and development of hyaluronan-functionalized polybenzofulvene nanoparticles as CD44 receptor mediated drug delivery system

NASA Astrophysics Data System (ADS)

Licciardi, Mariano; Scialabba, Cinzia; Giammona, Gaetano; Paolino, Marco; Razzano, Vincenzo; Grisci, Giorgio; Giuliani, Germano; Makovec, Francesco; Cappelli, Andrea

2017-06-01

A tri-component polymer brush (TCPB ), composed of a polybenzofulvene copolymer bearing low molecular weight hyaluronic acid (HA) on the surface of its cylindrical brush-like backbone and oligo-PEG fractions, was employed in the preparation of 350 nm nanostructured drug delivery systems capable of delivering the anticancer drug doxorubicin. The obtained drug delivery systems were characterized on the basis of drug loading and release, dimensions and zeta potential, morphology and in vitro cell activity, and uptake on three different human cell lines, namely the bronchial epithelial 16HBE, the breast adenocarcinoma MCF-7, and the colon cancer HCT116 cells. Finally, the ability of doxorubicin-loaded TCPB nanoparticles (DOXO-TCPB) to be internalized into cancer cells by CD44 receptor mediated uptake was assessed by means of uptake studies in HCT cells. These data were supported by anti-CD44-FITC staining assay. The proposed TCPB nanostructured drug delivery systems have many potential applications in nanomedicine, including cancer targeted drug delivery.

Carbachol regulation of rabbit ileal brush border Na+–H+ exchanger 3 (NHE3) occurs through changes in NHE3 trafficking and complex formation and is Src dependent

PubMed Central

Li, Xuhang; Zhang, Huiping; Cheong, Alice; Yueping Chen, Sharon Leu; Elowsky, Christian G; Donowitz, Mark

2004-01-01

The epithelial brush border membrane (BBM) Na+–H+ exchanger 3 (NHE3) is the major transport protein responsible for ileal electroneutral Na+ absorption. We have previously shown that ileal BBM NHE3 activity is rapidly inhibited by carbachol, an agonist that mimics cholinergic activation in digestion. In this study, we investigated the mechanisms involved in this NHE3 inhibition. Carbachol decreased the amount of ileal Na+ absorptive cell BBM NHE3 within 10 min of exposure. Based on OptiPrep gradient centrifugation, carbachol increased the amount of NHE3 in early endosomes and decreased the amount of NHE3 in BBM, consistent with effects on NHE3 trafficking. The decrease in BBM NHE3 occurred in the detergent-soluble BBM fraction with no change in the amount of NHE3 in the BBM detergent-resistant membranes. The size of BBM NHE3 complexes increased in carbachol-exposed ileum, as studied with sucrose gradient centrifugation. The NHE3 complex size increased in the total BBM, but did not change in the detergent-soluble fraction. This suggests that carbachol treatment enhanced the association of proteins with NHE3 complexes specifically in the detergent-resistant fraction of ileal BBM. NHERF2, α-actinin-4 and protein kinase C were among those NHE3-associated proteins because they were more efficiently coimmunoprecipitated from total BBM after carbachol treatment. Moreover, Src was involved in the carbachol-mediated inhibition since: (1) c-Src was rapidly activated in the detergent-resistant membranes by carbachol; and (2) carbachol inhibition of ileal Na+ absorption was completely abolished by the Src family inhibitor 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2). Moreover, the carbachol-induced increase in the size of NHE3-containing complexes was reversed by PP2. These data demonstrate that regulation of NHE3 activity by carbachol can be achieved at several interrelated levels: (1) the subcellular level, at which NHE3 is rapidly endocytosed from BBM to endocytic vesicles upon treatment with carbachol; (2) multiple BBM pools, in which carbachol selectively decreases the amount of NHE3 in the BBM detergent-soluble fraction but not the detergent-resistant membrane; and (3) the molecular level, at which NHE3 complex-associated proteins can be changed upon carbachol treatment, with carbachol leading to larger BBM NHE3 complexes and increased co-IP of NHERF2 with α-actinin-4 and activated PKC. The study further describes NHE3 presence simultaneously in multiple dynamic BBM pools in which NHE3 distribution and associated proteins are altered as part of carbachol-induced and Src-mediated rapid signal transduction, which decreases the amount of BBM NHE3 and thus inhibits NHE3 activity. PMID:14978207

Intestinal Alkaline Phosphatase: Potential Roles in Promoting Gut Health in Weanling Piglets and Its Modulation by Feed Additives - A Review.

PubMed

Melo, A D B; Silveira, H; Luciano, F B; Andrade, C; Costa, L B; Rostagno, M H

2016-01-01

The intestinal environment plays a critical role in maintaining swine health. Many factors such as diet, microbiota, and host intestinal immune response influence the intestinal environment. Intestinal alkaline phosphatase (IAP) is an important apical brush border enzyme that is influenced by these factors. IAP dephosphorylates bacterial lipopolysaccharides (LPS), unmethylated cytosine-guanosine dinucleotides, and flagellin, reducing bacterial toxicity and consequently regulating toll-like receptors (TLRs) activation and inflammation. It also desphosphorylates extracellular nucleotides such as uridine diphosphate and adenosine triphosphate, consequently reducing inflammation, modulating, and preserving the homeostasis of the intestinal microbiota. The apical localization of IAP on the epithelial surface reveals its role on LPS (from luminal bacteria) detoxification. As the expression of IAP is reported to be downregulated in piglets at weaning, LPS from commensal and pathogenic gram-negative bacteria could increase inflammatory processes by TLR-4 activation, increasing diarrhea events during this phase. Although some studies had reported potential IAP roles to promote gut health, investigations about exogenous IAP effects or feed additives modulating IAP expression and activity yet are necessary. However, we discussed in this paper that the critical assessment reported can suggest that exogenous IAP or feed additives that could increase its expression could show beneficial effects to reduce diarrhea events during the post weaning phase. Therefore, the main goals of this review are to discuss IAP's role in intestinal inflammatory processes and present feed additives used as growth promoters that may modulate IAP expression and activity to promote gut health in piglets.

Role of Tir and Intimin in the Virulence of Rabbit Enteropathogenic Escherichia coli Serotype O103:H2

PubMed Central

Marchès, Olivier; Nougayrède, Jean-Philippe; Boullier, Séverine; Mainil, Jacques; Charlier, Gérard; Raymond, Isabelle; Pohl, Pierre; Boury, Michèle; De Rycke, Jean; Milon, Alain; Oswald, Eric

2000-01-01

Attaching and effacing (A/E) rabbit enteropathogenic Escherichia coli (REPEC) strains belonging to serogroup O103 are an important cause of diarrhea in weaned rabbits. Like human EPEC strains, they possess the locus of enterocyte effacement clustering the genes involved in the formation of the A/E lesions. In addition, pathogenic REPEC O103 strains produce an Esp-dependent but Eae (intimin)-independent alteration of the host cell cytoskeleton characterized by the formation of focal adhesion complexes and the reorganization of the actin cytoskeleton into bundles of stress fibers. To investigate the role of intimin and its translocated coreceptor (Tir) in the pathogenicity of REPEC, we have used a newly constructed isogenic tir null mutant together with a previously described eae null mutant. When human HeLa epithelial cells were infected, the tir mutant was still able to induce the formation of stress fibers as previously reported for the eae null mutant. When the rabbit epithelial cell line RK13 was used, REPEC O103 produced a classical fluorescent actin staining (FAS) effect, whereas both the eae and tir mutants were FAS negative. In a rabbit ligated ileal loop model, neither mutant was able to induce A/E lesions. In contrast to the parental strain, which intimately adhered to the enterocytes and destroyed the brush border microvilli, bacteria of both mutants were clustered in the mucus without reaching and damaging the microvilli. The role of intimin and Tir was then analyzed in vivo by oral inoculation of weaned rabbits. Although both mutants were still present in the intestinal flora of the rabbits 3 weeks after oral inoculation, neither mutant strain induced any clinical signs or significant weight loss in the inoculated rabbits whereas the parental strain caused the death of 90% of the inoculated rabbits. Nevertheless, an inflammatory infiltrate was present in the lamina propria of the rabbits infected with both mutants, with an inflammatory response greater for the eae null mutant. In conclusion, we have confirmed the role of intimin in virulence, and we have shown, for the first time, that Tir is also a key factor in vivo for pathogenicity. PMID:10722617

Role of tir and intimin in the virulence of rabbit enteropathogenic Escherichia coli serotype O103:H2.

PubMed

Marchès, O; Nougayrède, J P; Boullier, S; Mainil, J; Charlier, G; Raymond, I; Pohl, P; Boury, M; De Rycke, J; Milon, A; Oswald, E

2000-04-01

Attaching and effacing (A/E) rabbit enteropathogenic Escherichia coli (REPEC) strains belonging to serogroup O103 are an important cause of diarrhea in weaned rabbits. Like human EPEC strains, they possess the locus of enterocyte effacement clustering the genes involved in the formation of the A/E lesions. In addition, pathogenic REPEC O103 strains produce an Esp-dependent but Eae (intimin)-independent alteration of the host cell cytoskeleton characterized by the formation of focal adhesion complexes and the reorganization of the actin cytoskeleton into bundles of stress fibers. To investigate the role of intimin and its translocated coreceptor (Tir) in the pathogenicity of REPEC, we have used a newly constructed isogenic tir null mutant together with a previously described eae null mutant. When human HeLa epithelial cells were infected, the tir mutant was still able to induce the formation of stress fibers as previously reported for the eae null mutant. When the rabbit epithelial cell line RK13 was used, REPEC O103 produced a classical fluorescent actin staining (FAS) effect, whereas both the eae and tir mutants were FAS negative. In a rabbit ligated ileal loop model, neither mutant was able to induce A/E lesions. In contrast to the parental strain, which intimately adhered to the enterocytes and destroyed the brush border microvilli, bacteria of both mutants were clustered in the mucus without reaching and damaging the microvilli. The role of intimin and Tir was then analyzed in vivo by oral inoculation of weaned rabbits. Although both mutants were still present in the intestinal flora of the rabbits 3 weeks after oral inoculation, neither mutant strain induced any clinical signs or significant weight loss in the inoculated rabbits whereas the parental strain caused the death of 90% of the inoculated rabbits. Nevertheless, an inflammatory infiltrate was present in the lamina propria of the rabbits infected with both mutants, with an inflammatory response greater for the eae null mutant. In conclusion, we have confirmed the role of intimin in virulence, and we have shown, for the first time, that Tir is also a key factor in vivo for pathogenicity.

Early Biochemical Effects of an Organic Mercury Fungicide on Infants: ``Dose Makes the Poison''

NASA Astrophysics Data System (ADS)

Gotelli, Carlos A.; Astolfi, Emilio; Cox, Christopher; Cernichiari, Elsa; Clarkson, Thomas W.

1985-02-01

Phenylmercury absorbed through the skin from contaminated diapers affected urinary excretion in infants in Buenos Aires. The effects were reversible and quantitatively related to the concentration of urinary mercury. Excretion of γ -glutamyl transpeptidase, an enzyme in the brush borders of renal tubular cells, increased in a dose-dependent manner when mercury excretion exceeded a ``threshold'' value. Urine volume also increased but at a higher threshold with respect to mercury. The results support the threshold concept of the systemic toxicity of metals. γ -Glutamyl transpeptidase is a useful and sensitive marker for preclinical effects of toxic metals.

Cigarette smoke suppresses Bik to cause epithelial cell hyperplasia and mucous cell metaplasia.

PubMed

Mebratu, Yohannes A; Schwalm, Kurt; Smith, Kevin R; Schuyler, Mark; Tesfaigzi, Yohannes

2011-06-01

Aberrant regulation of airway epithelial cell numbers in airways leads to increased mucous secretions in chronic lung diseases such as chronic bronchitis. Because the Bcl-2 family of proteins is crucial for airway epithelial homeostasis, identifying the players that reduce cigarette smoke (CS)-induced mucous cell metaplasia can help to develop effective therapies. To identify the Bcl-2 family of proteins that play a role in reducing CS-induced mucous cell metaplasia. We screened for dysregulated expression of the Bcl-2 family members. We identified Bik to be significantly reduced in bronchial brushings of patients with chronic epithelial cell hyperplasia compared with nondiseased control subjects. Reduced Bik but increased MUC5AC mRNA levels were also detected when normal human airway epithelial cells (HAECs) were exposed to CS or when autopsy tissues from former smokers with and without chronic bronchitis were compared. Similarly, exposure of C57Bl/6 mice to CS resulted in increased numbers of epithelial and mucous cells per millimeter of basal lamina, along with reduced Bik but increased Muc5ac expression, and this change was sustained even when mice were allowed to recover in filtered air for 8 weeks. Restoring Bik expression significantly suppressed CS-induced mucous cell metaplasia in differentiated primary HAEC cultures and in airways of mice in vivo. Bik blocked nuclear translocation of phospho-ERK1/2 to induce apoptosis of HAECs. The conserved Leu61 within Bik and ERK1/2 activation were essential to induce cell death in hyperplastic mucous cells. These studies show that CS suppresses Bik expression to block airway epithelia cell death and thereby increases epithelial cell hyperplasia in chronic bronchitis.

Cigarette Smoke Suppresses Bik To Cause Epithelial Cell Hyperplasia and Mucous Cell Metaplasia

PubMed Central

Mebratu, Yohannes A.; Schwalm, Kurt; Smith, Kevin R.; Schuyler, Mark; Tesfaigzi, Yohannes

2011-01-01

Rationale: Aberrant regulation of airway epithelial cell numbers in airways leads to increased mucous secretions in chronic lung diseases such as chronic bronchitis. Because the Bcl-2 family of proteins is crucial for airway epithelial homeostasis, identifying the players that reduce cigarette smoke (CS)-induced mucous cell metaplasia can help to develop effective therapies. Objectives: To identify the Bcl-2 family of proteins that play a role in reducing CS-induced mucous cell metaplasia. Methods: We screened for dysregulated expression of the Bcl-2 family members. Measurements and Main Results: We identified Bik to be significantly reduced in bronchial brushings of patients with chronic epithelial cell hyperplasia compared with nondiseased control subjects. Reduced Bik but increased MUC5AC mRNA levels were also detected when normal human airway epithelial cells (HAECs) were exposed to CS or when autopsy tissues from former smokers with and without chronic bronchitis were compared. Similarly, exposure of C57Bl/6 mice to CS resulted in increased numbers of epithelial and mucous cells per millimeter of basal lamina, along with reduced Bik but increased Muc5ac expression, and this change was sustained even when mice were allowed to recover in filtered air for 8 weeks. Restoring Bik expression significantly suppressed CS-induced mucous cell metaplasia in differentiated primary HAEC cultures and in airways of mice in vivo. Bik blocked nuclear translocation of phospho-ERK1/2 to induce apoptosis of HAECs. The conserved Leu61 within Bik and ERK1/2 activation were essential to induce cell death in hyperplastic mucous cells. Conclusions: These studies show that CS suppresses Bik expression to block airway epithelia cell death and thereby increases epithelial cell hyperplasia in chronic bronchitis. PMID:21317312

Cloning of oligopeptide transport carrier PepT1 and comparative analysis of PepT1 messenger ribonucleic acid expression in response to dietary nitrogen levels in yak () and indigenous cattle () on the Qinghai-Tibetan plateau.

PubMed

Wang, H C; Shi, F Y; Hou, M J; Fu, X Y; Long, R J

2016-08-01

The gastrointestinal lumen can directly absorb all di- and tripeptide protein degradation products, and oligopeptide absorption depends on the specific peptide transport carriers, which are located in gastrointestinal epithelial cells on the brush border membrane. Yak () use N more efficiently than cattle do, which implies that yak have a specific mechanism of nonprotein utilization including a peptide absorption mechanism. However, this mechanism has not been clarified. Our objective was to explore whether yak possess any adaptive mechanisms of peptide absorption to survive in the harsh foraging environment of the Qinghai-Tibetan plateau. Twelve castrated males of each of 2 genotypes, yak () and indigenous cattle (), were fed diets of various N levels. The yak PepT1 (yPepT1) cDNA was cloned in omasum epithelial tissue. Our results showed that the full-length yPepT1 cDNA contains 2,805 bp, and a 2,121-bp open reading frame encodes a putative protein of 707 AA residues. The yPepT1 AA sequence identified 5 putative extracellular N-glycosylation sites (Asn, Asn, Asn, Asn, and Asn), 2 putative intracellular protein kinase A sites (Ser and Thr), and 3 intracellular putative protein kinase C sites (Ser, Ser, and Ser). The yPepT1 AA sequence was 99, 95, 86, and 83% identical to PepT1 from cattle (), sheep (), pigs (), and humans (), respectively. The relative PepT1 mRNA expression for indigenous cattle was greater than yak in the rumen, omasum, duodenum, ileum, and liver ( < 0.001); however, it was lower in jejunum tissue ( < 0.01). The relative PepT1 mRNA expression in response to increasing dietary N for both genotypes were linear in the rumen and jejunum ( < 0.10); quadratic or cubic in the reticulum ( < 0.01); linear or quadratic in the duodenum, ileum, and liver ( ≤ 0.01); and linear, quadratic, or cubic in the omasum ( < 0.001). Moreover, there were significant interactions between genotype and dietary N in rumen, reticulum, omasum, duodenum, jejunum, ileum, and liver tissues. In conclusion, the PepT1 profile and expression in gastrointestinal epithelial cells of yak varied from those of cattle, implying that yak have evolved a peptide transport mechanism to adapt the environment of the Qinghai-Tibetan plateau.

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

The effect of big endothelin-1 in the proximal tubule of the rat kidney

PubMed Central

Beara-Lasić, Lada; Knotek, Mladen; Čejvan, Kenan; Jakšić, Ozren; Lasić, Zoran; Skorić, Boško; Brkljačić, Vera; Banfić, Hrvoje

1997-01-01

An obligatory step in the biosynthesis of endothelin-1 (ET-1) is the conversion of its inactive precursor, big ET-1, into the mature form by the action of specific, phosphoramidon-sensitive, endothelin converting enzyme(s) (ECE). Disparate effects of big ET-1 and ET-1 on renal tubule function suggest that big ET-1 might directly influence renal tubule function. Therefore, the role of the enzymatic conversion of big ET-1 into ET-1 in eliciting the functional response (generation of 1,2-diacylglycerol) to big ET-1 was studied in the rat proximal tubules.In renal cortical slices incubated with big ET-1, pretreatment with phosphoramidon (an ECE inhibitor) reduced tissue immunoreactive ET-1 to a level similar to that of cortical tissue not exposed to big ET-1. This confirms the presence and effectiveness of ECE inhibition by phosphoramidon.In freshly isolated proximal tubule cells, big ET-1 stimulated the generation of 1,2-diacylglycerol (DAG) in a time- and dose-dependent manner. Neither phosphoramidon nor chymostatin, a chymase inhibitor, influenced the generation of DAG evoked by big ET-1.Big ET-1-dependent synthesis of DAG was found in the brush-border membrane. It was unaffected by BQ123, an ETA receptor antagonist, but was blocked by bosentan, an ETA,B-nonselective endothelin receptor antagonist.These results suggest that the proximal tubule is a site for the direct effect of big ET-1 in the rat kidney. The effect of big ET-1 is confined to the brush-border membrane of the proximal tubule, which may be the site of big ET-1-sensitive receptors. PMID:9051300

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

Activation of intestinal brush border guanylate cyclase by aromatic disulphide compounds.

PubMed Central

elDeib, M M; Parker, C D; White, A A

1991-01-01

Guanylate cyclase in pig intestinal brush border membranes was stimulated by certain aromatic disulphides. The most effective were 6-thioguanine disulphide [(TGS)2], 6-mercaptopurine disulphide, 6,6'-dithiodinicotinic acid, 5,5'-dithiobis-(2-nitrobenzoic acid) and 5-carboxy-2-thiouracil disulphide. (TGS)2 stimulated activity 15-fold when present at 0.1 mM. The optimum concentration for each disulphide was different, and higher concentrations were inhibitory. There was no activation by alkyl disulphides or by N-ethylmaleimide. Activation by 50 microM-(TGS)2 was partially reversed by later addition of 0.1 mM-dithiothreitol, whereas activation by the Escherichia coli heat-stable enterotoxin STa was relatively unaffected. Pretreatment of the membranes with (TGS)2 produced a concentration-dependent inhibition of STa-stimulated activity, while stimulating basal activity, until the activities were equal at 50 microM. Activity was [Mg2+]-dependent, the optimal [Mg2+] progressively increasing as the enzyme was stimulated by (TGS)2, STa and Lubrol PX respectively. However, (TGS)2 pretreatment prevented the shift to higher [Mg2+]optima induced by STa or Lubrol alone. Substitution of Mn2+ for Mg2+ in the reaction elevated basal activity and eliminated by activation (TGS)2. (TGS)2 only inhibited Mn2(+)-dependent activity (both basal and stimulated). The affinity of 125I-STa for its receptor was slightly increased by (TGS)2. We propose that (TGS)2 undergoes thiol-disulphide exchange with at least three different protein thiols of decreasing reactivity. The first is associated with Mg2(+)-dependent activation, the second is associated with a tonic inhibition of activity and the third is associated with the catalytic activity, although probably not at the active site. PMID:1673335

Effect of proinflammatory interleukins on jejunal nutrient transport

PubMed Central

Hardin, J; Kroeker, K; Chung, B; Gall, D

2000-01-01

AIM—We examined the effect of proinflammatory and anti-inflammatory interleukins on jejunal nutrient transport and expression of the sodium-glucose linked cotransporter (SGLT-1).
METHODS—3-O-methyl glucose and L-proline transport rates were examined in New Zealand White rabbit stripped, short circuited jejunal tissue. The effects of the proinflammatory cytokines interleukin (IL)-1α, IL-6, and IL-8, IL-1α plus the specific IL-1 antagonist, IL-1ra, and the anti-inflammatory cytokine IL-10 were investigated. In separate experiments, passive tissue permeability was assessed and brush border SGLT-1 expression was measured by western blot in tissues exposed to proinflammatory interleukins.
RESULTS—The proinflammatory interleukins IL-6, IL-1α, and IL-8 significantly increased glucose absorption compared with control levels. This increase in glucose absorption was due to an increase in mucosal to serosal flux. IL-1α and IL-8 also significantly increased L-proline absorption due to an increase in absorptive flux. The anti-inflammatory IL-10 had no effect on glucose transport. The receptor antagonist IL-1ra blocked the ability of IL-1α to stimulate glucose transport. IL-8 had no effect on passive tissue permeability. SGLT-1 content did not differ in brush border membrane vesicles (BBMV) from control or interleukin treated tissue.
CONCLUSIONS—These findings suggest that intestinal inflammation and release of inflammatory mediators such as interleukins increase nutrient absorption in the gut. The increase in glucose transport does not appear to be due to changes in BBMV SGLT-1 content.


Keywords: glucose transport; small intestine; intestinal inflammation; inflammation PMID:10896908

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

Food grade titanium dioxide disrupts intestinal brush border microvilli in vitro independent of sedimentation.

PubMed

Faust, James J; Doudrick, Kyle; Yang, Yu; Westerhoff, Paul; Capco, David G

2014-06-01

Bulk- and nano-scale titanium dioxide (TiO2) has found use in human food products for controlling color, texture, and moisture. Once ingested, and because of their small size, nano-scale TiO2 can interact with a number of epithelia that line the human gastrointestinal tract. One such epithelium responsible for nutrient absorption is the small intestine, whose constituent cells contain microvilli to increase the total surface area of the gut. Using a combination of scanning and transmission electron microscopy it was found that food grade TiO2 (E171 food additive coded) included ∼25% of the TiO2 as nanoparticles (NPs; <100 nm), and disrupted the normal organization of the microvilli as a consequence of TiO2 sedimentation. It was found that TiO2 isolated from the candy coating of chewing gum and a commercially available TiO2 food grade additive samples were of the anatase crystal structure. Exposure to food grade TiO2 additives, containing nanoparticles, at the lowest concentration tested within this experimental paradigm to date at 350 ng/mL (i.e., 100 ng/cm(2) cell surface area) resulted in disruption of the brush border. Through the use of two independent techniques to remove the effects of gravity, and subsequent TiO2 sedimentation, it was found that disruption of the microvilli was independent of sedimentation. These data indicate that food grade TiO2 exposure resulted in the loss of microvilli from the Caco-2BBe1 cell system due to a biological response, and not simply a physical artifact of in vitro exposure.

Sulfate-bicarbonate exchange in brush-border membranes from rat renal cortex

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pritchard, J.B.

1987-02-01

Under Na/sup +/-free conditions /sup 35/SO/sub 4//sup 2 -/ uptake by rat renal brush-border membrane (BBM) vesicles could be driven by imposition of a HCO/sup -//sub 3/ gradient (in greater than out). The initial rate of /sup 35/SO/sub 4//sup 2 -/ uptake was stimulated 10-fold, and peak overshoot exceeded equilibrium uptake by 2-3 times. Cl/sup -/, SCN/sup -/, NO/sub 3//sup -/, I/sup -/, and OH/sup -/ were able to substitute for HCO/sub 3//sup -/. Divalent anions, including /sup 35/SO/sub 4//sup 2 -/ itself, were less effective as counterions. HCO/sub 3//sup -/-SO/sub 4//sup 2 -/ exchange was cis-inhibited by disulfonic stilbenes,more » ((SITS)(DIDS)), phloretin, Hg, and S/sub 2/O/sub 3//sup 2 -/. HCO/sub 3//sup -/-driven /sup 35/SO/sub 4//sup 2 -/ uptake was saturable, with an apparent K/sub m/ of 0.4 mM for SO/sub 4//sup 2 -/. Simultaneous imposition of Na/sup +/ and HCO/sub 3//sup -/ gradients produced approximately additive stimulation of /sup 35/SO/sub 4//sup 2 -/ uptake. The HCO/sub 3//sup -/-driven component of /sup 35/SO/sub 4//sup 2 -/ uptake, but not the component driven by Na/sup +/, was inhibited by SITS. Finally, Na/sup +/-driven SO/sub 4//sup 2 -/ accumulation could be reduced by imposing an out greater than in HCO/sub 3//sup +/ gradient, conditions accelerating exchange driven SO/sub 4//sup 2 -/ efflux. These findings indicate the presence of separate Na/sup +/-SO/sub 4//sup 2 -/ cotransport and SO /sub 4//sup 2 -/-anion exchange pathways in the same BBM vesicles.« less

Identification of a New cry1I-Type Gene as a Candidate for Gene Pyramiding in Corn To Control Ostrinia Species Larvae

PubMed Central

Zhao, Can; Abdelgaffar, Heba M.; Pan, Hongyu; Song, Fuping

2015-01-01

Pyramiding of diverse cry toxin genes from Bacillus thuringiensis with different modes of action is a desirable strategy to delay the evolution of resistance in the European corn borer (Ostrinia nubilalis). Considering the dependency of susceptibility to Cry toxins on toxin binding to receptors in the midgut of target pests, a diverse mode of action is commonly defined as recognition of unique binding sites in the target insect. In this study, we present a novel cry1Ie toxin gene (cry1Ie2) as a candidate for pyramiding with Cry1Ab or Cry1Fa in corn to control Ostrinia species larvae. The new toxin gene encodes an 81-kDa protein that is processed to a protease-resistant core form of approximately 55 kDa by trypsin digestion. The purified protoxin displayed high toxicity to Ostrinia furnacalis and O. nubilalis larvae but low to no activity against Spodoptera or heliothine species or the coleopteran Tenebrio molitor. Results of binding assays with 125I-labeled Cry1Ab toxin and brush border membrane vesicles from O. nubilalis larvae demonstrated that Cry1Ie2 does not recognize the Cry1Ab binding sites in that insect. Reciprocal competition binding assays with biotin-labeled Cry1Ie2 confirmed the lack of shared sites with Cry1Ab or Cry1Fa in O. nubilalis brush border membrane vesicles. These data support Cry1Ie2 as a good candidate for pyramiding with Cry1Ab or Cry1Fa in corn to increase the control of O. nubilalis and reduce the risk of resistance evolution. PMID:25795679

Structural and functional development of small intestine in intrauterine growth retarded porcine offspring born to gilts fed diets with differing protein ratios throughout pregnancy.

PubMed

Mickiewicz, M; Zabielski, R; Grenier, B; Le Normand, L; Savary, G; Holst, J J; Oswald, I P; Metges, C C; Guilloteau, P

2012-06-01

Protein level in the maternal diet plays a crucial role in fetal programming during pregnancy. Low or high protein level increases the risk of intrauterine growth retardation (IUGR). The aim of this study was to investigate the structural and functional development of the small intestine in piglets from sows fed a control (C, 12.1% protein), a high protein (HP, 30% protein), or a low protein (LP, 6.5% protein) diet during pregnancy. Newborns were classified as IUGR (birth weight ≤1.18 kg) and non-IUGR (birth weight >1.18 kg). The piglets were euthanized on postnatal day (PD)1, PD28 and PD188. The LP diet in non-IUGR neonates resulted in decreased body weight on PD1. The LP and HP diets resulted in both decreased body weight and delayed catch-up growth in the IUGR piglets. The HP and LP-diets increased the length of villi on PD1 in non-IUGRs but not in IUGRs. At birth, the expressions of Ki67 and active caspase 3 in mid-jejunum epithelium of HP and LP non-IUGR neonates were significantly lower as compared to C non-IUGRs whilst in IUGRs the respective expressions were as high as in C non-IUGRs. The postnatal dynamics of brush border enzyme activities and vacuolated enterocytes disappearance showed significant drop in enterocyte maturation in IUGR as compared to non-IUGR neonates. In conclusion, both HP and LP diets led to retarded development of non-IUGR piglets. In IUGR piglets both HP and LP diets resulted in delayed catch-up growth, without adaptive changes in brush border digestive enzymes.

Effect of dietary fatty acids on jejunal and ileal oleic acid uptake by rat brush border membrane vesicles.

PubMed

Prieto, R M; Stremmel, W; Sales, C; Tur, J A

1996-04-18

To test the effect of dietary fatty acids on fatty acid uptake, the influx kinetics of a representative long-chain fatty acid, 3H-oleic acid, in both the jejunum and ileum of rats has been studied using brush border membrane vesicles (BBMV). Animals were fed with semipurified diets containing 5 g fat/100 g diet, as corn oil (control group), safflower oil (unsaturated group) and coconut oil hydrogenated (saturated group). With increasing unbound oleate concentration in the medium, the three dietary groups showed saturable kinetics in both jejunal and ileal BBMV (controls: Vmax = 0.15 +/- 0.01 nmol x mg protein-1 x 5 min-1 and Km = 136 +/- 29.1 nmol for jejunum, and Vmax = 0.23 +/- 0.03 nmol x mg protein-1 x 5 min-1 and Km = 196 +/- 50.3 nmol for ileum; unsaturated: Vmax = 0.28 +/- 0.05 nmol x mg protein-1 x 5 min-1 and Km = 242.7 +/- 91.8 nmol for jejunum, and Vmax = 1.29 +/- 0.06 nmol x mg protein-1 x 5 min-1 and Km = 509.8 +/- 97.5 nmol for ileum; saturated: Vmax = 0.03 +/- 0.01 nmol x mg protein-1 x 5 min-1 and Km = 124.5 +/- 72.6 nmol for jejunum, and Vmax = 0.04 +/- 0.01 nmol x mg protein -1.5 min-1 and Km = 205.6 +/- 85.3 nmol for ileum). These results support the theory that feeding an isocaloric diet containing only unsaturated fatty acids enhanced oleic acid uptake, and feeding an isocaloric diet containing only saturated fatty acids decreased oleic acid uptake. The results obtained in the present work also show the adaptative ability of jejunum and ileum to the type of dietary fat.

Oral administration of thymoquinone mitigates the effect of cisplatin on brush border membrane enzymes, energy metabolism and antioxidant system in rat intestine.

PubMed

Shahid, Faaiza; Farooqui, Zeba; Abidi, Subuhi; Parwez, Iqbal; Khan, Farah

2017-10-01

Cisplatin (CP) is a widely used chemotherapeutic agent that elicits severe gastrointestinal toxicity. Nigella sativa, a member of family Ranunculaceae, is one of the most revered medicinal plant known for its numerous health benefits. Thymoquinone (TQ), a major bioactive component derived from the volatile oil of Nigella sativa seeds, has been shown to improve gastrointestinal functions in animal models of acute gastric/intestinal injury. In view of this, the aim of the present study was to investigate the protective effect of TQ on CP induced toxicity in rat intestine and to elucidate the mechanism underlying these effects. Rats were divided into four groups viz. control, CP, TQ and CP+TQ. Animals in CP+TQ and TQ groups were orally administered TQ (1.5mg/kg bwt) with and without a single intraperitoneal dose of CP (6mg/kg bwt) respectively. The effect of TQ was determined on CP induced alterations in the activities of brush border membrane (BBM), carbohydrate metabolism, and antioxidant defense enzymes in rat intestine. TQ administration significantly mitigated CP induced decline in the specific activities of BBM marker enzymes, both in the mucosal homogenates and in the BBM vesicles (BBMV) prepared from intestinal mucosa. Furthermore, TQ administration restored the redox and metabolic status of intestinal mucosal tissue in CP treated rats. The biochemical results were supported by histopathological findings that showed extensive damage to intestine in CP treated rats and markedly preserved intestinal histoarchitecture in CP and TQ co-treated group. The biochemical and histological data suggest a protective effect of TQ against CP-induced gastrointestinal damage. Thus, TQ may have a potential for clinical application to counteract the accompanying gastrointestinal toxicity in CP chemotherapy. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Studies on the uptake of fatty acids by brush border membranes of the rabbit intestine.

PubMed

Proulx, P; Aubry, H; Brglez, I; Williamson, D G

1985-04-01

Initial studies revealed that the uptake of palmitic acid and oleic acid into brush border membranes was similar when these were isolated from either whole small intestine, jejunum, or ileum. The uptake of these fatty acids was somewhat lower with membranes obtained from duodenum. Subsequent studies, all with membranes obtained from whole intestine, indicated an increase in binding with chain length of fatty acid of up to 16 carbons. Unsaturation decreased this uptake somewhat. Taurocholate and 1-palmitoyl lysolecithin had a moderate stimulatory effect on the binding of oleic acid and palmitic acid at concentrations of 10 and 0.5 mM, respectively, and inhibited at higher concentrations. Addition of 1.4 mM egg lecithin to the fatty acid - bile salt micelles, such that the lecithin - bile salt ratio was 0.2, decreased the uptake of fatty acids generally, but did not significantly affect the pattern of binding by membrane fractions isolated from different segments nor did it change the pattern of labelling when fatty acid chain length and unsaturation were varied. At lower concentrations, egg lecithin had little effect on the uptake of oleic acid, whereas dipalmitoyl phosphatidylcholine stimulated binding of both palmitic acid and oleic acid over the entire range of concentrations tested. Preincubation of the membranes with this saturated phospholipid stimulated the uptake of oleic acid, and addition of this choline lipid to the oleic acid - bile salt containing micelles did not substantially enhance fatty acid uptake in lipid-treated membranes. The binding of fatty acid was very rapid either in the presence or the absence of Ca2+, such that even in zero-time controls essentially equilibrium bindings were obtained.(ABSTRACT TRUNCATED AT 250 WORDS)

Identification of a New cry1I-Type Gene as a Candidate for Gene Pyramiding in Corn To Control Ostrinia Species Larvae.

PubMed

Zhao, Can; Jurat-Fuentes, Juan Luis; Abdelgaffar, Heba M; Pan, Hongyu; Song, Fuping; Zhang, Jie

2015-06-01

Pyramiding of diverse cry toxin genes from Bacillus thuringiensis with different modes of action is a desirable strategy to delay the evolution of resistance in the European corn borer (Ostrinia nubilalis). Considering the dependency of susceptibility to Cry toxins on toxin binding to receptors in the midgut of target pests, a diverse mode of action is commonly defined as recognition of unique binding sites in the target insect. In this study, we present a novel cry1Ie toxin gene (cry1Ie2) as a candidate for pyramiding with Cry1Ab or Cry1Fa in corn to control Ostrinia species larvae. The new toxin gene encodes an 81-kDa protein that is processed to a protease-resistant core form of approximately 55 kDa by trypsin digestion. The purified protoxin displayed high toxicity to Ostrinia furnacalis and O. nubilalis larvae but low to no activity against Spodoptera or heliothine species or the coleopteran Tenebrio molitor. Results of binding assays with (125)I-labeled Cry1Ab toxin and brush border membrane vesicles from O. nubilalis larvae demonstrated that Cry1Ie2 does not recognize the Cry1Ab binding sites in that insect. Reciprocal competition binding assays with biotin-labeled Cry1Ie2 confirmed the lack of shared sites with Cry1Ab or Cry1Fa in O. nubilalis brush border membrane vesicles. These data support Cry1Ie2 as a good candidate for pyramiding with Cry1Ab or Cry1Fa in corn to increase the control of O. nubilalis and reduce the risk of resistance evolution. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Morduchowicz, G.A.; Sheikh-Hamad, D.; Dwyer, B.E.

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 searchmore » 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.« less

Semi-automated discrimination of retinal pigmented epithelial cells in two-photon fluorescence images of mouse retinas.

PubMed

Alexander, Nathan S; Palczewska, Grazyna; Palczewski, Krzysztof

2015-08-01

Automated image segmentation is a critical step toward achieving a quantitative evaluation of disease states with imaging techniques. Two-photon fluorescence microscopy (TPM) has been employed to visualize the retinal pigmented epithelium (RPE) and provide images indicating the health of the retina. However, segmentation of RPE cells within TPM images is difficult due to small differences in fluorescence intensity between cell borders and cell bodies. Here we present a semi-automated method for segmenting RPE cells that relies upon multiple weak features that differentiate cell borders from the remaining image. These features were scored by a search optimization procedure that built up the cell border in segments around a nucleus of interest. With six images used as a test, our method correctly identified cell borders for 69% of nuclei on average. Performance was strongly dependent upon increasing retinosome content in the RPE. TPM image analysis has the potential of providing improved early quantitative assessments of diseases affecting the RPE.

Cell cycle progression is required for zebrafish somite morphogenesis but not segmentation clock function

PubMed Central

Zhang, Lixia; Kendrick, Christina; Jülich, Dörthe; Holley, Scott A.

2010-01-01

Summary Cell division, differentiation and morphogenesis are coordinated during embryonic development and frequently in disarray in pathologies such as cancer. Here, we present a zebrafish mutant that ceases mitosis at the beginning of gastrulation, but undergoes axis elongation and develops blood, muscle and a beating heart. We identify the mutation as being in early mitotic inhibitor 1 (emi1), a negative regulator of the Anaphase Promoting Complex, and utilize the mutant to examine the role of the cell cycle in somitogenesis. The mutant phenotype indicates that axis elongation during the segmentation period is substantially driven by cell migration. We find that the segmentation clock, which regulates somitogenesis, functions normally in the absence of cell cycle progression and observe that mitosis is a modest source of noise for the clock. Somite morphogenesis involves the epithelialization of the somite border cells around a core of mesenchyme. As in wild-type embryos, somite boundary cells are polarized along a Fibronectin matrix in emi1−/−. The mutants also display evidence of segment polarity. However, in the absence of a normal cell cycle, somites appear to hyper-epithelialize as the internal mesenchymal cells exit the core of the somite after initial boundary formation. Thus, cell cycle progression is not required during the segmentation period for segmentation clock function but is necessary for normal segmental arrangement of epithelial borders and internal mesenchymal cells. PMID:18480162

Genotoxic effects of X-rays on keratinized mucosa cells during panoramic dental radiography.

PubMed

Cerqueira, E M M; Meireles, J R C; Lopes, M A; Junqueira, V C; Gomes-Filho, I S; Trindade, S; Machado-Santelli, G M

2008-10-01

The aim of this study was to evaluate the genotoxic effects of X-rays on epithelial gingival cells during panoramic dental radiography using a differentiated protocol for the micronucleus test. 40 healthy individuals who underwent this procedure for diagnostic purposes on request from their dentists agreed to participate in this study. All of them answered a questionnaire before the examination. Epithelial gingival cells were obtained from the keratinized mucosa of the upper dental arcade by gentle scraping with a cervical brush immediately before exposure and 10 days later. Cytological preparations were stained according to the Feulgen-Rossenbeck reaction, counterstained with fast green 1% for 1 min and analysed under a light microscope. Micronuclei, nuclear projections (broken eggs) and degenerative nuclear alterations (pyknosis, karyolysis, karyorrhexis and condensed chromatin) were scored. The frequency of micronuclei was significantly higher after exposure (P < 0.05), as were the frequencies of nuclear alterations indicative of apoptosis (P < 0.001). These results indicate that X-ray radiation emitted during panoramic dental radiography induces a genotoxic effect on epithelial gingival cells that increases the frequency of chromosomal damage and nuclear alterations indicative of apoptosis.

Design and field results of a walk-through EDS

NASA Astrophysics Data System (ADS)

Wendel, Gregory J.; Bromberg, Edward E.; Durfee, Memorie K.; Curby, William A.

1997-01-01

A walk-through portal sampling module which incorporates active sampling has been developed. The module uses opposing wands which actively brush the subjects exterior clothing to disturb explosive traces. These traces are entrained in an air stream and transported to a High Speed GC- chemiluminescence explosives detection system. This combination provides automatic screening of passengers at rates of 10 per minute. The system exhibits sensitivity and selectivity which equals or betters that available from commercially available manual equipment. The systems has been developed for deployment at border crossings, airports and other security screening points. Detailed results of laboratory tests and airport field trials are reviewed.

Gadolinium and didymium (praseodymium/neodymium) cations as capture agents in lightmicroscopical histochemistry of acid and alkaline phosphatase.

PubMed

Halbhuber, K J; Zimmermann, N

1987-01-01

In previous papers, cerium and lanthanum based methods for light-microscopical detection of acid and alkaline phosphatase activity were proposed. In this paper, the usefulness of other lanthanide cations such as gadolinium and praseodymium/neodymium cations as capture agents in phosphatase histochemistry is tested. It is evident that phosphate ions were sufficiently trapped by these cations. According to the lead and silver multistep procedures earlier described it is possible to visualize alkaline phosphatase activity in the brush borders of the intestine or kidney as well as acid phosphatase activity in the lysosomes. These methods can be recommended.

Physical labeling of papillomavirus-infected, immortal, and cancerous cervical epithelial cells reveal surface changes at immortal stage.

PubMed

Swaminathan Iyer, K; Gaikwad, R M; Woodworth, C D; Volkov, D O; Sokolov, Igor

2012-06-01

A significant change of surface features of malignant cervical epithelial cells compared to normal cells has been previously reported. Here, we are studying the question at which progressive stage leading to cervical cancer the surface alteration happens. A non-traditional method to identify malignant cervical epithelial cells in vitro, which is based on physical (in contrast to specific biochemical) labelling of cells with fluorescent silica micron-size beads, is used here to examine cells at progressive stages leading to cervical cancer which include normal epithelial cells, cells infected with human papillomavirus type-16 (HPV-16), cells immortalized by HPV-16, and carcinoma cells. The study shows a statistically significant (at p < 0.01) difference between both immortal and cancer cells and a group consisting of normal and infected. There is no significant difference between normal and infected cells. Immortal cells demonstrate the signal which is closer to cancer cells than to either normal or infected cells. This implies that the cell surface, surface cellular brush changes substantially when cells become immortal. Physical labeling of the cell surface represents a substantial departure from the traditional biochemical labeling methods. The results presented show the potential significance of physical properties of the cell surface for development of clinical methods for early detection of cervical cancer, even at the stage of immortalized, premalignant cells.

Physical Labeling of Papillomavirus-Infected, Immortal, and Cancerous Cervical Epithelial Cells Reveal Surface Changes at Immortal Stage

PubMed Central

Iyer, K. Swaminathan; Gaikwad, R. M.; Woodworth, C. D.; Volkov, D. O.

2013-01-01

A significant change of surface features of malignant cervical epithelial cells compared to normal cells has been previously reported. Here, we are studying the question at which progressive stage leading to cervical cancer the surface alteration happens. A non-traditional method to identify malignant cervical epithelial cells in vitro, which is based on physical (in contrast to specific biochemical) labelling of cells with fluorescent silica micron-size beads, is used here to examine cells at progressive stages leading to cervical cancer which include normal epithelial cells, cells infected with human papillomavirus type-16 (HPV-16), cells immortalized by HPV-16, and carcinoma cells. The study shows a statistically significant (at p <0.01) difference between both immortal and cancer cells and a group consisting of normal and infected. There is no significant difference between normal and infected cells. Immortal cells demonstrate the signal which is closer to cancer cells than to either normal or infected cells. This implies that the cell surface, surface cellular brush changes substantially when cells become immortal. Physical labeling of the cell surface represents a substantial departure from the traditional biochemical labeling methods. The results presented show the potential significance of physical properties of the cell surface for development of clinical methods for early detection of cervical cancer, even at the stage of immortalized, pre-malignant cells. PMID:22351422

Epithelial cytoskeletal framework and nuclear matrix-intermediate filament scaffold: three-dimensional organization and protein composition.

PubMed

Fey, E G; Wan, K M; Penman, S

1984-06-01

Madin-Darby canine kidney (MDCK) cells grow as differentiated, epithelial colonies that display tissue-like organization. We examined the structural elements underlying the colony morphology in situ using three consecutive extractions that produce well-defined fractions for both microscopy and biochemical analysis. First, soluble proteins and phospholipid were removed with Triton X-100 in a physiological buffer. The resulting skeletal framework retained nuclei, dense cytoplasmic filament networks, intercellular junctional complexes, and apical microvillar structures. Scanning electron microscopy showed that the apical cell morphology is largely unaltered by detergent extraction. Residual desmosomes, as can be seen in thin sections, were also well-preserved. The skeletal framework was visualized in three dimensions as an unembedded whole mount that revealed the filament networks that were masked in Epon-embedded thin sections of the same preparation. The topography of cytoskeletal filaments was relatively constant throughout the epithelial sheet, particularly across intercellular borders. This ordering of epithelial skeletal filaments across contiguous cell boundaries was in sharp contrast to the more independent organization of networks in autonomous cells such as fibroblasts. Further extraction removed the proteins of the salt-labile cytoskeleton and the chromatin as separate fractions, and left the nuclear matrix-intermediate filament (NM-IF) scaffold. The NM-IF contained only 5% of total cellular protein, but whole mount transmission electron microscopy and immunofluorescence showed that this scaffold was organized as in the intact epithelium. Immunoblots demonstrate that vimentin, cytokeratins, desmosomal proteins, and a 52,000-mol-wt nuclear matrix protein were found almost exclusively in the NM-IF scaffold. Vimentin was largely perinuclear while the cytokeratins were localized at the cell borders. The 52,000-mol-wt nuclear matrix protein was confined to the chromatin-depleted matrix and the desmosomal proteins were observed in punctate polygonal arrays at intercellular junctions. The filaments of the NM-IF were seen to be interconnected, via the desmosomes, over the entire epithelial colony. The differentiated epithelial morphology was reflected in both the cytoskeletal framework and the NM-IF scaffold.

Epithelial cytoskeletal framework and nuclear matrix-intermediate filament scaffold: three-dimensional organization and protein composition

PubMed Central

1984-01-01

Madin-Darby canine kidney (MDCK) cells grow as differentiated, epithelial colonies that display tissue-like organization. We examined the structural elements underlying the colony morphology in situ using three consecutive extractions that produce well-defined fractions for both microscopy and biochemical analysis. First, soluble proteins and phospholipid were removed with Triton X-100 in a physiological buffer. The resulting skeletal framework retained nuclei, dense cytoplasmic filament networks, intercellular junctional complexes, and apical microvillar structures. Scanning electron microscopy showed that the apical cell morphology is largely unaltered by detergent extraction. Residual desmosomes, as can be seen in thin sections, were also well- preserved. The skeletal framework was visualized in three dimensions as an unembedded whole mount that revealed the filament networks that were masked in Epon-embedded thin sections of the same preparation. The topography of cytoskeletal filaments was relatively constant throughout the epithelial sheet, particularly across intercellular borders. This ordering of epithelial skeletal filaments across contiguous cell boundaries was in sharp contrast to the more independent organization of networks in autonomous cells such as fibroblasts. Further extraction removed the proteins of the salt-labile cytoskeleton and the chromatin as separate fractions, and left the nuclear matrix-intermediate filament (NM-IF) scaffold. The NM-IF contained only 5% of total cellular protein, but whole mount transmission electron microscopy and immunofluorescence showed that this scaffold was organized as in the intact epithelium. Immunoblots demonstrate that vimentin, cytokeratins, desmosomal proteins, and a 52,000-mol-wt nuclear matrix protein were found almost exclusively in the NM-IF scaffold. Vimentin was largely perinuclear while the cytokeratins were localized at the cell borders. The 52,000-mol-wt nuclear matrix protein was confined to the chromatin- depleted matrix and the desmosomal proteins were observed in punctate polygonal arrays at intercellular junctions. The filaments of the NM-IF were seen to be interconnected, via the desmosomes, over the entire epithelial colony. The differentiated epithelial morphology was reflected in both the cytoskeletal framework and the NM-IF scaffold. PMID:6202700

Reproductive functional anatomy and oestrous cycle pattern of the female brush-tailed porcupine (Atherurus africanus, Gray 1842) from Gabon.

PubMed

Mayor, P; López-Béjar, M; Jori, F; Fenech, M; López-Gatius, F

2003-07-15

In the present study, we examined certain features of the functional anatomy of the female genital tract of the wild brush-tailed porcupine (Atherurus africanus) to obtain data on the reproductive biology of this African forest rodent. Two consecutive experiments were performed. The aim of the first was to establish macroscopic and microscopic features of the genital organs, and to explore correlations between predominant ovarian structures and vaginal contents in 20 wild, mature females. In the second experiment, we inspected the external genitalia and vaginal smears of a further 10 females in captivity on a daily basis for 90 days. The uterus of the brush-tailed porcupine is bicornuate and composed of two separated uterine horns, a uterine body and cervix. The genital tract does not present a vaginal vestibule. Thus, there is no portion common to genital and urinary tracts. Females in the follicular phase of the oestrous cycle showed increased cornification of the vaginal epithelium and a high density of eosinophilic cells in vaginal smears. The vulva and vaginal opening were open, reddish and tumefacted. In luteal phase or in pregnancy, epithelial cornification and eosinophilic features were notably reduced and the vagina presented a pale, non-tumefacted vulva and a vaginal closure membrane. Females in captivity showed spontaneous cycles, a polyoestrous reproduction pattern and, based on features of the external genitalia and vaginal smears, their oestrous cycle length was 27.1+/-6.4 days (n=12).

Increased Fatty Acid Oxidation in Differentiated Proximal Tubular Cells Surviving a Reversible Episode of Acute Kidney Injury.

PubMed

Bataille, Aurélien; Galichon, Pierre; Chelghoum, Nadjim; Oumoussa, Badreddine Mohand; Ziliotis, Marie-Julia; Sadia, Iman; Vandermeersch, Sophie; Simon-Tillaux, Noémie; Legouis, David; Cohen, Raphaël; Xu-Dubois, Yi-Chun; Commereuc, Morgane; Rondeau, Eric; Le Crom, Stéphane; Hertig, Alexandre

2018-06-19

Fatty acid oxidation (FAO), the main source of energy produced by tubular epithelial cells in the kidney, was found to be defective in tubulo-interstitial samples dissected out in kidney biopsies from patients with chronic kidney disease (CKD). Experimental data indicated that this decrease was a strong determinant of renal fibrogenesis, hence a focus for therapeutic interventions. Nevertheless, whether persistently differentiated renal tubules, surviving in a pro-fibrotic environment, also suffer from a decrease in FAO, is currently unknown. To address this question, we isolated proximal tubules captured ex vivo on the basis of the expression of an intact brush border antigen (Prominin-1) in C57BL6/J mice subjected to a controlled, two-hit model of renal fibrosis (reversible ischemic acute kidney injury (AKI) or sham surgery, followed by angiotensin 2 administration). A transcriptomic high throughput sequencing was performed on total mRNA from these cells, and on whole kidneys. In contrast to mice subjected to sham surgery, mice with a history of AKI displayed histologically more renal fibrosis when exposed to angiotensin 2. High throughput RNA sequencing, principal component analysis and clustering showed marked consistency within experimental groups. As expected, FAO transcripts were decreased in whole fibrotic kidneys. Surprisingly, however, up- rather than down-regulation of metabolic pathways (oxidative phosphorylation, fatty acid metabolism, glycolysis, and PPAR signalling pathway) was a hallmark of the differentiated tubules captured from fibrotic kidneys. Immunofluorescence co-staining analysis confirmed that the expression of FAO enzymes was dependent of tubular trophicity. These data suggest that in differentiated proximal tubules energetic hyperactivity is promoted concurrently with organ fibrogenesis. © 2018 The Author(s). Published by S. Karger AG, Basel.

Guanylyl Cyclase C Is a Specific Marker for Differentiating Primary and Metastatic Ovarian Mucinous Neoplasms

PubMed Central

Ciocca, Vincenzo; Bombonati, Alessandro; Palazzo, Juan P.; Schulz, Stephanie; Waldman, Scott A.

2011-01-01

Distinguishing primary ovarian mucinous neoplasms from metastatic mucinous adenocarcinomas with ovarian involvement can be difficult, especially when characteristic gross and microscopic features are not present. CK7/CK20 expression appears to be more useful for distinguishing metastatic gastrointestinal adenocarcinomas from the lower tract. The addition of CDX2 for distinguishing metastatic upper gastrointestinal tract adenocarcinomas from primary ovarian mucinous neoplasms offers little advantage over CK7/CK20 coordinate expression. Guanylyl cyclase C (GCC) is a brush border membrane receptor for the endogenous peptides guanylin and uroguanylin, and the homologous diarrheagenic bacterial heat-stable enterotoxins that is selectively expressed by epithelial cells from the duodenum to the rectum, but not by normal epithelia of the stomach or esophagus, or normal extramucosal cells in humans. We studied 50 ovarian tumors: 27 primary ovarian mucinous neoplasms (7 cystadenomas, 10 borderline tumors, and 10 cystadenocarcinomas) and 23 metastatic mucinous adenocarcinomas with ovarian involvement (13 colorectal adenocarcinomas, 4 gastric adenocarcinomas, 6 appendiceal mucinous tumors (4 adenocarcinomas, 1 with neuroendocrine features, and 2 appendiceal mucinous cystadenomas). For primary ovarian mucinous neoplasms, 25 of 27 were negative for GCC. Twelve of thirteen cases of colorectal adenocarcinoma (except for 1 neuroendocrine adenocarcinoma) were positive for GCC. Three of four appendiceal mucinous adenocarcinomas were positive for GCC in both the primary and metastatic tumors (except for 1 neuroendocrine adenocarcinoma). Two of two appendiceal mucinous cystadenomas were positive for GCC. Of four cases of gastric adenocarcinoma with ovarian involvement, only one (primary tumor) exhibited focal GCC staining. These findings suggest GCC may be a useful marker for differentiating primary and secondary ovarian mucinous neoplasms. PMID:19694825

Loss of PDZ-adaptor protein NHERF2 affects membrane localization and cGMP- and [Ca2+]- but not cAMP-dependent regulation of Na+/H+ exchanger 3 in murine intestine

PubMed Central

Chen, Mingmin; Sultan, Ayesha; Cinar, Ayhan; Yeruva, Sunil; Riederer, Brigitte; Singh, Anurag Kumar; Li, Junhua; Bonhagen, Janina; Chen, Gang; Yun, Chris; Donowitz, Mark; Hogema, Boris; deJonge, Hugo; Seidler, Ursula

2010-01-01

Trafficking and regulation of the epithelial brush border membrane (BBM) Na+/H+ exchanger 3 (NHE3) in the intestine involves interaction with four different members of the NHERF family in a signal-dependent and possibly segment-specific fashion. The aim of this research was to study the role of NHERF2 (E3KARP) in intestinal NHE3 BBM localization and second messenger-mediated and receptor-mediated inhibition of NHE3. Immunolocalization of NHE3 in WT mice revealed predominant microvillar localization in jejunum and colon, a mixed distribution in the proximal ileum but localization near the terminal web in the distal ileum. The terminal web localization of NHE3 in the distal ileum correlated with reduced acid-activated NHE3 activity (fluorometrically assessed). NHERF2 ablation resulted in a shift of NHE3 to the microvilli and higher basal fluid absorption rates in the ileum, but no change in overall NHE3 protein or mRNA expression. Forskolin-induced NHE3 inhibition was preserved in the absence of NHERF2, whereas Ca2+ ionophore- or carbachol-mediated inhibition was abolished. Likewise, Escherichia coli heat stable enterotoxin peptide (STp) lost its inhibitory effect on intestinal NHE3. It is concluded that in native murine intestine, the NHE3 adaptor protein NHERF2 plays important roles in tethering NHE3 to a position near the terminal web and in second messenger inhibition of NHE3 in a signal- and segment-specific fashion, and is therefore an important regulator of intestinal fluid transport. PMID:20962002

Sulfate-chloride exchange by lobster hepatopancreas is regulated by pH-sensitive modifier sites

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cattey, M.A.; Ahearn, G.A.; Gerencser, G.A.

1991-03-15

{sup 35}SO{sub 4}{sup 2{minus}} uptake by Atlantic lobster (Homarus americanus) hepatopancreatic epithelial brush border membrane vesicles (BBMV) was stimulated by internal Cl{sup {minus}}, but not internal HCO{sub 3}{sup {minus}}, or external Na{sup +}. Sulfate-chloride exchange was stimulated by inside positive, and inhibited by inside negative, trans-membrane K diffusion potentials. {sup 35}SO{sub 4}{sup 2{minus}}-Cl{sup {minus}} exchange was strongly inhibited by 4,4{prime} diisothiocyanostilbene-2,2{prime}-disulfonic acid (DIDS), 4-acetamido-4{prime}-isotheocynaostilbene-2,2{prime}-disulfonic acid, (SITS), and thiosulfate. Chloride, bicarbonate, furosamide, and bumetanide slightly, yet significantly, cis-inhibited {sup 35}SO{sub 4}{sup 2{minus}}-Cl{sup {minus}} exchange. Altering bilateral pH from 8.0 to 5.4 stimulated {sup 35}SO{sub 4}{sup 2{minus}}-Cl{sup {minus}} exchange when vesicles weremore » loaded with Cl{sup {minus}}, but reduced bilateral pH alone or the presence of pH gradients did not affect {sup 35}SO{sub 4}{sup 2{minus}} transport in the absence of internal Cl{sup {minus}}. {sup 36}Cl uptake into SO{sub 4}{sup 2{minus}}-loaded BBMV was stimulated by an internal negative membrane potential and inhibited when the interior was electrically positive. A model is proposed which suggests that SO{sub 4}{sup 2{minus}}-Cl{sup {minus}} exchange is regulated by internal and external pH-sensitive modifier sites on the anion antiporter and by coupling to the electrogenic 2 Na{sup +}/1 H{sup +} antiporter and by coupling to the electrogenic 2 Na{sup +}/1 H{sup +} antiporter on the same membrane.« less

Congenital Sodium Diarrhea: A Form of Intractable Diarrhea, With a Link to Inflammatory Bowel Disease.

PubMed

Janecke, Andreas R; Heinz-Erian, Peter; Müller, Thomas

2016-08-01

Congenital diarrheal disorders (CDDs) represent a group of challenging clinical conditions for pediatricians because of the severity of the presentation and the broad range of possible differential diagnoses. CDDs arise from alterations in the transport of nutrients and electrolytes across the intestinal mucosa, from enterocyte and enteroendocrine cell differentiation and/or polarization defects, and from the modulation of the intestinal immune response. Advances were made recently in deciphering the etiology and pathophysiology of one of these disorders, congenital sodium diarrhea (CSD). CSD refers to an intractable diarrhea of intrauterine onset with high fecal sodium loss. CSD is clinically and genetically heterogeneous. A syndromic form of CSD features choanal and intestinal atresias as well as recurrent corneal erosions. Small bowel histology frequently detects an epithelial "tufting" dysplasia. It is autosomal recessively inherited, and caused by SPINT2 mutations. The nonsyndromic form of CSD can be caused by dominant activating mutations in GUCY2C, encoding intestinal receptor guanylate cyclase C (GC-C), and by autosomal recessive SLC9A3 loss-of-function mutations. SLC9A3 encodes Na/H antiporter 3, the major intestinal brush border Na/H exchanger, and a downstream target of GC-C. A number of patients with GUCY2C and SLC9A3 mutations developed inflammatory bowel disease. Both the number of recognized CDD forms as well as the number of underlying disease genes are gradually increasing. Knowledge of these CDD genes enables noninvasive, next-generation gene panel-based testing to facilitate an early diagnosis in CDD. Primary Na/H antiporter 3 and GC-C malfunction is implicated as a predisposition for inflammatory bowel disease in subset of patients.

Paracellular transport of avidin saturated or not with biotinylated cobalamin through Caco-2 cell epithelium monolayer.

PubMed

Guy, M; Pons, L; Namour, F; de Nonancourt, M; Michalski, J C; Hatier, R; Guéant, J L

2001-01-01

The cationic charge of molecules may promote their uptake across epithelia, which are rich in brush border anionic sites. The transport of unsaturated avidin and avidin saturated with a biotinylated compound was investigated across Caco-2 adenocarcinoma cell with fetal enterocyte phenotype. The unsaturated avidin and avidin saturated with either biotin or a biotinyl-cobalamin conjugate (biotinyl-Cbl) were iodinated to follow their transport through the cell monolayer. Their apparent permeability coefficient (Papp) and transepithelial pathway were determined and compared to those for control radiolabeled markers [3H]-mannitol, [125I]-beta-lactoglobulin and [57Co]-cobalamin/intrinsic factor (Cbl/IF). The Papp of [125I]-avidin estimated at 2.8 x 10(-7) +/- 0.08 cm/s was close to that for mannitol that uses paracellular pathway. The binding of biotin or biotin conjugate to avidin enhanced its tetrameric conformation. The Papp for [125I]-avidin/biotin and [125I]- avidin/biotinyl-Cbl were respectively increased by 2-fold, compared to that for [125I]-avidin and 4-fold, compared to that for [125I]-beta-lactoglobulin and [54Co]-Cbl/IF. The protein was not accumulated in the cell and was found in intact form in the basolateral side, after its transport across the monolayer. Chloroquine (0.66 micromol/ml) did not significantly decrease the Papp for [125I]-avidin/biotinyl-Cbl. Conversely it decreased by 80% the Papp for Cbl/IF, that uses transepithelial pathway. Avidin (either saturated or not with biotin and biotinyl-Cbl) was able to cross the monolayer of Caco-2 cell line through a paracellular pathway. This study pointed out the interest for using this protein as a shuttle for increasing the transport rate of biotinylated compounds through fetal epithelial barriers. Copyright 2001 S. Karger AG, Basel

β-Casein(94-123)-derived peptides differently modulate production of mucins in intestinal goblet cells.

PubMed

Plaisancié, Pascale; Boutrou, Rachel; Estienne, Monique; Henry, Gwénaële; Jardin, Julien; Paquet, Armelle; Léonil, Joëlle

2015-02-01

We recently reported the identification of a peptide from yoghurts with promising potential for intestinal health: the sequence (94-123) of bovine β-casein. This peptide, composed of 30 amino acid residues, maintains intestinal homoeostasis through production of the secreted mucin MUC2 and of the transmembrane-associated mucin MUC4. Our study aimed to search for the minimal sequence responsible for the biological activity of β-CN(94-123) by using several strategies based on (i) known bioactive peptides encrypted in β-CN(94-123), (ii) in silico prediction of peptides reactivity and (iii) digestion of β-CN(94-123) by enzymes of intestinal brush border membranes. The revealed sequences were tested in vitro on human intestinal mucus-producing HT29-MTX cells. We demonstrated that β-CN(108-113) (an ACE-inhibitory peptide) and β-CN(114-119) (an opioid peptide named neocasomorphin-6) up-regulated MUC4 expression whereas levels of the secreted mucins MUC2 and MUC5AC remained unchanged. The digestion of β-CN(94-123) by intestinal enzymes showed that the peptides β-CN(94-108) and β-CN(117-123) were present throughout 1·5 to 3 h of digestion, respectively. These two peptides raised MUC5AC expression while β-CN(117-123) also induced a decrease in the level of MUC2 mRNA and protein. In addition, this inhibitory effect was reproduced in airway epithelial cells. In conclusion, β-CN(94-123) is a multifunctional molecule but only the sequence of 30 amino acids has a stimulating effect on the production of MUC2, a crucial factor of intestinal protection.

Differential cystine and dibasic amino acid handling after loss of function of the amino acid transporter b0,+AT (Slc7a9) in mice.

PubMed

Di Giacopo, Andrea; Rubio-Aliaga, Isabel; Cantone, Alessandra; Artunc, Ferruh; Rexhepaj, Rexhep; Frey-Wagner, Isabelle; Font-Llitjós, Mariona; Gehring, Nicole; Stange, Gerti; Jaenecke, Isabel; Mohebbi, Nilufar; Closs, Ellen I; Palacín, Manuel; Nunes, Virginia; Daniel, Hannelore; Lang, Florian; Capasso, Giovambattista; Wagner, Carsten A

2013-12-15

Cystinuria is an autosomal recessive disease caused by mutations in SLC3A1 (rBAT) and SLC7A9 (b(0,+)AT). Gene targeting of the catalytic subunit (Slc7a9) in mice leads to excessive excretion of cystine, lysine, arginine, and ornithine. Here, we studied this non-type I cystinuria mouse model using gene expression analysis, Western blotting, clearance, and brush-border membrane vesicle (BBMV) uptake experiments to further characterize the renal and intestinal consequences of losing Slc7a9 function. The electrogenic and BBMV flux studies in the intestine suggested that arginine and ornithine are transported via other routes apart from system b(0,+). No remarkable gene expression changes were observed in other amino acid transporters and the peptide transporters in the intestine and kidney. Furthermore, the glomerular filtration rate (GFR) was reduced by 30% in knockout animals compared with wild-type animals. The fractional excretion of arginine was increased as expected (∼100%), but fractional excretions of lysine (∼35%), ornithine (∼16%), and cystine (∼11%) were less affected. Loss of function of b(0,+)AT reduced transport of cystine and arginine in renal BBMVs and completely abolished the exchanger activity of dibasic amino acids with neutral amino acids. In conclusion, loss of Slc7a9 function decreases the GFR and increases the excretion of several amino acids to a lesser extent than expected with no clear regulation at the mRNA and protein level of alternative transporters and no increased renal epithelial uptake. These observations indicate that transporters located in distal segments of the kidney and/or metabolic pathways may partially compensate for Slc7a9 loss of function.

VSL#3 probiotic upregulates intestinal mucosal alkaline sphingomyelinase and reduces inflammation.

PubMed

Soo, I; Madsen, K L; Tejpar, Q; Sydora, B C; Sherbaniuk, R; Cinque, B; Di Marzio, L; Cifone, M Grazia; Desimone, C; Fedorak, R N

2008-03-01

Alkaline sphingomyelinase, an enzyme found exclusively in bile and the intestinal brush border, hydrolyzes sphingomyelin into ceramide, sphingosine and sphingosine-1-phosphate, thereby inducing epithelial apoptosis. Reduced levels of alkaline sphingomyelinase have been found in premalignant and malignant intestinal epithelia and in ulcerative colitis tissue. Probiotic bacteria can be a source of sphingomyelinase. To determine the effect of VSL#3 probiotic therapy on mucosal levels of alkaline sphingomyelinase, both in a mouse model of colitis and in patients with ulcerative colitis. Interleukin-10 gene-deficient (IL10KO) and wild type control mice were treated with VSL#3 (10(9) colony-forming units per day) for three weeks, after which alkaline sphingomyelinase activity was measured in ileal and colonic tissue. As well, 15 patients with ulcerative colitis were treated with VSL#3 (900 billion bacteria two times per day for five weeks). Alkaline sphingomyelinase activity was measured through biopsies and comparison of ulcerative colitis disease activity index scores obtained before and after treatment. Lowered alkaline sphingomyelinase levels were seen in the colon (P=0.02) and ileum (P=0.04) of IL10KO mice, as compared with controls. Treatment of these mice with VSL#3 resulted in upregulation of mucosal alkaline sphingomyelinase activity in both the colon (P=0.04) and the ileum (P=0.01). VSL#3 treatment of human patients who had ulcerative colitis decreased mean (+/- SEM) ulcerative colitis disease activity index scores from 5.3+/-1.8946 to 0.70+/-0.34 (P=0.02) and increased mucosal alkaline sphingomyelinase activity. Mucosal alkaline sphingomyelinase activity is reduced in the intestine of IL10KO mice with colitis and in humans with ulcerative colitis. VSL#3 probiotic therapy upregulates mucosal alkaline sphingomyelinase activity.

Saccharomyces boulardii protease inhibits Clostridium difficile toxin A effects in the rat ileum.

PubMed Central

Castagliuolo, I; LaMont, J T; Nikulasson, S T; Pothoulakis, C

1996-01-01

Saccharomyces boulardii, a nonpathogenic yeast, is effective in treating some patients with Clostridium difficile diarrhea and colitis. We have previously reported that S. boulardii inhibits rat ileal secretion in response to C. difficile toxin A possibly by releasing a protease that digests the intestinal receptor for this toxin (C. Pothoulakis, C. P. Kelly, M. A. Joshi, N. Gao, C. J. O'Keane, I. Castagliuolo, and J. T. LaMont, Gastroenterology 104: 1108-1115, 1993). The aim of this study was to purify and characterize this protease. S. boulardii protease was partially purified by gel filtration on Sephadex G-50 and octyl-Sepharose. The effect of S. boulardii protease on rat ileal secretion, epithelial permeability, and morphology in response to toxin A was examined in rat ileal loops in vivo. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified S. boulardii protease revealed a major band at 54 kDa. Pretreatment of rat ileal brush border (BB) membranes with partially purified protease reduced specific toxin A receptor binding (by 26%). Partially purified protease digested the toxin A molecule and significantly reduced its binding to BB membranes in vitro (by 42%). Preincubation of toxin A with S. boulardii protease inhibited ileal secretion (46% inhibition, P < 0.01), mannitol permeability (74% inhibition, P < 0.01), and histologic damage caused by toxin A. Thus, S. boulardii protease inhibits the intestinal effects of C. difficile toxin A by proteolysis of the toxin and inhibition of toxin A binding to its BB receptor. Our results may be relevant to the mechanism by which S. boulardii exerts its protective effects in C. difficile infection in humans. PMID:8945570

Carbachol-mediated endocytosis of NHE3 involves a clathrin-independent mechanism requiring lipid rafts and Cdc42.

PubMed

Zachos, Nicholas C; Alamelumangpuram, Bharath; Lee, Luke J; Wang, Peng; Kovbasnjuk, Olga

2014-01-01

In intestinal epithelial cells, acute regulation of the brush border Na(+)/H(+) exchanger, NHE3, usually occurs by changes in endocytosis and/or exocytosis. Constitutive NHE3 endocytosis involves clathrin. Carbachol (CCH), which elevates intracellular Ca(2+) ([Ca(2+)]i), decreases NHE3 activity and stimulates endocytosis; however, the mechanism involved in calcium-mediated endocytosis of NHE3 is unclear. A pool of NHE3 resides in lipid rafts, which contributes to basal, but not cAMP-mediated, NHE3 trafficking, suggesting that an alternative mechanism exists for NHE3 endocytosis. Cdc42 was demonstrated to play an integral role in some cases of cholesterol-sensitive, clathrin-independent endocytosis. Therefore, the current study was designed to test the hypotheses that (1) clathrin-mediated endocytosis (CME) is involved in constitutive, but not CCH-mediated, endocytosis of NHE3, and (2) CCH-mediated endocytosis of NHE3 occurs through a lipid raft, activated Cdc42-dependent pathway that does not involve clathrin. The role of Cdc42 and lipid rafts on NHE3 activity and endocytosis were investigated in polarized Caco-2/BBe cells using pharmacological and shRNA knockdown approaches. Basal NHE3 activity was increased in the presence of CME blockers (chlorpromazine; K(+) depletion) supporting previous reports that constitutive NHE3 endocytosis is clathrin dependent. In contrast, CCH-inhibition of NHE3 activity was abolished in Caco-2/BBe cells treated with MβCD (to disrupt lipid rafts) as well as in Cdc42 knockdown cells but was unaffected by CME blockers. CCH-mediated inhibition of NHE3 activity is not dependent on clathrin and involves lipid rafts and requires Cdc42. © 2014 S. Karger AG, Basel.

Carbachol-Mediated Endocytosis of NHE3 Involves a Clathrin-Independent Mechanism Requiring Lipid Rafts and Cdc42

PubMed Central

Zachos, Nicholas C.; Alamelumangpuram, Bharath; Lee, Luke J.; Wang, Peng; Kovbasnjuk, Olga

2014-01-01

Background In intestinal epithelial cells, acute regulation of the brush border Na+/H+ exchanger, NHE3, usually occurs by changes in endocytosis and/or exocytosis. Constitutive NHE3 endocytosis involves clathrin. Carbachol (CCH), which elevates intracellular Ca2+ ([Ca2+]i), decreases NHE3 activity and stimulates endocytosis; however, the mechanism involved in calcium-mediated endocytosis of NHE3 is unclear. A pool of NHE3 resides in lipid rafts, which contributes to basal, but not cAMP-mediated, NHE3 trafficking, suggesting that an alternative mechanism exists for NHE3 endocytosis. Cdc42 was demonstrated to play an integral role in some cases of cholesterol-sensitive, clathrin-independent endocytosis. Therefore, the current study was designed to test the hypotheses that (1) clathrin-mediated endocytosis (CME) is involved in constitutive, but not CCH-mediated, endocytosis of NHE3, and (2) CCH-mediated endocytosis of NHE3 occurs through a lipid raft, activated Cdc42-dependent pathway that does not involve clathrin. Methods The role of Cdc42 and lipid rafts on NHE3 activity and endocytosis were investigated in polarized Caco-2/BBe cells using pharmacological and shRNA knockdown approaches. Results Basal NHE3 activity was increased in the presence of CME blockers (chlorpromazine; K+ depletion) supporting previous reports that constitutive NHE3 endocytosis is clathrin dependent. In contrast, CCH-inhibition of NHE3 activity was abolished in Caco-2/BBe cells treated with MβCD (to disrupt lipid rafts) as well as in Cdc42 knockdown cells but was unaffected by CME blockers. Conclusion CCH-mediated inhibition of NHE3 activity is not dependent on clathrin and involves lipid rafts and requires Cdc42. PMID:24713550

Modulation of ileal bile acid transporter (ASBT) activity by depletion of plasma membrane cholesterol: association with lipid rafts

PubMed Central

Annaba, Fadi; Sarwar, Zaheer; Kumar, Pradeep; Saksena, Seema; Turner, Jerrold R.; Dudeja, Pradeep K.; Gill, Ravinder K.; Alrefai, Waddah A.

2016-01-01

Apical sodium-dependent bile acid transporter (ASBT) represents a highly efficient conservation mechanism of bile acids via mediation of their active transport across the luminal membrane of terminal ileum. To gain insight into the cellular regulation of ASBT, we investigated the association of ASBT with cholesterol and sphingolipid-enriched specialized plasma membrane microdomains known as lipid rafts and examined the role of membrane cholesterol in maintaining ASBT function. Human embryonic kidney (HEK)-293 cells stably transfected with human ASBT, human ileal brush-border membrane vesicles, and human intestinal epithelial Caco-2 cells were utilized for these studies. Floatation experiments on Optiprep density gradients demonstrated the association of ASBT protein with lipid rafts. Disruption of lipid rafts by depletion of membrane cholesterol with methyl-β-cyclodextrin (MβCD) significantly reduced the association of ASBT with lipid rafts, which was paralleled by a decrease in ASBT activity in Caco-2 and HEK-293 cells treated with MβCD. The inhibition in ASBT activity by MβCD was blocked in the cells treated with MβCD-cholesterol complexes. Kinetic analysis revealed that MβCD treatment decreased the Vmax of the transporter, which was not associated with alteration in the plasma membrane expression of ASBT. Our study illustrates that cholesterol content of lipid rafts is essential for the optimal activity of ASBT and support the association of ASBT with lipid rafts. These findings suggest a novel mechanism by which ASBT activity may be rapidly modulated by alterations in cholesterol content of plasma membrane and thus have important implications in processes related to maintenance of bile acid and cholesterol homeostasis. PMID:18063707

Normal tubular regeneration and differentiation of the post-ischemic kidney in mice lacking vimentin.

PubMed Central

Terzi, F.; Maunoury, R.; Colucci-Guyon, E.; Babinet, C.; Federici, P.; Briand, P.; Friedlander, G.

1997-01-01

Proliferation and dedifferentiation of tubular cells are the hallmark of early regeneration after renal ischemic injury. Vimentin, a class III intermediate filament expressed only in mesenchymal cells of mature mammals, was shown to be transiently expressed in post-ischemic renal tubular epithelial cells. Vimentin re-expression was interpreted as a marker of cellular dedifferentiation, but its role in tubular regeneration after renal ischemia has also been hypothesized. This role was evaluated in mice bearing a null mutation of the vimentin gene. Expression of vimentin, proliferating cell nuclear antigen (a marker of cellular proliferation), and villin (a marker of differentiated brush-border membranes) was studied in wild-type (Vim+/+), heterozygous (Vim+/-), and homozygous (Vim-/-) mice subjected to transient ischemia of the left kidney. As expected, vimentin was detected by immunohistochemistry at the basal pole of proximal tubular cells from post-ischemic kidney in Vim+/+ and Vim+/- mice from day 2 to day 28. The expression of the reporter gene beta-galactosidase in Vim+/- and Vim-/- mice confirmed the tubular origin of vimentin. No compensatory expression of keratin could be demonstrated in Vim-/- mice. The intensity of proliferating cell nuclear antigen labeling and the pattern of villin expression were comparable in Vim-/-, Vim+/- and Vim+/+ mice at any time of the study. After 60 days, the structure of post-ischemic kidneys in Vim-/- mice was indistinguishable from that of normal non-operated kidneys in Vim+/+ mice. In conclusion, 1) the pattern of post-ischemic proximal tubular cell proliferation, differentiation, and tubular organization was not impaired in mice lacking vimentin and 2) these results suggest that the transient tubular expression of vimentin is not instrumental in tubular regeneration after renal ischemic injury. Images Figure 1 Figure 2 Figure 3 Figure 5 Figure 6 Figure 7 PMID:9094992

A fluorescence method for determination of glucose transport by intestinal BBMV of common carp.

PubMed

Yang, Li-Ping; Yan, Xiao; Zheng, Wen-Jia; Hu, Jun-Yi; Zhang, Yu-Ru; Qin, Chao-Bin; Meng, Xiao-Lin; Lu, Rong-Hua; Chen, Fang; Xie, Di-Zhi; Nie, Guo-Xing

2017-11-15

Epithelial brush-border membrane vesicles (BBMVs) were isolated from the intestine of common carp and studied systematically by enzyme activity, transmission electron microscopy and immunoblotting. The uptake time course and the substrate concentration effect were assessed, and then, the ability of phlorizin and cytochalasin B to inhibit uptake was analyzed. The results show that sucrase, alkaline phosphatase and Na + -K + -ATPase activities in these vesicles were enriched 7.94-, 6.74- and 0.42-fold, respectively, indicating a relatively pure preparation of apical membrane with little basolateral contamination. The vesicular structure was in complete closure, as confirmed by electron microscopy. The presence of SGLT1 on the BBMVs was confirmed by Western blot analysis. In the time course experiment, the glucose uptake by BBMVs in Na + medium displayed an initial accumulation (overshoot) at 5 min followed by a rapid return to equilibrium values at 60 min. Over the 2-NBDG concentration range selected, the external 2-NBDG concentration in NaSCN medium graphed as a curved line. Phlorizin and cytochalasin B had an obvious inhibitory effect on 2-NBDG transport in carp BBMVs, and the detected fluorescence intensity decreased. The inhibition rate in the 1000 μM group was the strongest at 64.18% and 63.61% of phlorizin and cytochalasin B, respectively, indicating the presence of carriers other than SGLT1. This study is the first to demonstrate that 2-NBDG can be used as a convenient and sensitive probe to detect glucose uptake in fish BBMVs. This technology will provide a convenient method to discover new effects and factors in glucose metabolism. Copyright © 2017. Published by Elsevier Inc.

Developmental and cell-specific expression of thyroid hormone transporters in the mouse cochlea.

PubMed

Sharlin, David S; Visser, Theo J; Forrest, Douglas

2011-12-01

Thyroid hormone is essential for the development of the cochlea and auditory function. Cochlear response tissues, which express thyroid hormone receptor β (encoded by Thrb), include the greater epithelial ridge and sensory epithelium residing inside the bony labyrinth. However, these response tissues lack direct blood flow, implying that mechanisms exist to shuttle hormone from the circulation to target tissues. Therefore, we investigated expression of candidate thyroid hormone transporters L-type amino acid transporter 1 (Lat1), monocarboxylate transporter (Mct)8, Mct10, and organic anion transporting polypeptide 1c1 (Oatp1c1) in mouse cochlear development by in situ hybridization and immunofluorescence analysis. L-type amino acid transporter 1 localized to cochlear blood vessels and transiently to sensory hair cells. Mct8 localized to the greater epithelial ridge, tympanic border cells underlying the sensory epithelium, spiral ligament fibrocytes, and spiral ganglion neurons, partly overlapping with the Thrb expression pattern. Mct10 was detected in a highly restricted pattern in the outer sulcus epithelium and weakly in tympanic border cells and hair cells. Organic anion transporting polypeptide 1c1 localized primarily to fibrocytes in vascularized tissues of the spiral limbus and spiral ligament and to tympanic border cells. Investigation of hypothyroid Tshr(-/-) mice showed that transporter expression was delayed consistent with retardation of cochlear tissue maturation but not with compensatory responses to hypothyroidism. The results demonstrate specific expression of thyroid hormone transporters in the cochlea and suggest that a network of thyroid hormone transport underlies cochlear development.

Basolateral junctions are sufficient to suppress epithelial invasion during Drosophila oogenesis.

PubMed

Szafranski, Przemyslaw; Goode, Scott

2007-02-01

Epithelial junctions play crucial roles during metazoan evolution and development by facilitating tissue formation, maintenance, and function. Little is known about the role of distinct types of junctions in controlling epithelial transformations leading to invasion of neighboring tissues. Discovering the key junction complexes that control these processes and how they function may also provide mechanistic insight into carcinoma cell invasion. Here, using the Drosophila ovary as a model, we show that four proteins of the basolateral junction (BLJ), Fasciclin-2, Neuroglian, Discs-large, and Lethal-giant-larvae, but not proteins of other epithelial junctions, directly suppress epithelial tumorigenesis and invasion. Remarkably, the expression pattern of Fasciclin-2 predicts which cells will invade. We compared the apicobasal polarity of BLJ tumor cells to border cells (BCs), an epithelium-derived cluster that normally migrates during mid-oogenesis. Both tumor cells and BCs differentiate a lateralized membrane pattern that is necessary but not sufficient for invasion. Independent of lateralization, derepression of motility pathways is also necessary, as indicated by a strong linear correlation between faster BC migration and an increased incidence of tumor invasion. However, without membrane lateralization, derepression of motility pathways is also not sufficient for invasion. Our results demonstrate that spatiotemporal patterns of basolateral junction activity directly suppress epithelial invasion by organizing the cooperative activity of distinct polarity and motility pathways.

A study of the early detection of insect infestations and density/distribution of host plants. [Rio Grande

NASA Technical Reports Server (NTRS)

Hart, W. G. (Principal Investigator); Ingle, S. J.; Davis, M. R.

1975-01-01

The author has identified the following significant results. With comparative observations of film types and seasonal influences on reflectance characteristics, many crop varieties can be identified. This study shows that citrus, sugar cane, brush, some winter vegetables, and grain crops could be identified. Vegetative patterns in border areas can be detected. This information can be useful in detecting avenues of entry of pest species and areas of stress that require vigilance in stopping the spread of destructive species. Influence of some environmental factors on crops that may be confused with pest injury, or related factors, can be detected and identified with Skylab data (S-190B).

Structural Studies of the Intestinal α-Glucosidases, Maltase-glucoamylase and Sucrase-isomaltase.

PubMed

Rose, David R; Chaudet, Marcia M; Jones, Kyra

2018-06-01

Maltase-glucoamylase and sucrase-isomaltase are enzymes in the brush-border membrane of the small intestinal lumen responsible for the breakdown of postamylase starch polysaccharides to release monomeric glucose. As such, they are critical players in healthy nutrition and their malfunction can lead to severe disorders. This review covers investigations of the structures and functions of these enzymes. Each consists of 2 enzyme domains of the glycoside hydrolase family GH31 classification, yet with somewhat differing enzymatic properties. Crystallographic structures of 3 of the domains have been published. Insights into substrate binding and specificity will be discussed, along with future lines of inquiry related to the enzymes' roles in disease and potential avenues for therapeutics.

Expression of the cystic fibrosis transmembrane conductance regulator gene in the respiratory tract of normal individuals and individuals with cystic fibrosis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Trapnell, B.C.; Chinshyan Chu; Paakko, P.K.

1991-08-01

The most common mutation of the cystic fibrosis transmembrane conductance regulator gene, CFTR, associated with the clinical disorder cystic fibrosis (CF) is called {Delta}Phe{sup 508}, a triple-base deletion resulting in loss of phenylalanine at residue 508 of the predicted 1480-amino acid CFTR protein. In the context that the lung is the major site of morbidity and mortality in CF, the authors evaluated airway epithelial cells for CFTR mRNA transcripts in normal individuals, normal-{Delta}Phe{sup 508} heterozygotes, and {Delta}Phe{sup 508} homozygotes to determine if the normal and {Delta}Phe{sup 508} CFTR alleles are expressed in the respiratory epithelium, to what extent they aremore » expressed, and whether there are relative differences in the expression of the normal and abnormal alleles at the mRNA level. Respiratory tract epithelial cells recovered by fiberoptic bronchoscopy with a cytology brush demonstrated CFTR mRNA transcripts with sequences appropriately reflecting the normal and {Delta}Phe{sup 508} CFTR alleles of the various study groups. CFTR gene expression quantified by limited polymerase chain reaction amplification showed that in normal individuals, CFTR mRNA transcripts are expressed in nasal, tracheal, and bronchial epithelial cells.« less

HIV Impairs Lung Epithelial Integrity and Enters the Epithelium to Promote Chronic Lung Inflammation.

PubMed

Brune, Kieran A; Ferreira, Fernanda; Mandke, Pooja; Chau, Eric; Aggarwal, Neil R; D'Alessio, Franco R; Lambert, Allison A; Kirk, Gregory; Blankson, Joel; Drummond, M Bradley; Tsibris, Athe M; Sidhaye, Venkataramana K

2016-01-01

Several clinical studies show that individuals with HIV are at an increased risk for worsened lung function and for the development of COPD, although the mechanism underlying this increased susceptibility is poorly understood. The airway epithelium, situated at the interface between the external environment and the lung parenchyma, acts as a physical and immunological barrier that secretes mucins and cytokines in response to noxious stimuli which can contribute to the pathobiology of chronic obstructive pulmonary disease (COPD). We sought to determine the effects of HIV on the lung epithelium. We grew primary normal human bronchial epithelial (NHBE) cells and primary lung epithelial cells isolated from bronchial brushings of patients to confluence and allowed them to differentiate at an air- liquid interface (ALI) to assess the effects of HIV on the lung epithelium. We assessed changes in monolayer permeability as well as the expression of E-cadherin and inflammatory modulators to determine the effect of HIV on the lung epithelium. We measured E-cadherin protein abundance in patients with HIV compared to normal controls. Cell associated HIV RNA and DNA were quantified and the p24 viral antigen was measured in culture supernatant. Surprisingly, X4, not R5, tropic virus decreased expression of E-cadherin and increased monolayer permeability. While there was some transcriptional regulation of E-cadherin, there was significant increase in lysosome-mediated protein degradation in cells exposed to X4 tropic HIV. Interaction with CXCR4 and viral fusion with the epithelial cell were required to induce the epithelial changes. X4 tropic virus was able to enter the airway epithelial cells but not replicate in these cells, while R5 tropic viruses did not enter the epithelial cells. Significantly, X4 tropic HIV induced the expression of intercellular adhesion molecule-1 (ICAM-1) and activated extracellular signal-regulated kinase (ERK). We demonstrate that HIV can enter airway epithelial cells and alter their function by impairing cell-cell adhesion and increasing the expression of inflammatory mediators. These observed changes may contribute local inflammation, which can lead to lung function decline and increased susceptibility to COPD in HIV patients.

Developmental and Cell-Specific Expression of Thyroid Hormone Transporters in the Mouse Cochlea

PubMed Central

Sharlin, David S.; Visser, Theo J.

2011-01-01

Thyroid hormone is essential for the development of the cochlea and auditory function. Cochlear response tissues, which express thyroid hormone receptor β (encoded by Thrb), include the greater epithelial ridge and sensory epithelium residing inside the bony labyrinth. However, these response tissues lack direct blood flow, implying that mechanisms exist to shuttle hormone from the circulation to target tissues. Therefore, we investigated expression of candidate thyroid hormone transporters L-type amino acid transporter 1 (Lat1), monocarboxylate transporter (Mct)8, Mct10, and organic anion transporting polypeptide 1c1 (Oatp1c1) in mouse cochlear development by in situ hybridization and immunofluorescence analysis. L-type amino acid transporter 1 localized to cochlear blood vessels and transiently to sensory hair cells. Mct8 localized to the greater epithelial ridge, tympanic border cells underlying the sensory epithelium, spiral ligament fibrocytes, and spiral ganglion neurons, partly overlapping with the Thrb expression pattern. Mct10 was detected in a highly restricted pattern in the outer sulcus epithelium and weakly in tympanic border cells and hair cells. Organic anion transporting polypeptide 1c1 localized primarily to fibrocytes in vascularized tissues of the spiral limbus and spiral ligament and to tympanic border cells. Investigation of hypothyroid Tshr−/− mice showed that transporter expression was delayed consistent with retardation of cochlear tissue maturation but not with compensatory responses to hypothyroidism. The results demonstrate specific expression of thyroid hormone transporters in the cochlea and suggest that a network of thyroid hormone transport underlies cochlear development. PMID:21878515

Mechanisms of zinc transport into pig small intestine brush-border membrane vesicles.

PubMed Central

Tacnet, F; Lauthier, F; Ripoche, P

1993-01-01

1. The purpose of the present work was to examine certain membrane transport mechanisms likely to carry zinc across the brush-border membrane of pig small intestine, isolated in a vesicular form. 2. In initial velocity conditions, saturation kinetics revealed a great effect of pH on zinc transport: optimal conditions were observed with an intravesicular pH of around 6.6 with or without a H+ gradient; however, this did not allow us to conclude the existence of a neutral exchange between Zn2+ and H+ ions. 3. By measuring 36Cl uptakes, the presence of the Cl(-)-HCO3- or Cl(-)-OH-antiporter with typical 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS) sensitivity was detected in vesicles; zinc did not alter this anionic exchange activity. A 65Zn time course, performed in conditions identical with those for 36Cl uptake, was DIDS insensitive and was greatly inhibited by an outward OH- gradient. This could argue against a transport of zinc as a complex with Cl- and HCO3- through the anion antiporter. 4. When external Cl- and HCO3- were replaced by SCN-, able to form a Zn(SCN)4(2-) complex, we observed a stimulating effect of outward HCO3- gradients on 65Zn uptake but neither DIDS nor diphenylamine-2-carboxylate (DPC) inhibited the transport in these conditions. This suggested that the intestinal anion antiporter was not a major route for zinc reabsorption. 5. The tripeptide Gly-Gly-His at low concentrations stimulated 65Zn uptake, then inhibited it in a dose-dependent manner either in the presence of an inward H+ gradient or in the presence of a membrane potential 'negative inside' or in both situations. These conditions are necessary for the active transport of the peptide and this strongly suggests that zinc can be transported as a [Gly-Gly-His-Zn] complex, utilizing the peptide carrier system. PMID:8229851

Heat-Induced Structural Changes Affect OVA-Antigen Processing and Reduce Allergic Response in Mouse Model of Food Allergy

PubMed Central

Wallner, Michael; Kverka, Miloslav; Kozakova, Hana; Srutkova, Dagmar; Klimesova, Klara; Sotkovsky, Petr; Palova-Jelinkova, Lenka; Ferreira, Fatima; Tuckova, Ludmila

2012-01-01

Background and Aims The egg protein ovalbumin (OVA) belongs to six most frequent food allergens. We investigated how thermal processing influences its ability to induce allergic symptoms and immune responses in mouse model of food allergy. Methodology/Principal Findings Effect of increased temperature (70°C and 95°C) on OVA secondary structure was characterized by circular dichroism and by the kinetics of pepsin digestion with subsequent HPLC. BALB/c mice were sensitized intraperitoneally and challenged with repeated gavages of OVA or OVA heated to 70°C (h-OVA). Levels of allergen-specific serum antibodies were determined by ELISA (IgA and IgGs) or by β-hexosaminidase release test (IgE). Specific activities of digestive enzymes were determined in brush border membrane vesicles of jejunal enterocytes. Cytokine production and changes in regulatory T cells in mesenteric lymph nodes and spleen were assessed by ELISA and FACS. Heating of OVA to 70°C caused mild irreversible changes in secondary structure compared to boiling to 95°C (b-OVA), but both OVA treatments led to markedly different digestion kinetics and Tregs induction ability in vitro, compared to native OVA. Heating of OVA significantly decreased clinical symptoms (allergic diarrhea) and immune allergic response on the level of IgE, IL-4, IL-5, IL-13. Furthermore, h-OVA induced lower activities of serum mast cell protease-1 and enterocyte brush border membrane alkaline phosphatase as compared to native OVA. On the other hand h-OVA stimulated higher IgG2a in sera and IFN-γ secretion by splenocytes. Conclusions Minor irreversible changes in OVA secondary structure caused by thermal processing changes both its digestion and antigenic epitopes formation, which leads to activation of different T cell subpopulations, induces shift towards Th1 response and ultimately reduces its allergenicity. PMID:22629361

Molecular transport machinery involved in orchestrating luminal acid-induced duodenal bicarbonate secretion in vivo

PubMed Central

Singh, Anurag Kumar; Liu, Yongjian; Riederer, Brigitte; Engelhardt, Regina; Thakur, Basant Kumar; Soleimani, Manoocher; Seidler, Ursula

2013-01-01

The duodenal villus brush border membrane expresses several ion transporters and/or channels, including the solute carrier 26 anion transporters Slc26a3 (DRA) and Slc26a6 (PAT-1), the Na+/H+ exchanger isoform 3 (NHE3), as well as the anion channels cystic fibrosis transmembrane conductance regulator (CFTR) and Slc26a9. Using genetically engineered mouse models lacking Scl26a3, Slc26a6, Slc26a9 or Slc9a3 (NHE3), the study was carried out to assess the role of these transporters in mediating the protective duodenal bicarbonate secretory response (DBS-R) to luminal acid; and to compare it to their role in DBS-R elicited by the adenylyl cyclase agonist forskolin. While basal DBS was reduced in the absence of any of the three Slc26 isoforms, the DBS-R to forskolin was not altered. In contrast, the DBS-R to a 5 min exposure to luminal acid (pH 2.5) was strongly reduced in the absence of Slc26a3 or Slc26a9, but not Slc26a6. CFTR inhibitor [CFTR(Inh)-172] reduced the first phase of the acid-induced DBS-R, while NHE3 inhibition (or knockout) abolished the sustained phase of the DBS-R. Luminal acid exposure resulted in the activation of multiple intracellular signalling pathways, including SPAK, AKT and p38 phosphorylation. It induced a biphasic trafficking of NHE3, first rapidly into the brush border membrane, followed by endocytosis in the later stage. We conclude that the long-lasting DBS-R to luminal acid exposure activates multiple duodenocyte signalling pathways and involves changes in trafficking and/or activity of CFTR, Slc26 isoforms Slc26a3 and Slc26a9, and NHE3. PMID:24018950

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. Copyright 2004 Wiley-Liss, Inc.

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 to dispose of endogenous cholesterol efficiently for therapeutic purposes. PMID:27023132

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 to dispose of endogenous cholesterol efficiently for therapeutic purposes.

Papillary Architecture and Functional Characterization of Mucosubstances in the Sheep Tongue.

PubMed

Erdoğan, Serkan; Sağsöz, Hakan

2018-04-30

This research aimed to reveal the general morphology and topographic distribution of lingual papillae, epithelial characteristics, mucosal structure, and glands with their mucin content in the sheep tongue, with consideration of species-specific characteristics. The tongues of ten sheep were analyzed for this purpose. Filiform and fungiform papillae existed within the borders of the ventral surface of the lingual apex. The majority of the filiform papillae had multiple secondary projections. Fungiform papillae were also seen on the lingual torus among lenticular papillae, as well as 6 to 10 circumvallate papillae arranged on its caudal border. The species-specific details of the general anatomical structure of the tongue were determined and, in general, the papillary organization in the sheep was similar to goats, while the papillary organization also was similar to features with deer species, specifically the filiform papilla from the mechanical papillae and fungiform papilla from the gustatory papillae. Neutral and weak sulfated mucins and N-acetyl sialomucins were located in seromucous glands, salivary duct epithelium and von Ebner's glands. Carboxylated acid mucins and N-acetyl sialomucins were not present in seromucous and von Ebner's glands. In seromucous glands, MUC1, MUC5AC and MUC6 localized only in epithelial cells of ducts, whereas MUC2 localized in both glandular and ductal epithelial cells. All MUCs were present in both von Ebner's glands and salivary ducts. We showed that this mucin composition, may serve as a physical barrier in the initial section of the digestive system. This article is protected by copyright. All rights reserved. © 2018 Wiley Periodicals, Inc.

Surface characteristics of isopod digestive gland epithelium studied by SEM.

PubMed

Millaku, Agron; Leser, Vladka; Drobne, Damjana; Godec, Matjaz; Torkar, Matjaz; Jenko, Monika; Milani, Marziale; Tatti, Francesco

2010-05-01

The structure of the digestive gland epithelium of a terrestrial isopod Porcellio scaber has been investigated by conventional scanning electron microscopy (SEM), focused ion beam-scanning electron microscopy (FIB/SEM), and light microscopy in order to provide evidence on morphology of the gland epithelial surface in animals from a stock culture. We investigated the shape of cells, extrusion of lipid droplets, shape and distribution of microvilli, and the presence of bacteria on the cell surface. A total of 22 animals were investigated and we found some variability in the appearance of the gland epithelial surface. Seventeen of the animals had dome-shaped digestive gland "normal" epithelial cells, which were densely and homogeneously covered by microvilli and varying proportions of which extruded lipid droplets. On the surface of microvilli we routinely observed sparsely distributed bacteria of different shapes. Five of the 22 animals had "abnormal" epithelial cells with a significantly altered shape. In three of these animals, the cells were much smaller, partly or completely flat or sometimes pyramid-like. A thick layer of bacteria was detected on the microvillous border, and in places, the shape and size of microvilli were altered. In two animals, hypertrophic cells containing large vacuoles were observed indicating a characteristic intracellular infection. The potential of SEM in morphological investigations of epithelial surfaces is discussed.

[Genetic hypophosphatemia: recent advances in physiopathogenic concept].

PubMed

Beraud, G; Perimenis, P; Velayoudom, Fr-L; Wemeau, J-L; Vantyghem, M-Chr

2005-04-01

Renal proximal tubular reabsorption of phosphate and intestinal absorption both regulate phosphate homeostasis. Brush-border membrane Npt2a cotransporter is the key element in proximal tubular P (i) reabsorption. Inactivating mutations of Npt2a cause bone demineralisation and urolithiasis. An excess of a phosphaturic factor, called "Phosphatonin", could modulate phosphate reabsorption by inhibition on Npt2a. Inactivating mutation of PHEX, an endopeptidase-membrane coding gene, is responsible for X-linked Hypophosphatemia (XLH), because of an impaired degradation of phosphatonine by PHEX product. Autosomic Dominant Hypophosphatemic Rickets (ADHR) is explained by a mutation preventing FGF23 (one of the best identified phosphatonines) from cleavage. According recent data, FGF23, MEPE (Matrix Extracellular Phosphoglycoprotein) et FRP4 (frizzled related protein-4) are 3 putative "phosphatonines".

Vaccination of sheep against haemonchosis with H11, a gut membrane-derived protective antigen from the adult parasite: prevention of the periparturient rise and colostral transfer of protective immunity.

PubMed

Andrews, S J; Hole, N J; Munn, E A; Rolph, T P

1995-07-01

Pregnant ewes were immunised with a fraction highly enriched in the membrane glycoprotein antigen H11, isolated from the intestinal brush border of adult Haemonchus contortus. Immunity induced by immunisation was able to abolish almost completely (98-99%) the worm egg output from pregnant ewes challenged with ca. 10,000 infective larvae of H. contortus during the last trimester. Furthermore, lambs born and reared on vaccinated ewes had substantial antibody levels to H11 derived from maternal transfer. This antibody conferred moderate protection against a bolus challenge of ca. 3000 infective larvae of H. contortus in 5-week-old lambs.

Metabolic stability of some tachykinin analogues to cell-surface peptidases: roles for endopeptidase-24.11 and aminopeptidase N.

PubMed

Medeiros, M D; Turner, A J

1995-01-01

The metabolism of several tachykinin antagonists by membrane peptidases has been examined. [beta Ala8]NKA(4-10) was not stabilized against degradation by endopeptidase-24.11 and this was the major activity in renal brush border membranes hydrolyzing this peptide. The antagonist MEN 10263 was much more resistant to hydrolysis by endopeptidase-24.11, although hydrolysis of the C-terminal Leu-Phe bond was detectable. Three other tachykinin receptor antagonists (MEN 10208, MEN 10207, and MEN 10376), by virtue of D-Trp substitutions, were rendered resistant to endopeptidase-24.11 but were still susceptible to aminopeptidase action. These studies provide further insight into design features necessary to produce metabolically stable peptide analogues.

On physical changes on surface of human cervical epithelial cells during cancer transformations

NASA Astrophysics Data System (ADS)

Sokolov, Igor; Dokukin, Maxim; Guz, Nataliia; Woodworth, Craig

2013-03-01

Physical changes of the cell surface of cells during transformation from normal to cancerous state are rather poorly studied. Here we describe our recent studies of such changes done on human cervical epithelial cells during their transformation from normal through infected with human papillomavirus type-16 (HPV-16), immortalized (precancerous), to cancerous cells. The changes were studied with the help of atomic force microscopy (AFM) and through the measurement of physical adhesion of fluorescent silica beads to the cell surface. Based on the adhesion experiments, we clearly see the difference in nonspecific adhesion which occurs at the stage of immortalization of cells, precancerous cells. The analysis done with the help of AFM shows that the difference observed comes presumably from the alteration of the cellular ``brush,'' a layer that surrounds cells and which consists of mostly microvilli, microridges, and glycocalyx. Further AFM analysis reveals the emergence of fractal scaling behavior on the surface of cells when normal cells turn into cancerous. The possible causes and potential significance of these observations will be discussed.

Chemical coding and chemosensory properties of cholinergic brush cells in the mouse gastrointestinal and biliary tract.

PubMed

Schütz, Burkhard; Jurastow, Innokentij; Bader, Sandra; Ringer, Cornelia; von Engelhardt, Jakob; Chubanov, Vladimir; Gudermann, Thomas; Diener, Martin; Kummer, Wolfgang; Krasteva-Christ, Gabriela; Weihe, Eberhard

2015-01-01

The mouse gastro-intestinal and biliary tract mucosal epithelia harbor choline acetyltransferase (ChAT)-positive brush cells with taste cell-like traits. With the aid of two transgenic mouse lines that express green fluorescent protein (EGFP) under the control of the ChAT promoter (EGFP (ChAT) ) and by using in situ hybridization and immunohistochemistry we found that EGFP (ChAT) cells were clustered in the epithelium lining the gastric groove. EGFP (ChAT) cells were numerous in the gall bladder and bile duct, and found scattered as solitary cells along the small and large intestine. While all EGFP (ChAT) cells were also ChAT-positive, expression of the high-affinity choline transporter (ChT1) was never detected. Except for the proximal colon, EGFP (ChAT) cells also lacked detectable expression of the vesicular acetylcholine transporter (VAChT). EGFP (ChAT) cells were found to be separate from enteroendocrine cells, however they were all immunoreactive for cytokeratin 18 (CK18), transient receptor potential melastatin-like subtype 5 channel (TRPM5), and for cyclooxygenases 1 (COX1) and 2 (COX2). The ex vivo stimulation of colonic EGFP (ChAT) cells with the bitter substance denatonium resulted in a strong increase in intracellular calcium, while in other epithelial cells such an increase was significantly weaker and also timely delayed. Subsequent stimulation with cycloheximide was ineffective in both cell populations. Given their chemical coding and chemosensory properties, EGFP (ChAT) brush cells thus may have integrative functions and participate in induction of protective reflexes and inflammatory events by utilizing ACh and prostaglandins for paracrine signaling.

Interferon-gamma increased epithelial barrier function via upregulating claudin-7 expression in human submandibular gland duct epithelium.

PubMed

Abe, Ayumi; Takano, Kenichi; Kojima, Takashi; Nomura, Kazuaki; Kakuki, Takuya; Kaneko, Yakuto; Yamamoto, Motohisa; Takahashi, Hiroki; Himi, Tetsuo

2016-06-01

Tight junctions (TJs) are necessary for salivary gland function and may serve as indicators of salivary gland epithelial dysfunction. IgG4-related disease (IgG4-RD) is a newly recognized fibro-inflammatory condition which disrupts the TJ associated epithelial barrier. The salivary glands are one of the most frequently involved organs in IgG4-RD, however, changes of the TJ associated epithelial barrier in salivary gland duct epithelium is poorly understood. Here, we investigated the regulation and function of TJs in human submandibular gland ductal epithelial cells (HSDECs) in normal and IgG4-RD. We examined submandibular gland (SMG) tissue from eight control individuals and 22 patients with IgG4-RD and established an HSDEC culture system. Immunohistochemistry, immunocytochemistry, western blotting, and measurement of transepithelial electrical resistance (TER) were performed. Claudin-4, claudin-7, occludin, and JAM-A were expressed at the apical side of the duct epithelium in submandibular gland (SMG) tissue and at the cell borders in HSDECs of normal and IgG4-RD. The expression and distribution of TJs in SMG tissue were not different in control individuals and patients with IgG4-RD in vivo and in vitro. Although interferon-gamma (IFNγ) generally disrupts the integrity and function of TJs, as manifested by decreased epithelial barrier function, IFNγ markedly increased the epithelial barrier function of HSDECs via upregulation of claudin-7 expression in HSDECs from patients with IgG4-RD. This is the first report showing an IFNγ-dependent increase in epithelial barrier function in the salivary gland duct epithelium. Our results provide insights into the functional significance of TJs in salivary gland duct epithelium in physiological and pathological conditions, including IgG4-RD.

A Comparative Study of Intratesticular Ductules in the Spermatogenically Active Testes of Shortfin Mako and Thresher Sharks.

PubMed

McClusky, Leon Mendel; Sulikowski, James

2016-10-01

This comparative study of the radial testes of sexually mature thresher sharks (Alopias vulpinus) and shortfin mako sharks (Isurus oxyrinchus) describes the histology of the three-tiered network of sperm-carrying ductules in the testis and the lymphomyeloid tissue associated with it, namely the epigonal organ. In both species, a testis → epigonal gradient was evident regarding the thickness of the ductule epithelial lining and subepithelial investment of connective tissue. Ductules straddling the testis-epigonal border often displayed luminal leukocytes and various signs of regression, including the progressive thickening of the ductule epithelial lining, dissolution of the cytoplasm, and loss of normal histoarchitecture. In Isurus, large amorphous areas formed due to the fusion of neighboring regressing ductules. The epigonal organ of Alopias additionally revealed circular degenerative sperm-containing, Hassall-like bodies with either a degenerate or cellular appearance, the latter the result of cell proliferative activity (as shown by proliferating cell nuclear antigen (PCNA) immunohistochemistry) in an expanding outer border comprising cells with intensely PCNA immunoreactive slender and oblong nuclei. The latter cells exhibited a periphery-to-center transformation of their nuclei, at which stage they were PCNA-negative and most likely in a terminally differentiated state as they phagocytized the cell debris in the degenerate core. Intermediate stages of these circular bodies were a rarity. The relationship between these degenerate bodies, and the common occurrence of blind pockets in the epithelial linings and non-apoptosis-related degenerate patches in the apical cytoplasmic regions of the irregular shaped ductules in Alopias is unclear, and needs further elucidation. Anat Rec, 299:1435-1448, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Stromal and epithelial cells react differentially to c-kit in fibroepithelial tumors of the breast.

PubMed

Logullo, Angela F; Nonogaki, Suely; Do Socorro Maciel, Maria; Mourão-Neto, Mário; Soares, Fernando Augusto

2008-01-01

The CD117 protein is a tyrosine-kinase receptor encoded by the c-kit gene that frequently bears activating mutations in gastrointestinal tumors. Conflicting findings regarding CD117 expression in other stromal tumors, including phyllodes tumors (PTs), have been reported in the literature. The purpose of this study was to evaluate c-kit expression in the stroma and epithelia of fibroepithelial breast tumors and its correlation with clinical pathological variables. Ninety-six fibroepithelial tumors of the breast, including 14 fibroadenomas (FAs), 12 juvenile FAs and 70 PTs, were classified according to stromal cellularity, atypia, epithelial hyperplasia, mitosis and borders into 45 benign (PTB), 17 borderline (PTBL) and 8 malignant (PTM) tumors. CD117 expression was identified in the stromal component in only two cases of PTBL. Overall, 38 cases (39.6%) showed positive CD117 in the epithelial component, including 20 FAs (10 regular, 10 juvenile) and 18 PTs (11 PTBs and 8 PTBLs). Other cases, including all PTMs, 6 FAs (4 regular, 2 juvenile), 34 PTBs and 10 PTBLs, showed no positivity in the epithelial component. Expression of c-kit did not correlate with diagnosis or malignancy (p>0.05). In conclusion, c-kit is expressed more often in the epithelial than in the stromal component in fibroepithelial tumors of the breast, and is associated with benign lesions.

Intestinal alkaline phosphatase prevents metabolic syndrome in mice.

PubMed

Kaliannan, Kanakaraju; Hamarneh, Sulaiman R; Economopoulos, Konstantinos P; Nasrin Alam, Sayeda; Moaven, Omeed; Patel, Palak; Malo, Nondita S; Ray, Madhury; Abtahi, Seyed M; Muhammad, Nur; Raychowdhury, Atri; Teshager, Abeba; Mohamed, Mussa M Rafat; Moss, Angela K; Ahmed, Rizwan; Hakimian, Shahrad; Narisawa, Sonoko; Millán, José Luis; Hohmann, Elizabeth; Warren, H Shaw; Bhan, Atul K; Malo, Madhu S; Hodin, Richard A

2013-04-23

Metabolic syndrome comprises a cluster of related disorders that includes obesity, glucose intolerance, insulin resistance, dyslipidemia, and fatty liver. Recently, gut-derived chronic endotoxemia has been identified as a primary mediator for triggering the low-grade inflammation responsible for the development of metabolic syndrome. In the present study we examined the role of the small intestinal brush-border enzyme, intestinal alkaline phosphatase (IAP), in preventing a high-fat-diet-induced metabolic syndrome in mice. We found that both endogenous and orally supplemented IAP inhibits absorption of endotoxin (lipopolysaccharides) that occurs with dietary fat, and oral IAP supplementation prevents as well as reverses metabolic syndrome. Furthermore, IAP supplementation improves the lipid profile in mice fed a standard, low-fat chow diet. These results point to a potentially unique therapy against metabolic syndrome in at-risk humans.

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

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 backbonemore » 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.« less

Restoration of CFTR function in patients with cystic fibrosis carrying the F508del-CFTR mutation.

PubMed

De Stefano, Daniela; Villella, Valeria R; Esposito, Speranza; Tosco, Antonella; Sepe, Angela; De Gregorio, Fabiola; Salvadori, Laura; Grassia, Rosa; Leone, Carlo A; De Rosa, Giuseppe; Maiuri, Maria C; Pettoello-Mantovani, Massimo; Guido, Stefano; Bossi, Anna; Zolin, Anna; Venerando, Andrea; Pinna, Lorenzo A; Mehta, Anil; Bona, Gianni; Kroemer, Guido; Maiuri, Luigi; Raia, Valeria

2014-01-01

Restoration of BECN1/Beclin 1-dependent autophagy and depletion of SQSTM1/p62 by genetic manipulation or autophagy-stimulatory proteostasis regulators, such as cystamine, have positive effects on mouse models of human cystic fibrosis (CF). These measures rescue the functional expression of the most frequent pathogenic CFTR mutant, F508del, at the respiratory epithelial surface and reduce lung inflammation in Cftr(F508del) homozygous mice. Cysteamine, the reduced form of cystamine, is an FDA-approved drug. Here, we report that oral treatment with cysteamine greatly reduces the mortality rate and improves the phenotype of newborn mice bearing the F508del-CFTR mutation. Cysteamine was also able to increase the plasma membrane expression of the F508del-CFTR protein in nasal epithelial cells from F508del homozygous CF patients, and these effects persisted for 24 h after cysteamine withdrawal. Importantly, this cysteamine effect after washout was further sustained by the sequential administration of epigallocatechin gallate (EGCG), a green tea flavonoid, both in vivo, in mice, and in vitro, in primary epithelial cells from CF patients. In a pilot clinical trial involving 10 F508del-CFTR homozygous CF patients, the combination of cysteamine and EGCG restored BECN1, reduced SQSTM1 levels and improved CFTR function from nasal epithelial cells in vivo, correlating with a decrease of chloride concentrations in sweat, as well as with a reduction of the abundance of TNF/TNF-alpha (tumor necrosis factor) and CXCL8 (chemokine [C-X-C motif] ligand 8) transcripts in nasal brushing and TNF and CXCL8 protein levels in the sputum. Altogether, these results suggest that optimal schedules of cysteamine plus EGCG might be used for the treatment of CF caused by the F508del-CFTR mutation.

Protective effect of Urtica dioica L. on renal ischemia/reperfusion injury in rat.

PubMed

Sayhan, Mustafa Burak; Kanter, Mehmet; Oguz, Serhat; Erboga, Mustafa

2012-12-01

Renal ischemia-reperfusion (I/R) injury may occur after renal transplantation, thoracoabdominal aortic surgery, and renal artery interventions. This study was designed to investigate the effect of Urtica dioica L. (UD), in I/R induced renal injury. A total of 32 male Sprague-Dawley rats were divided into four groups: control, UD alone, I/R and I/R + UD; each group contain 8 animals. A rat model of renal I/R injury was induced by 45-min occlusion of the bilateral renal pedicles and 24-h reperfusion. In the UD group, 3 days before I/R, UD (2 ml/kg/day intraperitoneal) was administered by gastric gavage. All animals were sacrificed at the end of reperfusion and kidney tissues samples were obtained for histopathological investigation in all groups. To date, no more histopathological changes on intestinal I/R injury in rats by UD treatment have been reported. Renal I/R caused severe histopathological injury including tubular damage, atrophy dilatation, loss of brush border and hydropic epithelial cell degenerations, renal corpuscle atrophy, glomerular shrinkage, markedly focal mononuclear cell infiltrations in the kidney. UD treatment significantly attenuated the severity of intestinal I/R injury and significantly lowered tubulointerstitial damage score than the I/R group. The number of PCNA and TUNEL positive cells in the control and UD alone groups was negligible. When kidney sections were PCNA and TUNEL stained, there was a clear increase in the number of positive cells in the I/R group rats in the renal cortical tissues. However, there is a significant reduction in the activity of PCNA and TUNEL in kidney tissue of renal injury induced by renal I/R with UD therapy. Our results suggest that administration of UD attenuates renal I/R injury. These results suggest that UD treatment has a protective effect against renal damage induced by renal I/R. This protective effect is possibly due to its ability to inhibit I/R induced renal damage, apoptosis and cell proliferation.

Study of the role of epididymal alpha-glucosidase in the fertility of male rats by the administration of the enzyme inhibitor castanospermine.

PubMed

Yeung, C H; Cooper, T G

1994-11-01

The activity of epididymal alpha-glucosidase in adult rats was rapidly suppressed to histochemically undetectable levels within 2 days by the continuous release of the enzyme inhibitor castanospermine via a peritoneal osmotic pump at a rate of 100-200 nmol h-1. It was established that mating activities overnight depleted 72% of the spermatozoa in the distal cauda, which was replenished in 2 days, and that fertility began to decline 3 weeks after efferent duct ligation. Male rats of proven mating proficiency and fertility were treated with castanospermine, or buffered saline as control, for up to 30 days and enzyme inhibition was confirmed at the end of treatment by histochemistry. Fertility was normal at the first mating test on day 7, significantly decreased at the second mating on day 9, but recovered in a stepwise manner at subsequent matings on days 12 and 14. Delaying the third mating until day 25 did not sustain the transient subfertility. However, prolonging sperm storage in the distal cauda epididymides and preventing replenishment with freshly matured spermatozoa, by efferent duct ligation for 14 days performed on day 15 during castanospermine administration, caused a decrease in fertility and a change in the kinematics of epididymal spermatozoa of the castanospermine-treated group. In control rats, binding of epididymal spermatozoa to Vicia faba, a lectin specific for glucose and glucosamine, and mannose and mannosamine residues, decreased from the proximal caput to the distal corpus coincident with the increase in alpha-glucosidase activity on the epithelial brush border. Lectin binding then increased in the cauda where enzyme activity was absent. However, castanospermine treatment did not significantly alter this binding profile. The findings suggest that epididymal alpha-glucosidase does not play a crucial role in the development of sperm fertilizing capacity, but may be involved in the preparation of spermatozoa for storage.

Goblet Cells and Mucus Types in the Digestive Intestine and Respiratory Intestine in Bronze Corydoras (Callichthyidae: Teleostei).

PubMed

Leknes, I L

2015-10-01

The structure and histochemical properties of the intestine in bronze corydoras (Corydoras aeneus), a stomach-containing teleost, are described, with emphasis on goblet cells and mucin types. The proximal intestine displayed a normal structure for teleosts, whereas the distal intestine was wide, translucent, thin-walled, richly vascularized and constantly filled with air, suggesting an important respiratory role. Goblet cells were common throughout the entire intestine and displayed a variable, but mainly faint metachromatic colour after toluidine blue. They were moderately coloured by alcian blue at both pH 2.5 and 0.2 and displayed no colour after periodic acid followed by Schiff's solution (PAS), but a distinct purple-brown colour after high iron diamine followed by alcian blue (pH 2.5). Together, these results suggest that the mucin in the intestine goblet cells consists mainly of sulphated proteoglycans. Further, the results from the present lectin and neuraminidase tests suggest that these mucins contain much N-acetylglucoseamines and some N-acetylgalactosamines and sialic acid, but seem to lack glucose and mannose. They also contain some galactose-N-acetylgalactosamines sequences, normally hidden by sialic acid. The distinct brush border and mucus layer on the epithelial cells in the respiratory intestine may indicate some digestive roles, such as absorption of water, ions and simple carbohydrates. As sulphated proteoglycans are tough and attract much water, this mucus may play important roles in the protection against mechanical and chemical damages and in the defence against micro-organisms throughout the entire intestine, but in the respiratory intestine it may impede significantly the oxygen uptake. However, as this part of the intestine usually contains no digesta, but is completely filled with air, frequently renewed by dry air from the atmosphere, and the main function of the mucus may be to protect the respiratory epithelium against a destroying and dangerous desiccation. © 2014 Blackwell Verlag GmbH.

Membrane peptidases in the pig choroid plexus and on other cell surfaces in contact with the cerebrospinal fluid.

PubMed Central

Bourne, A; Barnes, K; Taylor, B A; Turner, A J; Kenny, A J

1989-01-01

A comprehensive survey of 11 peptidases, all of which are markers for renal microvillar membranes, has been made in membrane fractions prepared from pig choroid plexus. Two fractionation schemes were explored, both depending on a MgCl2-precipitation step, the preferred one having advantages in speed and yield of the activities. The specific activities of the peptidases in the choroid-plexus membranes were, with the exception of carboxypeptidase M, lower than in renal microvillar membranes: those of aminopeptidase N, peptidyl dipeptidase A ('angiotensin-converting enzyme') and gamma-glutamyltransferase were 3-5-fold lower, those of aminopeptidase A and endopeptidase-24.11 were 12-15 fold lower, and those of dipeptidyl peptidase IV and aminopeptidase W were 50-70-fold lower. Carboxypeptidase M had a similar activity in both membranes. Alkaline phosphatase and (Na+ + K+)-activated ATPase were more active in the choroid-plexus membranes. No activity for microsomal dipeptidase, aminopeptidase P and carboxypeptidase P could be detected. Six of the peptidases and (Na+ + K+)-activated ATPase were also studied by immunoperoxidase histochemistry at light- and electron-microscopic levels. Endopeptidase-24.11 and (Na+ + K+)-activated ATPase were uniquely located on the brush border, and the other two peptidases appeared to be much more abundant on the endothelial lining of microvessels. Dipeptidyl peptidase IV and aminopeptidase W were also detected in microvasculature. Pial membranes associated with the brain and spinal cord also stained positively for endopeptidase-24.11, aminopeptidase N and peptidyl dipeptidase A. The immunohistochemical studies indicated the subcellular fractionation did not discriminate between membranes derived from epithelial cells (i.e. microvilli) and those from endothelial cells. The possible significance of these studies in relation to neuropeptide metabolism and the control of cerebrospinal fluid production is discussed. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. PMID:2655579

Bacillus thuringiensis Cry1Ia10 and Vip3Aa protein interactions and their toxicity in Spodoptera spp. (Lepidoptera).

PubMed

Bergamasco, V B; Mendes, D R P; Fernandes, O A; Desidério, J A; Lemos, M V F

2013-02-01

The polyphagous pests belonging to the genus Spodoptera are considered to be among the most important causes of damage and are widely distributed throughout the Americas'. Due to the extensive use of genetically modified plants containing Bacillus thuringiensis genes that code for insecticidal proteins, resistant insects may arise. To prevent the development of resistance, pyramided plants, which express multiple insecticidal proteins that act through distinct mode of actions, can be used. This study analyzed the mechanisms of action for the proteins Cry1Ia10 and Vip3Aa on neonatal Spodoptera frugiperda, Spodoptera albula, Spodoptera eridania and Spodoptera cosmioides larvae. The interactions of these toxins with receptors on the intestinal epithelial membrane were also analyzed by binding biotinylated toxins to brush border membrane vesicles (BBMVs) from the intestines of these insects. A putative receptor of approximately 65 kDa was found by ligand blotting in all of these species. In vitro competition assays using biotinylated proteins have indicated that Vip3Aa and Cry1Ia10 do not compete for the same receptor for S. frugiperda, S. albula and S. cosmioides and that Vip3Aa was more efficient than Cry1Ia10 when tested individually, by bioassays. A synergistic effect of the toxins in S. frugiperda, S. albula and S. cosmioides was observed when they were combined. However, in S. eridania, Cry1Ia10 and Vip3Aa might compete for the same receptor and through bioassays Cry1Ia10 was more efficient than Vip3Aa and showed an antagonistic effect when the proteins were combined. These results suggest that using these genes to develop pyramided plants may not prove effective in preventing the development of resistance in S. eridiana. Copyright © 2012 Elsevier Inc. All rights reserved.

Effect of Frequency of Brushing Teeth on Plaque and Calculus Accumulation, and Gingivitis in Dogs.

PubMed

Harvey, Colin; Serfilippi, Laurie; Barnvos, Donald

2015-01-01

The efficacy of brushing the teeth of beagle dogs in a randomized, controlled, blinded study design using a clearly-defined brushing technique was evaluated for 4 brushing frequencies: brushing daily, brushing every other day, brushing weekly and brushing every other week, compared with no brushing in a control group of dogs. All dogs were fed a standard dry kibble diet during the study. Standard plaque, calculus, and gingivitis indices were used to score the teeth. A 'clean tooth' model was used. No gingival or non-gingival lacerations or other signs of injury to oral tissues were found at the end of the 28 day trial period. Brushing more frequently had greater effectiveness in retarding accumulation of plaque and calculus, and reducing the severity of pre-existing gingivitis. Brushing daily or every other day produced statistically significant improved results compared with brushing weekly or every other week. Based on the results of this study, daily brushing is recommended.

Band-like arrangement of taste-like sensory cells at the gastric groove: evidence for paracrine communication.

PubMed

Eberle, Julia Anna-Maria; Richter, Patric; Widmayer, Patricia; Chubanov, Vladimir; Gudermann, Thomas; Breer, Heinz

2013-01-01

The discovery of taste-related elements within the gastrointestinal tract has led to a growing interest in the mechanisms and physiological significance of chemosensory monitoring of chymus composition. Previous work suggests that brush cells located in the "gastric groove," which parallels the "limiting ridge," a structure in rodents that divides the fundus from the corpus, are candidate sensory cells. A novel sectioning technique revealed that these cells are arranged in a palisade-like manner forming a band which borders the whole length of the corpus epithelium. Using transgenic PLCβ2 promoter-GFP mice and specific antibodies, we have demonstrated that most of these cells express gustducin, PLCβ2, and TRPM5; typical signaling proteins of gustatory sensory "type II" cells. These molecular features strongly suggest that the cells may be capable of sensing nutrient or non-nutrient constituents of the ingested food. Since there is no evidence that brush cells are endocrine cells, attempts were made to explore how such putative chemosensory cells might transmit the information to "effector" cells. It was found that most of the cells express the neuronal nitric oxide synthase (NOS) suggesting some paracrine interaction with adjacent cells. Moreover, they also express choline acetyltransferase (ChAT) as well as the vesicular protein SNAP25, indicating the potential for cholinergic transmission, possibly with subjacent enteric nerve fibers.

Band-like arrangement of taste-like sensory cells at the gastric groove: evidence for paracrine communication

PubMed Central

Eberle, Julia Anna-Maria; Richter, Patric; Widmayer, Patricia; Chubanov, Vladimir; Gudermann, Thomas; Breer, Heinz

2013-01-01

The discovery of taste-related elements within the gastrointestinal tract has led to a growing interest in the mechanisms and physiological significance of chemosensory monitoring of chymus composition. Previous work suggests that brush cells located in the “gastric groove,” which parallels the “limiting ridge,” a structure in rodents that divides the fundus from the corpus, are candidate sensory cells. A novel sectioning technique revealed that these cells are arranged in a palisade-like manner forming a band which borders the whole length of the corpus epithelium. Using transgenic PLCβ2 promoter-GFP mice and specific antibodies, we have demonstrated that most of these cells express gustducin, PLCβ2, and TRPM5; typical signaling proteins of gustatory sensory “type II” cells. These molecular features strongly suggest that the cells may be capable of sensing nutrient or non-nutrient constituents of the ingested food. Since there is no evidence that brush cells are endocrine cells, attempts were made to explore how such putative chemosensory cells might transmit the information to “effector” cells. It was found that most of the cells express the neuronal nitric oxide synthase (NOS) suggesting some paracrine interaction with adjacent cells. Moreover, they also express choline acetyltransferase (ChAT) as well as the vesicular protein SNAP25, indicating the potential for cholinergic transmission, possibly with subjacent enteric nerve fibers. PMID:23565094

Control of brushing variables for the in vitro assessment of toothpaste abrasivity using a novel laboratory model.

PubMed

Parry, Jason; Harrington, Edward; Rees, Gareth D; McNab, Rod; Smith, Anthony J

2008-02-01

Design and construct a tooth-brushing simulator incorporating control of brushing variables including brushing force, speed and temperature, thereby facilitating greater understanding of their importance in toothpaste abrasion testing methodologies. A thermostable orbital shaker was selected as a base unit and 16- and 24-specimen brushing rigs were constructed to fit inside, consisting of: a square bath partitioned horizontally to provide brushing channels, specimen holders for 25 mm diameter mounted specimens to fit the brushing channels and individually weighted brushing arms, able to support four toothbrush holders suspended over the brushing channels. Brush head holders consisted of individually weighted blocks of Delrin, or PTFE onto which toothbrush heads were fixed. Investigating effects of key design criteria involved measuring abrasion depths of polished human enamel and dentine. The brushing simulator demonstrated good reproducibility of abrasion on enamel and dentine across consecutive brushing procedures. Varying brushing parameters had a significant impact on wear results: increased brushing force demonstrated a trend towards increased wear, with increased reproducibility for greater abrasion levels, highlighting the importance of achieving sufficient wear to optimise accuracy; increasing brushing temperature demonstrated increased enamel abrasion for silica and calcium carbonate systems, which may be related to slurry viscosities and particle suspension; varying brushing speed showed a small effect on abrasion of enamel at lower brushing speed, which may indicate the importance of maintenance of the abrasive in suspension. Adjusting key brushing variables significantly affected wear behaviour. The brushing simulator design provides a valuable model system for in vitro assessment of toothpaste abrasivity and the influence of variables in a controlled manner. Control of these variables will allow more reproducible study of in vitro tooth wear processes.

Intra-Amniotic Administration (Gallus gallus) of Cicer arietinum and Lens culinaris Prebiotics Extracts and Duck Egg White Peptides Affects Calcium Status and Intestinal Functionality

PubMed Central

Hou, Tao; Glahn, Raymond P.; Tako, Elad

2017-01-01

Calcium (Ca) is one of the most abundant inorganic elements in the human body and has many important physiological roles. Prebiotics and bioactive peptides are two important substances used to promote calcium uptake. However, the difference in mechanisms of the calcium uptake from these two supplements is not clear. By using the Gallus gallus model and the intra-amniotic administration procedure, the aim of this study was to investigate whether Ca status, intestinal functionality, and health-promoting bacterial populations were affected by prebiotics extracted from chickpea and lentil, and duck egg white peptides (DPs). Eleven groups (non-injected; 18 MΩ H2O; 4 mmol/L CaCl2; 50 mg/mL chickpea + 4 mmol/L CaCl2; 50 mg/mL lentil + 4 mmol/L CaCl2; 40 mg/mL DPs + 4 mmol/L CaCl2; 5 mg/mL Val-Ser-Glu-Glu (VSEE) + 4 mmol/L CaCl2; 50 mg/mL chickpea; 50 mg/mL lentil; 40 mg/mL DPs; 5 mg/mL VSEE) were utilized. Upon hatch, blood, cecum, small intestine, liver and bone were collected for assessment of serum bone alkaline phosphate level (BALP), the relative abundance of intestinal microflora, expression of Ca-related genes, brush border membrane (BBM) functional genes, and liver and bone mineral levels, respectively. The BALP level increased in the presence of lentil, DPs and VSEE (p < 0.05). The relative abundance of probiotics increased significantly (p < 0.05) by VSEE + Ca and chickpea. The expression of CalbindinD9k (Ca transporter) increased (p < 0.05) in Ca, chickpea + Ca and lentil + Ca groups. In addition, the brush border membrane functionality genes expressions increased (p < 0.05) by the chickpea or lentil extracts. Prebiotics and DPs beneficially affected the intestinal microflora and duodenal villus surface area. This research expands the understanding of the prebiotics’ properties of chickpea and lentil extracts, and peptides’ effects on calcium metabolism and gut health. PMID:28754012

Intra-Amniotic Administration (Gallus gallus) of Cicer arietinum and Lens culinaris Prebiotics Extracts and Duck Egg White Peptides Affects Calcium Status and Intestinal Functionality.

PubMed

Hou, Tao; Kolba, Nikolai; Glahn, Raymond P; Tako, Elad

2017-07-21

Calcium (Ca) is one of the most abundant inorganic elements in the human body and has many important physiological roles. Prebiotics and bioactive peptides are two important substances used to promote calcium uptake. However, the difference in mechanisms of the calcium uptake from these two supplements is not clear. By using the Gallus gallus model and the intra-amniotic administration procedure, the aim of this study was to investigate whether Ca status, intestinal functionality, and health-promoting bacterial populations were affected by prebiotics extracted from chickpea and lentil, and duck egg white peptides (DPs). Eleven groups (non-injected; 18 MΩ H₂O; 4 mmol/L CaCl₂; 50 mg/mL chickpea + 4 mmol/L CaCl₂; 50 mg/mL lentil + 4 mmol/L CaCl₂; 40 mg/mL DPs + 4 mmol/L CaCl₂; 5 mg/mL Val-Ser-Glu-Glu (VSEE) + 4 mmol/L CaCl₂; 50 mg/mL chickpea; 50 mg/mL lentil; 40 mg/mL DPs; 5 mg/mL VSEE) were utilized. Upon hatch, blood, cecum, small intestine, liver and bone were collected for assessment of serum bone alkaline phosphate level (BALP), the relative abundance of intestinal microflora, expression of Ca-related genes, brush border membrane (BBM) functional genes, and liver and bone mineral levels, respectively. The BALP level increased in the presence of lentil, DPs and VSEE ( p < 0.05). The relative abundance of probiotics increased significantly ( p < 0.05) by VSEE + Ca and chickpea. The expression of CalbindinD9k (Ca transporter) increased ( p < 0.05) in Ca, chickpea + Ca and lentil + Ca groups. In addition, the brush border membrane functionality genes expressions increased ( p < 0.05) by the chickpea or lentil extracts. Prebiotics and DPs beneficially affected the intestinal microflora and duodenal villus surface area. This research expands the understanding of the prebiotics' properties of chickpea and lentil extracts, and peptides' effects on calcium metabolism and gut health.

Differentiation of F4 receptor profiles in pigs based on their mucin 4 polymorphism, responsiveness to oral F4 immunization and in vitro binding of F4 to villi.

PubMed

Nguyen, V U; Goetstouwers, T; Coddens, A; Van Poucke, M; Peelman, L; Deforce, D; Melkebeek, V; Cox, E

2013-03-15

F4(+) enterotoxigenic Escherichia coli (F4(+) ETEC) are an important cause of diarrhoea and mortality in piglets. F4(+) ETEC use their F4 fimbriae to adhere to specific receptors (F4Rs) on small intestinal brush borders, resulting in colonization of the small intestine. To prevent pigs from post-weaning diarrhoea, pigs should be vaccinated during the suckling period. Previously, we demonstrated that F4acR(+), but not F4acR(-) piglets could be orally immunized with purified F4 fimbriae resulting in a protective immunity against F4(+) ETEC infections, indicating that this immune response was F4R dependent. Recently, aminopeptidase N has been identified as a glycoprotein receptor important for this oral immune response. However, in some oral immunization experiments, a few F4acR(+) piglets did not show an antibody response upon oral immunization, suggesting additional receptors. Therefore, the binding profile of F4 to brush border membrane (glyco)proteins was determined for pigs differing in F4-specific antibody response upon oral immunization, in in vitro adhesion of F4(+)E. coli to small intestinal villi, and in Muc4 genotype. Six groups of pigs could be identified. Only two groups positive in all three assays showed two high molecular weight (MW) glycoprotein bands (>250kDa) suggesting that these high MW bands are linked to the MUC4 susceptible genotype. The fact that these bands were absent in the MUC4 resistant group which showed a positive immune response against F4 and was positive in the adhesion test confirm that at least one or perhaps more other F4Rs exist. Interestingly, two pigs that were positive in the villous adhesion assay did not show an immune response against F4 fimbriae. This suggests that a third receptor category might exist which allows the bacteria to adhere but does not allow effective immunization with soluble F4 fimbriae. Future research will be necessary to confirm or reveal the identity of these receptors. Copyright © 2012 Elsevier B.V. All rights reserved.

Adult bile duct strictures: differentiating benign biliary stenosis from cholangiocarcinoma.

PubMed

Nguyen Canh, Hiep; Harada, Kenichi

2016-12-01

Biliary epithelial cells preferentially respond to various insults under chronic pathological conditions leading to reactively atypical changes, hyperplasia, or the development of biliary neoplasms (such as biliary intraepithelial neoplasia, intraductal papillary neoplasm of the bile duct, and cholangiocarcinoma). Moreover, benign biliary strictures can be caused by a variety of disorders (such as IgG4-related sclerosing cholangitis, eosinophilic cholangitis, and follicular cholangitis) and often mimic malignancies, despite their benign nature. In addition, primary sclerosing cholangitis is a well-characterized precursor lesion of cholangiocarcinoma and many other chronic inflammatory disorders increase the risk of malignancies. Because of these factors and the changes in biliary epithelial cells, biliary strictures frequently pose a diagnostic challenge. Although the ability to differentiate neoplastic from non-neoplastic biliary strictures has markedly progressed with the advance in radiological modalities, brush cytology and bile duct biopsy examination remains effective. However, no single modality is adequate to diagnose benign biliary strictures because of the low sensitivity. Therefore, understanding the underlying causes by compiling the entire clinical, laboratory, and imaging data; considering the under-recognized causes; and collaborating between experts in various fields including cytopathologists with multiple approaches is necessary to achieve an accurate diagnosis.

Membrane receptor location defines receptor interaction with signaling proteins in a polarized epithelium.

PubMed

Amsler, K; Kuwada, S K

1999-01-01

Signal transduction from receptors is mediated by the interaction of activated receptors with proximate downstream signaling proteins. In polarized epithelial cells, the membrane is divided into subdomains: the apical and basolateral membranes. Membrane receptors may be present in one or both subdomains. Using a combination of immunoprecipitation and Western blot analyses, we tested the hypothesis that a tyrosine kinase growth factor receptor, epidermal growth factor receptor (EGFR), interacts with distinct signaling proteins when present at the apical vs. basolateral membrane of a polarized renal epithelial cell. We report here that tyrosine phosphorylation of phospholipase C-gamma (PLC-gamma) was induced only when basolateral EGFR was activated. In contrast, tyrosine phosphorylation of several other signaling proteins was increased by activation of receptor at either surface. All signaling proteins were distributed diffusely throughout the cytoplasm; however, PLC-gamma protein also displayed a concentration at lateral cell borders. These results demonstrate that in polarized epithelial cells the array of signaling pathways initiated by activation of a membrane receptor is defined, at least in part, by the membrane location of the receptor.

Brush head composition, wear profile, and cleaning efficacy: an assessment of three electric brush heads using in vitro methods.

PubMed

Kaiser, Eva; Meyners, Michael; Markgraf, Dirk; Stoerkel, Ulrich; von Koppenfels, Roxana; Adam, Ralf; Soukup, Martin; Wehrbein, Heinrich; Erbe, Christina

2014-01-01

The objective of this research was to evaluate a current store brand (SB) brush head for composition/physical characteristics, Wear Index (WI), and cleaning efficacy versus the previous SB brush head refill design (SB control) and the Oral-B Precision Clean brush head (positive control, PC). This research consisted of three parts: 1) Analytical analysis using Fourier Transform Infrared (FT-IR) spectrometry to evaluate the chemical composition of the current SB brush head bristles relative to the SB control. In addition, physical parameters such as bristle count and diameter were determined. 2) Wear Index (WI) investigation to determine the Wear Index scores of in vitro-aged brush heads at four weeks (one month) and 13 weeks (three months) by a trained investigator. To "age" the brush heads, a robot system was used as a new alternative in vitro method to simulate aging by consumer use. 3) Robot testing to determine the cleaning performance of in vitro-aged brush heads, comparing one month-aged current SB brush heads with the SB control (one and three months-aged) and the PC brush heads (three months-aged) in a standardized fashion. 1) FT-IR analysis revealed that the chemical composition of the current and control SB refill brush heads is identical. In terms of physical parameters, the current SB brush head has 12% more bristles and a slightly oval brush head compared to the round brush head of the SB control. 2) Wear Index analysis showed there was no difference in the one month-aged current SB brush head versus the one month-aged SB control (1.67 vs. 1.50, p = 0.65) or versus the three months-aged PC brush head (1.67 vs. 1.50, p = 0.65). The one month-aged current SB brush head demonstrated statistically significantly less wear than the three months-aged SB control (1.67 vs. 2.67, p = 0.01). 3) Analysis of cleaning efficacy shows that the one month-aged current SB brush head had improved cleaning performance over the one month-aged SB control brush head (p < 0.05), despite no statistically significant difference in wear. Both the one month-aged current and control SB brush heads showed statistically significantly lower cleaning performance compared to the three months-aged PC brush heads (p < 0.01). While the current SB brush head showed improved cleaning over the SB control, it demonstrated significantly lower durability and cleaning in comparison to the PC brush head. Dental professionals should be aware of these differences, both in durability and in cleaning performance, when recommending brush heads to their patients.

Virtual reality using guided imagery to create Kanji or Hiragana by computer graphics: I

NASA Astrophysics Data System (ADS)

Ishigame, Masaaki; Miura, Nozomu; Hosaka, Akiko

1997-04-01

We have been studying a kind of word-processor that is able to create Japanese characters, Kanji or Hiragana strings in the cursive style, using an electronic writing brush model. Int his paper, we describe in detail the operation characteristics of the electronic writing brush which we have proposed. We defined a touch shape pattern of the electronic writing brush as a form which is projected as a circle and a cone. The brush goes on certain points of the skeleton of the character figure which is given as skeleton data. The thickness of the line is determined by a diametric variable brush pressure. Our progressive action model can rotate the direction of the writing brush tip corresponding to the difference angle between the direction of brush tip and the direction of the brush movement, and also the softness of the writing brush to express the writing brush method called the side writing brush. The front side and back side of the writing brush can be expressed in a calligraphic drawing. With our technique we can draw characters in actual stroke order on a virtual computer plane as if they are written by an actual writing brush.

Tooth brushing habits in uninstructed adults--frequency, technique, duration and force.

PubMed

Ganss, C; Schlueter, N; Preiss, S; Klimek, J

2009-06-01

Professional recommendations for individual oral hygiene mostly include tooth brushing at least twice daily for 2-3 min with gentle force using the Bass technique or modifications of it. This study evaluated whether habitual tooth brushing actually meets these standards. Uninstructed adults (n = 103; mean age 31 +/- 6 years; 61 female, 42 male) with habitual manual tooth brushing were given a self-administered questionnaire about the frequency of brushing and a computer system recorded their brushing technique, duration and force. The majority (79.6%) of participants brushed twice daily. The mean brushing duration was 96.6 +/- 36.0 s, the mean brushing force was 2.3 +/- 0.7 N (max. 4.1 N), and no significant differences were found for quadrants. Most subjects (73.8%) brushed with circling, 8.7% with horizontal/scrubbing, 13.6% with horizontal/circling and 3.9% with vertical/sweeping movements. Modified Bass technique was not observed. When appropriate brushing habits were defined as brushing at least twice daily for 120 s with a brushing force of less than 3 N and with circling or vertical sweeping movements, only 25.2% of the participants fulfilled all criteria, emphasising the ongoing need for oral hygiene education.

A minimal model of epithelial tissue dynamics and its application to the corneal epithelium

NASA Astrophysics Data System (ADS)

Henkes, Silke; Matoz-Fernandez, Daniel; Kostanjevec, Kaja; Coburn, Luke; Sknepnek, Rastko; Collinson, J. Martin; Martens, Kirsten

Epithelial cell sheets are characterized by a complex interplay of active drivers, including cell motility, cell division and extrusion. Here we construct a particle-based minimal model tissue with only division/death dynamics and show that it always corresponds to a liquid state with a single dynamic time scale set by the division rate, and that no glassy phase is possible. Building on this, we construct an in-silico model of the mammalian corneal epithelium as such a tissue confined to a hemisphere bordered by the limbal stem cell zone. With added cell motility dynamics we are able to explain the steady-state spiral migration on the cornea, including the central vortex defect, and quantitatively compare it to eyes obtained from mice that are X-inactivation mosaic for LacZ.

Distribution of the cystine/glutamate antiporter system xc- in the brain, kidney, and duodenum.

PubMed

Burdo, Joseph; Dargusch, Richard; Schubert, David

2006-05-01

System x(c)(-), one of the main transporters responsible for central nervous system cystine transport, is comprised of two subunits, xCT and 4F2hc. The transport of cystine into cells is rate limiting for glutathione synthesis, the major antioxidant and redox cofactor in the brain. Alterations in glutathione status are prevalent in numerous neurodegenerative diseases, emphasizing the importance of proper cystine homeostasis. However, the distribution of xCT and 4F2hc within the brain and other areas has not been described. Using specific antibodies, both xCT and 4F2hc were localized predominantly to neurons in the mouse and human brain, but some glial cells were labeled as well. Border areas between the brain proper and periphery including the vascular endothelial cells, ependymal cells, choroid plexus, and leptomeninges were also highly positive for the system x(c)(-) components. xCT and 4F2hc are also present at the brush border membranes in the kidney and duodenum. These results indicate that system x(c)(-) is likely to play a role in cellular health throughout many areas of the brain as well as other organs by maintaining intracellular cystine levels, thereby resulting in low levels of oxidative stress.

Increasing operational life of brush-contact device in the turbine generator due to using lubricating molybdenum disulphide brushes

NASA Astrophysics Data System (ADS)

Izotov, A. I.; Fominykh, A. A.; Nikulin, S. V.; Prokoshev, D. K.; Legoti, A. B.; Timina, N. V.

2018-01-01

A way of reducing irregular current distribution in multi-brush systems of sliding current transfer with its wear reduction due to installing lubricating molybdenum disulphide brushes on slip rings to ensure a greasing nano-sized cover on the slip ring surface is proposed. The authors give the results of industrial tests estimated on the performance effectiveness of lubricating brushes on slip rings of the TBB-320-2UZ-type turbine generator. The results showed that the lubricating brushes reduce a) the wear of 6110 OM+M and EG2AF+M brushes by 1.2 and 2.1 times respectively, b) current distribution irregularity in parallel operating brushes due to stabilizing the contact arc, and c) the temperature of the electrical brush-contact device due to the friction reduction in brushes.

Ordered quasi-two-dimensional structure of nanoparticles in semiflexible ring polymer brushes under compression

NASA Astrophysics Data System (ADS)

Hua, Yunfeng; Deng, Zhenyu; Jiang, Yangwei; Zhang, Linxi

2017-06-01

Molecular dynamics simulations of a coarse-grained bead-spring model of ring polymer brushes under compression are presented. Flexible polymer brushes are always disordered during compression, whereas semiflexible polymer brushes tend to be ordered under sufficiently strong compression. Further, the polymer monomer density of the semiflexible polymer brush is very high near the brush surface, inducing a peak value of the free energy near the surface. Therefore, when nanoparticles are compressed in semiflexible ring polymer brushes, they tend to exhibit a closely packed single-layer structure between the brush surface and the impenetrable wall, and a quasi-two-dimensional ordered structure near the brush surface is formed under strong compression. These findings provide a new approach to designing responsive applications.

An Eight-Week Clinical Evaluation of an Oscillating-Rotating Power Toothbrush with a Brush Head Utilizing Angled Bristles Compared with a Sonic Toothbrush in the Reduction of Gingivitis and Plaque.

PubMed

Ccahuana-Vasquez, Renzo A; Conde, Erinn; Grender, Julie M; Cunningham, Pamela; Qaqish, Jimmy; Goyal, C Ram

2015-01-01

To evaluate and compare the efficacy of an oscillating-rotating (O-R) power toothbrush with a brush head utilizing angled bristles to a marketed sonic toothbrush in the reduction of plaque and gingivitis over an eight-week period. This study used a randomized, examiner-blind, single-center, two-treatment, parallel group, eight-week design. Subjects with mild-to-moderate plaque and gingivitis were evaluated for baseline whole mouth, gingival margin, and approximal plaque, gingivitis, and gingival bleeding. Clinical assessments were performed using the Modified Gingival Index, Gingival Bleeding Index, and the Rustogi Modified Navy Plaque Index. Subjects received either the O-R brush (Oral-B Professional Care 1000 [D16u] with Oral-B CrossAction brush head [EB50]) or the sonic brush (Sonicare DiamondClean with the standard DiamondClean brush head). Subjects brushed twice daily for two minutes per brushing with the assigned brush and a standard fluoride dentifrice for eight weeks before returning for plaque and gingivitis evaluations using the same methods. Prior to baseline and Week 8 measurements, participants abstained from oral hygiene for 12 hours. One hundred and forty-eight subjects completed the study; 75 in the O-R group and 73 in the sonic group. Both brushes demonstrated statistically significant reductions in plaque and gingivitis over the eight-week study period (p < 0.00 1). The O-R brush was statistically significantly more effective in reducing plaque and gingivitis than the sonic brush. Whole mouth, gingival margin, and approximal plaque reductions were 27.7%, 46.8%, and 29.3% greater, respectively, compared with the sonic brush, while the reductions in gingivitis, gingival bleeding, and number of bleeding sites were 34.6%, 36.4%, and 36.1% greater, respectively, for the O-R brush than for the sonic brush (p < 0.001 for all six measures). No adverse events were observed for either brush. The plaque and gingivitis reductions for the O-R power brush incorporating the angled-bristled brush head were significantly greater than for the sonic power brush.

Structure of Weakly Charged Polyelectrolyte Brushes: Monomer Density Profiles

NASA Astrophysics Data System (ADS)

Borisov, O. V.; Zhulina, E. B.

1997-03-01

The internal structure (the monomer density profiles) of weakly charged polyelectrolyte brushes of different morphologies has been analyzed on the basis of the self-consistent-field approach. In contrast to previous studies based on the local electroneutrality approximation valid for sufficiently strongly charged or densely grafted (“osmotic") brushes we consider the opposite limit of sparse brushes which are unable to retain the counterions inside the brush. We have shown that an exact analytical solution of the SCF-equations is available in the case of a planar brush. In contrast to Gaussian monomer density profile known for “osmotic" polyelectrolyte brushes we have found that weakly charged brushes are characterized by constant monomer density. At the same time free ends of grafted polyions are distributed throughout the brush. Thus, the structural cross-over between polyelectrolyte “mushrooms" and dense brush regimes is established.

NPC1L1 and Cholesterol Transport

PubMed Central

Betters, Jenna L.; Yu, Liqing

2010-01-01

The polytopic transmembrane protein, Niemann-Pick C1-Like 1 (NPC1L1), is enriched in the apical membrane of small intestine absorptive enterocytes where it mediates extracellular sterol transport across the brush border membrane. It is essential for intestinal sterol absorption and is the molecular target of ezetimibe, a potent cholesterol absorption inhibitor that lowers blood cholesterol in humans. NPC1L1 is also highly expressed in human liver. The hepatic function of NPC1L1 may be to limit excessive biliary cholesterol loss. NPC1L1-dependent sterol uptake seems to be a clathrin-mediated endocytic process and is regulated by cellular cholesterol content. Recently, NPC1L1 inhibition has been shown to have beneficial effects on components of the metabolic syndrome, such as obesity, insulin resistance, fatty liver, in addition to atherosclerosis. PMID:20307540

Structural and Functional Changes Associated with Normal and Abnormal Fundus Autofluorescence in Patients with Retinitis Pigmentosa

PubMed Central

Greenstein, Vivienne C.; Duncker, Tobias; Holopigian, Karen; Carr, Ronald E.; Greenberg, Jonathan; Tsang, Stephen H.; Hood, Donald C.

2013-01-01

Purpose To analyze the structure and visual function of regions bordering the hyperautofluorescent ring/arcs in retinitis pigmentosa (RP). Methods Twenty -one RP patients (21 eyes) with rings/arcs and 21 normals (21 eyes) were studied. Visual sensitivity in the central 10° was measured with microperimetry. Retinal structure was evaluated with spectral domain optical coherence tomography (SD-OCT). The distance from the fovea to disruption/loss of the inner outer segment (IS/OS) junction and thicknesses of the total receptor plus retinal pigment epithelial (RPE) complex (R+), and outer segment plus RPE complex (OS+) layers were measured. Results were compared to measurements of the distance from the fovea to the inner and outer borders of the ring/arc seen on fundus autofluorescence (FAF). Results Disruption/loss of the IS/OS junction occurred closer to the inner border of the ring/arc and it was closer to the fovea in 8 eyes. For 19 eyes, OS+ and R+ thicknesses were significantly decreased at locations closer to the fovea than the appearance of the inner border of hyperautofluorescence. Mean visual sensitivity was decreased inside, across and outside the ring/arc by 3.5 ± 3.8, 8.9 ± 4.8 and 17.0±2.4 dB respectively. Conclusions Structural and functional changes can occur inside the hyperfluorescent ring/arc in RP. PMID:21909055

Effects of desensitizing toothpastes on the permeability of dentin after different brushing times: An in vitro study.

PubMed

Yang, Meng; Lin, Hong; Jiang, Ruodan; Zheng, Gang

2016-12-01

To investigate the effects of three commercially available desensitizing toothpastes on dentin permeability, and compare the efficacy of each product for reducing dentin permeability in the short term according to the frequency and duration of usage. 100 dentin discs with no caries were prepared from freshly extracted human third molar teeth. The dentin discs were brushed with three desensitizing toothpastes or with a non-desensitizing toothpaste and distilled water, which served as control. The 100 dentin slices were randomly divided into two groups (n= 50): one group underwent continuous brushing (brushed for 3 minutes continuously), and the other group underwent discontinuous brushing (brushed three times, each time for 1 minute). Then, the two groups were divided into five subgroups (n = 10) for the five brushing applications. Dentin permeability was measured with a hydraulic permeability system before and after brushing. All desensitizing toothpastes reduced dentin permeability significantly after treatment. Sensodyne Repair and Protect (calcium sodium phosphosilicate) and discontinuous brushing reduced dentin permeability significantly compared with continuous brushing. Dentin permeability values showed no significant difference between the three toothpastes after 3 minutes of continuous brushing. When comparing the three toothpastes under discontinuous brushing conditions after 3 minutes, Sensodyne Repair and Protect (calcium sodium phosphosilicate) reduced dentin permeability significantly. Sensodyne Repair and Protect (calcium sodium phosphosilicate) and discontinuous brushing reduced dentin permeability significantly compared with continuous brushing. Moreover, brushing with Sensodyne Repair and Protect (calcium sodium phosphosilicate) resulted in the lowest dentin permeability compared with those of the other two toothpastes. These results indicated that Sensodyne Repair and Protect may relieve dentin hypersensitivity.

Interproximal plaque mass and fluoride retention after brushing and flossing--a comparative study of powered toothbrushing, manual toothbrushing and flossing.

PubMed

Sjögren, Karin; Lundberg, Ann-Britt; Birkhed, Dowen; Dudgeon, Douglas J; Johnson, Matthew R

2004-01-01

To determine differences in interproximal plaque mass and fluoride retention with different modes of toothbrushing and flossing. Forty-seven subjects in good health used four treatments in a four-period, randomized, crossover design: 1) manual brushing only; 2) manual brushing and daily flossing; 3) electric brushing using a rotational oscillation toothbrush; and 4) electric brushing using a sonic toothbrush. Subjects used a standard sodium fluoride dentifrice during the eight-day experimental periods and a fluoride-free dentifrice during the seven-day washout periods between treatments. Interproximal plaque samples were taken on Day 1 and on Day 8, weighed, and analyzed for fluoride content. The amount of interproximal plaque was lowest with sonic brushing, which left 43-65% less plaque than all other treatments. Manual brushing and flossing yielded less plaque than manual brushing alone and rotational oscillation brushing. Differences were statistically significant (p < 0.05) for treatment; there was no time effect on plaque mass. For fluoride retention, at Day 1 sonic brushing gave at least 54% more fluoride in the interproximal plaque than all other treatments, which was significant. All treatments demonstrated a significant increase in fluoride concentration with time except manual brushing and flossing, which showed a significant decrease. At Day 8, the fluoride concentration was significantly higher for sonic brushing than for manual brushing or rotational oscillation brushing by over 40%, and all treatments exhibited significantly greater fluoride than the manual brushing and flossing combination. The mode of toothbrushing may impact the amount of plaque retained interproximally and its fluoride concentration.

Application of tooth brushing behavior to active rest.

PubMed

Sadachi, Hidetoshi; Murakami, Yoshinori; Tonomura, Manabu; Yada, Yukihiro; Simoyama, Ichiro

2010-01-01

We evaluated the usefulness of tooth brushing with toothpaste as active rest using the flicker value as a physiological parameter and a subjective questionnaire as a psychological parameter. Seventeen healthy, right-handed subjects (12 males and 5 females) aged 22.5 +/- 1.5 yr (mean +/- standard deviation) were randomly divided into tooth brushing with toothpaste (N=9) and non-tooth brushing groups (N=8). The subjects performed a serial calculation task for 20 min using personal computers. Subsequently, the tooth brushing group brushed their teeth, and the flicker value and mood were compared before and after the tooth brushing. The flicker value significantly increased in the tooth brushing group compared with the non-tooth brushing group (p<0.05). Concerning the mood, in the tooth brushing group, the incidence of a "feeling of being refreshed" significantly increased (p<0.05), that of "concentration power" or a "feeling of clear-headedness" tended to increase (p<0.1), and that of "lassitude" or "sleepiness" significantly decreased (p<0.01). Somatosensory stimulation and intraoral tactile stimulation during tooth brushing activated cerebral activity, producing refreshing effects. These results suggest the applicability of tooth brushing to active rest.

On the role of PDZ domain-encoding genes in Drosophila border cell migration.

PubMed

Aranjuez, George; Kudlaty, Elizabeth; Longworth, Michelle S; McDonald, Jocelyn A

2012-11-01

Cells often move as collective groups during normal embryonic development and wound healing, although the mechanisms governing this type of migration are poorly understood. The Drosophila melanogaster border cells migrate as a cluster during late oogenesis and serve as a powerful in vivo genetic model for collective cell migration. To discover new genes that participate in border cell migration, 64 out of 66 genes that encode PDZ domain-containing proteins were systematically targeted by in vivo RNAi knockdown. The PDZ domain is one of the largest families of protein-protein interaction domains found in eukaryotes. Proteins that contain PDZ domains participate in a variety of biological processes, including signal transduction and establishment of epithelial apical-basal polarity. Targeting PDZ proteins effectively assesses a larger number of genes via the protein complexes and pathways through which these proteins function. par-6, a known regulator of border cell migration, was a positive hit and thus validated the approach. Knockdown of 14 PDZ domain genes disrupted migration with multiple RNAi lines. The candidate genes have diverse predicted cellular functions and are anticipated to provide new insights into the mechanisms that control border cell movement. As a test of this concept, two genes that disrupted migration were characterized in more detail: big bang and the Dlg5 homolog CG6509. We present evidence that Big bang regulates JAK/STAT signaling, whereas Dlg5/CG6509 maintains cluster cohesion. Moreover, these results demonstrate that targeting a selected class of genes by RNAi can uncover novel regulators of collective cell migration.

Social ranking effects on tooth-brushing behaviour.

PubMed

Maltby, John; Paterson, Kevin; Day, Liz; Jones, Ceri; Kinnear, Hayley; Buchanan, Heather

2016-05-01

A tooth-brushing social rank hypothesis is tested suggesting tooth-brushing duration is influenced when individuals position their behaviour in a rank when comparing their behaviour with other individuals. Study 1 used a correlation design, Study 2 used a semi-experimental design, and Study 3 used a randomized intervention design to examine the tooth-brushing social rank hypothesis in terms of self-reported attitudes, cognitions, and behaviour towards tooth-brushing duration. Study 1 surveyed participants to examine whether the perceived health benefits of tooth-brushing duration could be predicted from the ranking of each person's tooth-brushing duration. Study 2 tested whether manipulating the rank position of the tooth-brushing duration influenced participant-perceived health benefits of tooth-brushing duration. Study 3 used a longitudinal intervention method to examine whether messages relating to the rank positions of tooth-brushing durations causally influenced the self-report tooth-brushing duration. Study 1 demonstrates that perceptions of the health benefits from tooth-brushing duration are predicted by the perceptions of how that behaviour ranks in comparison to other people's behaviour. Study 2 demonstrates that the perceptions of the health benefits of tooth-brushing duration can be manipulated experimentally by changing the ranked position of a person's tooth-brushing duration. Study 3 experimentally demonstrates the possibility of increasing the length of time for which individuals clean their teeth by focusing on how they rank among their peers in terms of tooth-brushing duration. The effectiveness of interventions using social-ranking methods relative to those that emphasize comparisons made against group averages or normative guidelines are discussed. What is already known on this subject? Individual make judgements based on social rank information. Social rank information has been shown to influence positive health behaviours such as exercise. What does this study add? The health benefits of tooth-brushing are predicted by how tooth-brushing duration ranks within a distribution. Focussing on how teeth-cleaning duration ranks among others produces longer teeth-cleaning durations. © 2015 The British Psychological Society.

An "ex vivo model" contributing to the diagnosis and evaluation of new drugs in cystic fibrosis.

PubMed

Di Lullo, A M; Scorza, M; Amato, F; Comegna, M; Raia, V; Maiuri, L; Ilardi, G; Cantone, E; Castaldo, G; Iengo, M

2017-06-01

Cystic fibrosis (CF) is an autosomal recessive disease caused by mutations in the cystic fibrosis transmembrane regulator (CFTR) gene. About 2000 mutations have been described so far. We setup an ex vivo model of human nasal epithelial cells (HNECs) to study CF patients testing the effect of novel mutations and molecular therapies. We performed sampling (by brushing), followed by culture and analysis of HNECs using a series of molecular techniques. We performed 50 brushings from CF patients and controls. Using cultured cells, we: i) demonstrated the widely heterogeneous CFTR expression in patients and in controls; ii) defined the splicing effect of a CFTR mutation; iii) assessed the CFTR gating activity in patients bearing different mutations; iv) demonstrated that butyrate significantly enhances CFTR expression. Based on our data, we can conclude: 1) HNEC brushing is performed without anaesthesia and is well tolerated in all CF patients (children and adults); 2) HNECs can be preserved for up to 48 hours before culture allowings multicentre studies; 3) HNECs culture can be considered a suitable model to study the molecular effects of new CFTR gene mutations and/or uncertain meaning specific mutations of carriers; 4) an ex vivo model of HNECs may be used to evaluate, before human use, the effect of new drugs on patients' cells bearing specific CFTR mutations; 5) the methodology is adequate for a quantitative measurement, by fluorescence, of the CFTR gating activity of the HNECs from patients with different genotypes identifying: a) CF patients bearing two severe mutations with an activity < 10% (compared to controls - 100%); b) CF patients bearing at least a mild mutation with an activity of 10-20%; c) CF carriers (heterozygous subjects) with an activity between 40-70%. © Copyright by Società Italiana di Otorinolaringologia e Chirurgia Cervico-Facciale, Rome, Italy.

[Effect of a tooth-brushing education program on oral health of preschool children].

PubMed

Kang, Bok-Hee; Park, Sun-Nam; Sohng, Kyeong-Yae; Moon, Jung-Soon

2008-12-01

To examine the effect of tooth-brushing education on the oral health of preschoolers. A quasi-experimental design with a non-equivalent control group was used. Two kindergartens were selected and 39 preschoolers from one kindergarten were assigned to the experimental group with tooth-brushing education and 39 from the other kindergarten to the control group. The tooth-brushing education program included 1 session on oral health education, individual tooth-brushing instruction for 1 week and supervised tooth-brushing after lunch for 4 weeks. Oral health behavior including use of tooth paste, tooth-brushing time and method of tooth-brushing, plague, streptococcus mutans, lactobacillus and dental caries were measured before and after the education. Fisher's exact test, t-test and paired t-test with the Window SAS 9.1 program were used to analyze the data. A significant increase in the use of tooth paste, tooth-brushing time and the practice of correct tooth-brushing and a decrease in plague and development of dental caries were observed in the experimental group. This tooth-brushing education was partially effective in improving oral health of preschoolers.

Polyelectrolyte brushes on dielectric surfaces

NASA Astrophysics Data System (ADS)

Antila, Hanne; Luijten, Erik

When chains of charged polymers are grafted to a solid surface, a polyelectrolyte (PE) brush results. These types of PE assemblies have a wide range of applications ranging from fuel cells and switchable electrodes to drug delivery. Many of these applications stem from the ability of PE brushes to respond to external stimuli: the brush properties can be tuned, for example, by varying electric field, PE grafting density, pH, salt concentration or salt valency. Accordingly, deciphering the brush behavior under different conditions has been a subject of considerable experimental, theoretical, and computational research efforts. However, the effect of the dielectric properties of the substrate on the PE brush has received much less attention. We use coarse-grained molecular dynamics simulations to show how varying the dielectric mismatch between the solvent and the substrate can significantly affect the brush. We demonstrate how tuning this mismatch can either diminish or enhance the effects of other control parameters, such as pH, on the brush properties. Furthermore, we investigate how dielectric properties of the substrate affect the brush, and the ion distribution and mobility within the brush, when the brush is exposed to an electric field.

Effects of topical budesonide on epithelial restitution in vivo in guinea pig trachea.

PubMed Central

Erjefält, J. S.; Erjefält, I.; Sundler, F.; Persson, C. G.

1995-01-01

BACKGROUND--Continuous epithelial shedding and restitution processes may characterise the airways in diseases such as asthma. Epithelial restitution involves several humoral and cellular mechanisms that may potentially be affected by inhaled anti-asthma drugs. The present study examines the effect of a topical steroid on epithelial restitution in vivo in the guinea pig. METHODS--The airway epithelium was mechanically removed from well defined areas of guinea pig trachea without surgery and without damage to the basement membrane or bleeding. An anti-inflammatory dose of budesonide (1 mg) was administered repeatedly to the tracheal surface by local superfusion 24 hours before, at (0 hours), and 24 hours after the denudation. Migration of epithelial cells, formation of a plasma exudation-derived gel, and appearance of luminal leucocytes were recorded by scanning electron microscopy. Cell proliferation was visualised by bromodeoxyuridine immunohistochemistry and tissue neutrophils and eosinophils by enzyme histochemistry. RESULTS--Immediately after creation of the denuded zone ciliated and secretory cells on its border dedifferentiated, flattened out, and migrated speedily (mean (SE) 2.3 (0.3) micron/min) over the basement membrane. After 48 hours the entire denuded zone (800 microns wide) was covered by a tightly sealed epithelium; at this time increased proliferation was observed in new and old epithelium and subepithelial cells. Budesonide had no detectable effect on epithelial dedifferentiation, migration, sealing, or proliferation. Immediately after denudation and continuously during the migration phase plasma was extravasated creating a fibrinous gel rich in leucocytes, particularly neutrophils, over the denuded area. Budesonide had no effect on either the gel or the leucocyte density. CONCLUSIONS--These observations suggest that topical glucocorticoids may not interfere with a fast and efficient restitution of the epithelium in the airways. Images PMID:7570417

DNA decontamination of fingerprint brushes.

PubMed

Szkuta, Bianca; Oorschot, Roland A H van; Ballantyne, Kaye N

2017-08-01

Genetic profiling of DNA collected from fingerprints that have been exposed to various enhancement techniques is routine in many forensic laboratories. As a result of direct contact with fingermark residues during treatment, there is concern around the DNA contamination risk of dusting fingermarks with fingerprint brushes. Previous studies have demonstrated the potential for cross-contamination between evidentiary items through various mechanisms, highlighting the risk of using the same fingerprint brush to powder multiple surfaces within and between crime-scenes. Experiments were performed to assess the contamination risk of reused fingerprint brushes through the transfer of dried saliva and skin deposits from and to glass surfaces with new unused squirrel hair and fiberglass brushes. Additional new unused brushes and brushes previously used in casework were also tested for their ability to contaminate samples. In addition, the ability to eradicate DNA from used squirrel hair and fiberglass fingerprint brushes was assessed using a 1% sodium hypochlorite solution and a 5% solution of a commercially available alternative, Virkon. DNA profiling results from surfaces contacted by treated and untreated brushes were compared to determine the effectiveness of the devised cleaning protocol. Brush durability was also assessed over multiple wash/rinse/dry cycles with both agents. Varying amounts of DNA-containing material were collected and transferred by squirrel hair and fiberglass brushes, with detectability on the secondary surface dependent on the biological nature of the material being transferred. The impact of DNA contamination from dirty fingerprint brushes was most apparent in simulations involving the transfer of dried saliva and brushes previously used in casework, while minimal transfer of touch DNA was observed. Alarmingly, large quantities of DNA were found to reside on new unused squirrel hair brushes, while no DNA was detected on new unused fiberglass brushes or brushes sold as DNA-free. Squirrel hair brushes were easily and effectively cleaned with both hypochlorite and Virkon, with no evidence of DNA transfer between exhibits by treated brushes. Brushes were still deemed useable after multiple cleaning cycles with either agent. In contrast, fiberglass bristles became tangled and matted when wet and could not be cleaned effectively using either method. It is recommended they are disposed of following use. Each laboratory should consider their current circumstances before adapting a cleaning method. The implementation of a program to monitor the effectiveness of the cleaning regime is also advised. Copyright © 2017 Elsevier B.V. All rights reserved.

Real-Time In Vivo Imaging of Butterfly Wing Development: Revealing the Cellular Dynamics of the Pupal Wing Tissue

PubMed Central

Iwata, Masaki; Ohno, Yoshikazu; Otaki, Joji M.

2014-01-01

Butterfly wings are covered with regularly arranged single-colored scales that are formed at the pupal stage. Understanding pupal wing development is therefore crucial to understand wing color pattern formation. Here, we successfully employed real-time in vivo imaging techniques to observe pupal hindwing development over time in the blue pansy butterfly, Junonia orithya. A transparent sheet of epithelial cells that were not yet regularly arranged was observed immediately after pupation. Bright-field imaging and autofluorescent imaging revealed free-moving hemocytes and tracheal branches of a crinoid-like structure underneath the epithelium. The wing tissue gradually became gray-white, epithelial cells were arranged regularly, and hemocytes disappeared, except in the bordering lacuna, after which scales grew. The dynamics of the epithelial cells and scale growth were also confirmed by fluorescent imaging. Fluorescent in vivo staining further revealed that these cells harbored many mitochondria at the surface of the epithelium. Organizing centers for the border symmetry system were apparent immediately after pupation, exhibiting a relatively dark optical character following treatment with fluorescent dyes, as well as in autofluorescent images. The wing tissue exhibited slow and low-frequency contraction pulses with a cycle of approximately 10 to 20 minutes, mainly occurring at 2 to 3 days postpupation. The pulses gradually became slower and weaker and eventually stopped. The wing tissue area became larger after contraction, which also coincided with an increase in the autofluorescence intensity that might have been caused by scale growth. Examination of the pattern of color development revealed that the black pigment was first deposited in patches in the central areas of an eyespot black ring and a parafocal element. These results of live in vivo imaging that covered wide wing area for a long time can serve as a foundation for studying the cellular dynamics of living wing tissues in butterflies. PMID:24586829

Real-time in vivo imaging of butterfly wing development: revealing the cellular dynamics of the pupal wing tissue.

PubMed

Iwata, Masaki; Ohno, Yoshikazu; Otaki, Joji M

2014-01-01

Butterfly wings are covered with regularly arranged single-colored scales that are formed at the pupal stage. Understanding pupal wing development is therefore crucial to understand wing color pattern formation. Here, we successfully employed real-time in vivo imaging techniques to observe pupal hindwing development over time in the blue pansy butterfly, Junonia orithya. A transparent sheet of epithelial cells that were not yet regularly arranged was observed immediately after pupation. Bright-field imaging and autofluorescent imaging revealed free-moving hemocytes and tracheal branches of a crinoid-like structure underneath the epithelium. The wing tissue gradually became gray-white, epithelial cells were arranged regularly, and hemocytes disappeared, except in the bordering lacuna, after which scales grew. The dynamics of the epithelial cells and scale growth were also confirmed by fluorescent imaging. Fluorescent in vivo staining further revealed that these cells harbored many mitochondria at the surface of the epithelium. Organizing centers for the border symmetry system were apparent immediately after pupation, exhibiting a relatively dark optical character following treatment with fluorescent dyes, as well as in autofluorescent images. The wing tissue exhibited slow and low-frequency contraction pulses with a cycle of approximately 10 to 20 minutes, mainly occurring at 2 to 3 days postpupation. The pulses gradually became slower and weaker and eventually stopped. The wing tissue area became larger after contraction, which also coincided with an increase in the autofluorescence intensity that might have been caused by scale growth. Examination of the pattern of color development revealed that the black pigment was first deposited in patches in the central areas of an eyespot black ring and a parafocal element. These results of live in vivo imaging that covered wide wing area for a long time can serve as a foundation for studying the cellular dynamics of living wing tissues in butterflies.

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.

Efficient Energy Conversion by Grafting Nanochannels with End-charged Stimuli-responsive Polyelectrolyte Brush

NASA Astrophysics Data System (ADS)

Chen, Guang; Das, Siddhartha

2017-11-01

Polyelectrolyte (PE) brushes have aroused increasing attention in applications in energy conversion and chemical sensing due to the environmentally-responsive and designable nature. PE brushes are charged polymer chains densely grafted on solid-liquid interfaces. By designing copolymeric systems, one can localize the ionizable sites at the brush tip in order to get end-charged PE brushes. Such brushes demonstrate anomalous shrinking/swelling behaviors with tunable environmental parameters such as pH and salt concentration. In this study, we probe the conformation and electrostatics of such PE brush systems with various size, grafting density and charge distribution, and exploit the electrochemomechanical energy conversion capabilities of nanochannels grafted with such PE brush systems. Our results indicate that the presence of the end-charged PE brush layer can massively enhance the streaming potential mediated energy conversion efficiency, and the improvement is more significant in strongly ionic solution.

Inhibitory effect of hydrophilic polymer brushes on surface-induced platelet activation and adhesion.

PubMed

Zou, Yuquan; Lai, Benjamin F L; Kizhakkedathu, Jayachandran N; Brooks, Donald E

2010-12-08

Poly(N,N-dimethylacrylamide) (PDMA) brushes are successfully grown from unplasticized poly(vinyl chloride) (uPVC) by well-controlled surface-initiated atom transfer radical polymerization (SI-ATRP). Molecular weights of the grafted PDMA brushes vary from ≈ 35,000 to 2,170000 Da, while the graft density ranges from 0.08 to 1.13 chains · nm(-2). The polydispersity of the grafted PDMA brushes is controlled within 1.20 to 1.80. Platelet activation (expression of CD62) and adhesion studies reveal that the graft densities of the PDMA brushes play an important role in controlling interfacial properties. PDMA brushes with graft densities between 0.35 and 0.50 chains · nm(-2) induce a significantly reduced platelet activation compared to unmodified uPVC. Moreover, the surface adhesion of platelets on uPVC is significantly reduced by the densely grafted PDMA brushes. PDMA brushes that have high molecular weights lead to a relatively lower platelet activation compared to low-molecular-weight brushes. However, the graft density of the brush is more important than molecular weight in controlling platelet interactions with PVC. PDMA brushes do not produce any significant platelet consumption in platelet rich plasma. Up to a seven-fold decrease in the number of platelets adhered on high graft density brushes is observed compared to the bare PVC surface. Unlike the bare PVC, platelets do not form pseudopodes or change morphology on PDMA brush-coated surfaces. Copyright © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Enzymes of the γ-Glutamyl Cycle in the Ciliary Body and Lens

PubMed Central

Ross, Leonard L.; Barber, Lee; Tate, Suresh S.; Meister, Alton

1973-01-01

The enzymes of the γ-glutamyl cycle have been found in rabbit ciliary body and, except for 5-oxoprolinase, also in the ocular lens. Histochemical studies show that γ-glutamyl transpeptidase is localized mainly in the basal portions of the epithelial cells of the ciliary body; the findings are similar to those observed in the chloroid plexuses. The histochemical staining reaction in the ciliary epithelium is more intense than in the chloroid plexus, intestine, and kidney. γ-Glutamyl transpeptidase staining activity in the epithelium of the intestinal and renal proximal convoluted tubules is confined to the microvillus border. Moderate transpeptidase activity was found in the cytoplasm of nonpigmented epithelial cells of the iris at the posterior pupillary margin. The histochemical and enzyme activity studies are consistent with the thesis that the γ-glutamyl cycle functions in transport of amino acids across the blood-aqueous humor barrier. Images PMID:4152058

Effect of Salt Concentration on the pH Responses of Strong and Weak Polyelectrolyte Brushes.

PubMed

Zhang, Jian; Kou, Ran; Liu, Guangming

2017-07-11

Strong polyelectrolyte brushes (SPB) and weak polyelectrolyte brushes (WPB) have different origins with response to pH, which makes their pH-responsive properties sensitive to salt concentration in different ways. Herein, we have employed poly[2-(methacryloyloxy)ethyl trimethylammonium chloride] (PMETAC) and poly[2-(dimethylamino)ethyl methacrylate] (PDMAEMA) brushes as model systems for SPB and WPB, respectively, to investigate the effect of salt concentration on the pH responses of SPB and WPB using several surface-sensitive techniques. The pH-responsive properties of the PMETAC brushes are governed by the reorganization of the interchain hydrogen bonds between the grafted chains, whereas the pH response of the PDMAEMA brushes is controlled by the charge of the grafted chains. The response of the properties of the PMETAC brushes including hydration, conformation, and surface wettability becomes weaker with increasing salt concentration induced by the competitive adsorption of counterions to the brushes between OH - and Cl - . The weakening of the pH response of the PMETAC brushes is more remarkable at the relatively high pH values. The pH response of the PDMAEMA brushes also exhibits a salt-concentration dependence. As the salt concentration increases, the weakening of the pH response of the PDMAEMA brushes is attributed to the decrease in osmotic pressure within the brushes at relatively low pH values.

Ordering nanoparticles with polymer brushes

NASA Astrophysics Data System (ADS)

Cheng, Shengfeng; Stevens, Mark J.; Grest, Gary S.

2017-12-01

Ordering nanoparticles into a desired super-structure is often crucial for their technological applications. We use molecular dynamics simulations to study the assembly of nanoparticles in a polymer brush randomly grafted to a planar surface as the solvent evaporates. Initially, the nanoparticles are dispersed in a solvent that wets the polymer brush. After the solvent evaporates, the nanoparticles are either inside the brush or adsorbed at the surface of the brush, depending on the strength of the nanoparticle-polymer interaction. For strong nanoparticle-polymer interactions, a 2-dimensional ordered array is only formed when the brush density is finely tuned to accommodate a single layer of nanoparticles. When the brush density is higher or lower than this optimal value, the distribution of nanoparticles shows large fluctuations in space and the packing order diminishes. For weak nanoparticle-polymer interactions, the nanoparticles order into a hexagonal array on top of the polymer brush as long as the grafting density is high enough to yield a dense brush. An interesting healing effect is observed for a low-grafting-density polymer brush that can become more uniform in the presence of weakly adsorbed nanoparticles.

Ureteral retrograde brush biopsy

MedlinePlus

Biopsy - brush - urinary tract; Retrograde ureteral brush biopsy cytology; Cytology - ureteral retrograde brush biopsy ... should not be performed in people with a: Urinary tract infection Blockage at or below the biopsy site ...

Protocol for measurement of enamel loss from brushing with an anti-erosive toothpaste after an acidic episode.

PubMed

Dehghan, Mojdeh; Vieira Ozorio, Jose Estevam; Chanin, Simon; Tantbirojn, Daranee; Versluis, Antheunis; Garcia-Godoy, Franklin

2017-01-01

Tooth erosion from an acidic insult may be exacerbated by toothbrushing. The purposes of this study were to develop an in vitro methodology to measure enamel loss after brushing immediately following an acidic episode and to investigate the effect of brushing with an anti-erosive toothpaste. The null hypotheses tested were that tooth erosion after brushing with the toothpaste would not be different from brushing with water and that a 1-hour delay before brushing would not reduce tooth erosion. Forty bovine enamel slabs were embedded, polished, and subjected to baseline profilometry. Specimens were bathed in hydrochloric acid for 10 minutes to simulate stomach acid exposure before post-acid profilometry. Toothbrushing was then simulated with a cross-brushing machine and followed by postbrushing profilometry. Group 1 was brushed with water; group 2 was brushed with a 50:50 toothpaste-water slurry; and groups 3 and 4 were immersed in artificial saliva for 1 hour before brushing with water or the toothpaste slurry, respectively. The depth of enamel loss was analyzed and compared using 1-way analysis of variance and post hoc testing (α = 0.05). Greater enamel loss was measured in groups brushed with toothpaste than in groups brushed with water. One-hour immersion in artificial saliva significantly reduced enamel loss when teeth were brushed with water (group 3; P < 0.05) but not with toothpaste (group 4). This study established a protocol for measuring enamel loss resulting from erosion followed by toothbrush abrasion. The results confirmed the abrasive action of toothpaste on acid-softened enamel.

Comparison of Three types of Tooth Brushes on Plaque and Gingival Indices: A Randomized Clinical Trial.

PubMed

Moeintaghavi, Amir; Sargolzaie, Naser; Rostampour, Mehrnoosh; Sarvari, Sara; Kargozar, Sanaz; Gharaei, Shideh

2017-01-01

To compare clinical results of three types of manual tooth brushes on plaque removal efficacy and gingivitis. This study is a single blind randomized trial with crossover design which involved 30 periodontaly healthy individuals. Professional plaque removal and oral hygiene instruction were performed for all the participants in the first step of our study followed by asking them to avoid brushing for 2 days. Thereafter plaque and gingivitis scores were measured using plaque and gingival indices (PI and GI). Then subjects were instructed to use Pulsar tooth brush for a two-week period and then, GI and PI indices were assessed again. After passing one-week period for wash out, subjects didn't brush for 2 days and indices were recorded again. The same procedure was done for CrossAction, and Butler 411 tooth brushes respectively and at the end of the study these variables were analyzed using SPSS software ver.16. Repeated measurement ANOVA test was used to compare the scores between different brushes. Finding of this study reveals that using all three types of tooth brushes resulted in significant plaque and gingivitis reduction compared to baseline levels. Pulsar tooth brush was significantly more effective in diminishing PI and GI than Butler tooth brush (p=0.044 and 0.031 respectively). According to our findings all 3 types of tooth brushes are effective in reduction of plaque and gingivitis and this reduction is significantly greater for Pulsar tooth brush compared to Butler and CrossAction tooth brushes.

Self-Renewing Microns-Thick Biopolymer Brush Made of Hyaluronan under Active Synthesis

NASA Astrophysics Data System (ADS)

Wei, W.; Washburn, J.; Weigel, P.; Curtis, J. E.

Hyaluronan (HA) is a large anionic polysaccharide distributed throughout many vertebrate tissues. We introduce a technology to produce dynamic HA polymer brush interfaces. The strategy relies on the enzyme hyaluronan synthase (HA synthase), which synthesizes and extrudes HA polymers up to 20 microns in length. We show that interfaces decorated by HA synthase-rich membrane fragments robustly produce polymer brushes of predictable heights and concentration profiles. The brush thickness can be tuned by the duration of growth or the enzyme density in the membranes. The system is self-renewing in that old polymers desorb and new polymers are produced. The brush can also be replenished after enzymatic removal multiple times. The large extent of the polymer interface allows for characterization of the brush architecture and for studying dynamic processes inside the brush using optical microscopy. At low ionic strengths (1 mM), we measure one of the largest polymer brushes yet reported, an average of 7.8 microns thick. For applications that require a stable brush interface, we have covalently reinforced the HA to the surfaces, and demonstrated that the brush is stable for at least two months. This self-renewing, dynamic biopolymer brush has great potential as a new biomaterial for implants, antifouling, tissue engineering and drug delivery.

Surface friction of hydrogels with well-defined polyelectrolyte brushes.

PubMed

Ohsedo, Yutaka; Takashina, Rikiya; Gong, Jian Ping; Osada, Yoshihito

2004-08-03

Hydrogels of poly(2-hydroxyethyl methacrylate) (PHEMA) with well-defined polyelectrolyte brushes of poly(sodium 4-styrenesulfonate) (PNaSS) of various molecular weights were synthesized, keeping the distance between the polymer brushes constant at ca. 20 nm. The effect of polyelectrolyte brush length on the sliding friction against a glass plate, an electrorepulsive solid substrate, was investigated in water in a velocity range of 7.5 x 10(-5) to 7.5 x 10(-2) m/s. It is found that the presence of polymer brush can dramatically reduce the friction when the polymer brushes are short. With an increase in the length of the polymer brush, this drag reduction effect only works at a low sliding velocity, and the gel with long polymer brushes even shows a higher friction than that of a normal network gel at a high sliding velocity. The strong polymer length and sliding velocity dependence indicate a dynamic mechanism of the polymer brush effect.

Anthrax Cases Associated with Animal-Hair Shaving Brushes.

PubMed

Szablewski, Christine M; Hendricks, Kate; Bower, William A; Shadomy, Sean V; Hupert, Nathaniel

2017-05-01

During the First World War, anthrax cases in the United States and England increased greatly and seemed to be associated with use of new shaving brushes. Further investigation revealed that the source material and origin of shaving brushes had changed during the war. Cheap brushes of imported horsehair were being made to look like the preferred badger-hair brushes. Unfortunately, some of these brushes were not effectively disinfected and brought with them a nasty stowaway: Bacillus anthracis. A review of outbreak summaries, surveillance data, and case reports indicated that these cases originated from the use of ineffectively disinfected animal-hair shaving brushes. This historical information is relevant to current public health practice because renewed interest in vintage and animal-hair shaving brushes has been seen in popular culture. This information should help healthcare providers and public health officials answer questions on this topic.

Quantitative evaluation of toothbrush and arm-joint motion during tooth brushing.

PubMed

Inada, Emi; Saitoh, Issei; Yu, Yong; Tomiyama, Daisuke; Murakami, Daisuke; Takemoto, Yoshihiko; Morizono, Ken; Iwasaki, Tomonori; Iwase, Yoko; Yamasaki, Youichi

2015-07-01

It is very difficult for dental professionals to objectively assess tooth brushing skill of patients, because an obvious index to assess the brushing motion of patients has not been established. The purpose of this study was to quantitatively evaluate toothbrush and arm-joint motion during tooth brushing. Tooth brushing motion, performed by dental hygienists for 15 s, was captured using a motion-capture system that continuously calculates the three-dimensional coordinates of object's motion relative to the floor. The dental hygienists performed the tooth brushing on the buccal and palatal sides of their right and left upper molars. The frequencies and power spectra of toothbrush motion and joint angles of the shoulder, elbow, and wrist were calculated and analyzed statistically. The frequency of toothbrush motion was higher on the left side (both buccal and palatal areas) than on the right side. There were no significant differences among joint angle frequencies within each brushing area. The inter- and intra-individual variations of the power spectrum of the elbow flexion angle when brushing were smaller than for any of the other angles. This study quantitatively confirmed that dental hygienists have individual distinctive rhythms during tooth brushing. All arm joints moved synchronously during brushing, and tooth brushing motion was controlled by coordinated movement of the joints. The elbow generated an individual's frequency through a stabilizing movement. The shoulder and wrist control the hand motion, and the elbow generates the cyclic rhythm during tooth brushing.

Analysis of tooth brushing cycles.

PubMed

Tosaka, Yuki; Nakakura-Ohshima, Kuniko; Murakami, Nozomi; Ishii, Rikako; Saitoh, Issei; Iwase, Yoko; Yoshihara, Akihiro; Ohuchi, Akitsugu; Hayasaki, Haruaki

2014-11-01

The aim of this study was to demonstrate the effectiveness of an analysis of tooth brushing cycles using a system that measures tooth brushing motion and brushing force with an accelerometer and strain tension gage attached to a toothbrush. Mechanical plaque removal with a manual toothbrush remains the primary method of maintaining good oral hygiene for the majority of the population. Because toothbrush motion has not been fully understood, it should be clarified by analysis of tooth brushing cycles. Twenty healthy female dental hygienists participated in this study. Their tooth brushing motions were measured and analyzed using an American Dental Association-approved manual toothbrush to which a three-dimensional (3-D) accelerometer and strain tension gage were attached. 3-D motion and brushing force on the labial surface of the mandibular right central incisor and the lingual surface of the mandibular left first molar were measured, analyzed, and compared. Multilevel linear model analysis was applied to estimate variables and compare motion and forces related to the two tooth surfaces. The analysis of tooth brushing cycles was feasible, and significant differences were detected for durations and 3-D ranges of toothbrush motion as well as brushing force between the two tooth surfaces. The analysis used in this study demonstrated an ability to detect characteristics of tooth brushing motion, showing tooth brushing motion to change depending on the brushed location. These results also suggest that more detailed instructions might be required according to patient's oral condition.

Saccharomyces boulardii in childhood.

PubMed

Vandenplas, Yvan; Brunser, Oscar; Szajewska, Hania

2009-03-01

Probiotics are live microorganisms which confer a health benefit on the host. Saccharomyces boulardii, a yeast, has been found to be an effective probiotic in double-blind placebo-controlled randomized clinical studies. We reviewed the established mechanisms of actions and clinical efficacy in children of S. boulardii. The mechanisms of action of S. boulardii depend mainly on the inhibition of some bacterial toxins, anti-inflammatory effects, and on stimulating effects on the intestinal mucosa such as trophic effects on the brush border enzymes and immunostimulatory effects. At present, in pediatric populations, there is evidence that S. boulardii is beneficial for the treatment of acute gastroenteritis and the prevention of antibiotic-associated diarrhea. More data are needed in other indications such as traveller's diarrhea, Helicobacter pylori eradication, and inflammatory bowel disease. S. boulardii is a yeast strain that has been extensively studied in vitro and in vivo. Recent data have opened the door for new therapeutic indications.

Intestinal Alkaline Phosphatase Detoxifies Lipopolysaccharide and Prevents Inflammation in Response to the Gut Microbiota

PubMed Central

Bates, Jennifer M.; Akerlund, Janie; Mittge, Erika; Guillemin, Karen

2009-01-01

SUMMARY Vertebrates harbor abundant lipopolysaccharide (LPS) or endotoxin in their gut microbiota. Here we demonstrate that the brush border enzyme intestinal alkaline phosphatase (Iap), which dephosphorylates LPS, is induced during establishment of the microbiota and plays a crucial role in promoting mucosal tolerance to gut bacteria in zebrafish. We demonstrate that Iap deficient animals are hypersensitive to LPS toxicity through a mechanism mediated by Myd88 and Tumor Necrosis Factor Receptor (Tnfr). We further show that the endogenous microbiota establish the normal homeostatic level of neutrophils in the intestine through a process involving Myd88 and Tnfr. Iap deficient animals exhibit excessive intestinal neutrophil influx, similar to wild type animals exposed to LPS. When reared germ-free, however, the intestines of Iap deficient animals are devoid of neutrophils, demonstrating that Iap functions to prevent inflammatory responses to resident gut bacteria. PMID:18078689

A specific binding protein from Tenebrio molitor for the insecticidal toxin of Bacillus thuringiensis subsp. tenebrionis.

PubMed

Belfiore, C J; Vadlamudi, R K; Osman, Y A; Bulla, L A

1994-04-15

Biopesticides based on the bacterium Bacillus thuringiensis have attracted wide attention as safe alternatives to chemical insecticides. In this paper, we report, for the first time, the identification of a single binding protein from a coleopteran insect, Tenebrio molitor, that is specific for the cryIII toxin of B. thuringiensis. The protein appeared as a single band of 144 kDa on radioligand and immunoblots of total proteins extracted from brush border membrane vesicles of the midgut of T. molitor. Radiolabelled cryIIIA toxin bound to the protein with a Kd value of 17.5 nM and could be specifically blocked by unlabelled toxin but not by toxins from other subspecies of B. thuringiensis. This study lays the groundwork to clone the cryIIIA toxin binding protein and to determine the molecular mechanism(s) of toxin action.

Human epidermis is a novel site of phospholipase B expression.

PubMed

Maury, Eric; Prévost, Marie Claude; Nauze, Michel; Redoulès, Daniel; Tarroux, Roger; Charvéron, Marie; Salles, Jean Pierre; Perret, Bertrand; Chap, Hugues; Gassama-Diagne, Ama

2002-07-12

Phospholipase B (PLB) is an enzyme that displays both phospholipase A(2) and lysophospholipase activities. Analysis of human epidermis homogenates indicated the presence of a 97 kDa PLB protein, as well as a phospholipase A(2) activity, both being enriched in the soluble fraction. Immunolabelling and in situ hybridization experiments showed that this enzyme is expressed in the different layers of epidermis with an accumulation at the dermo-epidermis junction. RT-PCR data indicated that PLB is specifically expressed in natural and reconstructed epidermis. By 3'-RACE-PCR and screening of human genome databases, we obtained a 3600 bp cDNA coding for human PLB highly homologous to already described intestinal brush border PLBs. These data led us to conclude that the soluble PLB corresponds to a proteolytic cleavage of the membrane anchored protein. Altogether, our results provide the first characterization of human PLB which should play an important role in epidermal barrier function.

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.

A Laterally-Mobile Mixed Polymer/Polyelectrolyte Brush Undergoes a Macroscopic Phase Separation

NASA Astrophysics Data System (ADS)

Lee, Hoyoung; Park, Hae-Woong; Tsouris, Vasilios; Choi, Je; Mustafa, Rafid; Lim, Yunho; Meron, Mati; Lin, Binhua; Won, You-Yeon

2013-03-01

We studied mixed PEO and PDMAEMA brushes. The question we attempted to answer was: When the chain grafting points are laterally mobile, how will this lateral mobility influence the structure and phase behavior of the mixed brush? Two different model mixed PEO/PDMAEMA brush systems were prepared: a mobile mixed brush by spreading a mixture of two diblock copolymers, PEO-PnBA and PDMAEMA-PnBA, onto the air-water interface, and an inseparable mixed brush using a PEO-PnBA-PDMAEMA triblock copolymer having respective brush molecular weights matched to those of the diblock copolymers. These two systems were investigated by surface pressure-area isotherm, X-ray reflectivity and AFM imaging measurements. The results suggest that the mobile mixed brush undergoes a lateral macroscopic phase separation at high chain grafting densities, whereas the inseparable system is only microscopically phase separated under comparable brush density conditions. We also conducted an SCF analysis of the phase behavior of the mixed brush system. This analysis further supported the experimental findings. The macroscopic phase separation observed in the mobile system is in contrast to the microphase separation behavior commonly observed in two-dimensional laterally-mobile small molecule mixtures.

Ordering nanoparticles with polymer brushes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cheng, Shengfeng; Stevens, Mark J.; Grest, Gary S.

Ordering nanoparticles into a desired super-structure is often crucial for their technological applications. We use molecular dynamics simulations to study the assembly of nanoparticles in a polymer brush randomly grafted to a planar surface as the solvent evaporates. Initially, the nanoparticles are dispersed in a solvent that wets the polymer brush. After the solvent evaporates, the nanoparticles are either inside the brush or adsorbed at the surface of the brush, depending on the strength of the nanoparticle-polymer interaction. For strong nanoparticle-polymer interactions, a 2-dimensional ordered array is only formed when the brush density is finely tuned to accommodate a singlemore » layer of nanoparticles. When the brush density is higher or lower than this optimal value, the distribution of nanoparticles shows large fluctuations in space and the packing order diminishes. For weak nanoparticle-polymer interactions, the nanoparticles order into a hexagonal array on top of the polymer brush as long as the grafting density is high enough to yield a dense brush. As a result, an interesting healing effect is observed for a low-grafting-density polymer brush that can become more uniform in the presence of weakly adsorbed nanoparticles.« less

Ordering nanoparticles with polymer brushes

DOE PAGES

Cheng, Shengfeng; Stevens, Mark J.; Grest, Gary S.

2017-12-08

Ordering nanoparticles into a desired super-structure is often crucial for their technological applications. We use molecular dynamics simulations to study the assembly of nanoparticles in a polymer brush randomly grafted to a planar surface as the solvent evaporates. Initially, the nanoparticles are dispersed in a solvent that wets the polymer brush. After the solvent evaporates, the nanoparticles are either inside the brush or adsorbed at the surface of the brush, depending on the strength of the nanoparticle-polymer interaction. For strong nanoparticle-polymer interactions, a 2-dimensional ordered array is only formed when the brush density is finely tuned to accommodate a singlemore » layer of nanoparticles. When the brush density is higher or lower than this optimal value, the distribution of nanoparticles shows large fluctuations in space and the packing order diminishes. For weak nanoparticle-polymer interactions, the nanoparticles order into a hexagonal array on top of the polymer brush as long as the grafting density is high enough to yield a dense brush. As a result, an interesting healing effect is observed for a low-grafting-density polymer brush that can become more uniform in the presence of weakly adsorbed nanoparticles.« less

The effect of tooth brushing and flossing sequence on interdental plaque reduction and fluoride retention: A randomized controlled clinical trial.

PubMed

Mazhari, Fatemeh; Boskabady, Marzie; Moeintaghavi, Amir; Habibi, Atieh

2018-05-09

Mechanical plaque control methods such as brushing and flossing are highly recommended to remove dental plaque. The aim of this study is to evaluate the efficacy of the sequence of brushing and flossing on reducing interdental plaque and increasing fluoride retention in that area. This randomized controlled crossover trial was conducted on 25 dental students. After prophylaxis, they were asked to discontinue all forms of oral hygiene for 48 hours. The study was performed in two phases with two-week washout intervals. In one phase, they first brushed, then flossed (sequence 1: brush-floss group). In the other phase they initially used dental floss then brushed (sequence 2: floss-brush group). At each phase, dental plaque (using the Rustogi Modified Navy Plaque Index) and fluoride concentrations (using a fluoride ion specific electrode) were measured before and after flossing and brushing, and the dental plaque reduction and fluoride increase were compared between the two groups using the mixed model test. A significance level of 5% was selected. In the floss-brush group interdental and whole plaque was reduced significantly more than the brush-floss group (p = 0.001, p = 0.009 respectively). However, marginal plaque did not show any statistically significant difference between the two groups (p = 0.2). Fluoride concentrations in interdental plaque were significantly higher in the floss-brush group than the other group (p = 0.027). The results showed that flossing followed by brushing is preferred to brushing then flossing in order to reduce interdental plaque and increase fluoride concentration in interdental plaque. This article is protected by copyright. All rights reserved. © 2018 American Academy of Periodontology.

Targeting autophagy as a novel strategy for facilitating the therapeutic action of potentiators on ΔF508 cystic fibrosis transmembrane conductance regulator

PubMed Central

Luciani, Alessandro; Villella, Valeria Rachela; Esposito, Speranza; Gavina, Manuela; Russo, Ilaria; Silano, Marco; Guido, Stefano; Pettoello-Mantovani, Massimo; Carnuccio, Rosa; Scholte, Bob; De Matteis, Antonella; Maiuri, Maria Chiara; Raia, Valeria; Luini, Alberto; Kroemer, Guido; Maiuri, Luigi

2012-01-01

Channel activators (potentiators) of cystic fibrosis (CF) transmembrane conductance regulator (CFTR), can be used for the treatment of the small subset of CF patients that carry plasma membrane-resident CFTR mutants. However, approximately 90% of CF patients carry the misfolded ΔF508-CFTR and are poorly responsive to potentiators, because ΔF508-CFTR is intrinsically unstable at the plasma membrane (PM) even if rescued by pharmacological correctors. We have demonstrated that human and mouse CF airways are autophagy deficient due to functional sequestration of BECN1 and that the tissue transglutaminase-2 inhibitor, cystamine, or antioxidants restore BECN1-dependent autophagy and reduce SQSTM1/p62 levels, thus favoring ΔF508-CFTR trafficking to the epithelial surface. Here, we investigated whether these treatments could facilitate the beneficial action of potentiators on ΔF508-CFTR homozygous airways. Cystamine or the superoxide dismutase (SOD)/catalase-mimetic EUK-134 stabilized ΔF508-CFTR at the plasma membrane of airway epithelial cells and sustained the expression of CFTR at the epithelial surface well beyond drug withdrawal, overexpressing BECN1 and depleting SQSTM1. This facilitates the beneficial action of potentiators in controlling inflammation in ex vivo ΔF508-CFTR homozygous human nasal biopsies and in vivo in mouse ΔF508-CFTR lungs. Direct depletion of Sqstm1 by shRNAs in vivo in ΔF508-CFTR mice synergized with potentiators in sustaining surface CFTR expression and suppressing inflammation. Cystamine pre-treatment restored ΔF508-CFTR response to the CFTR potentiators genistein, Vrx-532 or Vrx-770 in freshly isolated brushed nasal epithelial cells from ΔF508-CFTR homozygous patients. These findings delineate a novel therapeutic strategy for the treatment of CF patients with the ΔF508-CFTR mutation in which patients are first treated with cystamine and subsequently pulsed with CFTR potentiators. PMID:22874563

Targeting autophagy as a novel strategy for facilitating the therapeutic action of potentiators on ΔF508 cystic fibrosis transmembrane conductance regulator.

PubMed

Luciani, Alessandro; Villella, Valeria Rachela; Esposito, Speranza; Gavina, Manuela; Russo, Ilaria; Silano, Marco; Guido, Stefano; Pettoello-Mantovani, Massimo; Carnuccio, Rosa; Scholte, Bob; De Matteis, Antonella; Maiuri, Maria Chiara; Raia, Valeria; Luini, Alberto; Kroemer, Guido; Maiuri, Luigi

2012-11-01

Channel activators (potentiators) of cystic fibrosis (CF) transmembrane conductance regulator (CFTR), can be used for the treatment of the small subset of CF patients that carry plasma membrane-resident CFTR mutants. However, approximately 90% of CF patients carry the misfolded ΔF508-CFTR and are poorly responsive to potentiators, because ΔF508-CFTR is intrinsically unstable at the plasma membrane (PM) even if rescued by pharmacological correctors. We have demonstrated that human and mouse CF airways are autophagy deficient due to functional sequestration of BECN1 and that the tissue transglutaminase-2 inhibitor, cystamine, or antioxidants restore BECN1-dependent autophagy and reduce SQSTM1/p62 levels, thus favoring ΔF508-CFTR trafficking to the epithelial surface. Here, we investigated whether these treatments could facilitate the beneficial action of potentiators on ΔF508-CFTR homozygous airways. Cystamine or the superoxide dismutase (SOD)/catalase-mimetic EUK-134 stabilized ΔF508-CFTR at the plasma membrane of airway epithelial cells and sustained the expression of CFTR at the epithelial surface well beyond drug withdrawal, overexpressing BECN1 and depleting SQSTM1. This facilitates the beneficial action of potentiators in controlling inflammation in ex vivo ΔF508-CFTR homozygous human nasal biopsies and in vivo in mouse ΔF508-CFTR lungs. Direct depletion of Sqstm1 by shRNAs in vivo in ΔF508-CFTR mice synergized with potentiators in sustaining surface CFTR expression and suppressing inflammation. Cystamine pre-treatment restored ΔF508-CFTR response to the CFTR potentiators genistein, Vrx-532 or Vrx-770 in freshly isolated brushed nasal epithelial cells from ΔF508-CFTR homozygous patients. These findings delineate a novel therapeutic strategy for the treatment of CF patients with the ΔF508-CFTR mutation in which patients are first treated with cystamine and subsequently pulsed with CFTR potentiators.

Shared Gene Expression Alterations in Nasal and Bronchial Epithelium for Lung Cancer Detection.

PubMed

2017-07-01

We previously derived and validated a bronchial epithelial gene expression biomarker to detect lung cancer in current and former smokers. Given that bronchial and nasal epithelial gene expression are similarly altered by cigarette smoke exposure, we sought to determine if cancer-associated gene expression might also be detectable in the more readily accessible nasal epithelium. Nasal epithelial brushings were prospectively collected from current and former smokers undergoing diagnostic evaluation for pulmonary lesions suspicious for lung cancer in the AEGIS-1 (n = 375) and AEGIS-2 (n = 130) clinical trials and gene expression profiled using microarrays. All statistical tests were two-sided. We identified 535 genes that were differentially expressed in the nasal epithelium of AEGIS-1 patients diagnosed with lung cancer vs those with benign disease after one year of follow-up ( P  < .001). Using bronchial gene expression data from the AEGIS-1 patients, we found statistically significant concordant cancer-associated gene expression alterations between the two airway sites ( P  < .001). Differentially expressed genes in the nose were enriched for genes associated with the regulation of apoptosis and immune system signaling. A nasal lung cancer classifier derived in the AEGIS-1 cohort that combined clinical factors (age, smoking status, time since quit, mass size) and nasal gene expression (30 genes) had statistically significantly higher area under the curve (0.81; 95% confidence interval [CI] = 0.74 to 0.89, P  = .01) and sensitivity (0.91; 95% CI = 0.81 to 0.97, P  = .03) than a clinical-factor only model in independent samples from the AEGIS-2 cohort. These results support that the airway epithelial field of lung cancer-associated injury in ever smokers extends to the nose and demonstrates the potential of using nasal gene expression as a noninvasive biomarker for lung cancer detection. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Restoration of CFTR function in patients with cystic fibrosis carrying the F508del-CFTR mutation

PubMed Central

Stefano, Daniela De; Villella, Valeria R; Esposito, Speranza; Tosco, Antonella; Sepe, Angela; Gregorio, Fabiola De; Salvadori, Laura; Grassia, Rosa; Leone, Carlo A; Rosa, Giuseppe De; Maiuri, Maria C; Pettoello-Mantovani, Massimo; Guido, Stefano; Bossi, Anna; Zolin, Anna; Venerando, Andrea; Pinna, Lorenzo A; Mehta, Anil; Bona, Gianni; Kroemer, Guido; Maiuri, Luigi; Raia, Valeria

2014-01-01

Restoration of BECN1/Beclin 1-dependent autophagy and depletion of SQSTM1/p62 by genetic manipulation or autophagy-stimulatory proteostasis regulators, such as cystamine, have positive effects on mouse models of human cystic fibrosis (CF). These measures rescue the functional expression of the most frequent pathogenic CFTR mutant, F508del, at the respiratory epithelial surface and reduce lung inflammation in CftrF508del homozygous mice. Cysteamine, the reduced form of cystamine, is an FDA-approved drug. Here, we report that oral treatment with cysteamine greatly reduces the mortality rate and improves the phenotype of newborn mice bearing the F508del-CFTR mutation. Cysteamine was also able to increase the plasma membrane expression of the F508del-CFTR protein in nasal epithelial cells from F508del homozygous CF patients, and these effects persisted for 24 h after cysteamine withdrawal. Importantly, this cysteamine effect after washout was further sustained by the sequential administration of epigallocatechin gallate (EGCG), a green tea flavonoid, both in vivo, in mice, and in vitro, in primary epithelial cells from CF patients. In a pilot clinical trial involving 10 F508del-CFTR homozygous CF patients, the combination of cysteamine and EGCG restored BECN1, reduced SQSTM1 levels and improved CFTR function from nasal epithelial cells in vivo, correlating with a decrease of chloride concentrations in sweat, as well as with a reduction of the abundance of TNF/TNF-alpha (tumor necrosis factor) and CXCL8 (chemokine [C-X-C motif] ligand 8) transcripts in nasal brushing and TNF and CXCL8 protein levels in the sputum. Altogether, these results suggest that optimal schedules of cysteamine plus EGCG might be used for the treatment of CF caused by the F508del-CFTR mutation. PMID:25350163

Study on brush of moving electrode type electrostatic precipitator (MEEP)

NASA Astrophysics Data System (ADS)

Zhao, Haibao; He, Yuzhong; Yao, Yuping

2018-02-01

MEEP was an efficient particle removal technology for coal plant and sintering machine. As the stability of brush in MEEP was relatively poor, the experiments was designed for the brush which was made by 0Cr18Ni9 stainless steel wire to find the failure mode and cause. Combining the results of the experiments, the failure models of brushes were different under different conditions and the brushes were suitable for being used in the condition of small diameter particles. And the life span of brushes can be more than 6 years.

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.

Short communication: use of a mechanical brush by Holstein dairy cattle around parturition.

PubMed

Newby, Nathalie C; Duffield, Todd F; Pearl, David L; Leslie, Ken E; LeBlanc, Stephen J; von Keyserlingk, Marina A G

2013-04-01

Grooming is a normal behavior that may contribute to relief of stress. The purpose of this study was to investigate the effect of access to a mechanical brush on auto-grooming behavior in parturient cows. The hypothesis was that cows would increase the total time spent scratching using the brush if they had access to a brush around the time of calving, whereas auto-grooming would be lower for the brush group compared with the no-brush group. The use of a mechanical brush was analyzed both in a group-housed pen (72 to 48 h before calving) and in an individual maternity pen (6h before to 6h after calving) in 16 multiparous Holstein cows. In the maternity pen, cows were randomly assigned to have access to the brush or not. The provision of a mechanical brush in the individual maternity pen did not change the amount of time spent auto-grooming but cows did use the brush before calving and after the calf was removed. Despite being housed in pens containing a brush, cows failed to use it when the calf was present. However, cows with access to a brush spent more time licking their calves in the first hour postcalving (β=8.7 min; 95% confidence interval: 1.5, 15.8) than cows that did not have access to a brush. Regardless of treatment, cows increased the time spent auto-grooming and scratching following separation of their newborn calf (β=1.4 min; 95% confidence interval: 0.46, 2.3 and β=0.07 min; 95% confidence interval: 0.02, 0.12, respectively). Further research is warranted to investigate possible benefits of mechanical brush devices at the time of calving, particularly for cows that experience difficult calving and require manual or mechanical assistance. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Structure of Polyelectrolyte Brushes in the Presence of Multivalent Counterions

DOE PAGES

Yu, Jing; Mao, Jun; Yuan, Guangcui; ...

2016-07-20

Polyelectrolyte brushes are of great importance to a wide range of fields, ranging from colloidal stabilization to responsive and tunable materials to lubrication. Here, we synthesized high-density polystyrenesulfonate (PSS) brushes using surface initiated atom transfer radical polymerization and performed neutron reflectivity (NR) and surface force measurements using a surface forces apparatus (SFA) to investigate the effect of monovalent Na +, divalent Ca 2+, Mg 2+, and Ba 2+, and trivalent Y 3+ counterions on the structure of the PSS brushes. NR and SFA results demonstrate that in monovalent salt solution the behavior of the PSS brushes agrees with scaling theorymore » well, exhibiting two distinct regimes: the osmotic and salted brush regimes. Introducing trivalent Y 3+ cations causes an abrupt shrinkage of the PSS brush due to the uptake of Y 3+ counterions. The uptake of Y 3+ counterions and shrinkage of the brush are reversible upon increasing the concentration of monovalent salt. Divalent cations, Mg 2+, Ca 2+, and Ba 2+, while all significantly affecting the structure of PSS brushes, show strong ion specific effects that are related to the specific interactions between the divalent cations and the sulfonate groups. Our results demonstrate that the presence of multivalent counterions, even at relatively low concentrations, can strongly affect the structure of polyelectrolyte brushes. Finally, the results also highlight the importance of ion specificity to the structure of polyelectrolyte brushes in solution.« less

Evaluation of the tooth brushing skills in children aged 6-12 years.

PubMed

Pujar, P; Subbareddy, V V

2013-08-01

To evaluate the tooth brushing skills in children aged between 6 and 12 years and its relation with age, type of grip used, duration of brushing and gender. A total of 105 children aged between 6 and 12 years were divided into seven groups and their plaque scores, type of grip used and duration of brushing were assessed. The most preferred grip was the distal oblique (58.1 %). Oblique, distal oblique and power grips were more efficient (plaque reduction up to 70 %). Plaque removal efficacy improved with age (57 % in 6-year-olds and 82 % in 12-year-old children). Plaque reduction was greater when the duration of brushing was longer (82 % plaque reduction when the brushing time was >2.5 min). Overall, the mean duration of tooth brushing in children aged 6-12 years was 1.71 min (103 s). Tooth brushing skills improved with age and the duration of tooth brushing made a significant difference to the oral hygiene status of the child. The type of grip used and gender did not influence tooth brushing ability in this group of children. Tooth brushing skills were low in children younger than 10 years of age. Hence, parental supervision is considered necessary and recommended until 10 years of age.

Lateral Structure Formation in Polyelectrolyte Brushes Induced by Multivalent Ions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Brettmann, Blair; Pincus, Philip; Tirrell, Matthew

2017-01-13

We provide a theoretical model for the collapse of polyelectrolyte brushes in the presence of multivalent ions, focusing on the formation of lateral inhomogeneties in the collapsed state. Polyelectrolyte brushes are important in a variety of applications, including stabilizing colloidal particles and lubricating surfaces. Many uses rely on the extension of the densely grafted polymer chains from the surface in the extended brush morphology. In the presence Extended Brush of multivalent ions, brushes are significantly shorter than in monovalent ionic solutions, which greatly affects their properties. We base our theoretical analysis on an analogous collapse of polyelectrolyte brushes in amore » poor solvent, providing an energy balance representation for pinned micelles and cylindrical bundles. The equilibrium brush heights predicted for these structures are of a similar magnitude to those measured experimentally. The formation of lateral structures can open new avenues for stimuli-responsive applications that rely on nanoscale pattern formation on surfaces.« less

Synthesis and Biomedical Applications of Poly((meth)acrylic acid) Brushes.

PubMed

Qu, Zhenyuan; Xu, Hong; Gu, Hongchen

2015-07-15

Poly((meth)acrylic acid) (P(M)AA) brushes possess a number of distinctive properties that are particularly attractive for biomedical applications. This minireview summarizes recent advances in the synthesis and biomedical applications of P(M)AA brushes and brushes containing P(M)AA segments. First, we review different surface-initiated polymerization (SIP) methods, with a focus on recent progress in the surface-initiated controlled/living radical polymerization (SI-CLRP) techniques used to generate P(M)AA brushes with a tailored structure. Next, we discuss biomolecule immobilization methods for P(M)AA brushes, including physical adsorption, covalent binding, and affinity interactions. Finally, typical biomedical applications of P(M)AA brushes are reviewed, and their performance is discussed based on their unique properties. We conclude that P(M)AA brushes are promising biomaterials, and more potential biomedical applications are expected to emerge with the further development of synthetic techniques and increased understanding of their interactions with biological systems.

Antifouling Properties of Fluoropolymer Brushes toward Organic Polymers: The Influence of Composition, Thickness, Brush Architecture, and Annealing.

PubMed

Wang, Zhanhua; Zuilhof, Han

2016-07-05

Fluoropolymer brushes are widely used to prevent nonspecific adsorption of commercial polymeric or biological materials due to their strongly hydrophobic character. Herein, a series of fluoropolymer brushes with different compositions, thicknesses and molecular architectures was prepared via surface-initiated atom transfer radical polymerization (ATRP). Subsequently, the antifouling properties of these fluoropolymer brushes against organic polymers were studied in detail using X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM) measurements and polystyrene as a representative fouling polymer. Among all of the molecular architectures studied, homopolymerized methacrylate-based fluoropolymer brushes (PMAF17) show the best antifouling properties. Annealing the fluoropolymer brushes improves the antifouling property dramatically due to the reregulated surface composition. These fluoropolymer brushes can be combined with, e.g., micro- and nanostructuring and other advanced materials properties to yield even better long-term antifouling behavior under harsh environments.

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.

Effects of short-term immersion and brushing with different denture cleansers on the roughness, hardness, and color of two types of acrylic resin.

PubMed

Panariello, Beatriz Helena Dias; Izumida, Fernanda Emiko; Moffa, Eduardo Buozi; Pavarina, Ana Claudia; Jorge, Janaina Habib; Giampaolo, Eunice Teresinha

2015-06-01

To investigate the cumulative effects of brushing (B) or immersion (I), using different cleansing agents, on the surface roughness, hardness and color stability of a heat-polymerized denture resin, Lucitone 550 (L), and a hard chairside reline resin, Tokuyama Rebase Fast II (T). A total of 316 specimens (10 x 2 mm) were fabricated. The specimens (n = 9) were divided into brushing or immersion groups according to the following agents: dentifrice/distilled water (D), 1% sodium hypochlorite (NaOCl), Corega Tabs (Pb), 1% chlorhexidine gluconate (Chx), and 0.2% peracetic acid (Ac). Brushing and immersion were tested independently. Assays were performed after 1, 3, 21, 45 and 90 brushing cycles or immersion of 10 seconds each. Data were evaluated statistically by repeated measures ANOVA. Tukey's honestly significant difference (HSD) post-hoc test was used to determine differences between means (α = 0.05). For L there was no statistically significant difference in roughness, except a significant decrease in roughness by brushing with D. T showed a significant effect on the roughness after 90 immersions with Ac. Hardness values decreased for L when specimens were immersed or brushed in NaOCl and Pb. The hardness of T decreased with increases in the repetitions (immersion or brushing), regardless of the cleaning method. Values of color stability for L resin showed significant color change after brushing with and immersion in Ac and Pb. Brushing with D exhibited a higher incidence of color change. For T there were no significant differences between cleaning agents and repetitions in immersion. A color change was noted after three brushings with the Ac, Chx, and D. Brushing with dentifrice decreased roughness of L. Immersion in or brushing with NaOCl and Pb decreased the hardness of L. For T, hardness decreased with increases in immersions or brushing. Color changes after the immersion in or brushing with cleaning agents were clinically acceptable according to National Bureau of Standards parameters for both resins.

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.

Voltage-induced swelling and deswelling of weak polybase brushes.

PubMed

Weir, Michael P; Heriot, Sasha Y; Martin, Simon J; Parnell, Andrew J; Holt, Stephen A; Webster, John R P; Jones, Richard A L

2011-09-06

We have investigated a novel method of remotely switching the conformation of a weak polybase brush using an applied voltage. Surface-grafted polyelectrolyte brushes exhibit rich responsive behavior and show great promise as "smart surfaces", but existing switching methods involve physically or chemically changing the solution in contact with the brush. In this study, high grafting density poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) brushes were grown from silicon surfaces using atom transfer radical polymerization. Optical ellipsometry and neutron reflectivity were used to measure changes in the profiles of the brushes in response to DC voltages applied between the brush substrate and a parallel electrode some distance away in the surrounding liquid (water or D(2)O). Positive voltages were shown to cause swelling, while negative voltages in some cases caused deswelling. Neutron reflectometry experiments were carried out on the INTER reflectometer (ISIS, Rutherford Appleton Laboratory, UK) allowing time-resolved measurements of polymer brush structure. The PDMAEMA brushes were shown to have a polymer volume fraction profile described by a Gaussian-terminated parabola both in the equilibrium and in the partially swollen states. At very high positive voltages (in this study, positive bias means positive voltage to the brush-bearing substrate), the brush chains were shown to be stretched to an extent comparable to their contour length, before being physically removed from the interface. Voltage-induced swelling was shown to exhibit a wider range of brush swelling states in comparison to pH switching, with the additional advantages that the stimulus is remotely controlled and may be fully automated. © 2011 American Chemical Society

Brushing force of manual and sonic toothbrushes affects dental hard tissue abrasion.

PubMed

Wiegand, Annette; Burkhard, John Patrik Matthias; Eggmann, Florin; Attin, Thomas

2013-04-01

This study aimed to determine the brushing forces applied during in vivo toothbrushing with manual and sonic toothbrushes and to analyse the effect of these brushing forces on abrasion of sound and eroded enamel and dentin in vitro. Brushing forces of a manual and two sonic toothbrushes (low and high frequency mode) were measured in 27 adults before and after instruction of the respective brushing technique and statistically analysed by repeated measures analysis of variance (ANOVA). In the in vitro experiment, sound and eroded enamel and dentin specimens (each subgroup n = 12) were brushed in an automatic brushing machine with the respective brushing forces using a fluoridated toothpaste slurry. Abrasion was determined by profilometry and statistically analysed by one-way ANOVA. Average brushing force of the manual toothbrush (1.6 ± 0.3 N) was significantly higher than for the sonic toothbrushes (0.9 ± 0.2 N), which were not significantly different from each other. Brushing force prior and after instruction of the brushing technique was not significantly different. The manual toothbrush caused highest abrasion of sound and eroded dentin, but lowest on sound enamel. No significant differences were detected on eroded enamel. Brushing forces of manual and sonic toothbrushes are different and affect their abrasive capacity. Patients with severe tooth wear and exposed and/or eroded dentin surfaces should use sonic toothbrushes to reduce abrasion, while patients without tooth wear or with erosive lesions confining only to enamel do not benefit from sonic toothbrushes with regard to abrasion.

A switchable polymer layer: Chain folding in end-charged polymer brushes

NASA Astrophysics Data System (ADS)

Heine, David; Wu, David T.

2001-03-01

We use a self-consistent field approximation to model the configurations of end-charged homopolymer and block copolymer brushes in response to an external electric field due to charges on the grafting surface. By varying the charge density on the grafting surface, we can cause the chains either to extend outward, greatly increasing the brush height, or to loop back to the grafting surface. We show that such a copolymer brush can present one block at the exposed surface in the extended state and present the other block in the retracted state. This occurs for both a solvated brush and a dry brush. We also compare these results to those of a modified Alexander-de Gennes model for the end-charged homopolymer brush.

Macroscopic lateral heterogeneity observed in a laterally mobile immiscible mixed polyelectrolyte-neutral polymer brush

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lee, Hoyoung; Tsouris, Vasilios; Lim, Yunho

We studied mixed poly(ethylene oxide) (PEO) and poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) brushes. The question we attempted to answer was: when the chain grafting points are laterally mobile, how will this lateral mobility influence the structure and phase behavior of the mixed brush? Three different model mixed PEO/PDMAEMA brush systems were prepared: (1) a laterally mobile mixed brush by spreading onto the air–water interface a mixture of poly(ethylene oxide)–poly(n-butyl acrylate) (PEO–PnBA) and poly(2-(dimethylamino)ethyl methacrylate)–poly(n-butyl acrylate) (PDMAEMA–PnBA) diblock copolymers (the specific diblock copolymers used will be denoted as PEO 113–PnBA 100 and PDMAEMA 118–PnBA 100, where the subscripts refer to the number-average degreesmore » of polymerization of the individual blocks), (2) a mobility-restricted (inseparable) version of the above mixed brush prepared using a PEO–PnBA–PDMAEMA triblock copolymer (denoted as PEO 113–PnBA 89–PDMAEMA 120) having respective brush molecular weights matched with those of the diblock copolymers, and (3) a different laterally mobile mixed PEO and PDMAEMA brush prepared from a PEO 113–PnBA 100 and PDMAEMA 200–PnBA 103 diblock copolymer combination, which represents a further more height-mismatched mixed brush situation than described in (1). These three mixed brush systems were investigated by surface pressure–area isotherm and X-ray (XR) reflectivity measurements. These experimental data were analyzed within the theoretical framework of a continuum self-consistent field (SCF) polymer brush model. The combined experimental and theoretical results suggest that the mobile mixed brush derived using the PEO 113–PnBA 100 and PDMAEMA 118–PnBA 100 combination (i.e., mixed brush System #1) undergoes a lateral macroscopic phase separation at high chain grafting densities, whereas the more height-mismatched system (System #3) is only microscopically phase separated under comparable brush density conditions even though the lateral mobility of the grafted chains is unrestricted. The macroscopic phase separation observed in the laterally mobile mixed brush system is in contrast with the microphase separation behavior commonly observed in two-dimensional laterally mobile charged small molecule mixtures. Further study is needed to determine the detailed morphologies of the macro- and microphase-separated mixed PEO/PDMAEMA brushes.« less

The effect of toothbrush bristle stiffness on nanohybrid surface roughness

NASA Astrophysics Data System (ADS)

Zairani, O.; Irawan, B.; Damiyanti, M.

2017-08-01

The surface of a restoration can be affected by toothpaste containing abrasive agents and the stiffness of toothbrush bristles. Objective: To identify the effect of toothbrush bristle stiffness on nanohybrid surface roughness. Methods: Sixteen nanohybrid specimens were separated into two groups. The first group was brushed using soft-bristle toothbrushes, and the second group was brushed using medium-bristle toothbrushes. Media such as aqua bides was used for brushing in both groups. Brushing was done 3 times for 5 minutes. Surface roughness was measured initially and at 5, 10, and 15 minutes using a surface roughness tester. Results: The results, tested with One-Way ANOVA and Independent Samples t Test, demonstrated that after brushing for 15 minutes, the soft-bristle toothbrush group showed a significantly different value (p < 0.05) of nanohybrid surface roughness. The group using medium-bristle toothbrushes showed the value of nano hybrid surface roughness significant difference after brushing for 10 minutes. Conclusion: Roughness occurs more rapidly when brushing with medium-bristle tooth brushes than when brushing with soft-bristle toothbrushes.

Early response of ponderosa pine to spacing and brush: observations on a 12-year-old plantation

Treesearch

William W. Oliver

1979-01-01

Ponderosa pine (Pinus ponderosa Laws.) was planted at five different spacings, from 6 by 6 to 18 by 18 feet, on a productive site in northern California. Spacing and brush effects on tree growth were evaluated both on plots where brush was allowed to develop and on plots kept free of brush. Competition between trees in brush-free plots began during...

Nonlinearity in the rotational dynamics of Haidinger's brushes

NASA Astrophysics Data System (ADS)

Rothmayer, Mark; Dultz, Wolfgang; Frins, Erna; Zhan, Qiwen; Tierney, Dennis; Schmitzer, Heidrun

2007-10-01

Haidinger's brushes are an entoptic effect of the human visual system that enables us to detect polarized light. However, individual perceptions of Haidinger's brushes can vary significantly. We find that the birefringence of the cornea influences the rotational motion and the contrast of Haidinger's brushes and may offer an explanation for individual differences. We have devised an experimental setup to simulate various phase shifts of the cornea and found a switching effect in the rotational dynamics of Haidinger's brushes. In addition, age related macular degeneration reduces the polarization effect of the macula and thus also leads to changes in the brush pattern.

Behavioral Determinants of Brushing Young Children's Teeth: Implications for Anticipatory Guidance

PubMed Central

Huebner, Colleen E.; Riedy, Christine A.

2010-01-01

Purpose The purposes of this study were to identify parents' motivation, support, and barriers to twice daily tooth-brushing of infants and preschool-age children and to discover new approaches to encourage this important health behavior. Methods Qualitative interviews were conducted with 44 rural parents about tooth-brushing habits and experiences. Results Forty of 44 parents reported that they had begun to brush their child's teeth; 24 (55%) reported brushing twice a day or more. Parents who brushed twice a day, vs less often, were more likely to describe specific skills to overcome barriers; they expressed high self-efficacy and held high self-standards for brushing. Parents who brushed their children's teeth less than twice daily were more likely to: hold false beliefs about the benefits of twice daily tooth-brushing; report little normative pressure or social support for the behavior; have lower self-standards; describe more external constraints; and offer fewer ideas to overcome barriers. Conclusions The findings support an integrative framework in which barriers and support for parents' twice daily brushing of their young children's teeth are multiple and vary among individuals. Knowledge of behavioral determinants specific to individual parents could strengthen anticipatory guidance and recommendations about at-home oral hygiene of young children. PMID:20298653

Analysis of carbon fiber brush loading in anodes on startup and performance of microbial fuel cells

NASA Astrophysics Data System (ADS)

Hutchinson, Adam J.; Tokash, Justin C.; Logan, Bruce E.

Flat carbon anodes placed near a cathode in a microbial fuel cell (MFC) are adversely affected by oxygen crossover, but graphite fiber brush anodes placed near the cathode produce high power densities. The impact of the brush size and electrode spacing was examined by varying the distance of the brush end from the cathode and solution conductivity in multiple MFCs. The startup time was increased from 8 ± 1 days with full brushes (all buffer concentrations) to 13 days (50 mM), 14 days (25 mM) and 21 days (8 mM) when 75% of the brush anode was removed. When MFCs were all first acclimated with a full brush, up to 65% of the brush material could be removed without appreciably altering maximum power. Electrochemical impedance spectroscopy (EIS) showed that the main source of internal resistance (IR) was diffusion resistance, which together with solution resistance reached 100 Ω. The IR using EIS compared well with that obtained using the polarization data slope method, indicating no major components of IR were missed. These results show that using full brush anodes avoids adverse effects of oxygen crossover during startup, although brushes are much larger than needed to sustain high power.

Potential effects of tooth-brushing on human dentin wear following exposure to acidic soft drinks.

PubMed

Choi, S; Park, K-H; Cheong, Y; Moon, S W; Park, Y-G; Park, H-K

2012-08-01

This study used scanning electron microscopy and atomic force microscopy to examine the short-term potential effects of brushing time and the start-time of tooth-brushing after demineralization on primary dentin wear in vitro. Thirty-six noncarious primary central incisors were assigned to 12 experimental groups. Exposure to cola drinks was used to initiate the demineralization process. Three brushing times (5, 15 and 30 s) and four start-times of brushing (0, 30, 60 and 120 min) after an erosive attack were used for the abrasion process. Tooth-brushing the softened dentin surface led to increases in the open tubular fraction and microstructural changes on the dentin surface. Brushing immediately after exposure to cola resulted in the greatest irreversible dentin loss, whereas brushing 60 or 120 min after pretreatment resulted in the least irreversible dentin loss. However, brushing time had no effect on the irreversible loss of dentin wear. Based on these experimental results, tooth-brushing should be performed at least 60 min after consuming a cola drink to achieve the desired tooth cleaning and avoid the introduction of surface lesions on dentin. © 2012 The Authors Journal of Microscopy © 2012 Wadsworth Center, New York State Department of Health.

Polyelectrolyte brushes: a novel stable lubrication system in aqueous conditions.

PubMed

Kobayashi, Motoyasu; Terada, Masami; Takahara, Atsushi

2012-01-01

Surface-initiated controlled radical copolymerizations of 2-dimethylaminoethyl methacrylate (DMAEMA), 2-(methacryloyloxy)ethyl phosphorylcholine (MPC), 2-(methacryloyloxy)ethyltrimethylammonium chloride) (MTAC), and 3-sulfopropyl methacrylate potassium salt (SPMK) were carried out on a silicon wafer and glass ball to prepare polyelectrolyte brushes with excellent water wettability. The frictional coefficient of the polymer brushes was recorded on a ball-on-plate type tribometer by linear reciprocating motion of the brush specimen at a selected velocity of 1.5 x 10(-3) m s-1 under a normal load of 0.49 N applied to the stationary glass ball (d = 10 mm) at 298 K. The poly(DMAEMA-co-MPC) brush partially cross-linked by bis(2-iodoethoxy)ethane maintained a relatively low friction coefficient around 0.13 under humid air (RH > 75%) even after 200 friction cycles. The poly(SPMK) brush revealed an extremely low friction coefficient around 0.01 even after 450 friction cycles. We supposed that the abrasion of the brush was prevented owing to the good affinity of the poly(SPMK) brush for water forming a water lubrication layer, and electrostatic repulsive interactions among the brushes bearing sulfonic acid groups. Furthermore, the poly(SPMK-co-MTAC) brush with a chemically cross-linked structure showed a stable low friction coefficient in water even after 1400 friction cycles under a normal load of 139 MPa, indicating that the cross-linking structure improved the wear resistance of the brush layer.

Schoolchildren's tooth brushing characteristics and oral hygiene habits assessed with video-recorded sessions at school and a questionnaire.

PubMed

Martignon, Stefania; González, María Clara; Tellez, Marisol; Guzmán, Adriana; Quintero, Ingrid K; Sáenz, Viviana; Martínez, Miguel; Mora, Angelica; Espinosa, Luis Fernando; Castiblanco, Gina A

2012-01-01

Tooth brushing habits become established during the first years of childhood and last throughout lifetime. To assess tooth-brushing characteristics, the procedure was videotaped at school and a questionnaire on oral hygiene knowledge, attitudes and practices was completed. A total 146 5- to 8-year-old low-SES schoolchildren from Bogotá participated. The median total tooth brushing time was 115 sec (75% Q3-178 sec; 25% Q1-83 sec). The median time the toothbrush was in the child's mouth was 89 sec (75% Q3-145 sec; 25% Q1-65 sec). Most children brushed their maxillary (97%), mandibular (95%), anterior (96%) and posterior (81%) teeth. The surfaces most often brushed were the buccal-anterior-maxillary (96%) and mandibular (94%) surfaces. The amount of toothpaste dispensed was 2/3 of toothbrush head in 51% of children. Most children spat (93%), used the mirror (78%), and rinsed their mouth (72%). The majority (97%) was confident that the toothbrushing session was effective. The questionnaire revealed the following: none of the children brush their teeth at school; only 34% is supervised by an adult during the tooth brushing procedure, and only 30% brush twice a day. The study shows overall positive findings of tooth brushing while being observed, in terms of time and use of toothpaste. These results, together with the poor oral-health status and toothbrushing habits reported at home, highly recommend incorporating daily-supervised school-based tooth brushing sessions with fluoride toothpaste.

Frequency of daily tooth brushing: predictors of change in 9- to 11-year old US children.

PubMed

Polk, D E; Geng, M; Levy, S; Koerber, A; Flay, B R

2014-09-01

To investigate whether an increase in daily tooth brushing frequency in children was predicted by either a) having a strong intention to brush twice a day or b) their parents receiving information about their new caries experience. Secondary data analyses were conducted on two waves of data from the Aban Aya Youth Project and the Iowa Fluoride Study. The Aban Aya study included 576 10- and 11-year olds from Chicago, Illinois. The Iowa Fluoride Study included a convenience sample of 709 babies born in Iowa. The present study includes those children at age 9. In both studies, reported daily tooth brushing frequency was assessed twice six months apart. In the Aban Aya data, compared with children with a weak intention at wave 1 to brush twice a day, children with a strong intention to brush twice a day were more likely to increase their brushing frequency by wave 2, OR 7.0, 95%CI 1.5,32.9. In the Iowa Fluoride Study, compared with children who did not have new caries at wave 1, children who had new caries experience were less likely to increase their brushing frequency by wave 2, OR 0.4, 95%CI 0.2,0.9. Strengthening intention to brush twice a day might increase children's brushing frequency. However, simply providing parents with information about new caries probably will not. Future studies should assess tooth brushing frequency, habit strength, intention, and situational cues at closely-spaced waves.

The effect of brushing with toothpaste containing nano calcium carbonate upon nanofill composite resin surface roughness

NASA Astrophysics Data System (ADS)

Ramadhani, A. M.; Herda, E.; Triaminingsih, S.

2017-08-01

This study aims to determine the effect of brushing with toothpaste containing nanocalcium carbonate on the roughness of nanofill composite resin surface. Brushing was conducted with 3 types of materials for 3 consecutive brushing periods of 10 minutes each. Surface roughness was measured using a surface-roughness tester and the results were analyzed using the repeated ANOVA and the one-way ANOVA test. The surface morphology was observed using SEM after 3 months’ worth of brushing with the 3 materials. It was found that the nanofill composite resin surface-roughness value increased significantly (p<0.005) after brushing with toothpaste containing nano calcium carbonate for 3 months, but the value was not as high as that obtained when brushing with other types of toothpaste.

The effect of dentifrice quantity and toothbrushing behaviour on oral delivery and retention of fluoride in vivo.

PubMed

Creeth, Jonathan; Zero, Domenick; Mau, Melissa; Bosma, Mary Lynn; Butler, Andrew

2013-12-01

While toothpaste F(-) concentration and rinsing regimen have well-characterised impacts on fluoride's effectiveness, other aspects of brushing regimen have much less well-established effects, in particular, dentifrice quantity and brushing duration. An in vivo study (n = 42) of oral fluoride delivery (i.e. oral disposition post-brushing), and retention (i.e. concentration of F(-) in saliva post-brushing, a known efficacy predictor), was performed to compare effects observed with those of dentifrice F(-) concentration and rinsing regimen. Subjects brushed with a NaF-silica dentifrice (Aquafresh Advanced, 1,150 ppm F(-) ) or a control dentifrice (250 ppm F(-) , same base), for 45, 60, 120 or 180 seconds with 0.5 or 1.5 g dentifrice, and rinsed with 15 ml water once or three times in a cross-over design. The F(-) concentration was measured in post-brushing expectorate, rinse and toothbrush washing samples, and in saliva between 5-120 minutes after brushing. Using 1.5 g versus 0.5 g dentifrice increased F(-) in all samples: oral retention of F(-) was almost doubled by this increase. Increasing duration of brushing had more complex effects. The amount of F(-) in the expectorate increased but decreased in both rinse and toothbrush washing samples. Oral F(-) retention increased, but only in the period 30-120 minutes after brushing. Over the ranges investigated, the order of importance on oral F(-) retention was: dentifrice F(-) concentration > quantity > rinsing regimen > brushing duration. Hence, increasing dentifrice quantity and, to a lesser extent, the duration of brushing, can elevate oral fluoride post-brushing. Evidence is accumulating that the importance of these variables to fluoride efficacy may have been underestimated. © 2013 FDI World Dental Federation.

Impact of brushing force on abrasion of acid-softened and sound enamel.

PubMed

Wiegand, A; Köwing, L; Attin, T

2007-11-01

The study aimed to analyse the effects of different brushing loads on abrasion of acid-softened and sound enamel surfaces. Sound and acid-softened surfaces of each 10 human enamel samples were submitted to brushing abrasion in an automatic brushing machine at 1.5 N (A), 2.5 N (B), 3.5 N (C) or 4.5 N (D) brushing load. Prior to abrasion, demineralisation of half of each enamel surface was performed by storage in hydrochloric acid (pH 2.0) for 60s. Brushing was carried out (1000 strokes) using a manual toothbrush and toothpaste slurry in a ratio of 1:3. Enamel loss was measured after 10, 20, 50, 100, 150, 200, 250, 300, 350 and 1000 brushing strokes (BS). Pre- and post-brushing values of Knoop indentation length (5 indentations each sample) were measured and mean enamel loss was calculated from the change in indentation depth. Within- and between-group comparisons were performed by ANOVA and t-test followed by Bonferroni-correction. Enamel loss of acid-softened surfaces was significantly influenced by the brushing load applied and was mostly significantly higher in group D (10-1000 BS: 225-462 nm) compared to A (10-1000 BS: 164-384), B (10-1000 BS: 175-370 nm) and C (10-1000 BS: 191-396 nm). Abrasion of acid-softened enamel was fourfold higher compared to sound surfaces. Sound enamel was significantly influenced by the brushing force at 20-200 brushing strokes only, but revealed no significant differences between groups A-D. Brushing load influences abrasion of briefly eroded enamel, but might be of minor importance for abrasion of sound enamel surfaces.

Effect of tooth brushing on gloss retention and surface roughness of five bulk-fill resin composites.

PubMed

O'Neill, Catherine; Kreplak, Laurent; Rueggeberg, Frederick A; Labrie, Daniel; Shimokawa, Carlos Alberto Kenji; Price, Richard Bengt

2018-01-01

To determine the effects of tooth brushing on five bulk-fill resin based composites (RBCs). Ten samples of Filtek Supreme Enamel (control), Filtek One Bulk Fill, Tetric EvoCeram Bulk Fill, SonicFill 2, SDR flow+, and Admira Fusion X-tra were light cured for 20 seconds using the Valo Grand curing light. After 24 hours storage in air at 37°C, specimens were brushed in a random order using Colgate OpticWhite dentifrice and a soft toothbrush. Surface gloss was measured prior to brushing, after 5,000, 10,000 and 15,000 back and forth brushing cycles. Surface roughness was measured after 15,000 brushing cycles using atomic force microscopy (AFM) and selected scanning electron microscope (SEM) images were taken. The data was examined using ANOVA and pair-wise comparisons using Scheffe's post-hoc multiple comparison tests (α = 0.05). Surface gloss decreased and the surface roughness increased after brushing. Two-way ANOVA showed that both the RBC and the number of brushing cycles had a significant negative effect on the gloss. One-way ANOVA showed that the RBC had a significant effect on the roughness after 15,000 brushing cycles. For both gloss and roughness, brushing had the least effect on the nano-filled control and nano-filled bulk-fill RBC, and the greatest negative effect on Admira Fusion X-tra. The SEM images provided visual agreement. There was an excellent linear correlation (R 2  = 0.98) between the logarithm of the gloss and roughness. After brushing, the bulk-fill RBCs were all rougher than the control nano-filled RBC. The nano-filled bulk-fill RBC was the least affected by brushing. Bulk-fill RBCs lose their gloss faster and become rougher than the nanofilled conventional RBC, Filtek Supreme Ultra. The nanofilled bulk-fill RBC was the least affected by tooth brushing. © 2017 Wiley Periodicals, Inc.

A randomized clinical trial evaluating gingivitis and plaque reduction of an oscillating-rotating power brush with a new brush head with angled bristles versus a marketed sonic brush with self-adjusting technology.

PubMed

Klukowska, Malgorzata; Grender, Julie M; Conde, Erinn; Ccahuana-Vasquez, Renzo Alberto; Ram Goyal, C

2014-08-01

To compare the efficacy of an oscillating-rotating power toothbrush with a novel brush head incorporating angled CrissCross bristles (Oral-B Pro 7000 SmartSeries and Oral-B CrossAction brush head) versus a marketed sonic toothbrush (Colgate ProClinical A1500 with the Triple Clean brush head) in the reduction of gingivitis and plaque over a 6-week period. This was a single center, randomized, open label, examiner-blind, 2-treatment, parallel group study. Study participants who met the entrance criteria were enrolled in the study and randomly assigned to one of the two toothbrush groups. Study participants brushed with their assigned toothbrush and a marketed fluoride dentifrice for 2 minutes twice daily at home for 6 weeks. Gingivitis and plaque were evaluated at baseline and Week 6. Gingivitis was assessed using the Modified Gingival Index (MGI) and Gingival Bleeding Index (GBI) and plaque was assessed using the Rustogi Modified Navy Plaque Index (RMNPI). Data was analyzed using the ANCOVA with baseline as the covariate. In total, 130 study participants were randomized to treatment resulting in 64 study participants per group completing the study. Both brushes produced statistically significant (P < 0.001) reductions in gingivitis and plaque measures relative to baseline. The oscillating-rotating,brush with the novel brush head demonstrated statistically significantly (P < 0.05) greater reductions in all gingivitis measures, as well as whole mouth and interproximal plaque measures, compared to the sonic toothbrush. The benefit for the oscillating- rotating brush over the sonic brush was 21.3% for gingivitis, 35.7% for gingival bleeding, 34.7% for number of bleeding sites, 17.4% for whole mouth plaque, and 21.2% for interproximal plaque. There were no adverse events reported or observed for either brush.

Toothbrush abrasivity in a long-term simulation on human dentin depends on brushing mode and bristle arrangement.

PubMed

Bizhang, Mozhgan; Schmidt, Ilka; Chun, Yong-Hee Patricia; Arnold, Wolfgang H; Zimmer, Stefan

2017-01-01

The aim of this study was to evaluate the susceptibility of dentin to brushing abrasion using four different toothbrushes (rotating-oscillating, sonic and two types of manual toothbrushes) with the same brushing forces. Dentin samples (n = 72) were selected from 72 impacted third molars. Half of the surface of dentin samples was covered with an adhesive tape, creating a protected and a freely exposed area in the same specimen. Brushing was performed with either a: sonic (Sonicare PowerUp, Philips GmbH, Hamburg, Germany), b: oscillating-rotating (Oral B Vitality Precisions Clean, Procter & Gamble, Schwalbach am Taunus, Germany) or two different manual toothbrushes c: flat trim brush head toothbrush (Dr. Best: Original, Glaxo-Smith-Kline, Bühl, Germany) and d: rippled-shaped brush head toothbrush (Blend-a-Dent, Complete V-Interdental, Blend-a-med, Schwalbach, Germany) in a custom made automatic brushing machine. The brushing force was set to 2 N and a whitening toothpaste (RDA = 150) was used. The simulation period was performed over a calculated period to mimic a brushing behavior of two times a day brushing for eight years and six months. Dentin loss was quantitatively determined by profilometry and statistically analyzed by Wilcoxon and Mann-Whitney-U Test (p < 0.05). The mean (standard deviation) surface loss was 21.03 (±1.26) μm for the sonic toothbrush, 15.71 (±0.85) μm for the oscillating-rotating toothbrush, 6.13 (±1.24) μm for the manual toothbrush with flat trim brush head and 2.50 (±0.43) μm for the manual toothbrush with rippled-shaped brush head. Differences between all groups were statistically significant at p<0.05. Using the same brushing force and a highly abrasive toothpaste, manual toothbrushes are significantly less abrasive compared to power toothbrushes for an 8.5-year simulation.

[Clinical relevance of tooth brushing in relation to dental caries].

PubMed

Pita-Fernández, Salvador; Pombo-Sánchez, Antonio; Suárez-Quintanilla, Juan; Novio-Mallón, Silvia; Rivas-Mundiña, Berta; Pértega-Díaz, Sonia

2010-07-01

To determine the impact and clinical relevance of tooth brushing on oral health. Prevalence study. Fontiñas Health Centre. (Santiago de Compostela, Spain). Prevalence study (n=281 children aged 5-14 years. Odontological examination according to WHO methodology, to determine the frequency of tooth brushing, frequency of sweet consumption and their impact on the prevalence of caries. Logistic regression and estimation of the relative prevalence difference (RPD) and the Number Needed to Treat in order to prevent one additional bad outcome (NNT). The children who never brush their teeth have a 40% (95% CI: 24.3%-57.8%) of early caries, while those who brush their teeth several times a day have 15.3% (95% CI: 9.4%-23.7%). An association between not brushing the teeth and caries in primary teeth (OR=2.3; 95% CI:1.05-5.3) was observed after adjusting for age, sweet consumption and visits to the dentist. The same occurred with final teeth (OR=3.9; 95% CI:1.4-10.3). The RPD was 62%(95% CI: 30%-79%), meaning that prevalence of caries is 62% lower in children who brush their teeth several times a day as compared to those who never brush their teeth. The NNT is 4 (95% CI: 2.4-14), so for every 4 children who brush their teeth several times a day, there is one less case of caries, compared to those who never brush their teeth. There is a dose-response relationship between prevalence of caries and brushing frequency. The same effect was observed with definitive caries: RPD=55% (95% CI:16%-76%), NNT=5 (95% CI:2.8-53.3). Tooth brushing is related to oral health, with a major clinical impact. The positive effect of tooth brushing was superior to that of a correct diet. 2009 Elsevier España, S.L. All rights reserved.

Toothbrush abrasivity in a long-term simulation on human dentin depends on brushing mode and bristle arrangement

PubMed Central

2017-01-01

Objective The aim of this study was to evaluate the susceptibility of dentin to brushing abrasion using four different toothbrushes (rotating-oscillating, sonic and two types of manual toothbrushes) with the same brushing forces. Methods Dentin samples (n = 72) were selected from 72 impacted third molars. Half of the surface of dentin samples was covered with an adhesive tape, creating a protected and a freely exposed area in the same specimen. Brushing was performed with either a: sonic (Sonicare PowerUp, Philips GmbH, Hamburg, Germany), b: oscillating-rotating (Oral B Vitality Precisions Clean, Procter & Gamble, Schwalbach am Taunus, Germany) or two different manual toothbrushes c: flat trim brush head toothbrush (Dr. Best: Original, Glaxo-Smith-Kline, Bühl, Germany) and d: rippled-shaped brush head toothbrush (Blend-a-Dent, Complete V-Interdental, Blend-a-med, Schwalbach, Germany) in a custom made automatic brushing machine. The brushing force was set to 2 N and a whitening toothpaste (RDA = 150) was used. The simulation period was performed over a calculated period to mimic a brushing behavior of two times a day brushing for eight years and six months. Dentin loss was quantitatively determined by profilometry and statistically analyzed by Wilcoxon and Mann-Whitney-U Test (p < 0.05). Results The mean (standard deviation) surface loss was 21.03 (±1.26) μm for the sonic toothbrush, 15.71 (±0.85) μm for the oscillating-rotating toothbrush, 6.13 (±1.24) μm for the manual toothbrush with flat trim brush head and 2.50 (±0.43) μm for the manual toothbrush with rippled-shaped brush head. Differences between all groups were statistically significant at p<0.05. Conclusion Using the same brushing force and a highly abrasive toothpaste, manual toothbrushes are significantly less abrasive compared to power toothbrushes for an 8.5—year simulation. PMID:28222156

``Smart'' Surfaces of Polymer Brushes

NASA Astrophysics Data System (ADS)

Wang, Qiang; Meng, Dong

2009-03-01

``Smart'' surfaces, also known as stimuli-responsive surfaces, can change their properties (e.g., wettability, adhesion, friction, elasticity, and biocompatibility) in response to external stimuli (e.g., temperature, pressure, light, solvent selectivity, ionic strength, type of salt, pH, applied electric field, etc.). In this work, we use numerical self-consistent field calculations to study in detail the structure and stimuli- responses of various polymer brushes, including (1) the thermo- response of PNIPAM brushes in water, (2) solvent-response of uncharged diblock copolymer brushes, and (3) the stimuli- response of charged two-component polymer brushes (including both the binary A/B brushes and diblock copolymer A-B brushes) to ionic strength, pH, and applied electric field. Among the many design parameters (e.g., chain lengths, grafting densities, A-B incompatibility, degree of ionization of charged polymers, etc.) we identify those that strongly affect the surface switchability. Such knowledge is useful to the experimental design of these smart polymer brushes for their applications.

Comparing solvophobic and multivalent induced collapse in polyelectrolyte brushes

DOE PAGES

Jackson, Nicholas E.; Brettmann, Blair K.; Vishwanath, Venkatram; ...

2017-02-03

Here, coarse-grained molecular dynamics enhanced by free-energy sampling methods is used to examine the roles of solvophobicity and multivalent salts on polyelectrolyte brush collapse. Specifically, we demonstrate that while ostensibly similar, solvophobic collapsed brushes and multivalent-ion collapsed brushes exhibit distinct mechanistic and structural features. Notably, multivalent-induced heterogeneous brush collapse is observed under good solvent polymer backbone conditions, demonstrating that the mechanism of multivalent collapse is not contingent upon a solvophobic backbone. Umbrella sampling of the potential of mean-force (PMF) between two individual brush strands confirms this analysis, revealing starkly different PMFs under solvophobic and multivalent conditions, suggesting the role ofmore » multivalent “bridging” as the discriminating feature in trivalent collapse. Structurally, multivalent ions show a propensity for nucleating order within collapsed brushes, whereas poor-solvent collapsed brushes are more disordered; this difference is traced to the existence of a metastable PMF minimum for poor solvent conditions, and a global PMF minimum for trivalent systems, under experimentally relevant conditions.« less

Comparing solvophobic and multivalent induced collapse in polyelectrolyte brushes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jackson, Nicholas E.; Brettmann, Blair K.; Vishwanath, Venkatram

Here, coarse-grained molecular dynamics enhanced by free-energy sampling methods is used to examine the roles of solvophobicity and multivalent salts on polyelectrolyte brush collapse. Specifically, we demonstrate that while ostensibly similar, solvophobic collapsed brushes and multivalent-ion collapsed brushes exhibit distinct mechanistic and structural features. Notably, multivalent-induced heterogeneous brush collapse is observed under good solvent polymer backbone conditions, demonstrating that the mechanism of multivalent collapse is not contingent upon a solvophobic backbone. Umbrella sampling of the potential of mean-force (PMF) between two individual brush strands confirms this analysis, revealing starkly different PMFs under solvophobic and multivalent conditions, suggesting the role ofmore » multivalent “bridging” as the discriminating feature in trivalent collapse. Structurally, multivalent ions show a propensity for nucleating order within collapsed brushes, whereas poor-solvent collapsed brushes are more disordered; this difference is traced to the existence of a metastable PMF minimum for poor solvent conditions, and a global PMF minimum for trivalent systems, under experimentally relevant conditions.« less

Repetitive switching for an electromagnetic rail gun

NASA Astrophysics Data System (ADS)

Gruden, J. M.

1983-12-01

Previous testing on a repetitive opening switch for inductive energy storage has proved the feasibility of the rotary switch concept. The concept consists of a rotating copper disk (rotor) with a pie-shaped insulator section and brushes which slide along each of the rotor surfaces. While on top of the copper surface, the brushes and rotor conduct current allowing the energy storage inductor to charge. When the brushes slide onto the insulator section, the current cannot pass through the rotor and is diverted into the load. This study investigates two new brush designs and a rotor modification designed to improve the current commutating capabilities of the switch. One brush design (fringe fiber) employs carbon fibers on the leading and trailing edge of the brush to increase the resistive commutating action as the switch opens and closes. The other brush design uses fingers to conduct current to the rotor surface, effectively increasing the number of brush contact points. The rotor modification was the placement of tungsten inserts at the copper-insulator interfaces.

Brush border enzyme-cleavable linkers: Evaluation for reducing renal uptake of radiolabeled prostate-specific membrane antigen inhibitors.

PubMed

Vaidyanathan, Ganesan; Kang, Choong Mo; McDougald, Darryl; Minn, Il; Brummet, Mary; Pomper, Martin G; Zalutsky, Michael R

2018-05-05

Radiolabeled, low-molecular-weight prostate-specific membrane antigen (PSMA) inhibitors based on the Glu-ureido pharmacophore show promise for the detection and treatment of castration-resistant prostate cancer; however, high renal retention of activity, related in part to overexpression of PSMA in kidneys can be problematic. The goal of the current study was to investigate the use of brush border enzyme-cleavable linkers as a strategy for reducing kidney activity levels from radiolabeled PSMA inhibitors. PSMA-769 (6), a derivative of the prototypical PSMA inhibitor (((S)‑1‑carboxy‑5‑(4‑iodobenzamido)pentyl)carbamoyl)glutamate (12) modified to contain a Gly-Tyr linker, and its protected tin precursor (11) were synthesized starting from the basic pharmacophore molecule Lys-urea-Glu. An analogue of 6 containing d‑tyrosine in lieu of l‑tyrosine (PSMA-769-d-tyrosine) and the corresponding tin precursor (d-11) also were synthesized. Both radioiodinated and 211 At-labeled 6 were synthesized by radiohalogenation of 11 and deprotection in situ. Similarly, radioiodinated d-6 was synthesized from d-11. Paired label biodistribution of [ 125 I]12 and [ 131 I]6 was performed in normal mice and in SCID mice bearing both PC3 PIP (PSMA+) and PC3 flu (PSMA-) subcutaneous prostate carcinoma xenografts. The biodistribution of [ 131 I]6 and [ 211 At]6 was also evaluated in this tumor model. Biodistribution of the two radioiodinated diastereomers of 6 was evaluated in normal mice and urine samples were analyzed for the presence of 4‑iodohippuric acid. Compounds [ 131 I]6 and [ 211 At]6 were synthesized from 11 in overall radiochemical yields of 32.5 ± 0.1% (n = 4) and 22% (n = 1), respectively; radiochemical purity was >95%. In normal mice, renal uptake of [ 131 I]6 was 1.4-, 2.8- and 161-fold lower than that seen for co-injected [ 125 I]12 at 1 h, 4 h and 21 h, respectively. In tumor-bearing mice, kidney uptake of [ 131 I]6 was similar to that for [ 125 I]12 (P > 0.05) at 1 h and 4 h but was 6- to 7-fold lower at 21 h; however, [ 131 I]6 uptake in PC3 PIP tumors was also lower than that seen for [ 125 I]12 at 21 h (12.6 ± 3.4%ID/g vs. 36.8 ± 12.4%ID/g). Uptake of [ 211 At]PSMA-769 in PC3 PIP tumors was slightly higher than that seen for [ 131 I]PSMA-769 at 4 h (9.6 ± 1.6%ID/g versus 7.8 ± 1.6%ID/g; P = 0.002); its uptake in a number of normal tissues also was higher. In normal mice, kidney uptake of [ 125 I]PSMA-769 at 4 h was about 73% of that seen for [ 131 I]PSMA-769-d-tyrosine. Activity in the urine of mice receiving [ 125 I]PSMA-769 contained mainly 4‑[ 125 I]iodohippuric acid while unmetabolized intact molecule was present in the case of [ 125 I]PSMA-769-d-tyrosine. Use of this brush border enzyme-cleavable linker reduced kidney uptake and resulted in improved tumor:kidney uptake ratios. Although further structural improvements are needed, this linker approach might be useful as a component in strategies for reducing renal uptake of radiolabeled PSMA inhibitors. Copyright © 2018 Elsevier Inc. All rights reserved.

AFM-based force spectroscopy on polystyrene brushes: effect of brush thickness on protein adsorption.

PubMed

Hentschel, Carsten; Wagner, Hendrik; Smiatek, Jens; Heuer, Andreas; Fuchs, Harald; Zhang, Xi; Studer, Armido; Chi, Lifeng

2013-02-12

Herein we present a study on nonspecific binding of proteins at highly dense packed hydrophobic polystyrene brushes. In this context, an atomic force microscopy tip was functionalized with concanavalin A to perform single-molecule force spectroscopy measurements on polystyrene brushes with thicknesses of 10 and 60 nm, respectively. Polystyrene brushes with thickness of 10 nm show an almost two times stronger protein adsorption than brushes with a thickness of 60 nm: 72 pN for the thinner and 38 pN for the thicker layer, which is in qualitative agreement with protein adsorption studies conducted macroscopically by fluorescence microscopy.

Design optimization of a brush turbine with a cleaner/water based solution

NASA Technical Reports Server (NTRS)

Kim, Rhyn H.

1995-01-01

Recently, a turbine-brush was analyzed based on the energy conservation and the force momentum equation with an empirical relationship of the drag coefficient. An equation was derived to predict the rotational speed of the turbine-brush in terms of the blade angle, number of blades, rest of geometries of the turbine-brush and the incoming velocity. Using the observed flow conditions, drag coefficients were determined. Based on the experimental values as boundary conditions, the turbine-brush flows were numerically simulated to understand first the nature of the flows, and to extend the observed drag coefficient to a flow without holding the turbine-brush.

Synthesis and characterization of poly(3-sulfopropylmethacrylate) brushes for potential antibacterial applications.

PubMed

Ramstedt, Madeleine; Cheng, Nan; Azzaroni, Omar; Mossialos, Dimitris; Mathieu, Hans Jörg; Huck, Wilhelm T S

2007-03-13

This article describes the aqueous atom transfer radical polymerization synthesis of poly(3-sulfopropylmethacrylate) brushes onto gold and Si/SiO2 surfaces in a controlled manner. The effect of Cu(I)/Cu(II) ratio was examined, and a quartz crystal microbalance was used to study the kinetics of the brush synthesis. The synthesized brushes displayed a thickness from a few nanometers to several hundred nanometers and were characterized using atomic force microscopy, ellipsometry, Fourier transform infrared spectroscopy (FTIR), contact angle measurements, and X-ray photoelectron spectroscopy (XPS). The as-synthesized sulfonate brushes had very good ion-exchange properties for the ions tested in this study, i.e., Na+, K+, Cu2+, and Ag+. FTIR and XPS show that the metal ions are coordinating to sulfonate moieties inside the brushes. The brushes were easily loaded with silver ions, and the effect of silver ion concentration on silver loading of the brush was examined. The silver-loaded brushes were shown to be antibacterial toward both gram negative and gram positive bacteria. The silver leaching was studied through leaching experiments into water, NaNO3, and NaCl (physiological medium). The results from these leaching experiments are compared and discussed in the article.

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

DOE PAGES

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

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

Comparison of Saudi child versus parent-report of child tooth-brushing practices.

PubMed

Salama, F; Abobakr, I; Al-Khodair, N; Al-Wakeel, M

2016-01-01

This cross-sectional study compared Saudi children's tooth-brushing practices as reported by children and parents and then matched the agreement of reports. A sample of 100 Saudi parents and their children of ages 8-12 years participated in this cross-sectional investigation. A self-administered structured questionnaire regarding the current tooth-brushing practice and habits at home was filled by the child. The same questionnaire was filled independently by parents. Mothers were more likely to teach children about tooth-brushing and fathers were minimally helping children during tooth-brushing. Mothers were the first to teach the children and approximately 33% of the children started brushing their teeth by age 5. Children and parents reported that children clean their teeth using a tooth-brush (73.74% and 76.77%), miswak (5.05% and 5.05%), or both (21.21% and 18.18%), respectively. There was some agreement between reported tooth-brushing practice of children and their parents. It is important to question both the child and parent regarding oral hygiene practice and compare their answers to get more broad knowledge about their practices. Approximately, one-third of the children started brushing their teeth by age 5, which differs from recommended oral hygiene practices.

In Vitro Assessment of the Abrasion Resistance of Two Types of Artificial Teeth Submitted to Brushing.

PubMed

Policastro, Vivian Barnabé; Giro, Gabriela; Leite, Andressa Rosa Perin; Mendoza-Marin, Danny Omar; Paleari, André Gustavo; Compagnoni, Marco Antonio; Pero, Ana Carolina

2016-08-01

To assess the effect of brushing with different solutions on the abrasion resistance of two types of acrylic resin teeth. Maxillary premolars from two types of acrylic teeth (Biotone and Biotone IPN) were divided into six groups (n = 12), according to the solution used during brushing: distilled water (control), coconut soap, or dentifrice. A mechanical brushing machine was used to simulate approximately 1 year of brushing (11,000 strokes). The weight loss (WL) of teeth was obtained from the difference between the initial (IW) and final weight (FW) of each specimen, and the mean of percentage of weight loss (PWL) was calculated for each group. Data were analyzed using the Kruskal-Wallis test, followed by Bonferroni's post-test comparison (α = 0.05). A statistically significant difference was found for the factor solution (p < 0.001). Brushing using dentifrice caused the highest values of weight loss (-0.50%), in comparison with the groups brushed with coconut soap (0.00%) or distilled water (0.00%). For both types of artificial teeth, brushing with dentifrice produced higher abrasion than brushing with coconut soap or water. © 2016 by the American College of Prosthodontists.

The efficacy of oral brush biopsy with computer-assisted analysis in identifying precancerous and cancerous lesions

PubMed Central

2011-01-01

Background Cancer of the oral cavity is the sixth most common malignancy reported worldwide and one with the highest mortality rate among all malignancies. There is a paucity of reliable diagnostic methods to detect early malignancies. This study was performed to evaluate the sensitivity and specificity of brush biopsy in identifying oral premalignant and malignant lesions. Methods Oral brush and scalpel biopsies were performed on 85 consecutive patients presenting with an oral lesion deemed to be minimally suspicious by clinical examination and the results were compared. Results Of 79 patients with adequate brush biopsy samples with matching scalpel biopsies, 27 revealed histopathologic evidence of dysplasia or carcinoma, 26 of which were independently identified with the oral brush biopsy (sensitivity: 96.3% - 95% CI, 87%-100%). 52 oral lesions did not reveal any histopathologic evidence of dysplasia or carcinoma and of these, brush biopsy reported 47 as "negative" and 5 as "atypical"(specificity of "positive" brush biopsy result is 100%- 95% CI, 93%-100%; specificity for "atypical" brush biopsy result is 90.4%- 95% CI, 82%-97%. The positive predictive value of an abnormal oral brush biopsy was 84% and the negative predictive value was 98%. Conclusion Our study demonstrated that the oral brush biopsy is an accurate test in identifying oral premalignant and malignant lesions, even if minimally suspicious. PMID:21864339

Effects of carbon brush anode size and loading on microbial fuel cell performance in batch and continuous mode

NASA Astrophysics Data System (ADS)

Lanas, Vanessa; Ahn, Yongtae; Logan, Bruce E.

2014-02-01

Larger scale microbial fuel cells (MFCs) require compact architectures to efficiently treat wastewater. We examined how anode-brush diameter, number of anodes, and electrode spacing affected the performance of the MFCs operated in fed-batch and continuous flow mode. All anodes were initially tested with the brush core set at the same distance from the cathode. In fed-batch mode, the configuration with three larger brushes (25 mm diameter) produced 80% more power (1240 mW m-2) than reactors with eight smaller brushes (8 mm) (690 mW m-2). The higher power production by the larger brushes was due to more negative and stable anode potentials than the smaller brushes. The same general result was obtained in continuous flow operation, although power densities were reduced. However, by moving the center of the smaller brushes closer to the cathode (from 16.5 to 8 mm), power substantially increased from 690 to 1030 mW m-2 in fed batch mode. In continuous flow mode, power increased from 280 to 1020 mW m-2, resulting in more power production from the smaller brushes than the larger brushes (540 mW m-2). These results show that multi-electrode MFCs can be optimized by selecting smaller anodes, placed as close as possible to the cathode.

Clinical validation of robot simulation of toothbrushing - comparative plaque removal efficacy

PubMed Central

2014-01-01

Background Clinical validation of laboratory toothbrushing tests has important advantages. It was, therefore, the aim to demonstrate correlation of tooth cleaning efficiency of a new robot brushing simulation technique with clinical plaque removal. Methods Clinical programme: 27 subjects received dental cleaning prior to 3-day-plaque-regrowth-interval. Plaque was stained, photographically documented and scored using planimetrical index. Subjects brushed teeth 33–47 with three techniques (horizontal, rotating, vertical), each for 20s buccally and for 20s orally in 3 consecutive intervals. The force was calibrated, the brushing technique was video supported. Two different brushes were randomly assigned to the subject. Robot programme: Clinical brushing programmes were transfered to a 6-axis-robot. Artificial teeth 33–47 were covered with plaque-simulating substrate. All brushing techniques were repeated 7 times, results were scored according to clinical planimetry. All data underwent statistical analysis by t-test, U-test and multivariate analysis. Results The individual clinical cleaning patterns are well reproduced by the robot programmes. Differences in plaque removal are statistically significant for the two brushes, reproduced in clinical and robot data. Multivariate analysis confirms the higher cleaning efficiency for anterior teeth and for the buccal sites. Conclusions The robot tooth brushing simulation programme showed good correlation with clinically standardized tooth brushing. This new robot brushing simulation programme can be used for rapid, reproducible laboratory testing of tooth cleaning. PMID:24996973

Use of a disclosed plaque visualization technique improved the self-performed, tooth brushing ability of primary schoolchildren.

PubMed

Chounchaisithi, Napa; Santiwong, Busayarat; Sutthavong, Sirikarn; Asvanit, Pompun

2014-02-01

Disclosing agents have a long history of use as an aid in children's tooth brushing instruction. However, their benefit when used to improve self-performed tooth brushing ability without any tooth brushing instruction has not been investigated. To evaluate the effect of disclosed plaque visualization on improving the self-performed, tooth brushing ability of primary school children. A cluster-randomized, crossover study was conducted in Nakhon Nayok province, Thailand. A total of 122 second-grade schoolchildren, aged 8-10 years old, from 12 schools were randomly divided into 2 groups. The first group was assigned to brush with disclosed plaque visualization, while the other group brushed without disclosed plaque visualization. One month later the groups switched procedures. Tooth brushing ability was evaluated by the subjects' reduction in patient hygiene performance (PHP) scores. The data were analyzed using repeated-measures analysis of variance, with significance set at p<0.05. Disclosed plaque visualization had a significant effect on improving the children's self-performed, tooth brushing ability in all areas of the mouth (p<0.001), particularly for anterior teeth, mandibular teeth, buccal surfaces, and areas adjacent to the gingival margin (p<0.001). Disclosed plaque visualization is a viable technique to improve children's self-performed tooth brushing ability, and could be used in school-based oral health promotion programs.

Clinical validation of robot simulation of toothbrushing--comparative plaque removal efficacy.

PubMed

Lang, Tomas; Staufer, Sebastian; Jennes, Barbara; Gaengler, Peter

2014-07-04

Clinical validation of laboratory toothbrushing tests has important advantages. It was, therefore, the aim to demonstrate correlation of tooth cleaning efficiency of a new robot brushing simulation technique with clinical plaque removal. Clinical programme: 27 subjects received dental cleaning prior to 3-day-plaque-regrowth-interval. Plaque was stained, photographically documented and scored using planimetrical index. Subjects brushed teeth 33-47 with three techniques (horizontal, rotating, vertical), each for 20s buccally and for 20s orally in 3 consecutive intervals. The force was calibrated, the brushing technique was video supported. Two different brushes were randomly assigned to the subject. Robot programme: Clinical brushing programmes were transfered to a 6-axis-robot. Artificial teeth 33-47 were covered with plaque-simulating substrate. All brushing techniques were repeated 7 times, results were scored according to clinical planimetry. All data underwent statistical analysis by t-test, U-test and multivariate analysis. The individual clinical cleaning patterns are well reproduced by the robot programmes. Differences in plaque removal are statistically significant for the two brushes, reproduced in clinical and robot data. Multivariate analysis confirms the higher cleaning efficiency for anterior teeth and for the buccal sites. The robot tooth brushing simulation programme showed good correlation with clinically standardized tooth brushing.This new robot brushing simulation programme can be used for rapid, reproducible laboratory testing of tooth cleaning.

Polymer Brushes Containing Sulfonated Sugar Repeat Units: Synthesis, Characterization and In Vitro Testing of Blood Coagulation Activation

PubMed Central

Ayres, N.; Holt, D. J.; Jones, C.F.; Corum, L. E.; Grainger, D. W.

2009-01-01

A new polymer brush chemistry containing sulfonated carbohydrate repeat units has been synthesized from silicon substrates using ATRP methods and characterized both in bulk and using surface analysis. The polymer brush was designed to act as a mimic for the naturally occurring sulfonated glycosaminoglycan, heparin, commonly used for modifying blood-contacting surfaces both in vitro and in vivo. Surface analysis showed conversion of brush saccharide precursor chemistry to the desired sulfonated polymer product. The sulfonated polymer brush surface was further analyzed using three conventional in vitro tests for blood compatibility -- plasma recalcification times, complement activation, and thrombin generation. The sulfonated polymer brush films on silicon oxide wafers exhibited better assay performance in these blood component assays than the unsulfonated sugar functionalized polymer brush in all tests performed. PMID:19859552

Methylation analysis of CMTM3 and DUSP1 gene promoters in high-quality brush hair in the Yangtze River delta white goat.

PubMed

Qiang, Wang; Guo, Haiyan; Li, Yongjun; Shi, Jianfei; Yin, Xiuyuan; Qu, Jingwen

2018-08-20

The Yangtze River delta white goat is the only goat breed that produces high-quality brush hair, which is specifically used in top-grade writing brushes. Previous studies have indicated that the CMTM3 and DUSP1 genes are involved in the growth and cycle of high-quality brush hair, and these genes are thought to be involved in the formation of high-quality brush hair traits. In this study, we investigated the relationship between methylation of CMTM3 and DUSP1 and such traits. The results indicated that the relative expression levels of the CMTM3 and DUSP1 genes were higher in non-high-quality brush hair than in high-quality brush hair. Furthermore, the CpG sites of the DUSP1 gene were not methylated, and the methylation level of CMTM3 was negatively correlated with the gene expression level. We believe that the DUSP1 gene regulates the formation of high-quality brush hair by non-methylated, and that methylation of the CMTM3 gene results in a decrease in its expression, causing an increase in the activity of the androgen receptor and the level of androgen. This high androgen level promotes the growth of high-quality brush hair. These study results provide a theoretical basis for further elucidating the molecular mechanism of the formation of high-quality brush hair characteristics, and provide scientific reference for the molecular breeding of high-quality brush hair. Copyright © 2018 Elsevier B.V. All rights reserved.

Attitudes and behavioural factors relating to toothbrushing and the use of fluoride toothpaste among caries-active Swedish adolescents - a questionnaire study.

PubMed

Nordström, Anna; Birkhed, Dowen

2017-10-01

The aim of this study was to identify attitudes and behaviour relating to fluoride toothpaste and toothbrushing habits among caries-active Swedish adolescents. This cross-sectional study is based on an earlier clinical, two-year toothpaste intervention study. At the last appointment, 206 adolescents (of 211) answered a questionnaire comprising nine semi-closed questions. The majority (93%) brushed their teeth every day, while 7% did so only occasionally. Most participants (77%) brushed twice a day, while 12% brushed just once a day. About half of those brushing just once a day forgot to brush in the evening. Similarly, more than half of the adolescents (53%) used 1 cm of toothpaste or less on their toothbrush. Moreover, 49% brushed for less than 2 min, 41% brushed for 2 min and 10% for more than 2 min. The majority (73%) rinsed with water after toothbrushing. A difference between boys and girls was also observed; 87% of the girls brushed twice a day, whereas only 67% of the boys did so and boys more frequently forgot in the evening. Attitudes and behaviour relating to fluoride toothpaste and toothbrushing habits among caries-active Swedish adolescents are still inadequate after two years of toothpaste intervention. There are several areas where improvements can be made, such as frequency of brushing, brushing time, amount of toothpaste and post-brushing procedures. The majority (81%) included 'fresh breath' as a reason for performing oral hygiene and this aspect can be used by dental staff in health promotion.

Effectiveness of Plaque Control with Novel Pediatric Oral Hygiene Need Station (Modified Oral Irrigation Device) as Compared with Manual Brushing and Flossing: Randomized Controlled Pilot Trial.

PubMed

Murthy, Prashanth Sadashiva; Shaik, Naseemoon; Deshmukh, Seema; Girish, M S

2018-01-01

Establishing good hygiene habits are valuable for present and future oral health. Below 6 years, tooth brushing should be performed by parents, as increasing dexterity and cognition may permit supervised brushing until the child is capable of independent brushing. The aim of the present study was to evaluate the effectiveness of modified oral irrigation device in children in terms of plaque control and to compare the effectiveness of plaque control with manual brushing with the modified oral irrigation device in children. A randomized clinical trial was performed on 12 subjects who were allocated to the two study groups. After obtaining the consent, the control group was instructed tooth brushing with regular pediatric commercially available toothbrush and the intervention group with modified oral irrigation device. Plaque scores in both groups were assessed pre- and post-brushing using modified navy plaque index. The data were subjected to Descriptive statistics and Paired t -test using SPSS version 22. Intragroup comparison of mean difference of plaque score in control group and intervention group pre- and post-brushing was statistically significant. Intergroup comparison of manual brushing group with modified oral irrigation group shows P < 0.05 was statistically significant. Within the limitation of the present study, it has been found novel pediatric oral hygiene need Station is more effective than manual brushing since it combined the effect of brushing, flossing (water floss), and rinsing in children simultaneously and at the same time did not demand any special motor skill.

Computer Simulations of Bottle Brushes: From Melts to Soft Networks

DOE PAGES

Cao, Zhen; Carrillo, Jan-Michael Y.; Sheiko, Sergei S.; ...

2015-07-13

We use a combination of Molecular dynamics simulations and analytical calculations, and study dens bottle-brush systems in a melt and network State. Analysis of our simulation results shows that bottle-brush macromolecules in melt behave as ideal chains with effective Kuhn length b K. Simulations show that the bottle-brush-induced bending rigidity is due to an entropy decrease caused by redistribution of the side chains upon backbone bending. The Kuhn length of the bottle:brushes increases with increasing the side-chain degree of polymerization n sc as b K proportional to n sc 0.46. Moreover, this model of bottle brush macromolecules is extended tomore » describe mechanical properties of bottle brush networks in linear and nonlinear deformation regimes. In the linear deformation regime, the network shear modulus scales with the degree of polymerization of the side chains as G 0 proportional to (n sc + 1) -1 as long as the ratio of the Kuhn length, b K, to the size of the fully extended bottle-brush backbone between cross-links, R-max, is smaller than unity, b K/R max << 1. Bottle-brush networks With b K/R max proportional to 1 demonstrate behavior similar to that of networks Of semiflexible chains with G 0 proportional to n sc -0.5. Finally, in the nonlinear network deformation regime, the deformation-dependent shear modulus is a universal function of the first strain invariant I 1 and bottle-brush backbone deformation ratio beta describing stretching ability of the bottle-brush backbone between cross-links.« less

Fluoroscopic percutaneous brush cytology, forceps biopsy and both in tandem for diagnosis of malignant biliary obstruction.

PubMed

Boos, Johannes; Yoo, Raphael J; Steinkeler, Jennifer; Ayata, Gamze; Ahmed, Muneeb; Sarwar, Ammar; Weinstein, Jeffrey; Faintuch, Salomao; Brook, Olga R

2018-02-01

To evaluate percutaneous brush cytology, forceps biopsy and a tandem procedure consisting of both, in the diagnosis of malignant biliary obstruction. A retrospective review of consecutive patients who underwent biliary brush cytology and/or forceps biopsy between 01/2010 and 09/2014 was performed. The cytology and pathology results were compared to the composite outcome (including radiological, pathological and clinical data). Cost for tandem procedure compared to brush cytology and forceps biopsy alone was calculated. A total of 232 interventions in 129 patients (70.8 ± 11.0 years) were included. Composite outcome showed malignancy in 94/129 (72.9%) patients. Sensitivity for brush cytology, forceps biopsy and tandem procedure was 40.6% (95% CI 32.6-48.7%), 42.7% (32.4-53.0%) and 55.8% (44.7-66.9%) with 100% specificity, respectively. There were 9/43 (20.9%) additional cancers diagnosed when forceps biopsy was performed in addition to brush cytology, while there were 13/43 (30.2%) more cancers diagnosed when brush cytology was performed in addition to forceps biopsy. Additional costs per additionally diagnosed malignancy if tandem approach is to be utilised in all cases was $704.96. Using brush cytology and forceps biopsy in tandem improves sensitivity compared to brush cytology and forceps biopsy alone in the diagnosis of malignant biliary obstruction. • Tandem procedure improves sensitivity compared to brush cytology and forceps biopsy. • Brush cytology may help to overcome "crush artefacts" from forceps biopsy. • The cost per diagnosed malignancy may warrant tandem procedure in all patients.

Experience of racism and tooth brushing among pregnant Aboriginal Australians: exploring psychosocial mediators.

PubMed

Ben, J; Jamieson, L M; Priest, N; Parker, E J; Roberts-Thomson, K F; Lawrence, H P; Broughton, J; Paradies, Y

2014-09-01

Despite burgeoning evidence regarding the pathways by which experiences of racism influence health outcomes, little attention has been paid to the relationship between racism and oral health-related behaviours in particular. We hypothesised that self-reported racism was associated with tooth brushing, and that this association was mediated by perceived stress and sense of control and moderated by social support. Data from 365 pregnant Aboriginal Australian women were used to evaluate tooth brushing behaviour, sociodemographic factors, psychosocial factors, general health, risk behaviours and racism exposure. Bivariate associations were explored and hierarchical logistic regression models estimated odds ratios (OR) and 95% confidence intervals (CI) for tooth brushing. Perceived stress and sense of control were examined as mediators of the association between self-reported racism and tooth brushing using binary mediation with bootstrapping. High levels of self-reported racism persisted as a risk indicator for tooth brushing (OR 0.51, 95%CI 0.27,0.98) after controlling for significant covariates. Perceived stress mediated the relationship between self-reported racism and tooth brushing: the direct effect of racism on tooth brushing was attenuated, and the indirect effect on tooth brushing was significant (beta coefficient -0.09; bias-corrected 95%CI -0.166,-0.028; 48.1% of effect mediated). Sense of control was insignificant as a mediator of the relationship between racism and tooth brushing. High levels of self-reported racism were associated with non-optimal tooth brushing behaviours, and perceived stress mediated this association among this sample of pregnant Aboriginal women.. Limitations and implications are discussed.

Frequency of daily tooth brushing: predictors of change in 9- to 11-year old US children

PubMed Central

Polk, D.E.; Geng, M.; Levy, S.; Koerber, A.; Flay, B.R.

2014-01-01

Objective To investigate whether an increase in daily tooth brushing frequency in children was predicted by either a) having a strong intention to brush twice a day or b) their parents receiving information about their new caries experience. Basic Research Design Secondary data analyses were conducted on two waves of data from the Aban Aya Youth Project and the Iowa Fluoride Study. Participants The Aban Aya study included 576 10- and 11-year olds from Chicago, Illinois. The Iowa Fluoride Study included a convenience sample of 709 babies born in Iowa. The present study includes those children at age 9. Main Outcome Measures In both studies, reported daily tooth brushing frequency was assessed twice six months apart. Results In the Aban Aya data, compared with children with a weak intention at wave 1 to brush twice a day, children with a strong intention to brush twice a day were more likely to increase their brushing frequency by wave 2, OR 7.0, 95%CI 1.5,32.9. In the Iowa Fluoride Study, compared with children who did not have new caries at wave 1, children who had new caries experience were less likely to increase their brushing frequency by wave 2, OR 0.4, 95%CI 0.2,0.9. Conclusions Strengthening intention to brush twice a day might increase children’s brushing frequency. However, simply providing parents with information about new caries probably will not. Future studies should assess tooth brushing frequency, habit strength, intention, and situational cues at closely-spaced waves. PMID:25300146

Three-dimensional telomere architecture of esophageal squamous cell carcinoma: comparison of tumor and normal epithelial cells.

PubMed

Sunpaweravong, S; Sunpaweravong, P; Sathitruangsak, C; Mai, S

2016-05-01

Telomeres are repetitive nucleotide sequences (TTAGGG)n located at the ends of chromosomes that function to preserve chromosomal integrity and prevent terminal end-to-end fusions. Telomere loss or dysfunction results in breakage-bridge-fusion cycles, aneuploidy, gene amplification and chromosomal rearrangements, which can lead to genomic instability and promote carcinogenesis. Evaluating the hypothesis that changes in telomeres contribute to the development of esophageal squamous cell carcinoma (ESCC) and to determine whether there are differences between young and old patients, we compared the three-dimensional (3D) nuclear telomere architecture in ESCC tumor cells with that of normal epithelial cells obtained from the same patient. Patients were equally divided by age into two groups, one comprising those less than 45 years of age and the other consisting of those over 80 years of age. Tumor and normal epithelial cells located at least 10 cm from the border of the tumor were biopsied in ESCC patients. Hematoxylin and eosin staining was performed for each sample to confirm and identify the cancer and normal epithelial cells. This study was based on quantitative 3D fluorescence in situ hybridization (Q-FISH), 3D imaging and 3D analysis of paraffin-embedded slides. The 3D telomere architecture data were computer analyzed using 100 nuclei per slide. The following were the main parameters compared: the number of signals (number of telomeres), signal intensity (telomere length), number of telomere aggregates, and nuclear volume. Tumor and normal epithelial samples from 16 patients were compared. The normal epithelial cells had more telomere signals and higher intensities than the tumor cells, with P-values of P < 0.0001 and P = 0.0078, respectively. There were no statistically significant differences in the numbers of telomere aggregates or the nuclear volumes between the tumor and normal epithelial cells. Secondary analyses examined the effects of age on 3D telomere architecture and found no statistically significant differences in any parameter tested between the young and old patients in either the tumor or epithelial cells. The 3D nuclear telomeric signature was able to detect differences in telomere architecture between the ESCC and normal epithelial tissues. However, there were no differences observed between the young and old patients. © 2015 International Society for Diseases of the Esophagus.

Characterization of Small DC Brushed and Brushless Motors

DTIC Science & Technology

2013-03-01

8 Figure 6. Torque vs. RPM for a SS7 -1.7-1 brushed motor...9 Figure 7. Efficiency vs. motor power output for SS7 -1.7-1 brushed motor...10 Figure 8. Efficiency vs. RPM for SS7 -1.7-1 brushed motor. ........................................................10 Figure 9

75 FR 21347 - Natural Bristle Paint Brushes From China

Federal Register 2010, 2011, 2012, 2013, 2014

2010-04-23

... Brushes From China AGENCY: United States International Trade Commission. ACTION: Scheduling of a full five-year review concerning the antidumping duty order on natural bristle paint brushes from China. SUMMARY... antidumping duty order on natural bristle paint brushes from China would be likely to lead to continuation or...

The Human Airway Epithelial Basal Cell Transcriptome

PubMed Central

Wang, Rui; Zwick, Rachel K.; Ferris, Barbara; Witover, Bradley; Salit, Jacqueline; Crystal, Ronald G.

2011-01-01

Background The human airway epithelium consists of 4 major cell types: ciliated, secretory, columnar and basal cells. During natural turnover and in response to injury, the airway basal cells function as stem/progenitor cells for the other airway cell types. The objective of this study is to better understand human airway epithelial basal cell biology by defining the gene expression signature of this cell population. Methodology/Principal Findings Bronchial brushing was used to obtain airway epithelium from healthy nonsmokers. Microarrays were used to assess the transcriptome of basal cells purified from the airway epithelium in comparison to the transcriptome of the differentiated airway epithelium. This analysis identified the “human airway basal cell signature” as 1,161 unique genes with >5-fold higher expression level in basal cells compared to differentiated epithelium. The basal cell signature was suppressed when the basal cells differentiated into a ciliated airway epithelium in vitro. The basal cell signature displayed overlap with genes expressed in basal-like cells from other human tissues and with that of murine airway basal cells. Consistent with self-modulation as well as signaling to other airway cell types, the human airway basal cell signature was characterized by genes encoding extracellular matrix components, growth factors and growth factor receptors, including genes related to the EGF and VEGF pathways. Interestingly, while the basal cell signature overlaps that of basal-like cells of other organs, the human airway basal cell signature has features not previously associated with this cell type, including a unique pattern of genes encoding extracellular matrix components, G protein-coupled receptors, neuroactive ligands and receptors, and ion channels. Conclusion/Significance The human airway epithelial basal cell signature identified in the present study provides novel insights into the molecular phenotype and biology of the stem/progenitor cells of the human airway epithelium. PMID:21572528

Preferential Generation of 15-HETE-PE Induced by IL-13 Regulates Goblet Cell Differentiation in Human Airway Epithelial Cells.

PubMed

Zhao, Jinming; Minami, Yoshinori; Etling, Emily; Coleman, John M; Lauder, Sarah N; Tyrrell, Victoria; Aldrovandi, Maceler; O'Donnell, Valerie; Claesson, Hans-Erik; Kagan, Valerian; Wenzel, Sally

2017-12-01

Type 2-associated goblet cell hyperplasia and mucus hypersecretion are well known features of asthma. 15-Lipoxygenase-1 (15LO1) is induced by the type 2 cytokine IL-13 in human airway epithelial cells (HAECs) in vitro and is increased in fresh asthmatic HAECs ex vivo. 15LO1 generates a variety of products, including 15-hydroxyeicosatetraenoic acid (15-HETE), 15-HETE-phosphatidylethanolamine (15-HETE-PE), and 13-hydroxyoctadecadienoic acid (13-HODE). In this study, we investigated the 15LO1 metabolite profile at baseline and after IL-13 treatment, as well as its influence on goblet cell differentiation in HAECs. Primary HAECs obtained from bronchial brushings of asthmatic and healthy subjects were cultured under air-liquid interface culture supplemented with arachidonic acid and linoleic acid (10 μM each) and exposed to IL-13 for 7 days. Short interfering RNA transfection and 15LO1 inhibition were applied to suppress 15LO1 expression and activity. IL-13 stimulation induced expression of 15LO1 and preferentially generated 15-HETE-PE in vitro, both of which persisted after removal of IL-13. 15LO1 inhibition (by short interfering RNA and chemical inhibitor) decreased IL-13-induced forkhead box protein A3 (FOXA3) expression and enhanced FOXA2 expression. These changes were associated with reductions in both mucin 5AC and periostin. Exogenous 15-HETE-PE stimulation (alone) recapitulated IL-13-induced FOXA3, mucin 5AC, and periostin expression. The results of this study confirm the central importance of 15LO1 and its primary product, 15-HETE-PE, for epithelial cell remodeling in HAECs.

Application of cytology and molecular biology in diagnosing premalignant or malignant oral lesions

PubMed Central

Mehrotra, Ravi; Gupta, Anurag; Singh, Mamta; Ibrahim, Rahela

2006-01-01

Early detection of a premalignant or cancerous oral lesion promises to improve the survival and the morbidity of patients suffering from these conditions. Cytological study of oral cells is a non-aggressive technique that is well accepted by the patient, and is therefore an attractive option for the early diagnosis of oral cancer, including epithelial atypia and squamous cell carcinoma. However its usage has been limited so far due to poor sensitivity and specificity in diagnosing oral malignancies. Lately it has re-emerged due to improved methods and it's application in oral precancer and cancer as a diagnostic and predictive method as well as for monitoring patients. Newer diagnostic techniques such as "brush biopsy" and molecular studies have been developed. Recent advances in cytological techniques and novel aspects of applications of scraped or exfoliative cytology for detecting these lesions and predicting their progression or recurrence are reviewed here. PMID:16556320

46 CFR 108.187 - Ventilation for brush type electric motors in classified spaces.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 46 Shipping 4 2010-10-01 2010-10-01 false Ventilation for brush type electric motors in classified... Ventilation for brush type electric motors in classified spaces. Ventilation for brush type electric motors in... Electrical Equipment in Hazardous Locations”, except audible and visual alarms may be used if shutting down...

77 FR 77037 - Procurement List; Additions

Federal Register 2010, 2011, 2012, 2013, 2014

2012-12-31

...-Compatible, 2'' x 8'' NSN: 7920-00-NIB-0545--Scrub Brush, Wire, Knuckle Guard, Long Handle, Ergonomic, 6'' x 1 1/8'', w/built-in scraper NSN: 7920-00-NIB-0547--Scrub Brush, Wire, Stainless, Ergonomic, 5'' NSN: 7920-00-NIB-0558--Scrub Brush, Wire, Black Tempered, Ergonomic, 5'' NSN: 7920-00-NIB-0563--Wire Brush...

DOE Office of Scientific and Technical Information (OSTI.GOV)

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

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

Facilitators and barriers to twice daily tooth brushing among children with special health care needs

PubMed Central

Campanaro, Maria; Huebner, Colleen E.; Davis, Beth Ellen

2015-01-01

Purpose The oral hygiene routines of preschool-age children with special health care needs (CSHCN) were examined to identify caregiver behaviors and beliefs associated with twice daily tooth brushing. Methods Ninety caregivers of CSHCN, ages 23 to 62 months, were interviewed to determine supports or barriers to tooth brushing. Results Ninety-eight percent of caregivers had begun brushing their child’s teeth and half reported brushing twice daily. Caregivers’ brushing skills and the availability of child-friendly supplies were associated with twice daily versus less frequent brushing (p = .02). Conclusions This study adds insight into the challenges of establishing daily oral health care for children who must rely on others for their care. The facilitators and barriers to tooth brushing by caregivers of CSHCN are similar to those noted previously among parents of typically developing children. Efforts to improve all caregivers’ oral hygiene skills are needed. For caregivers of CSHCN, oral health teaching opportunities may exist among professionals who provide ongoing medical care, special services and therapies. PMID:24252060

Facilitators and barriers to twice daily tooth brushing among children with special health care needs.

PubMed

Campanaro, Maria; Huebner, Colleen E; Davis, Beth Ellen

2014-01-01

The oral hygiene routines of preschool-age children with special health care needs (CSHCN) were examined to identify caregiver behaviors and beliefs associated with twice daily tooth brushing. Ninety caregivers of CSHCN, ages 23 to 62 months, were interviewed to determine supports or barriers to tooth brushing. Ninety-eight percent of caregivers had begun brushing their child's teeth and half reported brushing twice daily. Caregivers' brushing skills and the availability of child-friendly supplies were associated with twice daily versus less frequent brushing (p = .02). This study adds insight into the challenges of establishing daily oral health care for children who must rely on others for their care. The facilitators and barriers to tooth brushing by caregivers of CSHCN are similar to those noted previously among parents of typically developing children. Efforts to improve all caregivers' oral hygiene skills are needed. For caregivers of CSHCN, oral health teaching opportunities may exist among professionals who provide ongoing medical care, special services and therapies. © 2014 Special Care Dentistry Association and Wiley Periodicals, Inc.

TEMPORAL-SPATIAL ANALYSIS OF U.S.- MEXICO BORDER ENVIRONMENTAL FINE AND COARSE PM AIR SAMPLE EXTRACT ACTIVITY IN HUMAN BRONCHIAL EPITHELIAL CELLS

PubMed Central

Lauer, Fredine T.; Mitchell, Leah A.; Bedrick, Edward; McDonald, Jacob D.; Lee, Wen-Yee; Li, Wen-Whai; Olvera, Hector; Amaya, Maria A.; Berwick, Marianne; Gonzales, Melissa; Currey, Robert; Pingitore, Nicholas E.; Burchiel, Scott W.

2009-01-01

Particulate matter less than 10 μm (PM10) has been shown to be associated with aggravation of asthma and respiratory and cardiopulmonary morbidity. There is also great interest in the potential health effects of PM 2.5. Particulate matter (PM) varies in composition both spatially and temporally depending on the source, location and seasonal condition. El Paso County which lies in the Paso del Norte airshed is a unique location to study ambient air pollution due to three major points: the geological land formation, the relatively large population and the various sources of PM. In this study, dichotomous filters were collected from various sites in El Paso County every seven days for a period of one year. The sampling sites were both distant and near border crossings, which are near heavily populated areas with high traffic volume. Fine (PM2.5) and Coarse (PM10-2.5) PM filter samples were extracted using dichloromethane and were assessed for biologic activity and polycyclic aromatic (PAH) content. Three sets of marker genes human BEAS2B bronchial epithelial cells were utilized to assess the effects of airborne PAHs on biologic activities associated with specific biological pathways associated with airway diseases. These pathways included in inflammatory cytokine production (IL-6, IL-8), oxidative stress (HMOX-1, NQO-1, ALDH3A1, AKR1C1), and aryl hydrocarbon receptor (AhR)-dependent signaling (CYP1A1). Results demonstrated interesting temporal and spatial patterns of gene induction for all pathways, particularly those associated with oxidative stress, and significant differences in the PAHs detected in the PM10-2.5 and PM 2.5 fractions. Temporally, the greatest effects on gene induction were observed in winter months, which appeared to correlate with inversions that are common in the air basin. Spatially, the greatest gene expression increases were seen in extracts collected from the central most areas of El Paso which are also closest to highways and border crossings. PMID:19410595

Fine structure and development of the collar enamel in gars, Lepisosteus oculatus, Actinopterygii

NASA Astrophysics Data System (ADS)

Sasagawa, Ichiro; Ishiyama, Mikio; Yokosuka, Hiroyuki; Mikami, Masato

2008-06-01

The fine structure of collar enamel and the cells constituting the enamel organ during amelogenesis in Lepisosteus oculatus was observed by light, scanning electron and transmission electron microscopy. In the enamel, slender crystals were arranged perpendicular to the surface and the stripes that were parallel to the surface were observed, suggesting that the enamel in Lepisosteus shares common morphological features with that in sarcopterygian fish and amphibians. Ameloblasts containing developed Golgi apparatus, rough endoplasmic reticulum (rER) and secretory granules were found in the secretory stage. In the maturation stage, a ruffled border was not seen at the distal end of the ameloblasts, while many mitochondria and lysosome-like granules were obvious in the distal cytoplasm. The enamel organ consisted of the outer dental epithelial cells, stratum reticulum cells and ameloblasts, but there was no stratum intermedium. It is likely that the ameloblasts have less absorptive function in comparison with the inner dental epithelial cells facing cap enameloid.

Effectiveness of Plaque Control with Novel Pediatric Oral Hygiene Need Station (Modified Oral Irrigation Device) as Compared with Manual Brushing and Flossing: Randomized Controlled Pilot Trial

PubMed Central

Murthy, Prashanth Sadashiva; Shaik, Naseemoon; Deshmukh, Seema; Girish, M. S.

2018-01-01

Background: Establishing good hygiene habits are valuable for present and future oral health. Below 6 years, tooth brushing should be performed by parents, as increasing dexterity and cognition may permit supervised brushing until the child is capable of independent brushing. Aim and Objectives: The aim of the present study was to evaluate the effectiveness of modified oral irrigation device in children in terms of plaque control and to compare the effectiveness of plaque control with manual brushing with the modified oral irrigation device in children. Materials and Methods: A randomized clinical trial was performed on 12 subjects who were allocated to the two study groups. After obtaining the consent, the control group was instructed tooth brushing with regular pediatric commercially available toothbrush and the intervention group with modified oral irrigation device. Plaque scores in both groups were assessed pre- and post-brushing using modified navy plaque index. Results: The data were subjected to Descriptive statistics and Paired t-test using SPSS version 22. Intragroup comparison of mean difference of plaque score in control group and intervention group pre- and post-brushing was statistically significant. Intergroup comparison of manual brushing group with modified oral irrigation group shows P < 0.05 was statistically significant. Conclusion: Within the limitation of the present study, it has been found novel pediatric oral hygiene need Station is more effective than manual brushing since it combined the effect of brushing, flossing (water floss), and rinsing in children simultaneously and at the same time did not demand any special motor skill. PMID:29875555

Oral hygiene practices and risk of oral leukoplakia.

PubMed

Macigo, F G; Gathece, L W; Guthua, S W; Njeru, E K; Wagaiyu, E G; Mulli, T K

2006-04-01

To determine the influence of oral hygiene habits and practices on the risk of developing oral leukoplakia. Case control study. Githongo sublocation in Meru District. Eighty five cases and 141 controls identified in a house-to-house screening. The relative risk (RR) of oral leukoplakia increased gradually across the various brushing frequencies from the reference RR of 1.0 in those who brushed three times a day, to 7.6 in the "don't brush" group. The trend of increase was statistically significant (X2 for Trend : p = 0.001). The use of chewing stick as compared to conventional tooth brush had no significant influence on RR of oral leukoplakia. Non-users of toothpastes had a significantly higher risk of oral leukoplakia than users (RR = 1.8; 95% confidence levels (CI) = 1.4-2.5). Among tobacco smokers, the RR increased from 4.6 in those who brushed to 7.3 in those who did not brush. Among non-smokers, the RR of oral leukoplakia in those who did not brush (1.8) compared to those who brushed was also statistically significant (95% CL = 1.6-3.8). Failure to brush teeth and none use of toothpastes are significantly associated with the development of oral leukoplakia, while the choice of brushing tools between conventional toothbrush and chewing stick is not. In addition, failure to brush teeth appeared to potentiate the effect of smoking tobacco in the development of oral leukoplakia. Oral health education, instruction and motivation for the improvement of oral hygiene habits and practices; and therefore oral hygiene status, should be among the strategies used in oral leukoplakia preventive and control programmes.

Interdental cleaning and gingival injury potential of interdental toothbrushes.

PubMed

Vogel, Marc; Sener, Beatrice; Roos, M; Attin, Thomas; Schmidlin, Patrick R

2014-01-01

The effective cleaning of interdental spaces using toothbrushes is a challenge. The aim of the present in vitro study was to evaluate on the one hand the interdental cleaning efficiency and on the other hand the gingival injury potential of an electric single-headed sonic toothbrush (Water-pik) and two single-tufted manual toothbrushes (Curaprox 1009; Lactona Interdental Brush). Brushes were evaluated using a brushing device. Test dental casts (maxillary sextants) consisting of black teeth coated with white paint were brushed using standardized horizontal movements. Thereafter, black (i.e. cleaned) areas were measured planimetrically. The soft tissue injury potential was evaluated using front segments of porcine mandibles. In the same brushing device, test brushes were moved over the gingiva. Before and after each treatment, the porcine mucosa was stained with a plaque disclosing agent to visualize injured areas, which could then be measured planimetrically as well. These evaluations were each made after 15, 30, 60, and 120 seconds of brushing. The statistical analysis was performed using non-parametric Mann-Whitney tests, and the level of significance was set at 5%. The best cleaning performance of 46% across all interdental spaces assessed was found with the electric sonic toothbrush (Waterpik), while the performances of the manual brushes from Lactona and Curaprox were 14.8% and 5%, respectively. At each point of evaluation, the gingiva was injured most markedly by the powered sonic toothbrush (Waterpik), followed by the manual Curaprox brush. The smallest damage of the porcine gingiva was produced by the manual Lactona brush. When comparing the manual toothbrushes, the Lactona product revealed a better cleaning performance combined with a smaller injury potential than the Curaprox brush. Thus, the prophylactic goal to achieve high degrees of cleaning while producing minimal damage is important and should have priority when evaluating and selecting toothbrushes.

Efficacy of 3 toothbrush treatments on plaque removal in orthodontic patients assessed with digital plaque imaging: a randomized controlled trial.

PubMed

Erbe, Christina; Klukowska, Malgorzata; Tsaknaki, Iris; Timm, Hans; Grender, Julie; Wehrbein, Heinrich

2013-06-01

Good oral hygiene is a challenge for orthodontic patients because food readily becomes trapped around the brackets and under the archwires, and appliances are an obstruction to mechanical brushing. The purpose of this study was to compare plaque removal efficacy of 3 toothbrush treatments in orthodontic subjects. This was a replicate-use, single-brushing, 3-treatment, examiner-blind, randomized, 6-period crossover study with washout periods of approximately 24 hours between visits. Forty-six adolescent and young adult patients with fixed orthodontics from a university clinic in Germany were randomized, based on computer-generated randomization, to 1 of 3 treatments: (1) oscillating-rotating electric toothbrush with a specially designed orthodontic brush head (Oral-B Triumph, OD17; Procter & Gamble, Cincinnati, Ohio); (2) the same electric toothbrush handle with a regular brush head (EB25; Procter & Gamble); and (3) a regular manual toothbrush (American Dental Association, Chicago, Ill). The primary outcome was the plaque score change from baseline, which we determined using digital plaque image analysis. Forty-five subjects completed the study. The differences in mean plaque removal (95% confidence interval) between the electric toothbrush with an orthodontic brush head (6% [4.4%-7.6%]) or a regular brush head (3.8% [2.2%-5.3%]) and the manual toothbrush were significant (P <0.001). Plaque removal with the electric toothbrush with the orthodontic brush head was superior (2.2%; P = 0.007) to the regular brush head. No adverse events were seen. The electric toothbrush, with either brush head, demonstrated significantly greater plaque removal over the manual brush. The orthodontic brush head was superior to the regular head. Copyright © 2013 American Association of Orthodontists. Published by Mosby, Inc. All rights reserved.

Development and testing of a theory-based behavioural change intervention: a pilot investigation in a nursery school in a deprived area of Scotland.

PubMed

Gilinsky, A; Swanson, V; Merrett, M; Power, K; Marley, L

2012-03-01

Investigate the effect of a theory-based intervention on oral-health knowledge, attitudes and behaviours of early years staff (EYS), parents and nursery children. Qualitative research with staff and parents from eight nurseries through interviews/focus groups. An intervention was developed and piloted using pre-posttest design. Nurseries in deprived communities in Dundee, Scotland. 111 children aged 3-5 years attending nursery, including 79 parents and 8 nursery staff. Staff session targeted outcome expectancies, subjective norms and self-efficacy for tooth-brushing in nursery using information provision, modelling and goal-setting, followed by a three-week intervention. Parent-child dyads received a leaflet with instructions for goal-setting, planning and monitoring home brushing. Tooth-brushing self-monitoring materials (e.g. two-minute timer, diaries) were used and certificates provided in the nursery as rewards. EYS knowledge, attitudes and behaviours were assessed before and after the intervention using self-report questionnaires. Parents completed interviews assessing beliefs about tooth-brushing and their children's tooth-brushing behaviour at baseline and post-intervention follow-up. Significant improvements in staff knowledge, but not attitudes, self-efficacy, or nursery tooth-brushing were reported. Parent-child dyads completing the intervention were not more likely to report their child carried out twice-daily tooth-brushing at home. The intervention did not improve parents' intentions to brush their child's teeth twice a day or beliefs about the ease of twice-daily tooth-brushing. Only past behaviour significantly predicted posttest brushing. Parents who found brushing easier at baseline were more likely to complete the intervention. Recommendations are made regarding implementing psychological theory and methods into oral-health interventions.

Polymer brush coatings for DNA: fundamental polymer physics and nanofabrication applications

NASA Astrophysics Data System (ADS)

de Vries, Renko

Recombinant DNA technology allows for the production of precisely defined self-assembling protein-based polymers. So far, the major applications for such protein-based polymers have been self-assembling hydrogels and micellar structures with biomedical application. Inspired by minimal models for the self-ssembly of rod-shaped viruses such as the tobacco mosaic virus, I have developed protein-polymers that co-assemble with DNA into rod-shaped virus-like particles, and protein-polymers that provide brush coatings around single DNA molecules. In this presentation I will focus on the latter, showing that on the one hand brush coated DNA is a rich model system for exploring the physics of bottle-brush polymers, while on the other hand brush coatings of DNA can also play an important practical role in nanofabrication. A key problem in the physics of bottle-brush polymers that I will address is the scale-dependence of bottle-brush elasticity. For long-wavelength thermal deformations probed by AFM imaging I will demonstrate that there is significant stiffening due to the brush coating, while for short wavelength thermal deformations probed by force spectroscopy, we find that stiffening due to the brush coating disappears completely. DNA brush coatings can also play an important practical role in nanofabrication by acting as a compatibilizer between chemically different building blocks. I will explore the example of DNA origami in combination with gold nanoparticles: while Mg2+ ions and high concentrations of monovalent salts are crucial for the stability of DNA origami, such solution conditions are typically incompatible with the colloidal stability of gold nanoparticles.I will show how DNA brush coatings can dramatically enhance the yield of formation of isolated DNA-gold nanoparticle composite nanostructures.

Tooth brushing among 11- to 15-year-olds in Denmark: combined effect of social class and migration status.

PubMed

Bast, L S; Nordahl, H; Christensen, L B; Holstein, B E

2015-03-01

Regular tooth brushing in adolescence predicts stable tooth brushing habits later in life. Differences in tooth brushing habits by ethnic background and socioeconomic position have been suggested. We investigated migration status and social class in relation to infrequent tooth brushing both separately and combined. The study population was 11-15 year-olds chosen from a clustered random sample of schools. Univariate and multivariate logistic regression analyses estimated the separate and combined effects of migration status and social class on less than twice daily tooth brushing. 10,607 respondents: a response rate of 88.3%. Boys of lower social class had higher odds ratio (OR) of infrequent tooth brushing than girls: 1.98 (95% confidence interval 1.62-2.41) vs 1.80 (1.53-2.24). Immigrants and descendants had higher odds compared to adolescents of Danish origin: immigrant boys OR 1.39 (1.05-1.89), girls OR 1.92 (1.47-2.50); descendant boys OR 2.53 (1.97-3.27), girls OR 2.56 (2.02-3.35). Analyses of the combined effect of social class and migration status showed that the social gradient in tooth brushing habits observed among ethnic Danes cannot be found among groups of immigrants and descendants. The study shows that both non-Danish origin and low social class increases the risk of infrequent tooth brushing among school-aged children. The study calls for in depth analyses of the processes which influence young people's tooth brushing habits. Further, there is a need to strengthen the promotion of appropriate tooth brushing habits of minority and low social class youths.

Tooth-brushing behaviour in 6-12 year olds.

PubMed

Sandström, Anna; Cressey, Janet; Stecksén-Blicks, Christina

2011-01-01

A common clinical finding is that many schoolchildren display a nonacceptable oral hygiene. To evaluate the tooth-brushing behaviour in children aged 6-12 years. The study used a cross-sectional descriptive design. Children aged 6, 8, 10, and 12 years in an elementary school in a middle class area in Umeå, a city in northern Sweden, were invited and 82 (82%) consented. Visible plaque on buccal surfaces of incisors and canines was recorded from photographs of the participant's teeth before and after brushing using the scores of the Green and Vermillion Oral Hygiene Index. Brushing technique was recorded with a video camera. A questionnaire was used to collect data about oral hygiene habits at home. The ratio between the sum of plaque scores after and before brushing was statistically significantly higher in the 6-year-old group compared with the 10-year olds, (P < 0.05). There was a negative correlation between time spent for brushing and the ratio between the sum of plaque scores after and before brushing (r = -0.31, P < 0.01). The lowest correlation was displayed in the youngest age group (r = 0.07, P > 0.05). Six-year olds spent statistically significantly less time for brushing than older children (P < 0.05). Plaque removal from buccal surfaces from brushing was poor and averaged 19% for 6-year olds and 30% for older children. The results of brushing for children aged 8-12 years could benefit from increasing tooth-brushing time. Children could be given an increasing responsibility from 7 to 8 year of age but parental help is motivated up to 10 years of age. © 2010 The Authors. International Journal of Paediatric Dentistry © 2010 BSPD, IAPD and Blackwell Publishing Ltd.

A comparative evaluation of the in vitro penetration performance of the improved Crest complete toothbrush versus the Current Crest complete toothbrush, the Colgate Precision toothbrush and the Oral-B P40 toothbrush.

PubMed

Volpenhein, D W; Handel, S E; Hughes, T J; Wild, J

1996-01-01

Removal of plaque and debris from interproximal surfaces during toothbrushing has generally been difficult to achieve, in large part because traditional flat-bristled toothbrushes do not offer good interproximal penetration. As a result, a number of varying bristle designs have been developed, with the rippled-design brush shown to be particularly effective at removing interproximal plaque. Recently, an existing brush, the original Crest Complete, was modified to offer a more deeply rippled version. This study evaluated the interproximal penetration of four bristle designs: rippled pattern (original Crest Complete), deeper rippled pattern (improved Crest Complete), multi-level (Colgate Precision), and flat-tufted (Oral-B P40). The study used a previously reported in vitro model for determining interproximal penetration of manual toothbrushes (J Clin Dent 5:27-33, 1994). In order to effectively mimic the in-use characteristics of toothbrushing, this model is based on analysis of videotaped consumer brushing habits, tooth morphology, and in vivo plaque tenacity characteristics and uses the three most predominantly used brushing techniques (circular, up-and-down, and back-and-forth, with the brush held at both 45 and 90 degrees to the tooth surface). In addition, the model's brush stroke length, brush force, and brush speed are likewise based on analysis of consumer brushing patterns. The results of the study indicate that the new Crest Complete with deeper rippled bristles provided significantly superior (p < or = 0.05) interproximal penetration than the Colgate Precision and Oral-B brushes overall and for three of the four brush strokes tested. In addition, the new Crest Complete was found to provide significantly superior interproximal penetration to the original Crest Complete overall and in circular and up-and-down strokes, and the original Crest Complete provided superior overall interproximal penetration to the Colgate and Oral-B brushes.

A hybrid floating brush seal (HFBS) for improved sealing and wear performance in turbomachinery applications

NASA Astrophysics Data System (ADS)

Lattime, Scott Byran

A conceptually new type of seal has been developed for gas turbine applications which dramatically reduces wear and leakage associated with current labyrinth and brush seal technologies. The Hybrid Floating Brush Seal (HFBS) combines brush seal and film riding face seal technologies to create a hybrid seal that allows both axial and radial excursions of the sealed shaft, while simultaneously eliminating interface surface speeds (friction and heat) between the rotor and the brush material that characterize standard brush seal technology. A simple test rig was designed to evaluate feasibility of the HFBS under relatively low pressures and rotational speeds (50psig, 5krpm). A second test stand was created to study the effects of centrifugal force on bristle deflection. A third test facility was constructed for prototype development and extensive room temperature testing at moderate pressures and fairly high rotational speeds (100psig, 40krpm). This test rig also allowed the evaluation of the HFBS during axial movement of a rotating shaft. An analytical model to predict the effects of centrifugal force on the bristles of a rotating brush seal was developed. Room temperature analysis of the HFBS proved successful for relatively high operating rotational velocities at moderate pressures with very acceptable leakage rates for gas turbine engines. Brush seals were able to track rotor speeds up to 24krpm while maintaining sealing integrity. The HFBS's ability to function under axial shaft displacement and synchronous dynamic radial loading was also proven successful. Hydrodynamic performance of the face seal was proven to provide adequate stiffness and load carrying capacity to keep the brush seal from contacting the face seal at pressure drops across the brush of up to 100psi. Leakage performance over standard brush seal and labyrinth technology was quite dramatic. The HFBS showed its sealing advantage using much higher radial interference between the rotor and the bristle bore over standard brush seal designs. Experimental results of the HFBS showed leakage reductions of 6 to 7 times that of a standard brush seal at the same operating pressure ratios and rotational speed and an order of magnitude less than numerical predictions of a standard labyrinth seal. (Abstract shortened by UMI.)

Comparison between observed children's tooth brushing habits and those reported by mothers.

PubMed

Martins, Carolina C; Oliveira, Maria J; Pordeus, Isabela A; Paiva, Saul M

2011-09-03

Information bias can occur in epidemiological studies and compromise scientific outcomes, especially when evaluating information given by a patient regarding their own health. The oral habits of children reported by their mothers are commonly used to evaluate tooth brushing practices and to estimate fluoride intake by children. The aim of the present study was to compare observed tooth-brushing habits of young children using fluoridated toothpaste with those reported by mothers. A sample of 201 mothers and their children (aged 24-48 months) from Montes Claros, Brazil, took part in a cross-sectional study. At day-care centres, the mothers answered a self-administered questionnaire on their child's tooth-brushing habits. The structured questionnaire had six items with two to three possible answers. An appointment was then made with each mother/child pair at day-care centres. The participants were asked to demonstrate the tooth-brushing practice as usually performed at home. A trained examiner observed and documented the procedure. Observed tooth brushing and that reported by mothers were compared for overall agreement using Cohen's Kappa coefficient and the McNemar test. Cohen's Kappa values comparing mothers' reports and tooth brushing observed by the examiner ranged from poor-to-good (0.00-0.75). There were statistically significant differences between observed tooth brushing habits and those reported by mothers (p < 0.001). When observed by the examiner, the frequencies of dentifrice dispersed on all bristles (35.9%), children who brushed their teeth alone (33.8%) and those who did not rinse their mouths during brushing (42.0%) were higher than those reported by the mothers (12.1%, 18.9% and 6.5%, respectively; p < 0.001). In general, there was low agreement between observed tooth brushing and mothers' reports. Moreover, the different methods of estimation resulted in differences in the frequencies of tooth brushing habits, indicative of reporting bias. Data regarding children's tooth-brushing habits as reported by mothers should be considered with caution in epidemiological surveys on fluoridated dentifrice use and the risk of dental fluorosis.

Comparison between observed children's tooth brushing habits and those reported by mothers

PubMed Central

2011-01-01

Background Information bias can occur in epidemiological studies and compromise scientific outcomes, especially when evaluating information given by a patient regarding their own health. The oral habits of children reported by their mothers are commonly used to evaluate tooth brushing practices and to estimate fluoride intake by children. The aim of the present study was to compare observed tooth-brushing habits of young children using fluoridated toothpaste with those reported by mothers. Methods A sample of 201 mothers and their children (aged 24-48 months) from Montes Claros, Brazil, took part in a cross-sectional study. At day-care centres, the mothers answered a self-administered questionnaire on their child's tooth-brushing habits. The structured questionnaire had six items with two to three possible answers. An appointment was then made with each mother/child pair at day-care centres. The participants were asked to demonstrate the tooth-brushing practice as usually performed at home. A trained examiner observed and documented the procedure. Observed tooth brushing and that reported by mothers were compared for overall agreement using Cohen's Kappa coefficient and the McNemar test. Results Cohen's Kappa values comparing mothers' reports and tooth brushing observed by the examiner ranged from poor-to-good (0.00-0.75). There were statistically significant differences between observed tooth brushing habits and those reported by mothers (p < 0.001). When observed by the examiner, the frequencies of dentifrice dispersed on all bristles (35.9%), children who brushed their teeth alone (33.8%) and those who did not rinse their mouths during brushing (42.0%) were higher than those reported by the mothers (12.1%, 18.9% and 6.5%, respectively; p < 0.001). Conclusions In general, there was low agreement between observed tooth brushing and mothers' reports. Moreover, the different methods of estimation resulted in differences in the frequencies of tooth brushing habits, indicative of reporting bias. Data regarding children's tooth-brushing habits as reported by mothers should be considered with caution in epidemiological surveys on fluoridated dentifrice use and the risk of dental fluorosis. PMID:21888664

Ca2+-dependent localization of integrin-linked kinase to cell junctions in differentiating keratinocytes.

PubMed

Vespa, Alisa; Darmon, Alison J; Turner, Christopher E; D'Souza, Sudhir J A; Dagnino, Lina

2003-03-28

Integrin complexes are necessary for proper proliferation and differentiation of epidermal keratinocytes. Differentiation of these cells is accompanied by down-regulation of integrins and focal adhesions as well as formation of intercellular adherens junctions through E-cadherin homodimerization. A central component of integrin adhesion complexes is integrin-linked kinase (ILK), which can induce loss of E-cadherin expression and epithelial-mesenchymal transformation when ectopically expressed in intestinal and mammary epithelia. In cultured primary mouse keratinocytes, we find that ILK protein levels are independent of integrin expression and signaling, since they remain constant during Ca(2+)-induced differentiation. In contrast, keratinocyte differentiation is accompanied by marked reduction in kinase activity in ILK immunoprecipitates and altered ILK subcellular distribution. Specifically, ILK distributes in close apposition to actin fibers along intercellular junctions in differentiated but not in undifferentiated keratinocytes. ILK localization to cell-cell borders occurs independently of integrin signaling and requires Ca(2+) as well as an intact actin cytoskeleton. Further, and in contrast to what is observed in other epithelial cells, ILK overexpression in differentiated keratinocytes does not promote E-cadherin down-regulation and epithelial-mesenchymal transition. Thus, novel tissue-specific mechanisms control the formation of ILK complexes associated with cell-cell junctions in differentiating murine epidermal keratinocytes.

46 CFR 108.187 - Ventilation for brush type electric motors in classified spaces.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 46 Shipping 4 2011-10-01 2011-10-01 false Ventilation for brush type electric motors in classified... Ventilation for brush type electric motors in classified spaces. Ventilation for brush type electric motors in classified locations must meet N.F.P.A. 496-1974 “Standard for Purged and Pressurized Enclosures for...

Turbine with radial acting seal

DOEpatents

Eng, Darryl S; Ebert, Todd A

2016-11-22

A floating brush seal in a rim cavity of a turbine in a gas turbine engine, where the floating brush seal includes a seal holder in which the floating brush seal floats, and a expandable seal that fits within two radial extending seal slots that maintains a seal with radial displacement of the floating brush seal and the seal holder.

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.

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.

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.

Method for making a high current fiber brush collector

NASA Astrophysics Data System (ADS)

Scuro, S. J.

1986-05-01

An axial-type homopolar motor having high density, high current fiber brush collectors affording efficient, low contact resistance and low operating temperatures is discussed. The collectors include a ring of concentric row of brushes in equally spaced beveled holes soldered in place using a fixture for heating the ring to just below the solder melting point at a soldering iron for the local application of additional heat at each brush. Prior to soldering, an oxide film is formed on the surfaces of the brushes and ring, and the bevels are burnished to form a wetting surface. Flux applied with the solder at each bevel removes to an effective soldering depth the oxide film on the brushes and the holes.

KERATOACANTHOMA OF THE INFERIOR LIP: REVIEW AND REPORT OF CASE WITH SPONTANEOUS REGRESSION

PubMed Central

Ramos, Lara Maria A.; Cardoso, Sérgio V.; Loyola, Adriano M.; Rocha, Marcus A.; Durighetto, Antônio Francisco

2009-01-01

Keratoacanthoma (KA) is a self-limited benign epithelial proliferative lesion that eventually presents with very similar clinical features to squamous cell carcinoma. Many KA appear in the vermilion border of the lips and therefore dental professionals must be familiar of the disease. This article reports the case of a 40-year-old female patient presenting with an exophytic ulcerative tumor in her lower lip that resolved after incisional biopsy. Photographic documentation of the case is presented and topics that are relevant to the clinical management of the disease are addressed. PMID:19466263

A loss of profilin-1 in late-stage oral squamous cell carcinoma.

PubMed

Adami, Guy R; O'Callaghan, Thomas N; Kolokythas, Antonia; Cabay, Robert J; Zhou, Yalu; Schwartz, Joel L

2017-08-01

The genes for PFN1 and TMSB4 are both highly expressed in oral tissue and both encode actin monomer binding proteins thought to play a role in cell motility and possibly other crucial parts of tumor progression. Oral brush cytology of epithelium from oral squamous cell carcinoma (OSCC) was used to measure PFN1 and TMSB4 mRNA in OSCC, while immunohistochemical analysis of tissue was used to check protein levels. High but variable expression of mRNAs encoding these two proteins was observed suggesting they may contribute to tumor characteristics in a subset of OSCCs. Both proteins were highly expressed in normal appearing basal epithelium, in the cytoplasm, and perinuclear area, while expression was minimal in upper epithelial layers. In OSCCs, expression of these proteins varied. In tumors classified as later stage, based on size and/or lymph node involvement, PFN1 levels were lower in tumor epithelium. A control gene, KRT13, showed expression in normal differentiated basal and suprabasal oral mucosa epithelial cells and as reported was lost in OSCC cells. Loss of PFN1 in tumor cells has been associated with lymph node invasion and metastasis in other tumor types, strengthening the argument that the protein has the potential to be a tumor suppressor in late-stage OSCC. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Frequency of tooth brushing and associated factors in Mexican schoolchildren six to nine years of age.

PubMed

Casanova-Rosado, J F; Vallejos-Sánchez, A A; Minaya-Sánchez, M; Medina-Solís, C E; De La Rosa-Santillana, R; Márquez-Corona, M de L; Maupomé, G

2013-01-01

To determine the prevalence of daily tooth brushing and evaluate some variables associated. A cross-sectional study was carried out in 320 schoolchildren six to nine years old in Campeche, Mexico. Information on sociodemographic and socio-economic variables, oral hygiene practices and attitudes were collected through a questionnaire. The frequency of tooth brushing was categorized as "0" = fewer than seven times/week, "1" = at least once a day. In the analysis, nonparametric tests were used. Mean age was 6.99 +/- 1.00 years, 52.5% were boys. The prevalence of daily tooth brushing was 81.6%. In bivariate analysis, the prevalence of tooth brushing was higher (p < 0.05) among the children of mothers with higher schooling (9.80 years vs 8.47 years, p < 0.05), and in younger children (84.6% in 6-7-year olds vs 71.2% in 8-9-year olds, p < 0.05). A slight, non-significant association (p < 0.10) was noted between the current frequency of tooth brushing and an earlier age when the child first started brushing with toothpaste. There were no statistically significant differences (p > 0.05) in the frequency of tooth brushing by gender or by the mother's attitude toward the oral health of her child. The prevalence of daily tooth brushing was high compared to other studies. Mother's maximum level of schooling (as an indicator of socio-economic position) was associated with higher frequency of tooth brushing. Maternal characteristics are associated with the oral health behaviour of their children.

Parental Attitudes and Tooth Brushing Habits in Preschool Children in Mangalore, Karnataka: A Cross-sectional Study.

PubMed

Pullishery, Fawaz; Shenoy Panchmal, Ganesh; Shenoy, Rekha

2013-09-01

Adoption of consistent behavioral habits in childhood takes place at home, with the parents especially the mother, being the primary model for behavior. Tooth brushing habits which is learnt during early years of life, is deeply ingrained in the child's mind and it is expected that this leads to an adaptation of good oral hygiene in their later life. To assess the tooth brushing habits of preschool children and to determine the role and amount of supervision given to them by parents. A pretested self-designed questionnaire was used to collect information from parents of 130 preschool children in Anganwadi and Kindergarten in Mangalore. Statistical analysis was done and Chi-square test was used. Tooth brushing habits in these children was started at a mean age of 22.4 months (SD 8.4).62% of the preschool children used toothbrush and toothpaste for cleaning teeth and brushing habits were mainly (84%) introduced by mothers. Seventy-one percent of the children were cooperative when they were introduced to tooth brushing. Preschool children of Mangalore were introduced to tooth brushing at a mean age of 22.4 months. Mothers played a vital role in introducing and teaching the child how to brush. In children less than 10 months of age tooth brushing was not started at all. How to cite this article: Pullishery F, Panchmal GS, Shenoy R. Parental Attitudes and Tooth Brushing Habits in Preschool Children in Mangalore, Karnataka: A Cross-sectional Study. Int J Clin Pediatr Dent 2013;6(3):156-160.

Parental Attitudes and Tooth Brushing Habits in Preschool Children in Mangalore, Karnataka: A Cross-sectional Study

PubMed Central

Shenoy Panchmal, Ganesh; Shenoy, Rekha

2013-01-01

ABSTRACT Background: Adoption of consistent behavioral habits in childhood takes place at home, with the parents especially the mother, being the primary model for behavior. Tooth brushing habits which is learnt during early years of life, is deeply ingrained in the child's mind and it is expected that this leads to an adaptation of good oral hygiene in their later life. Objectives: To assess the tooth brushing habits of preschool children and to determine the role and amount of supervision given to them by parents. Study design: A pretested self-designed questionnaire was used to collect information from parents of 130 preschool children in Anganwadi and Kindergarten in Mangalore. Statistical analysis was done and Chi-square test was used. Results: Tooth brushing habits in these children was started at a mean age of 22.4 months (SD 8.4).62% of the preschool children used toothbrush and toothpaste for cleaning teeth and brushing habits were mainly (84%) introduced by mothers. Seventy-one percent of the children were cooperative when they were introduced to tooth brushing. Conclusion: Preschool children of Mangalore were introduced to tooth brushing at a mean age of 22.4 months. Mothers played a vital role in introducing and teaching the child how to brush. In children less than 10 months of age tooth brushing was not started at all. How to cite this article: Pullishery F, Panchmal GS, Shenoy R. Parental Attitudes and Tooth Brushing Habits in Preschool Children in Mangalore, Karnataka: A Cross-sectional Study. Int J Clin Pediatr Dent 2013;6(3):156-160. PMID:25206214

Tribological properties of hydrophilic polymer brushes under wet conditions.

PubMed

Kobayashi, Motoyasu; Takahara, Atsushi

2010-08-01

This article demonstrates a water-lubrication system using high-density hydrophilic polymer brushes consisting of 2,3-dehydroxypropyl methacrylate (DHMA), vinyl alcohol, oligo(ethylene glycol)methyl ether methacrylate, 2-(methacryloyloxy)ethyltrimethylammonium chloride (MTAC), 3-sulfopropyl methacrylate potassium salt (SPMK), and 2-methacryloyloxyethyl phosphorylcholine (MPC) prepared by surface-initiated controlled radical polymerization. Macroscopic frictional properties of brush surfaces were characterized by sliding a glass ball probe in water using a ball-on-plate type tribotester under the load of 0.1-0.49 N at the sliding velocity of 10(-5)-10(-1) m s(-1) at 298 K. A poly(DHMA) brush showed a relatively larger friction coefficient in water, whereas the polyelectrolyte brushes, such as poly(SPMK) and poly(MPC), revealed significantly low friction coefficients below 0.02 in water and in humid air conditions. A drastic reduction in the friction coefficient of polyelectrolyte brushes in aqueous solution was observed at around 10(-3)-10(-2) m s(-1) owing to the hydrodynamic lubrication effect, however, an increase in salt concentration in the aqueous solution led to the increase in the friction coefficients of poly(MTAC) and poly(SPMK) brushes. The poly(SPMK) brush showed a stable and low friction coefficient in water even after sliding over 450 friction cycles, indicating a good wear resistance of the brush film. Copyright 2010 The Japan Chemical Journal Forum and Wiley Periodicals, Inc.

Assessment of parental tooth-brushing following instruction with single-headed and triple-headed toothbrushes.

PubMed

Telishevesky, Yoel S; Levin, Liran; Ashkenazi, Malka

2012-01-01

The purpose of this study was to evaluate the effect of toothbrush design on the ability of parents to effectively brush their children's teeth. Parents of children (mean age=5.1±0.75 years old) from 4 kindergarten schools were randomly assigned to receive instruction in brushing their children's teeth using a manual single-headed toothbrush (2 schools) or a triple-headed toothbrush (2 schools). The parents' ability to brush their children's teeth was evaluated according to a novel toothbrush performing skill index (Ashkenazi Index), based on 2 criteria: (1) placement of the toothbrush on each tooth segment to be brushed ("reach"); and (2) completion of enough strokes on each segment ("stay"). One month after instruction, tooth-brushing ability was re-evaluated and plaque index of the children's teeth was assessed. One month after instruction, parents using the triple-headed toothbrush received significantly higher scores on the tooth-brushing performance index (~86%), than did those in the single-headed group (~61%; P=.001). The plaque index was significantly higher in the single-headed group (0.97±0.38) vs the triple-headed group (0.72±0.29; P<.01). The tooth-brushing performance index correlated negatively with the plaque index (P<.01). A triple-headed toothbrush promotes more consistent tooth-brushing by parents than does a single-headed toothbrush.

Chemical control of brush in ponderosa pine forests of central Oregon.

Treesearch

Walter G. Dahms

1961-01-01

The many acres of forest land that are occupied by brush in central Oregon represent a large-scale waste of timber-growing capacity and a major economic loss to the area. Although large brushfields devoid of tree growth present the most spectacular examples of loss, less obvious but equally important brush problems are common in established timber stands. Brush...

Spirit Has Flower Power

NASA Technical Reports Server (NTRS)

2004-01-01

The Mars Exploration Rover Spirit took this image with its hazard-avoidance camera on sol 86 (March 31, 2004), after the rover's rock abrasion tool had brushed for three minutes on each of six locations on the rock named 'Mazatzal' to create a flower-shaped mosaic.

The goal for this operation was to create a brushed area big enough for the miniature thermal emission spectrometer to capture within one of its pixels, which are 11 centimeters (4.3 inches) in diameter at the distance between the rock and the instrument. Because the rock abrasion tool creates individual brushed areas only about 5 centimeters (2 inches) in diameter, the team designed this six-location series of tool placements in order to brush 92 percent to 95 percent of the spectrometer's pixel size.

This operation was only the second time the rock abrasion tool has created a brushing mosaic. The first time was a three-spot brushing on the rock called 'Humphrey.' The brush was originally designed to be used as an aide during full grinding operations, however it has been very effective in brushing the top layer off of dusty martian rocks to allow scientists a multi-depth look into the rocks on Mars.

Regression modeling and prediction of road sweeping brush load characteristics from finite element analysis and experimental results.

PubMed

Wang, Chong; Sun, Qun; Wahab, Magd Abdel; Zhang, Xingyu; Xu, Limin

2015-09-01

Rotary cup brushes mounted on each side of a road sweeper undertake heavy debris removal tasks but the characteristics have not been well known until recently. A Finite Element (FE) model that can analyze brush deformation and predict brush characteristics have been developed to investigate the sweeping efficiency and to assist the controller design. However, the FE model requires large amount of CPU time to simulate each brush design and operating scenario, which may affect its applications in a real-time system. This study develops a mathematical regression model to summarize the FE modeled results. The complex brush load characteristic curves were statistically analyzed to quantify the effects of cross-section, length, mounting angle, displacement and rotational speed etc. The data were then fitted by a multiple variable regression model using the maximum likelihood method. The fitted results showed good agreement with the FE analysis results and experimental results, suggesting that the mathematical regression model may be directly used in a real-time system to predict characteristics of different brushes under varying operating conditions. The methodology may also be used in the design and optimization of rotary brush tools. Copyright © 2015 Elsevier Ltd. All rights reserved.

The effect of multivalent counterions to the structure of highly dense polystyrene sulfonate brushes

DOE PAGES

Yu, Jing; Mao, Jun; Yuan, Guangcui; ...

2016-02-23

Surface tethered polyelectrolyte brushes are scientifically interesting and technologically relevant to many applications, ranging from colloidal stabilization to responsive and tunable materials to lubrication. Many applications operate in environments containing multi-valent ions, media in which our scientific understanding is not yet well-developed. In this paper, we synthesized high-density polystyrene sulfonate (PSS) brushes via surface initiated atom-transfer radical polymerization, and performed neutron reflectivity (NR) measurements to investigate and compare the effects of mono-valent Rb + and tri-valent Y 3+ counterions to the structure of the densely tethered PSS brushes. Our NR results show that in mono-valent RbNO 3 solution, the densemore » PSS brush retained its full thickness up to a salt concentration of 1 M, whereas it immediately collapsed upon adding 1.67 mM of tri-valent Y 3+. Increasing the concentration of Y 3+ beyond this level did not lead to any significant further structure change of the PSS brush. Finally, our findings demonstrate that the presence of multi-valent counterions can significantly alter the structure of polyelectrolyte brushes, in a manner different from mono-valent ions, which has implications for the functionality of the brushes.« less

Design optimization of a brush turbine with a cleaner/water based solution

NASA Technical Reports Server (NTRS)

Kim, Rhyn H.

1994-01-01

Recently, a fluid turbine which has a brush attached to it has been designed and tested with water as fluid. The purpose of the turbine-brush is to clean up fouling in a tube. The Montreal Protocol prohibits the use of CFC products from refrigeration industry or from industry in general as a cleanser in 1996. Alternatives for the cleansers, devices or a combination of alternative devices with a cleanser should be found. One of the methods is to develop a device which cleans fouling with a cleaning medium. In this paper, we describe a turbine connected with a brush. However, the turbine with the brush should be simple and easy to install. This device is a combined small liquid turbine with a brush. The turbine is activated by the liquid flowing through the tube. Then the turbine turns the brush cleaning fouling along the tube. Based on the energy conservation and the Bernoulli equation along with an empirical relationship of drag force obtained from an experimental apparatus, a relationship of the rotational speed, the number of blades, and geometric variables of the turbine-brush was obtained. The predicted rotational speeds were compared with the experimental observations. Further work was recommended for improvements.

Stimuli Responsive Morphological Changes of Pnipa Polymer Brushes Synthesized on Silicon Substrate

NASA Astrophysics Data System (ADS)

Huda, Muhammad Nurul; Kabir, A. N. M. Hamidul

2013-08-01

High-density polymer brushes were grown from the silicon surface by atom transfer radical polymerization of Poly(N-isopropylacrylamide) (PNIPA) at different polymerization conditions. PNIPA brushes were prepared using Copper (I) Chloride/tris(2-(dimetylamino)ethyl)amine (Me6TREN) as a catalytic system in DMSO at 20°C. Free polymer formed during the brush formation was characterized by gel permeation chromatography. The grafting densities up to 0.52 chains/nm2 were obtained. The layer thickness of polymer brush increases with the increase of conversion of the monomer conversion as well as polymerization time. Atomic force microscopy and air bubble contact angle under pH solution were employed to study the surface morphology, reversible conformational changes of and stimulus-response behavior. PNIPA brushes exhibited a different nanomorphology after treatment with different pH solution. It also revealed a unique reversible wetting behavior with pH. The reversible properties of the PNIPA brushes can be used to regulate the adsorption of the sulfonated PS nanoparticles.

Grafting density effects, optoelectrical properties and nano-patterning of poly(para-phenylene) brushes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chen, Jihua; Alonzo, Jose; Yu, Xiang

2013-09-24

Well-defined conjugated polymers in confined geometries are challenging to synthesize and characterize, yet they are potentially useful in a broad range of organic optoelectronic devices such as transistors, light emitting diodes, solar cells, sensors, and nanocircuits. We report a systematic study of optoelectrical properties, grafting density effects, and nanopatterning of a model, end-tethered conjugated polymer system. Specifically, poly(para-phenylene) (PPP) brushes of various grafting density are created in situ by aromatizing well-defined, end-tethered poly(1,3-cyclohexadiene) (PCHD) “precursor brushes”. Furthermore, this novel precursor brush approach provides a convenient way to make and systematically control the grafting density of high molecular weight conjugated polymermore » brushes that would otherwise be insoluble. Finally, this allows us to examine how grafting density impacts the effective conjugation length of the conjugated PPP brushes and to adapt the fabrication method to develop spatially patterned conjugated brush systems, which is important for practical applications of conjugated polymer brushes.« less

The effect of brushing with nano calcium carbonate and calcium carbonate toothpaste on the surface roughness of nano-ionomer

NASA Astrophysics Data System (ADS)

Anisja, D. H.; Indrani, D. J.; Herda, E.

2017-08-01

Nanotechnology developments in dentistry have resulted in the development of nano-ionomer, a new restorative material. The surface roughness of restorative materials can increase bacteria adhesion and lead to poor oral hygiene. Abrasive agents in toothpaste can alter tooth and restorative material surfaces. The aim of this study is to identify the effect of brushing with nano calcium carbonate, and calcium carbonate toothpaste on surface roughness of nano-ionomer. Eighteen nano-ionomer specimens were brushed with Aquabidest (doubledistilled water), nano calcium carbonate and calcium carbonate toothpaste. Brushing lasted 30 minutes, and the roughness value (Ra) was measured after each 10 minute segment using a surface roughness tester. The data was analyzed using repeated ANOVA and one-way ANOVA test. The value of nano-ionomer surface roughness increased significantly (p<0.05) after 20 minutes of brushing with the nano calcium carbonate toothpaste. Brushing with calcium carbonate toothpaste leaves nano-ionomer surfaces more rugged than brushing with nano calcium carbonate toothpaste.

Drilling Deeper into tooth brushing skills: Is proactive interference an under-recognized factor in oral hygiene behavior change?

PubMed

Thavarajah, Rooban; Kumar, Madan; Mohandoss, Anusa Arunachalam; Vernon, Lance T

2015-09-01

Proper tooth brushing is a seemingly simple motor activity that can promote oral health. Applying health theories, such as the Information-Motivation-Behavioral Skills (IMB) model, Motivational Interviewing (MI) and Integrated Health Coaching (IHC), may help optimize tooth brushing technique in those with suboptimal skills. Some motor activities, including tooth brushing, may over time become rote and unconscious actions, such that an existing habit can inhibit new learning, i.e., exert proactive interference on learning the new skill. Proactive interference may impede the acquisition of new tooth brushing skills; thus, in this report, we: (1) Review how the habit of tooth brushing is formed; (2) Postulate how proactive interference could impede the establishment of proper tooth brushing retraining; (3) Discuss the merits of this hypothesis; and (4) Provide guidance for future work in this topic within the context of an approach to behavior change that integrates IMB, MI and IHC methodology.

Drilling Deeper into tooth brushing skills: Is proactive interference an under-recognized factor in oral hygiene behavior change?

PubMed Central

Thavarajah, Rooban; Kumar, Madan; Mohandoss, Anusa Arunachalam

2015-01-01

Proper tooth brushing is a seemingly simple motor activity that can promote oral health. Applying health theories, such as the Information-Motivation-Behavioral Skills (IMB) model, Motivational Interviewing (MI) and Integrated Health Coaching (IHC), may help optimize tooth brushing technique in those with suboptimal skills. Some motor activities, including tooth brushing, may over time become rote and unconscious actions, such that an existing habit can inhibit new learning, i.e., exert proactive interference on learning the new skill. Proactive interference may impede the acquisition of new tooth brushing skills; thus, in this report, we: (1) Review how the habit of tooth brushing is formed; (2) Postulate how proactive interference could impede the establishment of proper tooth brushing retraining; (3) Discuss the merits of this hypothesis; and (4) Provide guidance for future work in this topic within the context of an approach to behavior change that integrates IMB, MI and IHC methodology. PMID:26457238

The Art and Science of Polymer Brushes: Recent Developments in Patterning and Characterization Approaches.

PubMed

Panzarasa, Guido

2017-06-28

Polymer brushes are dense arrays of macromolecular chains tethered by one end at a surface. They are at the cutting edge of polymer nanotechnology since the dawn of controlled surface-initiated polymerization techniques unlocked new prospects for the synthesis of polymer brushes with tailorable properties. More recently, thanks to the growing interest in the use of brushes for the generation of functional surfaces, the need for advanced patterning and characterization approaches rapidly increased. Meeting these needs requires the contribution of experts from different disciplines: polymer chemistry, surface science, electrochemistry and particle physics. The focus of this review is to highlight recent developments in the field of polymer brushes, specifically the application of photocatalytic lithography as a versatile patterning strategy, the study of grafted-from polymer brushes by electrochemical methods and, most importantly, the introduction of positron annihilation spectroscopy as a powerful technique for the investigation of the structure of polymer brushes and of their composites with nanoparticles.

Friction, wear, and noise of slip ring and brush contacts for synchronous satellite use.

NASA Technical Reports Server (NTRS)

Lewis, N. E.; Cole, S. R.; Glossbrenner, E. W.; Vest, C. E.

1973-01-01

A program is being conducted for testing of slip rings for synchronous orbit application. Instrumentation systems necessary for monitoring electrical noise, friction, and brush wear at atmospheric pressure and at less than 50 nanotorr have been developed. A multiplex scheme necessary for the simultaneous recording of brush displacement, friction, and electrical noise has also been developed. Composite brushes consisting of silver-molybdenum disulfide-graphite and silver-niobium diselenide-graphite have been employed on rings of coin silver and rhodium plate. Brush property measurements made included measurement of density, electrical resistivity, shear strength, and microstructure.

[Daily living activities and oral condition among care facility residents with severe intellectual disabilities. Comparative analyses between residents receiving tooth-brushing assistance and those not receiving tooth-brushing assistance].

PubMed

Chiwata, Kaoru; Takeda, Fumi

2007-06-01

To clarify 1) differences in daily living activities and oral condition among care facility residents with severe intellectual disabilities and 2) chronological changes in daily living activities and oral condition for residents receiving tooth-brushing assistance and those never receiving tooth-brushing assistance. Subjects were 44 residents at a care facility for individuals with severe intellectual disabilities, who underwent dental screening in July 1994 and October 2003. At each time point, daily living activities, behavior during oral health guidance, behavior during dental health screening and oral condition were compared between residents receiving tooth-brushing assistance (assistance group) and those not receiving tooth-brushing assistance (independent group). Furthermore, chronological changes were analyzed for residents requiring assistance at both screenings, those requiring assistance only at the second screening, and those not requiring assistance at either screening. 1) In the assistance group, 100% and 36.4% of residents were unable to brush their teeth independently in 1994 and 2003, respectively. Significant differences between the assistance and independent groups were observed in all items of behavior during dental health screening in 1994, but not in 2003. No significant intergroup differences in oral condition were observed in 1994, but differences were seen in 2003; when compared to the assistance group, the number of lost teeth was significantly higher in the independent group, while the number of remaining teeth was lower. 2) Regarding changes over the nine-year period, a significantly greater proportion of residents not requiring assistance at either screening and those requiring assistance only at the second screening finally required assistance in bathing. As for oral condition, no significant changes in healthy teeth were observed in residents requiring assistance at both screening time points, while significant increases in dental caries and filled teeth and a significant decrease in the number of healthy teeth were observed in residents requiring assistance only at the second screening and those not requiring assistance at either screening. Over the nine-year period, the subjects of tooth-brushing assistance changed, and assistance was given to those able to brush their teeth independently in addition to those unable to brush their teeth independently. The number of healthy teeth did not change in residents receiving tooth-brushing assistance during this period, but in residents never receiving tooth-brushing assistance, decrease was noted. Therefore, even for individuals able to brush their teeth independently, some form of tooth-brushing assistance is needed to sufficiently prevent oral diseases.

Effects of whitening dentifrice on yttria-stabilized tetragonal zirconia polycrystal surfaces after simulating brushing.

PubMed

Pinelli, Lígia Antunes Pereira; Gimenes Olbera, Amanda Caroline; Candido, Lucas Miguel; Miotto, Larissa Natiele; Antonio, Selma Gutierrez; Fais, Laiza Maria Grassi

2017-01-01

The changes that occur after brushing yttria-stabilized tetragonal zirconia polycrystal (Y-TZP) are unknown. These changes may favor the retention of microorganisms and chemisorption of water, impairing its longevity. The purpose of this in vitro study was to evaluate the effects of a whitening dentifrice on Y-TZP surfaces after simulating 10 years of brushing. Seventy-two bar-shaped specimens (20×4×1.2 mm) were divided into 4 groups: storage in distilled water (SW, control), brushing with distilled water (BW), brushing with dentifrice (BD), and brushing with whitening dentifrice (BWD). Brushing was conducted using a linear brushing machine (878400 cycles, 0.98 N, soft toothbrush). The mean roughness (Ra) was analyzed with a profilometer and the superficial topography with scanning electron microscopy (SEM) at baseline and after treatment. Crystalline phases were characterized using x-ray diffraction. Baseline and posttreatment Ra were analyzed using the 1-way ANOVA and Tukey HSD multiple comparison test; the paired t test was used for intragroup comparison (all α=.05). The Ra (μm) means (before/after treatment) were SW 0.28/0.28; BW 0.32/0.31; BD 0.28/0.36; BWD 0.30/0.20. No statistically significant difference was found for Ra at baseline (P=.108) than for posttreatment results (P<.001); the BD group had higher Ra values when compared with baseline (P=.019); the BWD group had the lowest values (P<.001). The BD surfaces showed pronounced scratches and detachment of the surface, while BWD showed smoother surfaces; similar crystallographic results among groups were observed. Brushing Y-TZP with conventional dentifrice increased roughness, while brushing with whitening dentifrice reduced roughness. Neither dentifrice changed the crystallographic phases after brushing. Copyright © 2016 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

Fine needle aspiration of secondary synovial sarcoma of the thyroid gland.

PubMed

Murro, Diana; Slade, Jamie Macagba; Syed, Sahr; Gattuso, Paolo

2015-11-01

Synovial sarcomas (SS) of the head and neck region are extremely rare and arise in only 5% of cases. We present a case of secondary SS of the thyroid originally diagnosed as medullary carcinoma on fine needle aspiration (FNA). A 41-year-old man presented with several weeks of dysphonia and a left thyroid mass. FNA of the thyroid nodule showed a cellular smear composed of loosely cohesive oval to spindle-shaped cells with irregular nuclear borders, finely granular chromatin, and inconspicuous nucleoli. The patient was diagnosed with medullary carcinoma and underwent a total thyroidectomy. Intro-operatively, the mass was found to arise from the tracheoesophageal groove with spread to the left thyroid. Microscopic examination of the thyroid tumor revealed a dense spindle cell proliferation with abundant mitoses, scant cords and nests of epithelial cells and foci of necrosis. The spindle cells were positive for bcl2 and vimentin and the epithelial cells were positive for cytokeratin 8/18 and epithelial membrane antigen (EMA). Both spindle and epithelial cells were negative for thyroglobulin, calcitonin, synaptophysin and chromogranin. Fluorescence in situ hybridization (FISH) demonstrated translocation (X;18)(p11;q11), confirming the diagnosis of SS. The patient underwent a total laryngopharyngoesophagectomy with subsequent adjuvant therapy and is currently disease free. Only 6 cases of histologically confirmed primary SS of the thyroid have been reported. To the best of our knowledge, this is the first case of FISH-confirmed secondary SS of the thyroid and also the first case of SS arising from the tracheoesophageal groove. © 2015 Wiley Periodicals, Inc.

Gingival abrasion and recession in manual and oscillating-rotating power brush users.

PubMed

Rosema, N A M; Adam, R; Grender, J M; Van der Sluijs, E; Supranoto, S C; Van der Weijden, G A

2014-11-01

To assess gingival recession (GR) in manual and power toothbrush users and evaluate the relationship between GR and gingival abrasion scores (GA). This was an observational (cross-sectional), single-centre, examiner-blind study involving a single-brushing exercise, with 181 young adult participants: 90 manual brush users and 91 oscillating-rotating power brush users. Participants were assessed for GR and GA as primary response variables. Secondary response variables were the level of gingival inflammation, plaque score reduction and brushing duration. Pearson correlation was used to describe the relationship between number of recession sites and number of abrasions. Prebrushing (baseline) and post-brushing GA and plaque scores were assessed and differences analysed using paired tests. Two-sample t-test was used to analyse group differences; ancova was used for analyses of post-brushing changes with baseline as covariate. Overall, 97.8% of the study population had at least one site of ≥1 mm of gingival recession. For the manual group, this percentage was 98.9%, and for the power group, this percentage was 96.7% (P = 0.621). Post-brushing, the power group showed a significantly smaller GA increase than the manual group (P = 0.004); however, there was no significant correlation between number of recession sites and number of abrasions for either group (P ≥ 0.327). Little gingival recession was observed in either toothbrush user group; the observed GR levels were comparable. Lower post-brushing gingival abrasion levels were seen in the power group. There was no correlation between gingival abrasion as a result of brushing and the observed gingival recession following use of either toothbrush. © 2014 The Authors International Journal of Dental Hygiene Published by John Wiley & Sons Ltd.

Synergy of brushing mode and antibacterial use on in vivo biofilm formation.

PubMed

Jongsma, Marije A; van de Lagemaat, Marieke; Busscher, Henk J; Geertsema-Doornbusch, Gesinda I; Atema-Smit, Jelly; van der Mei, Henny C; Ren, Yijin

2015-12-01

Orthodontic, multi-strand retention-wires are used as a generalized model for oral retention sites to investigate whether biofilm left-behind after powered toothbrushing in-vivo enabled better penetration of antibacterials as compared with manual brushing. 2-cm multi-strand, stainless-steel retention-wires were placed in brackets bonded bilaterally in the upper arches of 10-volunteers. Volunteers used NaF-sodium-lauryl-sulphate-containing toothpaste and antibacterial, triclosan-containing toothpaste supplemented or not with an essential-oils containing mouthrinse. Opposite sides of the dentition including the retention-wires, were brushed manually or with a powered toothbrush. Health-care-regimens were maintained for 1-week, after which wires were removed and oral biofilm was collected. When powered toothbrushing was applied, slightly less bacteria were collected than after manual brushing, regardless whether an antibacterial-regimen was used or not. Powered-toothbrushing combined with antibacterial-regimens yielded lower biofilm viability than manual brushing, indicating better antibacterial penetration into biofilm left-behind after powered brushing. Major shifts in biofilm composition, with a decrease in prevalence of both cariogenic species and periodontopathogens, were induced after powered brushing using an antibacterial-regimen. Oral biofilm left-behind after powered brushing in-vivo enabled better penetration of antibacterials than after manual brushing. Mechanical removal of oral biofilm is important for prevention of dental pathologies, but biofilm is always left-behind, such as in fissures, buccal pits, interproximal areas and gingival margins and around orthodontic appliances. Use of antibacterial toothpastes or mouthrinses can contribute to removal or killing of biofilm bacteria, but biofilm structure hampers antibacterial penetration. A synergy between brushing mode and antibacterial-regimen applied exists with clinically demonstrable effects. Copyright © 2015 Elsevier Ltd. All rights reserved.

Compression induced phase transition of nematic brush: A mean-field theory study

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tang, Jiuzhou; Zhang, Xinghua, E-mail: zhangxh@bjtu.edu.cn; Yan, Dadong, E-mail: yandd@bnu.edu.cn

2015-11-28

Responsive behavior of polymer brush to the external compression is one of the most important characters for its application. For the flexible polymer brush, in the case of low grafting density, which is widely studied by the Gaussian chain model based theory, the compression leads to a uniform deformation of the chain. However, in the case of high grafting density, the brush becomes anisotropic and the nematic phase will be formed. The normal compression tends to destroy the nematic order, which leads to a complex responsive behaviors. Under weak compression, chains in the nematic brush are buckled, and the bendingmore » energy and Onsager interaction give rise to the elasticity. Under deep compression, the responsive behaviors of the nematic polymer brush depend on the chain rigidity. For the compressed rigid polymer brush, the chains incline to re-orientate randomly to maximize the orientational entropy and its nematic order is destroyed. For the compressed flexible polymer brush, the chains incline to fold back to keep the nematic order. A buckling-folding transition takes place during the compressing process. For the compressed semiflexible brush, the chains are collectively tilted to a certain direction, which leads to the breaking of the rotational symmetry in the lateral plane. These responsive behaviors of nematic brush relate to the properties of highly frustrated worm-like chain, which is hard to be studied by the traditional self-consistent field theory due to the difficulty to solve the modified diffusion equation. To overcome this difficulty, a single chain in mean-field theory incorporating Monte Carlo simulation and mean-field theory for the worm-like chain model is developed in present work. This method shows high performance for entire region of chain rigidity in the confined condition.« less

Gingival abrasion and recession in manual and oscillating–rotating power brush users

PubMed Central

Rosema, NAM; Adam, R; Grender, JM; Van der Sluijs, E; Supranoto, SC; Van der Weijden, GA

2014-01-01

Objective To assess gingival recession (GR) in manual and power toothbrush users and evaluate the relationship between GR and gingival abrasion scores (GA). Methods This was an observational (cross-sectional), single-centre, examiner-blind study involving a single-brushing exercise, with 181 young adult participants: 90 manual brush users and 91 oscillating–rotating power brush users. Participants were assessed for GR and GA as primary response variables. Secondary response variables were the level of gingival inflammation, plaque score reduction and brushing duration. Pearson correlation was used to describe the relationship between number of recession sites and number of abrasions. Prebrushing (baseline) and post-brushing GA and plaque scores were assessed and differences analysed using paired tests. Two-sample t-test was used to analyse group differences; ancova was used for analyses of post-brushing changes with baseline as covariate. Results Overall, 97.8% of the study population had at least one site of ≥1 mm of gingival recession. For the manual group, this percentage was 98.9%, and for the power group, this percentage was 96.7% (P = 0.621). Post-brushing, the power group showed a significantly smaller GA increase than the manual group (P = 0.004); however, there was no significant correlation between number of recession sites and number of abrasions for either group (P ≥ 0.327). Conclusions Little gingival recession was observed in either toothbrush user group; the observed GR levels were comparable. Lower post-brushing gingival abrasion levels were seen in the power group. There was no correlation between gingival abrasion as a result of brushing and the observed gingival recession following use of either toothbrush. PMID:24871587

The effect of food location, heat load, and intrusive medical procedures on brushing activity in dairy cows.

PubMed

Mandel, R; Whay, H R; Nicol, C J; Klement, E

2013-10-01

Animals allocate time and effort to a range of core (e.g., sleeping, feeding, drinking) and "luxury" (e.g., playing, exploring) activities. A luxury activity is characterized by low resilience and, as such, will be reduced when time or energy resources are limited, including under conditions of stress or discomfort. One seemingly luxurious activity available to cows on an increasing number of dairy farms is rubbing against an automated brush. The current study examined the effect of distance from food, heat load, and an intrusive medical procedure (i.e., artificial insemination and transrectal pregnancy examination) on the resilience of brush usage. The probability of using the brush decreased significantly when food was located distantly from the brush (mean=0.53) compared with days when food was located closer to the brush (mean=0.81). Brush usage also decreased at high temperature and humidity levels, with an average decrease of 0.062 brushing events for an increase of 1 temperature-humidity index unit (95% confidence interval=-0.93-0.030). In addition, a significant reduction of approximately 50% in brushing activity was observed on days of artificial insemination compared with the preceding 3d and the following 3d. These findings show that brush usage is a low resilience activity that reduces under a range of conditions. It may thus have the potential to be used as an indicator of a range of health and welfare problems in cows. Further research should be conducted to assess the sensitivity and specificity of this suggested tool and its possible contribution to the early detection of morbidity. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

A comparative evaluation of the in vitro penetration performance of the improved Crest Complete toothbrush versus the Colgate Total toothbrush and the Oral-B Advantage toothbrush.

PubMed

Volpenhein, D W; Hartman, W L

1996-01-01

Removal of plaque and debris from interproximal surfaces during toothbrushing has generally been difficult to achieve, in large part because traditional flat-bristled toothbrushes do not offer good interproximal penetration. As a result, a number of varying bristle designs have been developed, with the rippled-design brush shown to be particularly effective at removing interproximal plaque. Recently, an existing rippled brush, the original Crest Complete, was modified to offer longer rippled outer tufts to clean along the gumline more effectively. Therefore, this study evaluated the overall and gumline interproximal penetration of three bristle designs: rippled, raised pattern (Improved Crest Complete); and two multi-level patterns (Colgate Total and Oral-B Advantage). The study used a previously reported in vitro model for determining interproximal penetration of manual toothbrushes (J Clin Dent 5:27-33, 1994). In order to effectively mimic the in-use characteristics of toothbrushing, this model is based on analysis of videotaped consumer brushing habits, tooth morphology, and in vivo plaque tenacity characteristics, and uses the three most predominantly used brushing techniques (circular, up-and-down, and back-and-forth with the brush held at both 45 degrees and 90 degrees to the tooth surface). In addition, the model's brush stroke length, brush force, and brush speed are likewise based on an analysis of consumer brushing patterns. The results of this study indicate that the Improved Crest Complete with longer rippled outer bristles provided significantly superior (p < 0.05) interproximal penetration overall and at the gumline than the Colgate Total and Oral-B Advantage brushes.

Factors Affecting Oral Hygiene and Tooth Brushing in Preschool Children, Shiraz/Iran.

PubMed

S, Shaghaghian; M, Zeraatkar

2017-06-01

Inadequate tooth brushing and inappropriate oral hygiene can lead to dental caries, the most common chronic diseases of childhood with several side effects. To evaluate factors affecting on preschool children's oral hygiene and tooth brushing in Shiraz, Iran. In this cross-sectional study, we selected 453 children registered in Shiraz kindergartens in 2013 by randomized cluster sampling. The children's tooth brushing and oral hygiene were assessed using a reliable and valid questionnaire and Simplified Debris Index (DI-S), respectively. A dental student examined all the children in each kindergarten to determine their DI-S. The relationship between the children's demographic variables and their oral hygiene and tooth brushing status were evaluated. Tooth brushing for 272 children (71.2%) had been started after the age of 2 years. The teeth in 96 children (24.2%) had been brushed lower than once daily. The mean of the children's DI-S was 1.19 ± (0.77). The DI-S of only 126 children (31.8%) was found to be good and very good. After controlling the effect of confounding factors, we found that the children's tooth brushing frequency was significantly associated with the number of children in the family and mothers' employment status. The age at which tooth brushing had been started was significantly associated with the fathers' education. Furthermore, the DI-S was associated with children's age, number of the children in the family, and their mothers' education. Oral hygiene and tooth brushing of the preschool children were not in a desirable status. Interventional procedures, especially educational programs, are recommended for children and their parents. These programs seem to be more necessary for older children, low socioeconomic families, and families with more than one child.

Does tooth brushing influence the development and progression of non-inflammatory gingival recession? A systematic review.

PubMed

Rajapakse, P Sunethra; McCracken, Giles I; Gwynnett, Erika; Steen, Nick D; Guentsch, Arndt; Heasman, Peter A

2007-12-01

The aim of this systematic review was to produce the best available evidence and pool appropriate data to evaluate the effect of tooth brushing on the initiation and progression of non-inflammatory gingival recession. A protocol was developed a priori for the question: "Do factors associated with tooth brushing predict the development and progression of non-inflammatory gingival recession in adults?" The search covered six electronic databases between January 1966 and July 2005. Hand searching included searches of the Journal of Clinical Periodontology, Journal of Periodontal Research and the Journal of Periodontology. Bibliographies of narrative reviews, conference proceedings and relevant texts known to the authors were also searched. Inclusion of titles, abstracts and ultimately full texts was based on consensus between three reviewers. The full texts of 29 papers were read and 18 texts were eligible for inclusion. One abstract from EuroPerio 5 reported a randomized-controlled clinical trial [Level I evidence] in which the authors concluded that the toothbrushes significantly reduced recessions on buccal tooth surfaces over 18 months. Of the remaining 17 observational studies, two concluded that there appeared to be no relationship between tooth brushing frequency and gingival recession. Eight studies reported a positive association between tooth brushing frequency and recession. Other potential risk factors were duration of tooth brushing, brushing force, frequency of changing the toothbrush, brush (bristle) hardness and tooth brushing technique. None of the observational studies satisfied all the specified criteria for quality appraisal and a valid appraisal of the quality of the randomized-controlled trial was not possible. The data to support or refute the association between tooth brushing and gingival recession are inconclusive.

Nanostructured bio-functional polymer brushes.

PubMed

Padeste, Celestino; Farquet, Patrick; Potzner, Christian; Solak, Harun H

2006-01-01

Structured poly(glycidyl methracrylate) (poly-GMA) brushes have been grafted onto flexible fluoro-polymer films using a radiation grafting process. The reactive epoxide of poly-GMA provides the basis for a versatile biofunctionalization of the grafted brushes. Structure definition by extreme ultraviolet (EUV) exposure allowed nanometer-scale resolution of periodic patterns. By variation of the exposure dose the height of the grafted structures can be adapted in a wide range. Derivatization of the grafted brushes included reaction with various amines with different side chains, hydrolysis of the epoxide to diols to increase protein resistance and introduction of ionic groups to yield poly-electrolytes. As an example for biofunctionalization, biotin was linked to the grafted brush and biofunctionality was demonstrated in a competitive biotin-streptavidin assay. In this article we also present a brief review of other approaches to obtain structured biofunctional polymer brushes.

Shunt attachment and method for interfacing current collection systems

DOEpatents

Denney, P.E.; Iyer, N.C.; Hannan, W.F. III.

1992-12-08

A composite brush to shunt attachment wherein a volatile component of a composite but mostly metallic brush, used for current collection purposes, does not upon welding or brazing, adversely affect the formation of the interfacial bond with a conductive shunt which carries the current from the zone of the brush. The brush to shunt attachment for a brush material of copper-graphite composite and a shunt of copper, or substituting silver for copper as an alternative, is made through a hot isostatic pressing (HIP). The HIP process includes applying high pressure and temperature simultaneously at the brush to shunt interface, after it has been isolated or canned in a metal casing in which the air adjacent to the interface has been evacuated and the interfacial area has been sealed before the application of pressure and temperature. 6 figs.

Shunt attachment and method for interfacing current collection systems

DOEpatents

Denney, Paul E.; Iyer, Natraj C.; Hannan, III, William F.

1992-01-01

A composite brush to shunt attachment wherein a volatile component of a composite but mostly metallic brush, used for current collection purposes, does not upon welding or brazing, adversely affect the formation of the interfacial bond with a conductive shunt which carries the current from the zone of the brush. The brush to shunt attachment for a brush material of copper-graphite composite and a shunt of copper, or substituting silver for copper as an alternative, is made through a hot isostatic pressing (HIP). The HIP process includes applying high pressure and temperature simultaneously at the brush to shunt interface, after it has been isolated or canned in a metal casing in which the air adjacent to the interface has been evacuated and the interfacial area has been sealed before the application of pressure and temperature.

Surface roughness of composite resin veneer after application of herbal and non-herbal toothpaste

NASA Astrophysics Data System (ADS)

Nuraini, S.; Herda, E.; Irawan, B.

2017-08-01

The aim of this study was to find out the surface roughness of composite resin veneer after brushing. In this study, 24 specimens of composite resin veneer are divided into three subgroups: brushed without toothpaste, brushed with non-herbal toothpaste, and brushed with herbal toothpaste. Brushing was performed for one set of 5,000 strokes and continued for a second set of 5,000 strokes. Roughness of composite resin veneer was determined using a Surface Roughness Tester. The results were statistically analyzed using Kruskal-Wallis nonparametric test and Post Hoc Mann-Whitney. The results indicate that the highest difference among the Ra values occurred within the subgroup that was brushed with the herbal toothpaste. In conclusion, the herbal toothpaste produced a rougher surface on composite resin veneer compared to non-herbal toothpaste.

Effect of saliva and blood contamination after etching upon the shear bond strength between composite resin and enamel

NASA Astrophysics Data System (ADS)

Armadi, A. S.; Usman, M.; Suprastiwi, E.

2017-08-01

The aim of this study was to find out the surface roughness of composite resin veneer after brushing. In this study, 24 specimens of composite resin veneer are divided into three subgroups: brushed without toothpaste, brushed with non-herbal toothpaste, and brushed with herbal toothpaste. Brushing was performed for one set of 5,000 strokes and continued for a second set of 5,000 strokes. Roughness of composite resin veneer was determined using a Surface Roughness Tester. The results were statistically analyzed using Kruskal-Wallis nonparametric test and Post Hoc Mann-Whitney. The results indicate that the highest difference among the Ra values occurred within the subgroup that was brushed with the herbal toothpaste. In conclusion, the herbal toothpaste produced a rougher surface on composite resin veneer compared to non-herbal toothpaste.

Friendship Network and Dental Brushing Behavior among Middle School Students: An Agent Based Modeling Approach.

PubMed

Sadeghipour, Maryam; Khoshnevisan, Mohammad Hossein; Jafari, Afshin; Shariatpanahi, Seyed Peyman

2017-01-01

By using a standard questionnaire, the level of dental brushing frequency was assessed among 201 adolescent female middle school students in Tehran. The initial assessment was repeated after 5 months, in order to observe the dynamics in dental health behavior level. Logistic Regression model was used to evaluate the correlation among individuals' dental health behavior in their social network. A significant correlation on dental brushing habits was detected among groups of friends. This correlation was further spread over the network within the 5 months period. Moreover, it was identified that the average brushing level was improved within the 5 months period. Given that there was a significant correlation between social network's nodes' in-degree value, and brushing level, it was suggested that the observed improvement was partially due to more popularity of individuals with better tooth brushing habit. Agent Based Modeling (ABM) was used to demonstrate the dynamics of dental brushing frequency within a sample of friendship network. Two models with static and dynamic assumptions for the network structure were proposed. The model with dynamic network structure successfully described the dynamics of dental health behavior. Based on this model, on average, every 43 weeks a student changes her brushing habit due to learning from her friends. Finally, three training scenarios were tested by these models in order to evaluate their effectiveness. When training more popular students, considerable improvement in total students' brushing frequency was demonstrated by simulation results.

Tooth brushing frequency in Mexican schoolchildren and associated socio-demographic, socioeconomic, and dental variables

PubMed Central

Casanova-Rosado, Alejandro José; Medina-Solís, Carlo Eduardo; Casanova-Rosado, Juan Fernando; Vallejos-Sánchez, Ana Alicia; Minaya-Sánchez, Mirna; Mendoza-Rodríguez, Martha; Márquez-Rodríguez, Sonia; Maupomé, Gerardo

2014-01-01

Background Tooth brushing (with fluoridated toothpaste) is the most cost-effective intervention in dentistry and it is widely recommended to preserve good oral health. We aimed to determine the frequency of tooth brushing and the variables associated with this practice in schoolchildren living in southeast Mexico. Material/Methods A cross-sectional study was carried out in 1644 schoolchildren, 6 to 13 years old. Questionnaires with socio-demographic, socioeconomic, and dental variables were administered to mothers/guardians of children. The dependent variable was the frequency of tooth brushing, which was categorized as 0 = tooth brushing less than once a day and 1 = tooth brushing at least once a day. A logistic regression model was used to evaluate the final results. Results Mean age was 9.06±2.02 years and 49.1% were girls. The prevalence of tooth brushing at least once a day was 49.8%. In the multivariate model, characteristics related to tooth brushing frequency (p<0.05) were: older age (OR=1.11), being female (OR=1.64), having a larger family (OR=0.87), having had a visit to a dentist during the year preceding the study (OR=1.37), and having had fluoride applications by a professional (OR=1.39). Conclusions The results suggested that different variables (demographic, socioeconomic and dental) are associated with the frequency of tooth brushing. Family size (proxy variable for socioeconomic status) may indicate certain oral health inequalities in this population. PMID:24907713

Tooth brushing frequency in Mexican schoolchildren and associated socio-demographic, socioeconomic, and dental variables.

PubMed

Casanova-Rosado, Alejandro José; Medina-Solís, Carlo Eduardo; Casanova-Rosado, Juan Fernando; Vallejos-Sánchez, Ana Alicia; Minaya-Sánchez, Mirna; Mendoza-Rodríguez, Martha; Márquez-Rodríguez, Sonia; Maupomé, Gerardo

2014-06-07

Tooth brushing (with fluoridated toothpaste) is the most cost-effective intervention in dentistry and it is widely recommended to preserve good oral health. We aimed to determine the frequency of tooth brushing and the variables associated with this practice in schoolchildren living in southeast Mexico. A cross-sectional study was carried out in 1644 schoolchildren, 6 to 13 years old. Questionnaires with socio-demographic, socioeconomic, and dental variables were administered to mothers/guardians of children. The dependent variable was the frequency of tooth brushing, which was categorized as 0 = tooth brushing less than once a day and 1 = tooth brushing at least once a day. A logistic regression model was used to evaluate the final results. Mean age was 9.06±2.02 years and 49.1% were girls. The prevalence of tooth brushing at least once a day was 49.8%. In the multivariate model, characteristics related to tooth brushing frequency (p<0.05) were: older age (OR=1.11), being female (OR=1.64), having a larger family (OR=0.87), having had a visit to a dentist during the year preceding the study (OR=1.37), and having had fluoride applications by a professional (OR=1.39). The results suggested that different variables (demographic, socioeconomic and dental) are associated with the frequency of tooth brushing. Family size (proxy variable for socioeconomic status) may indicate certain oral health inequalities in this population.

The lid wiper and muco-cutaneous junction anatomy of the human eyelid margins: an in vivo confocal and histological study

PubMed Central

Knop, Erich; Knop, Nadja; Zhivov, Andrey; Kraak, Robert; Korb, Donald R; Blackie, Caroline; Greiner, Jack V; Guthoff, Rudolf

2011-01-01

The inner border of the eyelid margin is critically important for ocular surface integrity because it guarantees the thin spread of the tear film. Its exact morphology in the human is still insufficiently known. The histology in serial sections of upper and lower lid margins in whole-mount specimens from 10 human body donors was compared to in vivo confocal microscopy of eight eyes with a Heidelberg retina-tomograph (HRT II) and attached Rostock cornea module. Behind the posterior margin of the Meibomian orifices, the cornified epidermis stopped abruptly and was replaced by a continuous layer of para-keratinized (pk) cells followed by discontinuous pk cells. The pk cells covered the muco-cutaneous junction (MCJ), the surface of which corresponded to the line of Marx (0.2–0.3 mm wide). Then a stratified epithelium with a conjunctival structure of cuboidal cells, some pk cells, and goblet cells formed an epithelial elevation of typically about 100 μm initial thickness (lid wiper). This continued for 0.3–1.5 mm and formed a slope. The MCJ and lid wiper extended all along the lid margin from nasal to temporal positions in the upper and lower lids. Details of the epithelium and connective tissue were also detectable using the Rostock cornea module. The human inner lid border has distinct zones. Due to its location and morphology, the epithelial lip of the lid wiper appears a suitable structure to spread the tear film and is distinct from the MCJ/line of Marx. Better knowledge of the lid margin appears important for understanding dry eye disease and its morphology can be analysed clinically by in vivo confocal microscopy. PMID:21413985

Design and Testing of High Performance Brushes.

DTIC Science & Technology

1982-03-01

specific types, namely metal fiber brushes and met- al foil brushes, have been constructed and tested, and a theory has been developed which accounts for...combining electric- al measurements with mechanical investigations and optical observations. Since we now have reason to rely confidently on the theory of...reasonable requirements. packing densities between a few % and up to 20%, have been made of various materi- Brushes with gold, platinum, niobium als by the

Polyelectrolyte brushes grafted from cellulose nanocrystals using Cu-mediated surface-initiated controlled radical polymerization.

PubMed

Majoinen, Johanna; Walther, Andreas; McKee, Jason R; Kontturi, Eero; Aseyev, Vladimir; Malho, Jani Markus; Ruokolainen, Janne; Ikkala, Olli

2011-08-08

Herein we report the synthesis of cellulose nanocrystals (CNCs) grafted with poly(acrylic acid) (PAA) chains of different lengths using Cu-mediated surface initiated-controlled radical polymerization (SI-CRP). First, poly(tert-butylacrylate) (PtBA) brushes were synthesized; then, subsequent acid hydrolysis was used to furnish PAA brushes tethered onto the CNC surfaces. The CNCs were chemically modified to create initiator moieties on the CNC surfaces using chemical vapor deposition (CVD) and continued in solvent phase in DMF. A density of initiator groups of 4.6 bromine ester groups/nm(2) on the CNC surface was reached, suggesting a dense functionalization and a promising starting point for the controlled/living radical polymerization. The SI-CRP of tert-butylacrylate proceeded in a well-controlled manner with the aid of added sacrificial initiator, yielding polymer brushes with polydispersity values typically well below 1.12. We calculated the polymer brush grafting density to almost 0.3 chains/nm(2), corresponding to high grafting densities and dense polymer brush formation on the nanocrystals. Successful rapid acid hydrolysis to remove the tert-butyl groups yielded pH-responsive PAA-polyelectrolyte brushes bound to the CNC surface. Individually dispersed rod-like nanoparticles with brushes of PtBA or PAA were clearly visualized by AFM and TEM imaging.

The theory of reasoned action in describing tooth brushing, dental caries and diabetes adherence among diabetic patients.

PubMed

Syrjälä, Anna-Maija H; Niskanen, Mirka C; Knuuttila, Matti L E

2002-05-01

Preventive oral health behaviour is important among diabetic patients, as it has been found that dental diseases and insulin-dependent diabetes mellitus (IDDM) have some psychological and biological factors in common. The aim here was to analyze the variables of Ajzen and Fishbein's theory of reasoned action to explain the reported frequency of tooth brushing, dental caries, HbA1c level and diabetes adherence. Cross-sectional data were gathered from 149 IDDM patients by means of a quantitative questionnaire, clinical examination and patient records. The results showed that a firmer intention to brush the teeth was related to a higher reported frequency of tooth brushing (p < 0.001). The attitude to and the subjective norm of tooth brushing were related to the intention to brush(p < 0.001) and to the reported frequency of tooth brushing. A better dental attitude was related to better diabetes adherence (p = 0.002) and fewer decayed surfaces (p = 0.01), and a firmer intention to brush the teeth was related to a lower HbA1c level (p = 0.015). Our results suggest that in oral health promotion among diabetic patients, both subjective norm and attitude are important and that diabetes adherence may be influenced by promoting dental attitude.

Combined carbon mesh and small graphite fiber brush anodes to enhance and stabilize power generation in microbial fuel cells treating domestic wastewater

NASA Astrophysics Data System (ADS)

Wu, Shijia; He, Weihua; Yang, Wulin; Ye, Yaoli; Huang, Xia; Logan, Bruce E.

2017-07-01

Microbial fuel cells (MFCs) need to have a compact architecture, but power generation using low strength domestic wastewater is unstable for closely-spaced electrode designs using thin anodes (flat mesh or small diameter graphite fiber brushes) due to oxygen crossover from the cathode. A composite anode configuration was developed to improve performance, by joining the mesh and brushes together, with the mesh used to block oxygen crossover to the brushes, and the brushes used to stabilize mesh potentials. In small, fed-batch MFCs (28 mL), the composite anode produced 20% higher power densities than MFCs using only brushes, and 150% power densities compared to carbon mesh anodes. In continuous flow tests at short hydraulic retention times (HRTs, 2 or 4 h) using larger MFCs (100 mL), composite anodes had stable performance, while brush anode MFCs exhibited power overshoot in polarization tests. Both configurations exhibited power overshoot at a longer HRT of 8 h due to lower effluent CODs. The use of composite anodes reduced biomass growth on the cathode (1.9 ± 0.2 mg) compared to only brushes (3.1 ± 0.3 mg), and increased coulombic efficiencies, demonstrating that they successfully reduced oxygen contamination of the anode and the bio-fouling of cathode.

The Effects of Tooth Brushing on Whole Salivary Flow Rate in Older Adults

PubMed Central

Trottier, K.; Garrick, R.; Mascarenhas, T.; Jang, Y.

2018-01-01

Objectives (1) To determine whether manual (MTB), or electric, tooth brushing (ETB) modulates whole salivary flow rate in older adults who are free of systemic disease. (2) To determine the duration of the brushing-related modulation of salivary flow rate. (3) To compare salivary flow rate modulation associated with MTB and ETB. Method Twenty-one adults aged 60 years and older participated in two experimental sessions during which they used a manual, or electric, toothbrush to brush their teeth, tongue, and palate. Whole salivary flow rates were determined using the draining method before, during, and after brushing. Differences in salivary flow rates across time periods, and between conditions, were examined using paired samples t-tests applying a Holm-Bonferroni sequential procedure (pcorr < 0.0045). The relationship between tooth brushing and age with respect to maximum salivary flow rate increase was examined using Pearson's correlation coefficient (p < 0.05). Results/Conclusion Whole salivary flow rates increased during, and for up to 5 minutes following, tooth brushing in adults aged 60 years and older who were free of systemic disease. The salivary effects of MTB and ETB were not significantly different. A moderate, positive correlation was observed between tooth-brushing-related maximum salivary flow rate increase and age. PMID:29682540

Expression and effect of inhibition of aminopeptidase-A during nephrogenesis.

PubMed

Dijkman, Henry B P M; Assmann, Karel J M; Steenbergen, Eric J; Wetzels, Jack F M

2006-02-01

Aminopeptidase-A (APA) is a metalloprotease that cleaves N-terminal aspartyl and glutamyl residues from peptides. Its best-known substrate is angiotensin II (Ang II), the most active compound of the renin-angiotensin system (RAS). The RAS is involved in renal development. Most components of the RAS system are expressed in the developing kidney. Thus far, APA has not been studied in detail. In the present study we have evaluated the expression of APA at the protein, mRNA, and enzyme activity (EA) level in the kidney during nephrogenesis. Furthermore, we have studied the effect of inhibiting APA EA by injection of anti-APA antibodies into 1-day-old mice. APA expression was observed from the comma stage onwards, predominantly in the developing podocytes and brush borders of proximal tubular cells. Notably, APA was absent in the medulla or the renal arterioles. Inhibition of APA EA caused temporary podocyte foot-process effacement, suggesting a minimum role for APA during nephrogenesis.

Study of enzyme activities in the descending part of the duodenum in patients of duodenal ulcer.

PubMed

Fernandes, V L C; Bhasin, D K; Rana, S V

2006-03-01

The aim of this study was to measure the levels of lactase, sucrase, maltase, leucine amino peptidase and gamma-glutamyl transpeptidase in duodenum of Indian patients with duodenal ulcer. The effect of duodenum inflammation on these brush border enzymes has also been analysed in this study. Levels of lactase, sucrase, maltase, leucine amino peptidase and gamma-glutamyl transpeptidase were assessed in duodenal biopsies of 20 patients of duodenal ulcer and 20 non-ulcer dyspepsia. The duodenal biopsy specimens were also examined histopathologically for presence or absence of inflammation. There was no significant difference (p>0.05) in the activity of above mentioned enzyme levels in both the groups. Only levels of gamma-glutamyl transpeptidase were significantly decreased in patients of duodenal ulcer with duodenal inflammation. This study shows that only the levels of gamma-glutamyl transpeptidase were significantly decreased in patients of duodenal ulcer with inflammation but no change in duodenal enzymes due to duodenal ulcer as compared to non-ulcer dyspepsia.

Effect of andrographolide on cysteamine-induced duodenal ulcer in rats.

PubMed

Panneerselvam, Saranya; Arumugam, Geetha; Karthikeyan, Narmadha Selvamathy Selvaperumal Munis

2011-09-01

The aim of this study was to evaluate the gastroprotective efficacy of andrographolide isolated from Andrographis paniculata in rats induced with duodenal ulcers. Duodenal ulcers were induced by cysteamine administration in rats pretreated with 3 mg kg⁻¹ BW day⁻¹ of andrographolide for 30 days. Ulcer score, myeloperoxidase activity, TBARS level, GSH/GSSG ratio and enzyme antioxidants were measured in the duodenal tissue. Brush border and basolateral membranes were isolated to assay sucrase, maltase, alkaline phosphatase and total ATPases. Ulcer score was significantly minimised in rats pretreated with andrographolide. Elevation in myeloperoxidase and TBARS levels were found to be minimised significantly due to andrographolide treatment. Membrane-bound enzyme activities and the thiol redox status of glutathione were significantly maintained in duodenal mucosa of rats that received andrographolide. This study reveals that the major component of A. paniculata, andrographolide, has potent antiulcer properties that are most likely caused by minimising inflammatory changes, counteracting free radical formation and maintaining the thiol redox status in the duodenum.

The Diverse Forms of Lactose Intolerance and the Putative Linkage to Several Cancers

PubMed Central

Amiri, Mahdi; Diekmann, Lena; von Köckritz-Blickwede, Maren; Naim, Hassan Y.

2015-01-01

Lactase-phlorizin hydrolase (LPH) is a membrane glycoprotein and the only β-galactosidase of the brush border membrane of the intestinal epithelium. Besides active transcription, expression of the active LPH requires different maturation steps of the polypeptide through the secretory pathway, including N- and O-glycosylation, dimerization and proteolytic cleavage steps. The inability to digest lactose due to insufficient lactase activity results in gastrointestinal symptoms known as lactose intolerance. In this review, we will concentrate on the structural and functional features of LPH protein and summarize the cellular and molecular mechanism required for its maturation and trafficking. Then, different types of lactose intolerance are discussed, and the molecular aspects of lactase persistence/non-persistence phenotypes are investigated. Finally, we will review the literature focusing on the lactase persistence/non-persistence populations as a comparative model in order to determine the protective or adverse effects of milk and dairy foods on the incidence of colorectal, ovarian and prostate cancers. PMID:26343715

The Diverse Forms of Lactose Intolerance and the Putative Linkage to Several Cancers.

PubMed

Amiri, Mahdi; Diekmann, Lena; von Köckritz-Blickwede, Maren; Naim, Hassan Y

2015-08-28

Lactase-phlorizin hydrolase (LPH) is a membrane glycoprotein and the only β-galactosidase of the brush border membrane of the intestinal epithelium. Besides active transcription, expression of the active LPH requires different maturation steps of the polypeptide through the secretory pathway, including N- and O-glycosylation, dimerization and proteolytic cleavage steps. The inability to digest lactose due to insufficient lactase activity results in gastrointestinal symptoms known as lactose intolerance. In this review, we will concentrate on the structural and functional features of LPH protein and summarize the cellular and molecular mechanism required for its maturation and trafficking. Then, different types of lactose intolerance are discussed, and the molecular aspects of lactase persistence/non-persistence phenotypes are investigated. Finally, we will review the literature focusing on the lactase persistence/non-persistence populations as a comparative model in order to determine the protective or adverse effects of milk and dairy foods on the incidence of colorectal, ovarian and prostate cancers.

The Green Gut: Chlorophyll Degradation in the Gut of Spodoptera littoralis.

PubMed

Badgaa, Amarsanaa; Büchler, Rita; Wielsch, Natalie; Walde, Marie; Heintzmann, Rainer; Pauchet, Yannik; Svatos, Ales; Ploss, Kerstin; Boland, Wilhelm

2015-11-01

Chlorophylls, the most prominent natural pigments, are part of the daily diet of herbivorous insects. The spectrum of ingested and digested chlorophyll metabolites compares well to the pattern of early chlorophyll-degradation products in senescent plants. Intact chlorophyll is rapidly degraded by proteins in the front- and midgut. Unlike plants, insects convert both chlorophyll a and b into the corresponding catabolites. MALDI-TOF/MS imaging allowed monitoring the distribution of the chlorophyll catabolites along the gut of Spodoptera littoralis larvae. The chlorophyll degradation in the fore- and mid-gut is strongly pH dependent, and requires alkaline conditions. Using LC-MS/MS analysis we identified a lipocalin-type protein in the intestinal fluid of S. littoralis homolog to the chlorophyllide a binding protein from Bombyx mori. Widefield and high-resolution autofluorescence microscopy revealed that the brush border membranes are covered with the chlorophyllide binding protein tightly bound via its GPI-anchor to the gut membrane. A function in defense against gut microbes is discussed.

Identification of Escherichia coli F4ac-binding proteins in porcine milk fat globule membrane

PubMed Central

Novakovic, Predrag; Huang, Yanyun Y.; Lockerbie, Betty; Shahriar, Farshid; Kelly, John; Gordon, John R.; Middleton, Dorothy M.; Loewen, Matthew E.; Kidney, Beverly A.; Simko, Elemir

2015-01-01

F4ac-positive enterotoxigenic Escherichia coli (ETEC) must attach to the intestinal mucosa to cause diarrhea in piglets. Prevention of bacterial attachment to the intestinal mucosa is the most effective defense against ETEC-induced diarrhea. Porcine milk fat globule membranes (MFGM) were shown to be able to inhibit attachment of ETEC to the intestinal brush border; however, the specific components of porcine MFGM that inhibited attachment of ETEC to enterocytes were not identified. Accordingly, the purpose of this study was to identify F4ac-binding MFGM proteins by overlay Western blot and affinity chromatography. The proteome of porcine MFGM was characterized and the following F4ac-binding proteins were detected by overlay Western blot and affinity chromatography: lactadherin, butyrophilin, adipophilin, acyl-CoA synthetase 3, and fatty acid-binding protein 3. The biological function of these proteins was not investigated but it is possible that their interaction with F4ac fimbria interferes with bacterial attachment and colonization. PMID:25852227

Localization of a renal sodium-phosphate cotransporter gene to human chromosome 5q35

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kos, C.H.; Tenenhouse, H.S.; Tihy, F.

1994-01-01

Several Mendelian disorders of renal phosphate reabsorption, associated with hypophosphatemia and bone disease, have been described. These include X-linked hypophosphatemia (XLH), hereditary hypophosphatemic rickets with hypercalciuria, hypophosphatemic bone disease, and autosomal dominant and autosomal recessive hypophosphatemic rickets. The underlying mechanisms for renal phosphate wasting in these disorders remain unknown. The proximal tubule is the major site of renal phosphate reabsorption. Thus, mutations in genes that participate in the transepithelial transport of phosphate in this segment of the nephron may be responsible for these disorders. Recently, a cDNA encoding a renal proximal tubular, brush-border membrane Na[sup +]-phosphate cotransporter (NaP[sub i]-3) wasmore » cloned from human kidney cortex. As a first step in establishing whether mutations in the NaP[sub i]-3 gene are the cause of inherited disorders in phosphate homeostasis, the authors sought to determine its chromosomal localization. 9 refs., 1 fig.« less

A monoclonal IgM smooth muscle antibody reactive with fibroblast stress fibres produced by immunization with Treponema pallidum.

PubMed Central

Strugnell, R A; Underwood, J R; Clarke, F M; Pedersen, J S; Chalmers, P J; Faine, S; Toh, B H

1983-01-01

A monoclonal IgM smooth muscle antibody secreted by a hybrid (MMI-1) of mouse plasmacytoma NS-1 with spleen cells from mouse immunized with Treponema pallidum was detected by indirect immunofluorescence tests on frozen tissue sections and on acetone fixed monolayers of rat and human fibroblasts. The antibody did not react with acetone fixed smears of T. pallidum but reacted with smooth muscle fibres and with striations of skeletal and cardiac muscle. In non-muscle cells, the antibody stained liver in a 'polygonal' pattern, thymus with accentuated staining of the thymic medulla, renal glomeruli and the brush border and peritubular fibrils of renal tubules. In fibroblast monolayers, the antibody stained stress fibres in an interrupted pattern. Immunoblotting with muscle proteins and the antibody showed labelling of a 100K molecule. The cellular distribution of the mouse monoclonal antibody is similar to that obtained with anti-actin antibody suggesting that the corresponding antigen may be an actin binding protein. Images Fig. 3 PMID:6347470

Systemic and anti-neuronal auto-antibodies in patients with paraneoplastic neurological disease.

PubMed

Moll, J W; Hooijkaas, H; van Goorbergh, B C; Roos, L G; Henzen-Logmans, S C; Vecht, C J

1996-01-01

Sera from 23 patients with paraneoplastic disease of the central nervous system (PNS) were examined for the presence of anti-neuronal (anti-Hu, anti-Yo/PCA) and anti-Ri) and systemic auto-antibodies, including antibodies against DNA, centromeres, nRNP, Sm antigen, Scl-70, Ro(SS-A), La(SS-B), mitochondria, thyroid antigens, parietal calls, brush border antigen and rheumatoid factor. As controls, sera from 33 patients with small cell lung cancer, 33 with ovarian cancer and 7 with breast cancer and from 107 aged-matched healthy persons were used. Systemic auto-antibodies were found in 52% of patients with paraneoplastic neurological syndromes compared with only 16% (P = 0.001) in the control group with cancer only and 15% in the group of healthy controls. The relatively high percentage of systemic auto-antibodies in patients with PNS indicates that there is a genetic susceptibility to the development of auto-immune phenomena. This may provide an explanation for the relatively rare occurrence of PNS in patients with cancer.

Intestinal alkaline phosphatase detoxifies lipopolysaccharide and prevents inflammation in zebrafish in response to the gut microbiota.

PubMed

Bates, Jennifer M; Akerlund, Janie; Mittge, Erika; Guillemin, Karen

2007-12-13

Vertebrates harbor abundant lipopolysaccharide (LPS) in their gut microbiota. Alkaline phosphatases can dephosphorylate and detoxify the endotoxin component of LPS. Here, we show that expression of the zebrafish intestinal alkaline phosphatase (Iap), localized to the intestinal lumen brush border, is induced during establishment of the gut microbiota. Iap-deficient zebrafish are hypersensitive to LPS toxicity and exhibit the excessive intestinal neutrophil influx characteristic of wild-type zebrafish exposed to LPS. Both of these Iap mutant phenotypes are dependent on Myd88 and Tumor Necrosis Factor Receptor (Tnfr), proteins also involved in LPS sensitivity in mammals. When reared germ-free, the intestines of Iap-deficient zebrafish are devoid of neutrophils. Together, these findings demonstrate that the endogenous microbiota establish the normal homeostatic level of neutrophils in the zebrafish intestine through a process involving Iap, Myd88, and Tnfr. Thus, by preventing inflammatory responses, Iap plays a crucial role in promoting mucosal tolerance to resident gut bacteria.

Mechanisms involved in the intestinal digestion and absorption of dietary vitamin A.

PubMed

Harrison, E H; Hussain, M M

2001-05-01

Dietary retinyl esters are hydrolyzed in the intestine by the pancreatic enzyme, pancreatic triglyceride lipase (PTL), and intestinal brush border enzyme, phospholipase B. Recent work on the carboxylester lipase (CEL) knockout mouse suggests that CEL may not be involved in dietary retinyl ester digestion. The possible roles of the pancreatic lipase-related proteins (PLRP) 1 and 2 and other enzymes require further investigation. Unesterified retinol is taken up by the enterocytes, perhaps involving both diffusion and protein-mediated facilitated transport. Once in the cell, retinol is complexed with cellular retinol-binding protein type 2 (CRBP2) and the complex serves as a substrate for reesterification of the retinol by the enzyme lecithin:retinol acyltransferase (LRAT). Retinol not bound to CRBP2 is esterified by acyl-CoA acyltransferase (ARAT). The retinyl esters are incorporated into chylomicrons, intestinal lipoproteins that transport other dietary lipids such as triglycerides, phospholipids, and cholesterol. Chylomicrons containing newly absorbed retinyl esters are then secreted into the lymph.

Role of human placental apical membrane transporters in the efflux of glyburide, rosiglitazone, and metformin

PubMed Central

HEMAUER, Sarah J.; PATRIKEEVA, Svetlana L.; NANOVSKAYA, Tatiana N.; HANKINS, Gary D.V.; AHMED, Mahmoud S.

2010-01-01

Objective Substrates of placental efflux transporters could compete for a single transporter, which could result in an increase in the transfer of each substrate to the fetal circulation. Our aim was to determine the role of placental transporters in the biodisposition of oral hypoglycemic drugs that could be used as monotherapy or in combination therapy for gestational diabetes. Study design Inside-out brush border membrane vesicles from term placentas were used to determine the efflux of glyburide, rosiglitazone, and metformin by P-gp, Breast Cancer Resistance Protein (BCRP), and Multidrug Resistance Protein (MRP1). Results Glyburide was transported by MRP1 (43 ± 4%); BCRP (25 ± 5%); and P-gp (9 ± 5%). Rosiglitazone was transported predominantly by P-gp (71 ± 26%). Metformin was transported by P-gp (58 ± 20%) and BCRP (25 ± 14%). Conclusion Multiple placental transporters contribute to efflux of glyburide, rosiglitazone, and metformin. Administration of drug combinations could lead to their competition for efflux transporters. PMID:20350646

Thermomechanical Properties and Glass Dynamics of Polymer-Tethered Colloidal Particles and Films.

PubMed

Cang, Yu; Reuss, Anna N; Lee, Jaejun; Yan, Jiajun; Zhang, Jianan; Alonso-Redondo, Elena; Sainidou, Rebecca; Rembert, Pascal; Matyjaszewski, Krzysztof; Bockstaller, Michael R; Fytas, George

2017-11-14

Polymer-tethered colloidal particles (aka "particle brush materials") have attracted interest as a platform for innovative material technologies and as a model system to elucidate glass formation in complex structured media. In this contribution, Brillouin light scattering is used to sequentially evaluate the role of brush architecture on the dynamical properties of brush particles in both the individual and assembled (film) state. In the former state, the analysis reveals that brush-brush interactions as well as global chain relaxation sensitively depend on grafting density; i.e., more polymer-like behavior is observed in sparse brush systems. This is interpreted to be a consequence of more extensive chain entanglement. In contrast, the local relaxation of films does not depend on grafting density. The results highlight that relaxation processes in particle brush-based materials span a wider range of time and length scales as compared to linear chain polymers. Differentiation between relaxation on local and global scale is necessary to reveal the influence of molecular structure and connectivity on the aging behavior of these complex systems.

PRDA-2 and 3 Brush Seal Development Programs at EG and G

NASA Technical Reports Server (NTRS)

Loewenthal, Robert G.

1996-01-01

EG&G Mechanical Components Technology Group R&D completed a Brush Seal Development Program under PRDA-2 in late 1992. We started the Advanced Brush Seal Development program, under PRDA-3, in 1993 and will complete it in 1996. Both programs have been funded by the United States Air Force. In the first program, we made significant gains in the area of tribopairs (bristle materials vs. shaft coatings) and the 'Low Hysteresis' design for brush seals. These were reported in two AIAA Propulsion Conference papers, and the 'Low Hysteresis' design has been patented. Seals were delivered for test in an Air Force demonstrator at Allison. In PRDA-3, goals are to increase the pressure sealing capability, and the surface speeds and temperatures at which brush seals can be used. We have conducted part of the design and testing and have tested brush seals successfully at more severe conditions than in the previous program. We are continuing with the program, and will complete it in time to furnish brush seals for an Air Force Demonstrator test in 1997.

Assembly and Characterization of Well Defined High Molecular Weight Poly(p-phenylene) Polymer Brushes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Alonzo Calderon, Jose E; Kilbey, II, S Michael; Ankner, John Francis

2011-01-01

The assembly and characterization of well-defined, end-tethered poly(p-phenylene) (PPP) brushes having high molecular weight, low polydispersity and high 1,4-stereoregularity are presented. The PPP brushes are formed using a precursor route that relies on either self-assembly or spin coating of high molecular weight (degrees of polymerizations 54, 146, and 238) end-functionalized poly(1,3-cyclohexadiene) (PCHD) chains from benzene solutions onto silicon or quartz substrates, followed by aromatization of the end-attached PCHD chains on the surface. The approach allows the thickness (grafting density) of the brushes to be easily varied. The dry brushes before and after aromatization are characterized by ellipsometry, atomic force microscopy,more » grazing angle attenuated total reflectance Fourier transform infrared spectroscopy, and UV-Vis spectroscopy. The properties of the PPP brushes are compared with those of films made using oligo-paraphenylenes and with ab initio density functional theory simulations of optical properties. Our results suggest conversion to fully aromatized, end-tethered PPP polymer brushes having effective conjugation lengths of 5 phenyl units.« less

Endoscopic transpapillary brush cytology and forceps biopsy in patients with hilar cholangiocarcinoma.

PubMed

Weber, Andreas; von Weyhern, Claus; Fend, Falko; Schneider, Jochen; Neu, Bruno; Meining, Alexander; Weidenbach, Hans; Schmid, Roland M; Prinz, Christian

2008-02-21

To evaluate the sensitivity of brush cytology and forceps biopsy in a homogeneous patient group with hilar cholangiocarcinoma. Brush cytology and forceps biopsy were routinely performed in patients with suspected malignant biliary strictures. Fifty-eight consecutive patients undergoing endoscopic retrograde cholangio-pancreatography (ERCP) including forceps biopsy and brush cytology in patients with hilar cholangiocarcinoma between 1995-2005. Positive results for malignancy were obtained in 24/58 patients (41.4%) by brush cytology and in 31/58 patients (53.4%) by forceps biopsy. The combination of both techniques brush cytology and forceps biopsy resulted only in a minor increase in diagnostic sensitivity to 60.3% (35/58 patients). In 20/58 patients (34.5%), diagnosis were obtained by both positive cytology and positive histology, in 11/58 (19%) by positive histology (negative cytology) and only 4/58 patients (6.9%) were confirmed by positive cytology (negative histology). Brush cytology and forceps biopsy have only limited sensitivity for the diagnosis of malignant hilar tumors. In our eyes, additional diagnostic techniques should be evaluated and should become routine in patients with negative cytological and histological findings.

Nanoscale evaluation of lubricity on well-defined polymer brush surfaces using QCM-D and AFM.

PubMed

Kitano, Kazuhiko; Inoue, Yuuki; Matsuno, Ryosuke; Takai, Madoka; Ishihara, Kazuhiko

2009-11-01

For preparing a "highly lubricated biointerface", which has both excellent lubricity and biocompatibility, we investigated the factors responsible for resistance to friction during polymer grafting. We prepared poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC), poly(2-hydroxyethyl methacrylate) (PHEMA), and poly(methyl methacrylate) (PMMA) brush layers with high graft density and well-controlled thickness using atom transfer radical polymerization (ATRP). We measured the water absorptivity in the polymer brush layers and the viscoelasticity of the polymer-hydrated layers using a quartz crystal microbalance with dissipation monitoring (QCM-D) measurements. The PMPC brush layer had the highest water absorptivity, while the PMPC-hydrated layer had the highest fluidity. The friction properties of the polymer brush layers were determined in air, water, and toluene by atomic force microscopy (AFM). The friction on each polymer brush decreased only when a good solvent was chosen for each polymer. In conclusion, the brush layer possessing high water absorptivity and fluidity in water contributes to reduce friction. PMPC grafting is an effective and promising method for obtaining highly lubricated biointerfaces.

Polyelectrolyte brushes in mixed ionic medium studied via intermolecular forces

NASA Astrophysics Data System (ADS)

Farina, Robert; Laugel, Nicolas; Pincus, Philip; Tirrell, Matthew

2011-03-01

The vast uses and applications of polyelectrolyte brushes make them an attractive field of research especially with the growing interest in responsive materials. Polymers which respond via changes in temperature, pH, and ionic strength are increasingly being used for applications in drug delivery, chemical gating, etc. When polyelectrolyte brushes are found in either nature (e.g., surfaces of cartilage and mammalian lung interiors) or commercially (e.g., skin care products, shampoo, and surfaces of medical devices) they are always surrounded by mixed ionic medium. This makes the study of these brushes in varying ionic environments extremely relevant for both current and future potential applications. The polyelectrolyte brushes in this work are diblock co-polymers of poly-styrene sulfonate (N=420) and poly-t-butyl styrene (N=20) which tethers to a hydrophobic surface allowing for a purely thermodynamic study of the polyelectrolyte chains. Intermolecular forces between two brushes are measured using the SFA. As multi-valent concentrations are increased, the brushes collapse internally and form strong adhesion between one another after contact (properties not seen in a purely mono-valent environment).

Comparison of the clinical effects of white brand toothbrushes versus Vitis Suave®

PubMed Central

Segarra-Vidal, Marta; Lucas-Alcahuz, Eduardo; López-Roldán, Andrés; Gil-Loscos, Francisco; Alpiste-Illueca, Francisco

2015-01-01

Background There has been an increase in the use of white label manual toothbrushes and a greater increase in inquiries for discomfort of the gingiva and mucosa. Material and Methods A randomized, double-blind, cross-over clinical trial was made of four white brand toothbrushes versus a control brush (Vitis Suave®), with the recording of plaque index, bleeding upon probing, and gingival abrasions following utilization of the different brushes. Results All the brushes except Deliplus® were equally effective in terms of plaque removal (p<0.05). Vitis Suave® and Veckia® were the brushes associated to the greatest increase in minor abrasions (p<0.01), while Veckia®, Carrefour® and Deluxe® significantly increased the number of medium intensity abrasions (p<0.05). These brushes also increased the number of large abrasions, though statistical significance was not reached in this case. Conclusions The white brand brushes proved effective in controlling bacterial plaque, but were associated to more intense soft tissue abrasion. Key words:Gingival abrasions, manual tooth brushing, white brand, clinical effects. PMID:26535095

Covariates of Tooth-brushing Frequency in Low-income African Americans From Grades 5 to 8

PubMed Central

Koerber, A.; Graumlich, S.; Punwani, I.C.; Berbaum, M.L.; Burns, J.L.; Levy, S.R.; Cowell, J.M.; Flay, B.R.

2009-01-01

Purpose The purpose of this study was to examine tooth-brushing frequency in 575 urban and nearby suburban African American children as part of a comprehensive risk-reduction study for students at high risk for violence, drugs, school delinquency, and unsafe sexual behaviors to determine which covariates predicted tooth-brushing frequency. Methods Students were surveyed 5 times, from the beginning of grade 5 and the end of each year through grade 8, and parents were surveyed at the beginning of grade 5. Peer influence, importance of being liked, self-esteem, attitudes towards tooth-brushing, oral health knowledge, self-efficacy, parental attitudes, and other covariates were examined for the ability to predict self-reporting of tooth-brushing frequency. Results In the fifth grade, peer influence, the importance of being liked, and physical self-esteem were the significant predictors, and peer influence continued to predict tooth-brushing in the eighth grade. Oral health knowledge and parental influence were not significant. Conclusion Peer influence is an important factor in tooth-brushing behavior in metropolitan African American preadolescent children. PMID:17249434

Covariates of tooth-brushing frequency in low-income African Americans from grades 5 to 8.

PubMed

Koerber, A; Graumlich, S; Punwani, I C; Berbaum, M L; Burns, J L; Levy, S R; Cowell, J M; Flay, B R

2006-01-01

The purpose of this study was to examine tooth-brushing frequency in 575 urban and nearby suburban African American children as part of a comprehensive risk-reduction study for students at high risk for violence, drugs, school delinquency, and unsafe sexual behaviors to determine which covariates predicted tooth-brushing frequency. Students were surveyed 5 times, from the beginning of grade 5 and the end of each year through grade 8, and parents were surveyed at the beginning of grade 5. Peer influence, importance of being liked, self-esteem, attitudes towards tooth-brushing, oral health knowledge, self-efficacy, parental attitudes, and other covariates were examined for the ability to predict self-reporting of tooth-brushing frequency. In the fifth grade, peer influence, the importance of being liked, and physical self-esteem were the significant predictors, and peer influence continued to predict tooth-brushing in the eighth grade. Oral health knowledge and parental influence were not significant. Peer influence is an important factor in tooth-brushing behavior in metropolitan African American preadolescent children.

Magnetically Activated Self-Cleaning Membranes

DTIC Science & Technology

2010-06-01

potassium phthalimide was reacted with the brushes to attach the phthalimide group to the end of the brushes. Hydralization then yields a primary amine...Synthesis reaction For the first step of the Gabriel Synthesis, 1 g potassium phthalimide was dissolved into 20 mL of ethanol. This solution (4.5 mL...resulting polyacrylic acid (pAA) brushes 12 through an amide linkage. Grafting of pAA brushes was confirmed using two types of spectroscopy

Phase behavior of ternary polymer brushes

DOE PAGES

Simocko, Chester K.; Frischknecht, Amalie L.; Huber, Dale L.

2016-01-07

Ternary polymer brushes consisting of polystyrene, poly(methyl methacrylate), and poly(4-vinylpyridine) have been synthesized. These brushes laterally phase separate into several distinct phases and can be tailored by altering the relative polymer composition. Self-consistent field theory has been used to predict the phase diagram and model both the horizontal and vertical phase behavior of the polymer brushes. As a result, all phase behaviors observed experimentally correlate well with the theoretical model.

Single molecule spectroscopy reveals heterogeneous transport mechanisms for molecular ions in a polyelectrolyte polymer brush.

PubMed

Reznik, Carmen; Estillore, Nicel; Advincula, Rigoberto C; Landes, Christy F

2009-11-05

Single molecule polarization and fluorescence correlation spectroscopy were used to evaluate heterogeneous transport mechanisms of molecular ions within supported polyelectrolyte brushes. Modes of diffusive transport include periods of significantly restricted rotational motion, often maintained over tens of milliseconds; periods of fast molecular rotation; and occasional adsorption of fluorescent probe molecules in the brush. The studies reveal rapid switching between orientational states during each observed mode of motion. Through quantitative analysis of state occupation times, the rate constants for transitions from weakly associated to strongly associated states were extracted. Additionally, the pH dependence of the ion transport rates in the brush exhibits an abrupt, rather than continuous, trend. These single molecule studies demonstrate the presence of dynamic anisotropic interactions between the charged molecular probe and the polymer brush and provide experimental evidence of stimuli responsive switchable transport functionality in the polyelectrolyte brush.

Tandem catalysis for the preparation of cylindrical polypeptide brushes.

PubMed

Rhodes, Allison J; Deming, Timothy J

2012-11-28

Here, we report a method for synthesis of cylindrical copolypeptide brushes via N-carboxyanhydride (NCA) polymerization utilizing a new tandem catalysis approach that allows preparation of brushes with controlled segment lengths in a straightforward, one-pot procedure requiring no intermediate isolation or purification steps. To obtain high-density brush copolypeptides, we used a "grafting from" approach where alloc-α-aminoamide groups were installed onto the side chains of NCAs to serve as masked initiators. These groups were inert during cobalt-initiated NCA polymerization and gave allyloxycarbonyl-α-aminoamide-substituted polypeptide main chains. The alloc-α-aminoamide groups were then activated in situ using nickel to generate initiators for growth of side-chain brush segments. This use of stepwise tandem cobalt and nickel catalysis was found to be an efficient method for preparation of high-chain-density, cylindrical copolypeptide brushes, where both the main chains and side chains can be prepared with controlled segment lengths.

Real-time x-ray scattering study of the initial growth of organic crystals on polymer brushes

DOE Office of Scientific and Technical Information (OSTI.GOV)

An, Sung Yup; Ahn, Kwangseok; Kim, Doris Yangsoo

2014-04-21

We studied the early-stage growth structures of pentacene organic crystals grown on polymer brushes using real-time x-ray scattering techniques. In situ x-ray reflectivity and atomic force microscopy analyses revealed that at temperatures close to the glass transition temperature of polymer brush, the pentacene overlayer on a polymer brush film showed incomplete condensation and 3D island structures from the first monolayer. A growth model based on these observations was used to quantitatively analyze the real-time anti-Bragg x-ray scattering intensities measured during pentacene growth to obtain the time-dependent layer coverage of the individual pentacene monolayers. The extracted total coverage confirmed significant desorptionmore » and incomplete condensation in the pentacene films deposited on the polymer brushes. These effects are ascribed to the change in the surface viscoelasticity of the polymer brushes around the glass transition temperature.« less

Microsecond MD Simulations of Nano-patterned Polymer Brushes on Self-Assembled Monolayers

NASA Astrophysics Data System (ADS)

Buie, Creighton; Qiu, Liming; Cheng, Kwan; Park, Soyeun

2010-03-01

Nano-patterned polymer brushes end-grafted onto self-assembled monolayers have gained increasing research interests due to their unique thermodynamic properties and their chemical and biomedical applications in colloids, biosensing and tissue engineering. So far, the interactions between the polymer brushes with the surrounding environments such as the floor and solvent at the nanometer length scale and microsecond time scale are still difficult to obtained experimentally and computationally. Using a Coarse-Grained MD approach, polymer brushes of different monomeric lengths, grafting density and hydrophobicity of the monomers grafted on self-assembled monolayers and in explicit solvent were studied. Molecular level information, such as lateral diffusion, transverse height and volume contour of the brushes, were calculated from our microsecond-MD simulations. Our results demonstrated the significance of the hydration of the polymer in controlling the conformational arrangement of the polymer brushes.

In Situ Infrared Ellipsometry for Protein Adsorption Studies on Ultrathin Smart Polymer Brushes in Aqueous Environment

DOE PAGES

Kroning, Annika; Furchner, Andreas; Aulich, Dennis; ...

2015-02-10

The protein-adsorbing and -repelling properties of various smart nanometer-thin polymer brushes with high potential for biosensing and biomedical applications are studied by in-situ infrared-spectroscopic ellipsometry (IRSE). IRSE as a highly sensitive non-destructive technique allows us to investigate protein adsorption on polymer brushes in aqueous environment in dependence of external stimuli like temperature and pH. These stimuli are, for instance, relevant in switchable mixed brushes containing poly(N-isopropyl acrylamide) and poly(acrylic acid), respectively. We use such brushes as model surfaces for controlling protein adsorption of human serum albumin and human fibrinogen. IRSE can distinguish between polymer-specific vibrational bands, which yield insights intomore » the hydration state of the brushes, and changes in the protein-specific amide bands, which are related to changes of the protein secondary structure.« less

Importance of filament diameter when using bass brushing technique.

PubMed

Vowles, A D; Wade, A B

1977-08-01

A comparative study using a crossover experimental construction was made of the effectiveness of brushes containing 6/10 nylon filaments of 0.132 mm mean diameter using a Bass technique with those containing filaments of the same type of nylon but 0.280 mm diameter. Each type of brush was used for a 2-week period. Even though the finer filament brushes contained more than three times as many filaments, they were inferior in cleaning achievement to the brushes with the broader filaments. The difference was particularly marked on the facial aspect, but was largely nullified lingually. The effectiveness of the Bass technique in the gingival zone demonstrated in a previous investigation when brushes containing filaments of 0.18 and 0.20 mm were used, was not found in this investigation. It is concluded that filament diameter is critical in achieving effective cleaning using the Bass technique.

Influence of cutting time on brush response: implications for herbivory in linear (transportation) corridors.

PubMed

Rea, Roy V; Child, Kenneth N; Spata, David P; MacDonald, Douglas

2007-08-01

An experiment was conducted to determine the influence the time of brush-cutting can have on plant regrowth and attractiveness to herbivores that browse in linear corridors. The influence of cutting time on leaf flush and senescence, shoot morphometry, and biomass was measured for 3 consecutive years after initial brush-cutting. Results indicate that morphological and phenological attributes of three woody deciduous plants were influenced by the timing of brush-cutting for up to 3 years after initial cutting. Brush-cutting generally stimulated plants to produce larger than normal shoots and delay leaf senescence. The degree to which plants were affected, however, varied with the timing of initial cutting and the species in question. Generally, plants cut later in the year resprouted more vigorously and were taller in the third year after cutting but produced less overall biomass than when cut earlier. In the years following brush-cutting, plants cut earlier flushed leaves earlier in the spring but delayed leaf senescence in the fall when compared to uncut controls. Results of these trials suggest that brush-cutting time influences plant response and several plant attributes known to influence plant attractiveness to moose and other herbivores. We therefore recommend that roadside and railside vegetation management plans consider the influence of cutting time on plant regrowth. Such considerations can ensure that brush is cut to reduce the attractiveness of plant regrowth in these linear corridors, reduce the utilization of such brush by herbivores, and, as such, mitigate collision risk between motorists and herbivores such as moose.

Plaque-left-behind after brushing: intra-oral reservoir for antibacterial toothpaste ingredients.

PubMed

Otten, Marieke P T; Busscher, Henk J; Abbas, Frank; van der Mei, Henny C; van Hoogmoed, Chris G

2012-10-01

Plaque is never fully removed by brushing and may act as a reservoir for antibacterial ingredients, contributing to their substantive action. This study investigates the contribution of plaque-left-behind and saliva towards substantivity of three antibacterial toothpastes versus a control paste without antibacterial claims. First, volunteers brushed 2 weeks with a control or antibacterial toothpaste. Next, plaque and saliva samples were collected 6 and 12 h after brushing and bacterial concentrations and viabilities were measured. The contributions of plaque and saliva towards substantivity were determined by combining control plaques with experimental plaque or saliva samples and subsequently assessing their viabilities. Bacterial compositions in the various plaque and saliva samples were compared using denaturing gradient gel electrophoresis. The viabilities of plaques after brushing with Colgate-Total® and Crest-Pro-Health® were smaller than of control plaques and up to 12 h after brushing with Crest-Pro-Health® plaques still contained effective, residual antibacterial activity against control plaques. No effective, residual antibacterial activity could be measured in saliva samples after brushing. There was no significant difference in bacterial composition of plaque or saliva after brushing with the different toothpastes. Plaque-left-behind after mechanical cleaning contributes to the substantive action of an antibacterial toothpaste containing stannous fluoride (Crest-Pro-Health®). The absorptive capacity of plaque-left-behind after brushing is of utmost clinical importance, since plaque is predominantly left behind in places where its removal and effective killing matter most. Therewith this study demonstrates a clear and new beneficial effect of the use of antibacterial toothpastes.

A profilometry-based dentifrice abrasion Method for V8 brushing machines. Part I: Introduction to RDA-PE.

PubMed

White, Donald J; Schneiderman, Eva; Colón, Ellen; St John, Samuel

2015-01-01

This paper describes the development and standardization of a profilometry-based method for assessment of dentifrice abrasivity called Radioactive Dentin Abrasivity - Profilometry Equivalent (RDA-PE). Human dentine substrates are mounted in acrylic blocks of precise standardized dimensions, permitting mounting and brushing in V8 brushing machines. Dentin blocks are masked to create an area of "contact brushing." Brushing is carried out in V8 brushing machines and dentifrices are tested as slurries. An abrasive standard is prepared by diluting the ISO 11609 abrasivity reference calcium pyrophosphate abrasive into carboxymethyl cellulose/glycerin, just as in the RDA method. Following brushing, masked areas are removed and profilometric analysis is carried out on treated specimens. Assessments of average abrasion depth (contact or optical profilometry) are made. Inclusion of standard calcium pyrophosphate abrasive permits a direct RDA equivalent assessment of abrasion, which is characterized with profilometry as Depth test/Depth control x 100. Within the test, the maximum abrasivity standard of 250 can be created in situ simply by including a treatment group of standard abrasive with 2.5x number of brushing strokes. RDA-PE is enabled in large part by the availability of easy-to-use and well-standardized modern profilometers, but its use in V8 brushing machines is enabled by the unique specific conditions described herein. RDA-PE permits the evaluation of dentifrice abrasivity to dentin without the requirement of irradiated teeth and infrastructure for handling them. In direct comparisons, the RDA-PE method provides dentifrice abrasivity assessments comparable to the gold industry standard RDA technique.

Glucose monitoring using a polymer brush modified polypropylene hollow fiber-based hydraulic flow sensor.

PubMed

Fortin, Nicolas; Klok, Harm-Anton

2015-03-04

Tight regulation of blood glucose levels of diabetic patients requires durable and robust continuous glucose sensing schemes. This manuscript reports the fabrication of ultrathin, phenylboronic acid (PBA) functionalized polymer brushes that swell upon glucose binding and which were integrated as the sensing interface in a new polypropylene hollow fiber (PPHF)-based hydraulic flow glucose sensor prototype. The polymer brushes were prepared via surface-initiated atom transfer radical polymerization of sodium methacrylate followed by postpolymerization modification with 3-aminophenyl boronic acid. In a first series of experiments, the glucose-response of PBA-functionalized poly(methacrylic acid) (PMAA) brushes grafted from planar silicon surfaces was investigated by quartz crystal microbalance with dissipation (QCM-D) and atomic force microscopy (AFM) experiments. The QCM-D experiments revealed a more or less linear change of the frequency shift for glucose concentrations up to ∼10 mM and demonstrated that glucose binding was completely reversible for up to seven switching cycles. The AFM experiments indicated that glucose binding was accompanied by an increase in the film thickness of the PBA functionalized PMAA brushes. The PBA functionalized PMAA brushes were subsequently grafted from the surface of PPHF membranes. The hydraulic permeability of these porous fibers depends on the thickness and swelling of the PMAA brush coating. PBA functionalized brush-coated PPHFs showed a decrease in flux upon exposure to glucose, which is consistent with swelling of the brush coating. Because they avoid the use of enzymes and do not rely on an electrochemical transduction scheme, these PPHF-based hydraulic flow sensors could represent an interesting alternative class of continuous glucose sensors.

Prebrushing rinse with water on plaque removal: a split-mouth design.

PubMed

Van der Sluijs, E; Slot, D E; Hennequin-Hoenderdos, N L; Van Leeuwen, Mpc; Van der Weijden, G A

2017-11-01

The aim was to evaluate whether there is an additional beneficial effect on dental plaque removal of rinsing the oral cavity with water before toothbrushing. In total, 48 non-dental, systemically healthy participants ≥18 years were included in this randomized controlled clinical trial using a split-mouth design. The participants were requested to refrain from any form of oral hygiene for 48 h. First dental plaque scores (PI) were assessed full mouth. Two randomly assigned contra-lateral quadrants were brushed. Next the participant rinsed for 1 min with 15 ml water. Subsequently, the opposite two contra-lateral quadrants were brushed. Brushing was performed without toothpaste. Subsequently the second full-mouth PI assessment was performed. The brushing and rinsing procedure was performed under supervision and brushing time was tracked by a timer, each quadrant was brushed for 30 s. For the buccal, lingual, and approximal surfaces and tooth type, a subanalysis was performed. At baseline there was no statistically significantly difference between the two sets of contra-lateral quadrants. When a water rinse was used before toothbrushing the PI-score was reduced by 58%. If water rinse was used post-brushing the PI-score reduced by 57%. The difference of 0.04 in mean plaque index score reduction between the two brushing regimens was not significant(P = 0.162). When a 2 min brushing exercise was performed, on average more than 55% dental plaque was removed. Prerinsing with water did not contribute significantly to toothbrush efficacy. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Stages and transitions in the development of tooth brushing skills in children of Mexican immigrant families: a qualitative study.

PubMed

Benadof, Dafna; Polk, Deborah; Documet, Patricia

2015-01-01

Compared with white children, the oral health of Latino children in the United States is much worse. One factor contributing to oral health is tooth brushing. Few studies have addressed the formation of the tooth brushing habit in children, and only one of them studied a Latino population. The purpose of this study is to explore the development of the tooth brushing habit in children of Mexican immigrant families and develop hypothesis based on its results. This is an exploratory qualitative study, with a case study design based on 20 in-depth interviews. Participants were Mexican immigrant mothers living in Pittsburgh and Philadelphia, PA. Participants had at least one child six-years-old or younger. Interviews were recorded, transcribed verbatim, and analyzed using qualitative analysis procedures. Four stages were identified in the tooth brushing learning process: initiation and entirely dependent tooth brushing, assisted tooth brushing, road to tooth brushing independence, and independent tooth brushing. Two factors influenced parents' teaching approaches: parents' perceptions of their child's achievement of physical, cognitive, and motor developmental milestones and parents' knowledge about oral hygiene. We identified four distinct stages and found evidence to hypothesize that transitions from one stage to the next are triggered not by the age of the child but by parents' knowledge about oral hygiene and their perceptions of their child's achievement of physical, cognitive, and motor developmental milestones. Future quantitative research studies should be conducted to test this hypothesis in larger groups of Latinos as well as other ethnic groups. © 2015 American Association of Public Health Dentistry.

Unique Tuft Test Facility Dramatically Reduces Brush Seal Development Costs

NASA Technical Reports Server (NTRS)

Fellenstein, James A.

1997-01-01

Brush seals have been incorporated in the latest turbine engines to reduce leakage and improve efficiency. However, the life of these seals is limited by wear. Studies have shown that optimal sealing characteristics for a brush seal occur before the interference fit between the brush and shaft is excessively worn. Research to develop improved tribopairs (brush and coating) with reduced wear and lower friction has been hindered by the lack of an accurate, low-cost, efficient test methodology. Estimated costs for evaluating a new material combination in an engine company seal test program are on the order of $100,000. To address this need, the NASA Lewis Research Center designed, built, and validated a unique, innovative brush seal tuft tester that slides a single tuft of brush seal wire against a rotating shaft under controlled loads, speeds, and temperatures comparable to those in turbine engines. As an initial screening tool, the brush seal tuft tester can tribologicaly evaluate candidate seal materials for 1/10th the cost of full-scale seal tests. Previous to the development of the brush seal tuft tester facility, most relevant tribological data had been obtained from full-scale seal tests conducted primarily to determine seal leakage characteristics. However, from a tribological point of view, these tests included the confounding effects of varying contact pressures, bristle flaring, high-temperature oxidation, and varying bristle contact angles. These confounding effects are overcome in tuft testing. The interface contact pressures can be either constant or varying depending on the tuft mounting device, and bristle wear can be measured optically with inscribed witness marks. In a recent cooperative program with a U.S. turbine engine manufacturer, five metallic wire candidates were tested against a plasma-sprayed Nichrome-bonded chrome carbide. The wire materials used during this collaboration were either nickel-chrome- or cobaltchrome-based superalloys. These tests corroborated full-scale seal test results and provided insight into previously untested combinations. As the cycle temperature for improved efficiency turbine engines increases, new brush seal materials combinations must be considered. Future brush seal tuft testing will include both metallic and ceramic bristles versus commercial and NASA-developed shaft coatings. The ultimate goal of this work is to expand the current data base so that seal designers can tailor brush seal materials to specific applications.

Lamellipodin and the Scar/WAVE complex cooperate to promote cell migration in vivo

PubMed Central

Law, Ah-Lai; Vehlow, Anne; Kotini, Maria; Dodgson, Lauren; Soong, Daniel; Theveneau, Eric; Bodo, Cristian; Taylor, Eleanor; Navarro, Christel; Perera, Upamali; Michael, Magdalene; Dunn, Graham A.; Bennett, Daimark; Mayor, Roberto

2013-01-01

Cell migration is essential for development, but its deregulation causes metastasis. The Scar/WAVE complex is absolutely required for lamellipodia and is a key effector in cell migration, but its regulation in vivo is enigmatic. Lamellipodin (Lpd) controls lamellipodium formation through an unknown mechanism. Here, we report that Lpd directly binds active Rac, which regulates a direct interaction between Lpd and the Scar/WAVE complex via Abi. Consequently, Lpd controls lamellipodium size, cell migration speed, and persistence via Scar/WAVE in vitro. Moreover, Lpd knockout mice display defective pigmentation because fewer migrating neural crest-derived melanoblasts reach their target during development. Consistently, Lpd regulates mesenchymal neural crest cell migration cell autonomously in Xenopus laevis via the Scar/WAVE complex. Further, Lpd’s Drosophila melanogaster orthologue Pico binds Scar, and both regulate collective epithelial border cell migration. Pico also controls directed cell protrusions of border cell clusters in a Scar-dependent manner. Taken together, Lpd is an essential, evolutionary conserved regulator of the Scar/WAVE complex during cell migration in vivo. PMID:24247431

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sun, Liang; Akgun, Bulent; Narayanan, Suresh

Swollen polymer brushes are found in many systems where the brush is intended to mediate interactions with the surroundings. The surface height fluctuations of planar polystyrene brushes (0.04 – 0.63 chains/nm2) highly swollen in toluene vapor are so strongly slowed by the tethering of the chains that they are unobservable in the current experimental window of length and time. This is the case despite the fact that the segmental dynamics of the brush chains should be very fast due to the substantial plasticization by the solvent. With respect to thermally stimulated fluctuations, the surfaces of these swollen brushes are solid-likemore » on time scales and length scales pertinent to many practical applications.« less

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.

Autonomous multifunctional nanobrushes-autonomous materials

NASA Astrophysics Data System (ADS)

Ghasemi-Nejhad, Mehrdad N.; Tius, Marcus A.

2007-04-01

In this work, taking advantage of carbon nanotubes' small size, and exceptional mechanical, chemical and electrical properties, we report on a series of nano-synthesis procedures that combine conventional chemical vapor deposition and selective substrate area growth followed by chemical functionalizations to fabricate functionalized nano-brushes from aligned carbon nanotube arrays and chemically selective functional groups. The high aspect ratio and small dimension, mechanical stability and flexibility, surface chemical and adhesive characteristics of carbon nanotubes provide opportunities to create nano-brushes with selected chemical functionalities. The nano-brushes are made from aligned multi-walled carbon nanotube bristles grafted onto long SiC fiber handles in various configurations and functionalized with various chemical functional groups. These nano-brushes can easily be manipulated physically, either manually or with the aid of motors. Here, we explain the autonomous characteristics of the functionalized nano-brushes employing functional chemical groups such that the nano-brush can potentially collect various metal particles, ions, and contaminants from liquid solutions and the air environment, autonomously. These functionalized multiwalled carbon nanotube based nano-brushes can work swiftly in both liquid and air environments. With surface modification and functionalization, the nanotube nano-brushes can potentially become a versatile nano-devices in many chemical and biological applications, where they can autonomously pick up the particles they encounter since they can be chemically programmed to function as Autonomous Chemical Nano Robots (ACNR).

Spectroscopic study of a DNA brush synthesized in situ by surface initiated enzymatic polymerization.

PubMed

Khan, M Nuruzzaman; Tjong, Vinalia; Chilkoti, Ashutosh; Zharnikov, Michael

2013-08-29

We used a combination of synchrotron-based X-ray photoelectron spectroscopy (XPS) and angle-resolved near-edge X-ray absorption fine structure (NEXAFS) spectroscopy to study the chemical integrity, purity, and possible internal alignment of single-strand (ss) adenine deoxynucleotide (poly(A)) DNA brushes. The brushes were synthesized by surface-initiated enzymatic polymerization (SIEP) on a 25-mer of adenine self-assembled monolayer (SAM) on gold (A25-SH), wherein the terminal 3'-OH of the A25-SH serve as the initiation sites for SIEP of poly(A). XPS and NEXAFS spectra of poly(A) brushes were found to be almost identical to those of A25-SH initiator, with no unambiguous traces of contamination. Apart from the well-defined chemical integrity and contamination-free character, the brushes were found to have a high degree of orientational order, with an upright orientation of individual strands, despite their large thickness up to ~55 nm, that corresponds to a chain length of at least several hundred nucleotides for individual ssDNA molecules. The orientational order exhibited by these poly(A) DNA brushes, mediated presumably by base stacking, was found to be independent of the brush thickness as long as the packing density was high enough. The well-defined character and orientational ordering of the ssDNA brushes make them a potentially promising system for different applications.

The dynamics of complex formation between amylose brushes on gold and fatty acids by QCM-D.

PubMed

Cao, Zheng; Tsoufis, Theodoros; Svaldo-Lanero, Tiziana; Duwez, Anne-Sophie; Rudolf, Petra; Loos, Katja

2013-10-14

Amylose brushes were synthesized by enzymatic polymerization with glucose-1-phosphate as monomer and rabbit muscle phosphorylase b as catalyst on gold-covered surfaces of a quartz crystal microbalance. Fourier transform infrared (FT-IR) spectra confirmed the presence of the characteristic absorption peaks of amylose between 3100 cm(-1) and 3500 cm(-1). The thickness of the amylose brushes-measured by Spectroscopic Ellipsometry--can be tailored from 4 to 20 nm, depending on the reaction time. The contour length of the stretched amylose chains on gold surfaces has been evaluated by single molecule force spectroscopy, and a total chain length of about 20 nm for 16.2 nm thick amylose brushes was estimated. X-ray photoelectron spectroscopy (XPS) was employed to characterize the amylose brushes before and after the adsorption of fatty acids. The dynamics of inclusion complex formation between amylose brushes and two fatty acids (octanoic acid and myristic acid) with different chain length was investigated as a function of time using a quartz crystal microbalance with dissipation monitoring (QCM-D) immersed in the liquid phase. QCM-D signals including the frequency and dissipation shifts elucidated the effects of the fatty acid concentration, the solvent types, the chain length of the fatty acids and the thickness of the amylose brushes on the dynamics of fatty acid molecule adsorption on the amylose brush-modified sensor surfaces.

Mechanism of induction of fibroblast to corneal endothelial cell.

PubMed

Jiang, Yan; Fu, Wei-Cai; Zhang, Lin

2014-08-01

To explore mechanism of nduction of fibroblast to corneal endothelial cell. Rabbit conjunctiva fibroblasts were used as feeder cells, rabbit oral mucosa epithelial cells were used as seed cells, and human denuded amniotic membrane was used as carrier to establish tissue engineering corneal endothelium. The transformation effect was observed. As concentration of mitomycin C increased, cell survival rate gradually decreased, cell proliferation was obviously inhibited when concentration≥25 μg/mL; 5 days after being treated by 5 μg/mL mitomycin C, cell body was enlarged and extended without cell fusion, however after being treated by 0.5 μg/mL mitomycin C, cell body was significantly proliferated and gradually fused; after 3 weeks of culture, stratified epithelium appeared on rabbit oral mucosa epithelial cells, differentiation layers were 4-5 and were well differentiated, the morphology was similar to corneal endothelial cells; Under electron microscope, surface layer of cells were polygonal, tightly connected to another with microvilli on the border, there was hemidesmosome between basal cells and human denuded amniotic membrane. Fibroblast cells have the potential of multi-directional differentiation, effective induction can promote emergence of intercellular desmosomes between seed cells and emergence of epithelial surface microvilli, and differentiate to the corneal endothelial cell. However, clinical application still needs more research and safety evaluation. Copyright © 2014 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

Efficacy of plaque removal and learning effect of a powered and a manual toothbrush.

PubMed

Lazarescu, D; Boccaneala, S; Illiescu, A; De Boever, J A

2003-08-01

Subjects with high plaque and gingivitis scores can profit most from the introduction of new manual or powered tooth brushes. To improve their hygiene, not only the technical characteristics of new brushes but also the learning effect in efficient handling are of importance. : The present study compared the efficacy in plaque removal of an electric and a manual toothbrush in a general population and analysed the learning effect in efficient handling. Eighty healthy subjects, unfamiliar with electric brushes, were divided into two groups: group 1 used the Philips/Jordan HP 735 powered brush and group 2 used a manual brush, Oral-B40+. Plaque index (PI) and gingival bleeding index (GBI) were assessed at baseline and at weeks 3, 6, 12 and 18. After each evaluation, patients abstained from oral hygiene for 24 h. The next day a 3-min supervised brushing was performed. Before and after this brushing, PI was assessed for the estimation of the individual learning effect. The study was single blinded. Over the 18-week period, PI reduced gradually and statistically significantly (p<0.001) in group 1 from 2.9 (+/-0.38) to 1.5 (+/-0.24) and in group 2 from 2.9 (+/-0.34) to 2.2 (+/-0.23). From week 3 onwards, the difference between groups was statistically significant (p<0.001). The bleeding index decreased in group 1 from 28% (+/-17%) to 7% (+/-5%) (p<0.001) and in group 2 from 30% (+/-12%) to 12% (+/-6%) (p<0.001). The difference between groups was statistically significant (p<0.001) from week 6 onwards. The learning effect, expressed as the percentage of plaque reduction after 3 min of supervised brushing, was 33% for group 1 and 26% for group 2 at week 0. This percentage increased at week 18 to 64% in group 1 and 44% in group 2 (difference between groups statistically significant: p<0.001). The powered brush was significantly more efficient in removing plaque and improving gingival health than the manual brush in the group of subjects unfamiliar with electric brushes. There was also a significant learning effect that was more pronounced with the electric toothbrush.