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Sample records for cell wall anchor

  1. Restrictive glycosylphosphatidylinositol anchor synthesis in cwh6/gpi3 yeast cells causes aberrant biogenesis of cell wall proteins.

    OpenAIRE

    Vossen, J.H.; Müller, W. H.; Lipke, P N; Klis, F. M.

    1997-01-01

    We previously reported that the defects in the Saccharomyces cerevisiae cwh6 Calcofluor white-hypersensitive cell wall mutant are caused by a mutation in SPT14/GPI3, a gene involved in glycosylphosphatidylinositol (GPI) anchor biosynthesis. Here we describe the effect of cwh6/spt14/gpi3 on the biogenesis of cell wall proteins. It was found that the release of precursors of cell wall proteins from the endoplasmic reticulum (ER) was retarded. This was accompanied by proliferation of ER structur...

  2. Comparison of cell wall proteins of Saccharomyces cerevisiae as anchors for cell surface expression of heterologous proteins.

    OpenAIRE

    van der Vaart, J. M.; te Biesebeke, R; Chapman, J.W.; Toschka, H Y; Klis, F M; Verrips, C. T.

    1997-01-01

    The carboxyl-terminal regions of five cell wall proteins (Cwp1p, Cwp2p, Ag alpha 1p, Tip1p, and Flo1p) and three potential cell wall proteins (Sed1p, YCR89w, and Tir1p) all proved capable of immobilizing alpha-galactosidase in the cell wall of Saccharomyces cerevisiae. The fraction of the total amount of fusion protein that was localized to the cell wall varied depending on the anchor domain used. The highest proportion of cell wall incorporation was achieved with Cwp2p, Ag alpha 1p, or Sed1p...

  3. Cell wall antibiotics provoke accumulation of anchored mCherry in the cross wall of Staphylococcus aureus.

    Directory of Open Access Journals (Sweden)

    Wenqi Yu

    Full Text Available A fluorescence microscopy method to directly follow the localization of defined proteins in Staphylococcus was hampered by the unstable fluorescence of fluorescent proteins. Here, we constructed plasmid (pCX encoded red fluorescence (RF mCherry (mCh hybrids, namely mCh-cyto (no signal peptide and no sorting sequence, mCh-sec (with signal peptide, and mCh-cw (with signal peptide and cell wall sorting sequence. The S. aureus clones targeted mCh-fusion proteins into the cytosol, the supernatant and the cell envelope respectively; in all cases mCherry exhibited bright fluorescence. In staphylococci two types of signal peptides (SP can be distinguished: the +YSIRK motif SP(lip and the -YSIRK motif SP(sasF. mCh-hybrids supplied with the +YSIRK motif SP(lip were always expressed higher than those with -YSIRK motif SP(sasF. To study the location of the anchoring process and also the influence of SP type, mCh-cw was supplied on the one hand with +YSIRK motif (mCh-cw1 and the other hand with -YSIRK motif (mCh-cw2. MCh-cw1 preferentially localized at the cross wall, while mCh-cw2 preferentially localized at the peripheral wall. Interestingly, when treated with sub-lethal concentrations of penicillin or moenomycin, both mCh-cw1 and mCh-cw2 were concentrated at the cross wall. The shift from the peripheral wall to the cross wall required Sortase A (SrtA, as in the srtA mutant this effect was blunted. The effect is most likely due to antibiotic mediated increase of free anchoring sites (Lipid II at the cross wall, the substrate of SrtA, leading to a preferential incorporation of anchored proteins at the cross wall.

  4. Cell Wall Anchoring of the Campylobacter Antigens to Lactococcus lactis.

    Science.gov (United States)

    Kobierecka, Patrycja A; Olech, Barbara; Książek, Monika; Derlatka, Katarzyna; Adamska, Iwona; Majewski, Paweł M; Jagusztyn-Krynicka, Elżbieta K; Wyszyńska, Agnieszka K

    2016-01-01

    Campylobacter jejuni is the most frequent cause of human food-borne gastroenteritis and chicken meat is the main source of infection. Recent studies showed that broiler chicken immunization against Campylobacter should be the most efficient way to lower the number of human infections by this pathogen. Induction of the mucosal immune system after oral antigen administration should provide protective immunity to chickens. In this work we tested the usefulness of Lactococcus lactis, the most extensively studied lactic acid bacterium, as a delivery vector for Campylobacter antigens. First we constructed hybrid protein - CjaA antigen presenting CjaD peptide epitopes on its surface. We showed that specific rabbit anti-rCjaAD serum reacted strongly with both CjaA and CjaD produced by a wild type C. jejuni strain. Next, rCjaAD and CjaA were fused to the C-terminus of the L. lactis YndF containing the LPTXG motif. The genes expressing these proteins were transcribed under control of the L. lactis Usp45 promoter and their products contain the Usp45 signal sequences. This strategy ensures a cell surface location of both analyzed proteins, which was confirmed by immunofluorescence assay. In order to evaluate the impact of antigen location on vaccine prototype efficacy, a L. lactis strain producing cytoplasm-located rCjaAD was also generated. Animal experiments showed a decrease of Campylobacter cecal load in vaccinated birds as compared with the control group and showed that the L. lactis harboring the surface-exposed rCjaAD antigen afforded greater protection than the L. lactis producing cytoplasm-located rCjaAD. To the best of our knowledge, this is the first attempt to employ Lactic Acid Bacteria (LAB) strains as a mucosal delivery vehicle for chicken immunization. Although the observed reduction of chicken colonization by Campylobacter resulting from vaccination was rather moderate, the experiments showed that LAB strains can be considered as an alternative vector to

  5. Sortase A substrate specificity in GBS pilus 2a cell wall anchoring.

    Directory of Open Access Journals (Sweden)

    Francesca Necchi

    Full Text Available Streptococcus agalactiae, also referred to as Group B Streptococcus (GBS, is one of the most common causes of life-threatening bacterial infections in infants. In recent years cell surface pili have been identified in several Gram-positive bacteria, including GBS, as important virulence factors and promising vaccine candidates. In GBS, three structurally distinct types of pili have been discovered (pilus 1, 2a and 2b, whose structural subunits are assembled in high-molecular weight polymers by specific class C sortases. In addition, the highly conserved housekeeping sortase A (SrtA, whose main role is to link surface proteins to bacterial cell wall peptidoglycan by a transpeptidation reaction, is also involved in pili cell wall anchoring in many bacteria. Through in vivo mutagenesis, we demonstrate that the LPXTG sorting signal of the minor ancillary protein (AP2 is essential for pilus 2a anchoring. We successfully produced a highly purified recombinant SrtA (SrtA(ΔN40 able to specifically hydrolyze the sorting signal of pilus 2a minor ancillary protein (AP2-2a and catalyze in vitro the transpeptidation reaction between peptidoglycan analogues and the LPXTG motif, using both synthetic fluorescent peptides and recombinant proteins. By contrast, SrtA(ΔN40 does not catalyze the transpeptidation reaction with substrate-peptides mimicking sorting signals of the other pilus 2a subunits (the backbone protein and the major ancillary protein. Thus, our results add further insight into the proposed model of GBS pilus 2a assembly, in which SrtA is required for pili cell wall covalent attachment, acting exclusively on the minor accessory pilin, representing the terminal subunit located at the base of the pilus.

  6. Comparison of cell wall proteins of Saccharomyces cerevisiae as anchors for cell surface expression of heterologous proteins.

    Science.gov (United States)

    Van der Vaart, J M; te Biesebeke, R; Chapman, J W; Toschka, H Y; Klis, F M; Verrips, C T

    1997-01-01

    The carboxyl-terminal regions of five cell wall proteins (Cwp1p, Cwp2p, Ag alpha 1p, Tip1p, and Flo1p) and three potential cell wall proteins (Sed1p, YCR89w, and Tir1p) all proved capable of immobilizing alpha-galactosidase in the cell wall of Saccharomyces cerevisiae. The fraction of the total amount of fusion protein that was localized to the cell wall varied depending on the anchor domain used. The highest proportion of cell wall incorporation was achieved with Cwp2p, Ag alpha 1p, or Sed1p as an anchor. Although 80% of these fusion proteins were incorporated in the cell wall, the total production of alpha-galactosidase-Ag alpha 1p was sixfold lower than that of alpha-galactosidase-Cwp2p and eightfold lower than that of alpha-galactosidase-Sed1p. Differences in mRNA levels were not responsible for this discrepancy, nor was an intracellular accumulation of alpha-galactosidase-Ag alpha 1p detectable. A lower translation efficiency of the alpha-galactosidase-AG alpha 1 fusion construct is most likely to be responsible for the low level of protein production. alpha-Galactosidase immobilized by the carboxyl-terminal 67 amino acids of Cwp2p was most effective in the hydrolysis of the high-molecular-weight substrate guar gum from Cyamopsis tetragonoloba. This indicates that the use of a large anchoring domain does not necessarily result in a better exposure of the immobilized enzyme to the exterior of the yeast cell. PMID:9023939

  7. The role of Listeria monocytogenes cell wall surface anchor protein LapB in virulence, adherence, and intracellular replication

    Science.gov (United States)

    Lmof2365_2117 is a Listeria monocytogenes putative cell wall surface anchor protein with a conserved domain found in collagen binding proteins. We constructed a deletion mutation in lmof2365_2117 in serotype 4b strain F2365, evaluated its virulence, and determined its ability to adhere and invade co...

  8. Characterisation of a cell wall-anchored protein of Staphylococcus saprophyticus associated with linoleic acid resistance

    Directory of Open Access Journals (Sweden)

    King Nathan P

    2012-01-01

    Full Text Available Abstract Background The Gram-positive bacterium Staphylococcus saprophyticus is the second most frequent causative agent of community-acquired urinary tract infections (UTI, accounting for up to 20% of cases. A common feature of staphylococci is colonisation of the human skin. This involves survival against innate immune defenses including antibacterial unsaturated free fatty acids such as linoleic acid which act by disrupting bacterial cell membranes. Indeed, S. saprophyticus UTI is usually preceded by perineal skin colonisation. Results In this study we identified a previously undescribed 73.5 kDa cell wall-anchored protein of S. saprophyticus, encoded on plasmid pSSAP2 of strain MS1146, which we termed S. saprophyticus surface protein F (SssF. The sssF gene is highly prevalent in S. saprophyticus clinical isolates and we demonstrate that the SssF protein is expressed at the cell surface. However, unlike all other characterised cell wall-anchored proteins of S. saprophyticus, we were unable to demonstrate a role for SssF in adhesion. SssF shares moderate sequence identity to a surface protein of Staphylococcus aureus (SasF recently shown to be an important mediator of linoleic acid resistance. Using a heterologous complementation approach in a S. aureus sasF null genetic background, we demonstrate that SssF is associated with resistance to linoleic acid. We also show that S. saprophyticus strains lacking sssF are more sensitive to linoleic acid than those that possess it. Every staphylococcal genome sequenced to date encodes SssF and SasF homologues. Proteins in this family share similar predicted secondary structures consisting almost exclusively of α-helices in a probable coiled-coil formation. Conclusions Our data indicate that SssF is a newly described and highly prevalent surface-localised protein of S. saprophyticus that contributes to resistance against the antibacterial effects of linoleic acid. SssF is a member of a protein family

  9. PbsP, a cell wall-anchored protein that binds plasminogen to promote hematogenous dissemination of group B Streptococcus.

    Science.gov (United States)

    Buscetta, Marco; Firon, Arnaud; Pietrocola, Giampiero; Biondo, Carmelo; Mancuso, Giuseppe; Midiri, Angelina; Romeo, Letizia; Galbo, Roberta; Venza, Mario; Venza, Isabella; Kaminski, Pierre-Alexandre; Gominet, Myriam; Teti, Giuseppe; Speziale, Pietro; Trieu-Cuot, Patrick; Beninati, Concetta

    2016-07-01

    Streptococcus agalactiae (Group B Streptococcus or GBS) is a leading cause of invasive infections in neonates whose virulence is dependent on its ability to interact with cells and host components. We here characterized a surface protein with a critical function in GBS pathophysiology. This adhesin, designated PbsP, possesses two Streptococcal Surface Repeat domains, a methionine and lysine-rich region, and a LPXTG cell wall-anchoring motif. PbsP mediates plasminogen (Plg) binding both in vitro and in vivo and we showed that cell surface-bound Plg can be activated into plasmin by tissue plasminogen activator to increase the bacterial extracellular proteolytic activity. Absence of PbsP results in a decreased bacterial transmigration across brain endothelial cells and impaired virulence in a murine model of infection. PbsP is conserved among the main GBS lineages and is a major plasminogen adhesin in non-CC17 GBS strains. Importantly, immunization of mice with recombinant PbsP confers protective immunity. Our results indicate that GBS have evolved different strategies to recruit Plg which indicates that the ability to acquire cell surface proteolytic activity is essential for the invasiveness of this bacterium. PMID:26888569

  10. Role of the Group B antigen of Streptococcus agalactiae: a peptidoglycan-anchored polysaccharide involved in cell wall biogenesis.

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    Élise Caliot

    Full Text Available Streptococcus agalactiae (Group B streptococcus, GBS is a leading cause of infections in neonates and an emerging pathogen in adults. The Lancefield Group B carbohydrate (GBC is a peptidoglycan-anchored antigen that defines this species as a Group B Streptococcus. Despite earlier immunological and biochemical characterizations, the function of this abundant glycopolymer has never been addressed experimentally. Here, we inactivated the gene gbcO encoding a putative UDP-N-acetylglucosamine-1-phosphate:lipid phosphate transferase thought to catalyze the first step of GBC synthesis. Indeed, the gbcO mutant was unable to synthesize the GBC polymer, and displayed an important growth defect in vitro. Electron microscopy study of the GBC-depleted strain of S. agalactiae revealed a series of growth-related abnormalities: random placement of septa, defective cell division and separation processes, and aberrant cell morphology. Furthermore, vancomycin labeling and peptidoglycan structure analysis demonstrated that, in the absence of GBC, cells failed to initiate normal PG synthesis and cannot complete polymerization of the murein sacculus. Finally, the subcellular localization of the PG hydrolase PcsB, which has a critical role in cell division of streptococci, was altered in the gbcO mutant. Collectively, these findings show that GBC is an essential component of the cell wall of S. agalactiae whose function is reminiscent of that of conventional wall teichoic acids found in Staphylococcus aureus or Bacillus subtilis. Furthermore, our findings raise the possibility that GBC-like molecules play a major role in the growth of most if not all beta-hemolytic streptococci.

  11. Improved methods for binding acma-type protein anchor fusions yo cell-wall material of micro-organisms

    NARCIS (Netherlands)

    Leenhouts, Cornelis; Ramasamy, R.; Steen, Anton; Kok, Jan; Buist, Girbe; Kuipers, Oscar

    2002-01-01

    The invention provides a method for improving binding of a proteinaceous substance to cell-wall material of a Gram-positive bacterium, said substance comprising an AcmA cell wall binding domain or homolog or functional derivative thereof, said method comprising treating said cell-wall material with

  12. The Analysis Stability of Anchor Retaining Wall

    International Nuclear Information System (INIS)

    The construction of anchored retaining walls reach every day in the field of Civil Engineering especially in public works. Their dimensioning and stability are the axes of research for geotechnical. The rule is to reduce the active forces of the slide and increase the effective normal stress on the rupture surface. So that, we anchored tied-back (constituted by steel cables) in the stable ground located under the failure surface and we apply at the top a traction force. This effort can be distributed over the ground surface by means of small plates or massive reinforced concrete. The study of the stability of anchored retaining wall was also performed by using software GEO4. Many cases can be solved using analytical solutions available in the group GEO4 program, but for our standard model solution studied analytically proved unsatisfactory so we used a numerical analysis based on the method of finite element in this program. The results obtained by numerical study were interpreted to identify the precision numerical predictions. Moreover these methods were useful and economics in the realization of reinforced slopes by tied-buck. (author)

  13. Inactivation of the srtA Gene in Streptococcus gordonii Inhibits Cell Wall Anchoring of Surface Proteins and Decreases In Vitro and In Vivo Adhesion

    Science.gov (United States)

    Bolken, Tové C.; Franke, Christine A.; Jones, Kevin F.; Zeller, Gloria O.; Jones, C. Hal; Dutton, Emma K.; Hruby, Dennis E.

    2001-01-01

    The srtA gene product, SrtA, has been shown to be required for cell wall anchoring of protein A as well as virulence in the pathogenic bacterium Staphylococcus aureus. There are five major mechanisms for displaying proteins at the surface of gram-positive bacteria (P. Cossart and R. Jonquieres, Proc. Natl. Acad. Sci. USA 97:5013–5015, 2000). However, since many of the known surface proteins of gram-positive bacteria are believed to be exported and anchored via the sortase pathway, it was of interest to determine if srtA plays a similar role in other gram-positive bacteria. To that end, the srtA gene in the human oral commensal organism Streptococcus gordonii was insertionally inactivated. The srtA mutant S. gordonii exhibited a marked reduction in quantity of a specific anchored surface protein. Furthermore, the srtA mutant had reduced binding to immobilized human fibronectin and had a decreased ability to colonize the oral mucosa of mice. Taken together, these results suggest that the activity of SrtA plays an important role in the biology of nonpathogenic as well as pathogenic gram-positive cocci. PMID:11119491

  14. Cell Wall

    OpenAIRE

    Jamet, Elisabeth; Canut, Hervé; Boudart, Georges; Albenne, Cécile; Pont-Lezica, Rafael F

    2008-01-01

    This chapter covers our present knowledge of cell wall proteomics highlighting the distinctive features of cell walls and cell wall proteins in relation to problems encountered for protein extraction, separation and identification. It provides clues to design strategies for efficient cell wall proteomic studies. It gives an overview of the kinds of proteins that have yet been identified: the expected proteins vs the identified proteins. Finally, the new vision of the cell wall proteome, and t...

  15. Role of BGT-1 and BGT-2, two predicted GPI-anchored glycoside hydrolases/glycosyltransferases, in cell wall remodeling in Neurospora crassa.

    Science.gov (United States)

    Martínez-Núñez, Leonora; Riquelme, Meritxell

    2015-12-01

    Neurospora crassa BGT-1 (NCU06381) and BGT-2 (NCU09175) are two putative glycoside hydrolases (GHs) with additional predicted glycosyltransferase activity and binding sites for a glycosyl phosphatidyl inositol (GPI) anchor that would facilitate their attachment to the plasma membrane (PM). To discern their role in key morphogenetic events during vegetative development of N. crassa, BGT-1 and BGT-2 were labeled with the green fluorescent protein (GFP). The gfp was inserted immediately after the signal peptide sequence, within the bgt-1 encoding sequence, or directly before the GPI-binding site in the case of bgt-2. Both BGT-1-GFP and BGT-2-GFP were observed at the PM of the hyphal apical dome, excluding the foremost apical region and the Spitzenkörper (Spk), where chitin and β-1,3-glucan synthases have been previously found. These and previous studies suggest a division of labor of the cell wall synthesizing machinery at the hyphal dome: at the very tip, glucans are synthesized by enzymes that accumulate at the Spk, before getting incorporated into the PM, whereas at the subtending zone below the apex, glucans are presumably hydrolyzed, producing amenable ends for further branching and crosslinking with other cell wall polymers. Additionally, BGT-1-GFP and BGT-2-GFP were observed at the leading edge of new developing septa, at unreleased interconidial junctions, at conidial poles, at germling and hyphal fusion sites, and at sites of branch emergence, all of them processes that seemingly involve cell wall remodeling. Even though single and double mutant strains for the corresponding genes did not show a drastic reduction of growth rate, bgt-2Δ and bgt-1Δ::bgt-2Δ strains exhibited an increased resistance to the cell wall stressors calcofluor white (CW) and congo red (CR) than the reference strain, which suggests they present significant architectural changes in their cell wall. Furthermore, the conidiation defects observed in the mutants indicate a significant

  16. Role of the group B antigen of Streptococcus agalactiae a peptidoglycan-anchored polysaccharide involved in cell wall biogenesis : a Peptidoglycan-Anchored Polysaccharide involved in cell wall biogenesis

    OpenAIRE

    Élise Caliot; Shaynoor Dramsi; Marie-Pierre Chapot-Chartier; Pascal Courtin; Saulius Kulakauskas; Christine Péchoux; Patrick Trieu-Cuot; Michel-Yves Mistou

    2012-01-01

    Streptococcus agalactiae (Group B streptococcus, GBS) is a leading cause of infections in neonates and an emerging pathogen in adults. The Lancefield Group B carbohydrate (GBC) is a peptidoglycan-anchored antigen that defines this species as a Group B Streptococcus. Despite earlier immunological and biochemical characterizations, the function of this abundant glycopolymer has never been addressed experimentally. Here, we inactivated the gene gbcO encoding a putative UDP-N-acetylglucosamine-1-...

  17. Two novel, putatively cell wall-associated and glycosylphosphatidylinositol-anchored α-glucanotransferase enzymes of Aspergillus niger

    NARCIS (Netherlands)

    Kaaij, R.M. van der; Yuan, X.L.; Franken, A.; Ram, A.F.J.; Punt, P.J.; Maarel, M.J.E.C. van der; Dijkhuizen, L.

    2007-01-01

    In the genome sequence of Aspergillus niger CBS 513.88, three genes were identified with high similarity to fungal α-amylases. The protein sequences derived from these genes were different in two ways from all described fungal α-amylases: they were predicted to be glycosylphosphatidylinositol anchor

  18. Considerations on the design of through-wall anchors

    International Nuclear Information System (INIS)

    Connections to existing buildings are often the most difficult planning challenge for the realization of construction measures in case of piping system replacements in nuclear power plants. This is due to restricted space or limited load reserves of the building structure. Usually the realization of support connections to the existing buildings is achieved by anchor bolts. But in critical cases the preferred alternative solution uses through-wall anchors. Up to now uniform assessment thresholds are not available, no technical guidelines or regulations for construction variants exist. Through-wall anchors allow significantly higher load capacities for tensile and shear loads but require enhanced planning and realization efforts.

  19. Field Behavior of A High Anchored Reinforced Earth Wall

    Directory of Open Access Journals (Sweden)

    Faisal H. Ali

    2008-01-01

    Full Text Available Since the invention of Reinforced Earth by the French architect Hendri Vidal in 1966, numerous reinforced soil walls have been designed and constructed all over the world. In this study the focus was on a particular type of reinforced wall called Nehemiah wall which differed from the Vidal type in the sense that instead of steel strips, the reinforcing elements consisted of steel bars with anchor blocks attached at the free ends. A full scale high anchored reinforced earth wall was constructed and instrumented to capture the essential behavior of the wall. Two sections of the wall were monitored where at one of the sections polystyrene foam was inserted at the back face of the wall panel to allow for lateral deformation to take place which means that the facing was less flexible in the transverse direction. The lateral deformation, axial forces along the reinforcing bars and settlement were monitored and measured for both cases and the results were compared and discussed.

  20. Comparative genome-based identification of a cell wall-anchored protein from Lactobacillus plantarum increases adhesion of Lactococcus lactis to human epithelial cells

    OpenAIRE

    ZHANG, BO; Zuo, Fanglei; Yu, Rui; Zeng, Zhu; Ma, Huiqin; Chen, Shangwu

    2015-01-01

    Adhesion to host cells is considered important for Lactobacillus plantarum as well as other lactic acid bacteria (LAB) to persist in human gut and thus exert probiotic effects. Here, we sequenced the genome of Lt. plantarum strain NL42 originating from a traditional Chinese dairy product, performed comparative genomic analysis and characterized a novel adhesion factor. The genome of NL42 was highly divergent from its closest neighbors, especially in six large genomic regions. NL42 harbors a t...

  1. Physical and numerical modelling of earth pressure on anchored sheet pile walls in sand

    DEFF Research Database (Denmark)

    Krogsbøll, Anette Susanne; Fuglsang, Leif D

    The influence of wall flexibility on earth pressure, bending moments and failure modes is studied. Numerical models are compared to results from model tests carried out in a geotechnical centrifuge. The back-fill is dry sand and failure is introduced by allowing the wall to rotate around the anchor......-model showed the right behaviour in pre-failure as well as failure for both flexible and stiff walls, whereas the MC-model showed some shortcomings when stiff walls were modelled....

  2. Cell wall sorting signals in surface proteins of gram-positive bacteria.

    OpenAIRE

    Schneewind, O; Mihaylova-Petkov, D; Model, P

    1993-01-01

    Staphylococcal protein A is anchored to the cell wall, a unique cellular compartment of Gram-positive bacteria. The sorting signal sufficient for cell wall anchoring consists of an LPXTG motif, a C-terminal hydrophobic domain and a charged tail. Homologous sequences are found in many surface proteins of Gram-positive bacteria and we explored the universality of these sequences to serve as cell wall sorting signals. We show that several signals are able to anchor fusion proteins to the staphyl...

  3. Cell wall sorting of lipoproteins in Staphylococcus aureus.

    OpenAIRE

    Navarre, W W; Daefler, S; Schneewind, O

    1996-01-01

    Many surface proteins are thought to be anchored to the cell wall of gram-positive organisms via their C termini, while the N-terminal domains of these molecules are displayed on the bacterial surface. Cell wall anchoring of surface proteins in Staphylococcus aureus requires both an N-terminal leader peptide and a C-terminal cell wall sorting signal. By fusing the cell wall sorting of protein A to the C terminus of staphylococcal beta-lactamase, we demonstrate here that lipoproteins can also ...

  4. Assay and heterologous expression in Pichia pastoris of plant cell wall type-II membrane anchored glycosyltransferases

    DEFF Research Database (Denmark)

    Petersen, Bent; Egelund, Jack; Damager, Iben;

    2009-01-01

    Two Arabidopsis xylosyltransferases, designated RGXT1 and RGXT2, were recently expressed in Baculovirus transfected insect cells and by use of the free sugar assay shown to catalyse transfer of D-xylose from UDP-alpha-D-xylose to L-fucose and derivatives hereof. We have now examined expression of...... fractions. When incubated with 0.5 M L-fucose and UDP-D-xylose all four RGXT1 and RGXT2 variants catalyzed transfer of D-xylose onto L-fucose with estimated turnover numbers between 0.15 and 0.3 sec(-1), thus demonstrating that a free C-terminus is not required for activity. N- and O-glycanase treatment...

  5. Decoding Cytoskeleton-Anchored and Non-Anchored Receptors from Single-Cell Adhesion Force Data.

    Science.gov (United States)

    Sariisik, Ediz; Popov, Cvetan; Müller, Jochen P; Docheva, Denitsa; Clausen-Schaumann, Hauke; Benoit, Martin

    2015-10-01

    Complementary to parameters established for cell-adhesion force curve analysis, we evaluated the slope before a force step together with the distance from the surface at which the step occurs and visualized the result in a two-dimensional density plot. This new tool allows detachment steps of long membrane tethers to be distinguished from shorter jumplike force steps, which are typical for cytoskeleton-anchored bonds. A prostate cancer cell line (PC3) immobilized on an atomic-force-microscopy sensor interacted with three different substrates: collagen-I (Col-I), bovine serum albumin, and a monolayer of bone marrow-derived stem cells (SCP1). To address PC3 cells' predominant Col-I binding molecules, an antibody-blocking β1-integrin was used. Untreated PC3 cells on Col-I or SCP1 cells, which express Col-I, predominantly showed jumps in their force curves, while PC3 cells on bovine-serum-albumin- and antibody-treated PC3 cells showed long membrane tethers. The probability density plots thus revealed that β1-integrin-specific interactions are predominately anchored to the cytoskeleton, while the nonspecific interactions are mainly membrane-anchored. Experiments with latrunculin-A-treated PC3 cells corroborated these observations. The plots thus reveal details of the anchoring of bonds to the cell and provide a better understanding of receptor-ligand interactions. PMID:26445433

  6. Protein transport across the cell wall of monoderm Gram-positive bacteria

    OpenAIRE

    Forster, Brian M.; Marquis, Hélène

    2012-01-01

    In monoderm (single membrane) Gram-positive bacteria, the majority of secreted proteins are first translocated across the cytoplasmic membrane into the inner wall zone. For a subset of these proteins, final destination is within the cell envelope either as membrane-anchored or cell wall-anchored proteins, whereas another subset of proteins is destined to be transported across the cell wall into the extracellular milieu. Although the cell wall is a porous structure, there is evidence that, for...

  7. Breaking anchored droplets in a microfluidic Hele-Shaw cell

    CERN Document Server

    Amselem, Gabriel; Gallaire, François; Baroud, Charles N

    2015-01-01

    We study microfluidic self digitization in Hele-Shaw cells using pancake droplets anchored to surface tension traps. We show that above a critical flow rate, large anchored droplets break up to form two daughter droplets, one of which remains in the anchor. Below the critical flow velocity for breakup the shape of the anchored drop is given by an elastica equation that depends on the capillary number of the outer fluid. As the velocity crosses the critical value, the equation stops admitting a solution that satisfies the boundary conditions; the drop breaks up in spite of the neck still having finite width. A similar breaking event also takes place between the holes of an array of anchors, which we use to produce a 2D array of stationary drops in situ.

  8. Characterization of the Sclerotinia sclerotiorum cell wall proteome.

    Science.gov (United States)

    Liu, Longzhou; Free, Stephen J

    2016-08-01

    We used a proteomic analysis to identify cell wall proteins released from Sclerotinia sclerotiorum hyphal and sclerotial cell walls via a trifluoromethanesulfonic acid (TFMS) digestion. Cell walls from hyphae grown in Vogel's glucose medium (a synthetic medium lacking plant materials), from hyphae grown in potato dextrose broth and from sclerotia produced on potato dextrose agar were used in the analysis. Under the conditions used, TFMS digests the glycosidic linkages in the cell walls to release intact cell wall proteins. The analysis identified 24 glycosylphosphatidylinositol (GPI)-anchored cell wall proteins and 30 non-GPI-anchored cell wall proteins. We found that the cell walls contained an array of cell wall biosynthetic enzymes similar to those found in the cell walls of other fungi. When comparing the proteins in hyphal cell walls grown in potato dextrose broth with those in hyphal cell walls grown in the absence of plant material, it was found that a core group of cell wall biosynthetic proteins and some proteins associated with pathogenicity (secreted cellulases, pectin lyases, glucosidases and proteases) were expressed in both types of hyphae. The hyphae grown in potato dextrose broth contained a number of additional proteins (laccases, oxalate decarboxylase, peroxidase, polysaccharide deacetylase and several proteins unique to Sclerotinia and Botrytis) that might facilitate growth on a plant host. A comparison of the proteins in the sclerotial cell wall with the proteins in the hyphal cell wall demonstrated that sclerotia formation is not marked by a major shift in the composition of cell wall protein. We found that the S. sclerotiorum cell walls contained 11 cell wall proteins that were encoded only in Sclerotinia and Botrytis genomes. PMID:26661933

  9. Anchor reinforcements

    Energy Technology Data Exchange (ETDEWEB)

    Levkovich, P.Ye.; Sal' nikov, V.K.; Savich, N.S.

    1980-11-30

    An anchor reinforcement includes an anchor shaft, an elastic jig with a separator wall having compartments placed parallely along it and filled with reinforcement material and a device for destroying the jig wall separator. To quickly put the anchor in place and increase the reliability of the reinforcement by mixing the reinforcement material components better, the device for destroying the jig separator wall was made in the shape of a shovel, fastened to the anchor shaft and the separator wall has a longitudinal hollow for holding the anchor shaft/shovel.

  10. Bacterial glycobiology: rhamnose-containing cell wall polysaccharides in Gram-positive bacteria.

    OpenAIRE

    Mistou, Michel-Yves; Sutcliffe, Iain; van Sorge, Nina

    2016-01-01

    The composition of the Gram-positive cell wall is typically described as containing peptidoglycan, proteins and essential secondary cell wall structures called teichoic acids, which comprise approximately half of the cell wall mass. The cell walls of many species within the genera Streptococcus, Enterococcus and Lactococcus contain large amounts of the sugar rhamnose, which is incorporated in cell wall-anchored polysaccharides (CWP) that possibly function as homologues of well-studied wall te...

  11. Study of Individual Characteristic Abdominal Wall Thickness Based on Magnetic Anchored Surgical Instruments

    Directory of Open Access Journals (Sweden)

    Ding-Hui Dong

    2015-01-01

    Full Text Available Background: Magnetic anchored surgical instruments (MASI, relying on magnetic force, can break through the limitations of the single port approach in dexterity. Individual characteristic abdominal wall thickness (ICAWT deeply influences magnetic force that determines the safety of MASI. The purpose of this study was to research the abdominal wall characteristics in MASI applied environment to find ICAWT, and then construct an artful method to predict ICAWT, resulting in better safety and feasibility for MASI. Methods: For MASI, ICAWT is referred to the thickness of thickest point in the applied environment. We determined ICAWT through finding the thickest point in computed tomography scans. We also investigated the traits of abdominal wall thickness to discover the factor that can be used to predict ICAWT. Results: Abdominal wall at C point in the middle third lumbar vertebra plane (L3 is the thickest during chosen points. Fat layer thickness plays a more important role in abdominal wall thickness than muscle layer thickness. "BMI-ICAWT" curve was obtained based on abdominal wall thickness of C point in L3 plane, and the expression was as follow: f(x = P1 × x 2 + P2 × x + P3, where P1 = 0.03916 (0.01776, 0.06056, P2 = 1.098 (0.03197, 2.164, P3 = −18.52 (−31.64, −5.412, R-square: 0.99. Conclusions: Abdominal wall thickness of C point at L3 could be regarded as ICAWT. BMI could be a reliable predictor of ICAWT. In the light of "BMI-ICAWT" curve, we may conveniently predict ICAWT by BMI, resulting a better safety and feasibility for MASI.

  12. Automatic Thickness and Volume Estimation of Sprayed Concrete on Anchored Retaining Walls from Terrestrial LIDAR Data

    Science.gov (United States)

    Martínez-Sánchez, J.; Puente, I.; GonzálezJorge, H.; Riveiro, B.; Arias, P.

    2016-06-01

    When ground conditions are weak, particularly in free formed tunnel linings or retaining walls, sprayed concrete can be applied on the exposed surfaces immediately after excavation for shotcreting rock outcrops. In these situations, shotcrete is normally applied conjointly with rock bolts and mesh, thereby supporting the loose material that causes many of the small ground falls. On the other hand, contractors want to determine the thickness and volume of sprayed concrete for both technical and economic reasons: to guarantee their structural strength but also, to not deliver excess material that they will not be paid for. In this paper, we first introduce a terrestrial LiDAR-based method for the automatic detection of rock bolts, as typically used in anchored retaining walls. These ground support elements are segmented based on their geometry and they will serve as control points for the co-registration of two successive scans, before and after shotcreting. Then we compare both point clouds to estimate the sprayed concrete thickness and the expending volume on the wall. This novel methodology is demonstrated on repeated scan data from a retaining wall in the city of Vigo (Spain), resulting in a rock bolts detection rate of 91%, that permits to obtain a detailed information of the thickness and calculate a total volume of 3597 litres of concrete. These results have verified the effectiveness of the developed approach by increasing productivity and improving previous empirical proposals for real time thickness estimation.

  13. Cell Wall Biology: Perspectives from Cell Wall Imaging

    Institute of Scientific and Technical Information of China (English)

    Kieran J.D.Lee; Susan E.Marcus; J.Paul Knox

    2011-01-01

    Polysaccharide-rich plant cell walls are important biomaterials that underpin plant growth,are major repositories for photosynthetically accumulated carbon,and,in addition,impact greatly on the human use of plants. Land plant cell walls contain in the region of a dozen major polysaccharide structures that are mostly encompassed by cellulose,hemicelluloses,and pectic polysaccharides. During the evolution of land plants,polysaccharide diversification appears to have largely involved structural elaboration and diversification within these polysaccharide groups. Cell wall chemistry is well advanced and a current phase of cell wall science is aimed at placing the complex polysaccharide chemistry in cellular contexts and developing a detailed understanding of cell wall biology. Imaging cell wall glycomes is a challenging area but recent developments in the establishment of cell wall molecular probe panels and their use in high throughput procedures are leading to rapid advances in the molecular understanding of the spatial heterogeneity of individual cell walls and also cell wall differences at taxonomic levels. The challenge now is to integrate this knowledge of cell wall heterogeneity with an understanding of the molecular and physiological mechanisms that underpin cell wall properties and functions.

  14. Cell wall proteomics of crops

    OpenAIRE

    Komatsu, Setsuko; Yanagawa, Yuki

    2013-01-01

    Cell wall proteins play key roles in cell structure and metabolism, cell enlargement, signal transduction, responses to environmental stress, and many other physiological events. Agricultural crops are often used for investigating stress tolerance because cultivars with differing degrees of tolerance are available. Abiotic and biotic stress factors markedly influence the geographical distribution and yields of many crop species. Crop cell wall proteomics is of particular importance for improv...

  15. Phosphatidylinositol anchor of HeLa cell alkaline phosphatase

    International Nuclear Information System (INIS)

    Alkaline phosphatase from cancer cells, HeLa TCRC-1, was biosynthetically labeled with either 3H-fatty acids or [3H]ethanolamine as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography of immunoprecipitated material. Phosphatidylinositol-specific phospholipase C (PI-PLC) released a substantial proportion of the 3H-fatty acid label from immunoaffinity-purified alkaline phosphatase but had no effect on the radioactivity of [3H]ethanolamine-labeled material. PI-PLC also liberated catalytically active alkaline phosphatase from viable cells, and this could be selectively blocked by monoclonal antibodies to alkaline phosphatase. However, the alkaline phosphatase released from 3H-fatty acid labeled cells by PI-PLC was not radioactive. By contrast, treatment with bromelain removed both the 3H-fatty acid and the [3H]ethanolamine label from purified alkaline phosphatase. Subtilisin was also able to remove the [3H]ethanolamine label from the purified alkaline phosphatase. The 3H radioactivity in alkaline phosphatase purified from [3H]ethanolamine-labeled cells comigrated with authentic [3H]ethanolamine by anion-exchange chromatography after acid hydrolysis. The data suggest that the 3H-fatty acid and [3H]ethanolamine are covalently attached to the carboxyl-terminal segment since bromelain and subtilisin both release alkaline phosphatase from the membrane by cleavage at that end of the polypeptide chain. The data are consistent with findings for other proteins recently shown to be anchored in the membrane through a glycosylphosphatidylinositol structure and indicate that a similar structure contributes to the membrane anchoring of alkaline phosphatase

  16. Phosphatidylinositol anchor of HeLa cell alkaline phosphatase

    Energy Technology Data Exchange (ETDEWEB)

    Jemmerson, R.; Low, M.G.

    1987-09-08

    Alkaline phosphatase from cancer cells, HeLa TCRC-1, was biosynthetically labeled with either /sup 3/H-fatty acids or (/sup 3/H)ethanolamine as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography of immunoprecipitated material. Phosphatidylinositol-specific phospholipase C (PI-PLC) released a substantial proportion of the /sup 3/H-fatty acid label from immunoaffinity-purified alkaline phosphatase but had no effect on the radioactivity of (/sup 3/H)ethanolamine-labeled material. PI-PLC also liberated catalytically active alkaline phosphatase from viable cells, and this could be selectively blocked by monoclonal antibodies to alkaline phosphatase. However, the alkaline phosphatase released from /sup 3/H-fatty acid labeled cells by PI-PLC was not radioactive. By contrast, treatment with bromelain removed both the /sup 3/H-fatty acid and the (/sup 3/H)ethanolamine label from purified alkaline phosphatase. Subtilisin was also able to remove the (/sup 3/H)ethanolamine label from the purified alkaline phosphatase. The /sup 3/H radioactivity in alkaline phosphatase purified from (/sup 3/H)ethanolamine-labeled cells comigrated with authentic (/sup 3/H)ethanolamine by anion-exchange chromatography after acid hydrolysis. The data suggest that the /sup 3/H-fatty acid and (/sup 3/H)ethanolamine are covalently attached to the carboxyl-terminal segment since bromelain and subtilisin both release alkaline phosphatase from the membrane by cleavage at that end of the polypeptide chain. The data are consistent with findings for other proteins recently shown to be anchored in the membrane through a glycosylphosphatidylinositol structure and indicate that a similar structure contributes to the membrane anchoring of alkaline phosphatase.

  17. Domain Walls and Anchoring Transitions Mimicking Nematic Biaxiality in the Oxadiazole Bent-Core Liquid Crystal C7

    OpenAIRE

    Kim, Young-Ki; Cukrov, Greta; Xiang, Jie; Shin, Sung-Tae; Lavrentovich, Oleg D.

    2015-01-01

    We investigate the origin of secondary disclinations that were recently described as a new evidence of a biaxial nematic phase in an oxadiazole bent-core thermotropic liquid crystal C7. With an assortment of optical techniques such as polarizing optical microscopy, LC PolScope, and fluorescence confocal polarizing microscopy, we demonstrate that the secondary disclinations represent non-singular domain walls formed in an uniaxial nematic during the surface anchoring transition, in which surfa...

  18. The Chlamydomonas cell wall: characterization of the wall framework

    OpenAIRE

    1985-01-01

    The cell wall of the biflagellate alga Chlamydomonas reinhardtii is a multilayered, extracellular matrix composed of carbohydrates and 20-25 polypeptides. To learn more about the forces responsible for the integrity of this cellulose-deficient cell wall, we have begun studies to identify and characterize the framework of the wall and to determine the effects of the cell wall-degrading enzyme, lysin, on framework structure and protein composition. In these studies we used walls released into t...

  19. A glycophospholipid membrane anchor acts as an apical targeting signal in polarized epithelial cells

    OpenAIRE

    1989-01-01

    Glycosyl-phosphatidylinositol- (GPI) anchored proteins contain a large extracellular protein domain that is linked to the membrane via a glycosylated form of phosphatidylinositol. We recently reported the polarized apical distribution of all endogenous GPI-anchored proteins in the MDCK cell line (Lisanti, M. P., M. Sargiacomo, L. Graeve, A. R. Saltiel, and E. Rodriguez-Boulan. 1988. Proc. Natl. Acad. Sci. USA. 85:9557-9561). To study the role of this mechanism of membrane anchoring in targeti...

  20. DEX-1 and DYF-7 establish sensory dendrite length by anchoring dendritic tips during cell migration

    OpenAIRE

    Heiman, Maxwell G.; Shaham, Shai

    2009-01-01

    Cells are devices whose structures delimit function. For example, in the nervous system, neuronal and glial shapes dictate paths of information flow. To understand how cells acquire their shapes, we examined the formation of a sense organ in C. elegans. Using time-lapse imaging, we found that sensory dendrites form by stationary anchoring of dendritic tips during cell-body migration. A genetic screen identified DEX-1 and DYF-7, extracellular proteins required for dendritic tip anchoring, whic...

  1. Cell Wall Integrity Signaling in Saccharomyces cerevisiae

    OpenAIRE

    Levin, David E.

    2005-01-01

    The yeast cell wall is a highly dynamic structure that is responsible for protecting the cell from rapid changes in external osmotic potential. The wall is also critical for cell expansion during growth and morphogenesis. This review discusses recent advances in understanding the various signal transduction pathways that allow cells to monitor the state of the cell wall and respond to environmental challenges to this structure. The cell wall integrity signaling pathway controlled by the small...

  2. Back wall solar cell

    Science.gov (United States)

    Brandhorst, H. W., Jr. (Inventor)

    1978-01-01

    A solar cell is disclosed which comprises a first semiconductor material of one conductivity type with one face having the same conductivity type but more heavily doped to form a field region arranged to receive the radiant energy to be converted to electrical energy, and a layer of a second semiconductor material, preferably highly doped, of opposite conductivity type on the first semiconductor material adjacent the first semiconductor material at an interface remote from the heavily doped field region. Instead of the opposite conductivity layer, a metallic Schottky diode layer may be used, in which case no additional back contact is needed. A contact such as a gridded contact, previous to the radiant energy may be applied to the heavily doped field region of the more heavily doped, same conductivity material for its contact.

  3. Synthesis of subnanometer-diameter vertically aligned single-walled carbon nanotubes with copper-anchored cobalt catalysts

    Science.gov (United States)

    Cui, Kehang; Kumamoto, Akihito; Xiang, Rong; An, Hua; Wang, Benjamin; Inoue, Taiki; Chiashi, Shohei; Ikuhara, Yuichi; Maruyama, Shigeo

    2016-01-01

    We synthesize vertically aligned single-walled carbon nanotubes (VA-SWNTs) with subnanometer diameters on quartz (and SiO2/Si) substrates by alcohol CVD using Cu-anchored Co catalysts. The uniform VA-SWNTs with a nanotube diameter of 1 nm are synthesized at a CVD temperature of 800 °C and have a thickness of several tens of μm. The diameter of SWNTs was reduced to 0.75 nm at 650 °C with the G/D ratio maintained above 24. Scanning transmission electron microscopy energy-dispersive X-ray spectroscopy (EDS-STEM) and high angle annular dark field (HAADF-STEM) imaging of the Co/Cu bimetallic catalyst system showed that Co catalysts were captured and anchored by adjacent Cu nanoparticles, and thus were prevented from coalescing into a larger size, which contributed to the small diameter of SWNTs. The correlation between the catalyst size and the SWNT diameter was experimentally clarified. The subnanometer-diameter and high-quality SWNTs are expected to pave the way to replace silicon for next-generation optoelectronic and photovoltaic devices.We synthesize vertically aligned single-walled carbon nanotubes (VA-SWNTs) with subnanometer diameters on quartz (and SiO2/Si) substrates by alcohol CVD using Cu-anchored Co catalysts. The uniform VA-SWNTs with a nanotube diameter of 1 nm are synthesized at a CVD temperature of 800 °C and have a thickness of several tens of μm. The diameter of SWNTs was reduced to 0.75 nm at 650 °C with the G/D ratio maintained above 24. Scanning transmission electron microscopy energy-dispersive X-ray spectroscopy (EDS-STEM) and high angle annular dark field (HAADF-STEM) imaging of the Co/Cu bimetallic catalyst system showed that Co catalysts were captured and anchored by adjacent Cu nanoparticles, and thus were prevented from coalescing into a larger size, which contributed to the small diameter of SWNTs. The correlation between the catalyst size and the SWNT diameter was experimentally clarified. The subnanometer-diameter and high

  4. Physical and numerical modelling of earth pressure on anchored sheet pile walls in sand

    DEFF Research Database (Denmark)

    Krogsbøll, Anette Susanne; Fuglsang, Leif D

    The influence of wall flexibility on earth pressure, bending moments and failure modes is studied. Numerical models are compared to results from model tests carried out in a geotechnical centrifuge. The back-fill is dry sand and failure is introduced by allowing the wall to rotate around the anch...

  5. Physical and numerical modelling of earth pressure on anchored sheet pile walls in sand

    DEFF Research Database (Denmark)

    Krogsbøll, Anette Susanne; Fuglsang, Leif D

    2006-01-01

    The influence of wall flexibility on earth pressure, bending moments and failure modes is studied. Numerical models are compared to results from model tests carried out in a geotechnical centrifuge. The back-fill is dry sand and failure is introduced by allowing the wall to rotate around the anch...

  6. GPI-anchored proteins do not reside in ordered domains in the live cell plasma membrane

    OpenAIRE

    Sevcsik, Eva; Brameshuber, Mario; Fölser, Martin; Weghuber, Julian; Honigmann, Alf; Schütz, Gerhard J.

    2015-01-01

    The organization of proteins and lipids in the plasma membrane has been subject of a long-lasting debate. Membrane rafts of higher lipid chain order were proposed to mediate protein interactions, but have thus far not been directly observed. Here, we use protein micropatterning combined with single-molecule tracking to put current models to the test: we rearranged lipid-anchored raft proteins (glycosylphosphatidylinositol(GPI)-anchored mGFP) directly in the live cell plasma membrane and measu...

  7. GPI-anchored proteins do not reside in ordered domains in the live cell plasma membrane.

    OpenAIRE

    Sevcsik, E.; Brameshuber, M.; Fölser, M.; Weghuber, J.; Honigmann, A.; Schütz, G

    2015-01-01

    The organization of proteins and lipids in the plasma membrane has been the subject of a long-lasting debate. Membrane rafts of higher lipid chain order were proposed to mediate protein interactions, but have thus far not been directly observed. Here we use protein micropatterning combined with single-molecule tracking to put current models to the test: we rearranged lipid-anchored raft proteins (glycosylphosphatidylinositol(GPI)-anchored-mGFP) directly in the live cell plasma membrane and me...

  8. Anchors away: contribution of a glycolipid anchor to bacterial invasion of host cells

    OpenAIRE

    Baron, Miriam J.; Kasper, Dennis L.

    2005-01-01

    Group B Streptococcus (GBS) is an important cause of infections, including meningitis. The molecular events underlying its pathogenesis are poorly understood. A study in this issue of the JCI reports that the GBS invasion-associated gene (iagA) contributes to meningeal infection and virulence by facilitating invasion of the cells that compose the blood-brain barrier and of other host cells. The mechanism involved most likely relates to the gene product’s role in synthesis of a glycolipid anch...

  9. Synthesis of subnanometer-diameter vertically aligned single-walled carbon nanotubes with copper-anchored cobalt catalysts.

    Science.gov (United States)

    Cui, Kehang; Kumamoto, Akihito; Xiang, Rong; An, Hua; Wang, Benjamin; Inoue, Taiki; Chiashi, Shohei; Ikuhara, Yuichi; Maruyama, Shigeo

    2016-01-21

    We synthesize vertically aligned single-walled carbon nanotubes (VA-SWNTs) with subnanometer diameters on quartz (and SiO2/Si) substrates by alcohol CVD using Cu-anchored Co catalysts. The uniform VA-SWNTs with a nanotube diameter of 1 nm are synthesized at a CVD temperature of 800 °C and have a thickness of several tens of μm. The diameter of SWNTs was reduced to 0.75 nm at 650 °C with the G/D ratio maintained above 24. Scanning transmission electron microscopy energy-dispersive X-ray spectroscopy (EDS-STEM) and high angle annular dark field (HAADF-STEM) imaging of the Co/Cu bimetallic catalyst system showed that Co catalysts were captured and anchored by adjacent Cu nanoparticles, and thus were prevented from coalescing into a larger size, which contributed to the small diameter of SWNTs. The correlation between the catalyst size and the SWNT diameter was experimentally clarified. The subnanometer-diameter and high-quality SWNTs are expected to pave the way to replace silicon for next-generation optoelectronic and photovoltaic devices. PMID:26690843

  10. Accelerating forward genetics for cell wall deconstruction

    OpenAIRE

    Vidaurre, Danielle; Bonetta, Dario

    2012-01-01

    The elucidation of the genes involved in cell wall synthesis and assembly remains one of the biggest challenges of cell wall biology. Although traditional genetic approaches, using simple yet elegant screens, have identified components of the cell wall, many unknowns remain. Exhausting the genetic toolbox by performing sensitized screens, adopting chemical genetics or combining these with improved cell wall imaging, hold the promise of new gene discovery and function. With the recent introduc...

  11. Moss cell walls: structure and biosynthesis

    OpenAIRE

    Alison W. Roberts; Eric M Roberts; Haigler, Candace H.

    2012-01-01

    The genome sequence of the moss Physcomitrella patens has stimulated new research examining the cell wall polysaccharides of mosses and the glycosyl transferases that synthesize them as a means to understand fundamental processes of cell wall biosynthesis and plant cell wall evolution. The cell walls of mosses and vascular plants are composed of the same classes of polysaccharides, but with differences in side chain composition and structure. Similarly, the genomes of P. patens and angiosperm...

  12. DEX-1 and DYF-7 establish sensory dendrite length by anchoring dendritic tips during cell migration.

    Science.gov (United States)

    Heiman, Maxwell G; Shaham, Shai

    2009-04-17

    Cells are devices whose structures delimit function. For example, in the nervous system, neuronal and glial shapes dictate paths of information flow. To understand how cells acquire their shapes, we examined the formation of a sense organ in C. elegans. Using time-lapse imaging, we found that sensory dendrites form by stationary anchoring of dendritic tips during cell-body migration. A genetic screen identified DEX-1 and DYF-7, extracellular proteins required for dendritic tip anchoring, which act cooperatively at the time and place of anchoring. DEX-1 and DYF-7 contain, respectively, zonadhesin and zona pellucida domains, and DYF-7 self-associates into multimers important for anchoring. Thus, unlike other dendrites, amphid dendritic tips are positioned by DEX-1 and DYF-7 without the need for long-range guidance cues. In sequence and function, DEX-1 and DYF-7 resemble tectorins, which anchor stereocilia in the inner ear, suggesting that a sensory dendrite anchor may have evolved into part of a mechanosensor. PMID:19344940

  13. The cell wall of Fusarium oxysporum

    NARCIS (Netherlands)

    Schoffelmeer, EAM; Klis, FM; Sietsma, JH; Cornelissen, BJC

    1999-01-01

    Sugar analysis of isolated cell walls from three formae speciales of Fusarium oxysporum showed that they contained not only glucose and (N-acetyl)-glucosamine, but also mannose, galactose, and uronic acids, presumably originating from cell wall glycoproteins. Cell wall glycoproteins accounted for 50

  14. Unique aspects of the grass cell wall

    Science.gov (United States)

    Grasses are amongst the most important crops worldwide, and the composition of their cell walls is critical for uses as food, feed, and energy crops. Grass cell walls differ dramatically from dicot cell walls in terms of the major structural polysaccharides present, how those polysaccharides are lin...

  15. Shape dynamics of growing cell walls

    OpenAIRE

    Banerjee, Shiladitya; Scherer, Norbert F.; Dinner, Aaron R.

    2015-01-01

    We introduce a general theoretical framework to study the shape dynamics of actively growing and remodeling surfaces. Using this framework we develop a physical model for growing bacterial cell walls and study the interplay of cell shape with the dynamics of growth and constriction. The model allows us to derive constraints on cell wall mechanical energy based on the observed dynamics of cell shape. We predict that exponential growth in cell size requires a constant amount of cell wall energy...

  16. Shape dynamics of growing cell walls

    CERN Document Server

    Banerjee, Shiladitya; Dinner, Aaron R

    2015-01-01

    We introduce a general theoretical framework to study the shape dynamics of actively growing and remodeling surfaces. Using this framework we develop a physical model for growing bacterial cell walls and study the interplay of cell shape with the dynamics of growth and constriction. The model allows us to derive constraints on cell wall mechanical energy based on the observed dynamics of cell shape. We predict that exponential growth in cell size requires a constant amount of cell wall energy to be dissipated per unit volume. We use the model to understand and contrast growth in bacteria with different shapes such as spherical, ellipsoidal, cylindrical and toroidal morphologies. Coupling growth to cell wall constriction, we predict a discontinuous shape transformation, from partial constriction to cell division, as a function of the chemical potential driving cell-wall synthesis. Our model for cell wall energy and shape dynamics relates growth kinetics with cell geometry, and provides a unified framework to d...

  17. PRACTICAL EARTH ANCHORING TECHNOLOGY FOR STRENGTHENING SLOPE WALL PROTECTION (CASE STUDY: DG. SIRUA ROAD POST CONSTRUCTION FAILURE WITH SKYHOOK ??? GRID BEAM TECHNOLOGY IN MAKASSAR CITY ??? INDONESIA)

    OpenAIRE

    Samang, Lawalenna; Hino, Takenori; Harianto, Tri; Ramli, Muhammad Isran; Walenna, Muhammad Akbar

    2013-01-01

    The application of soil anchoring system and grid beam method of strengthening protection wall post construction failure was observed in this research. The construction of the slope wall protection is situated to suit the existence of barrier made for aqueduct. Based on the numerical simulation on scenario of potential causes of slope failure, it indicates several phenomena such as the stability of barrier foundation experiences drastic drop. Scrutinizing the result of investigation and ideal...

  18. Determination of azimuthal anchoring strength in twisted nematic liquid crystal cells using heterodyne polarimeter.

    Science.gov (United States)

    Yu, Tsung-Chih; Lo, Yu-Lung; Huang, Rei-Rong

    2010-09-27

    Two external-field-free methods are presented for measuring the azimuthal anchoring strength in twisted nematic liquid crystal (TNLC) cells. For asymmetrical TNLC samples, the twist angle is derived from the phase of the detected signal in a phase-sensitive heterodyne polarimeter and is then used to calculate the weak anchoring strength directly. The measurement resolution which is found to be about 0.01 μJ/m(2) makes the present method sensitive enough for the LC-based bio-sensing application. Using the proposed method, the weak azimuthal anchoring strength of a composite liquid crystal mixture (40% LCT-061153 + 60% MJO-42761) in contact with a plasma-alignment layer is found to be 7.19 μJ/m(2). For symmetrical TNLC samples, the liquid crystals are injected into a wedge cell, and the two-dimensional distributions of the twist angle and cell gap are extracted from the detected phase distribution using a genetic algorithm (GA). The azimuthal anchoring strength is then obtained by applying a fitting technique to the twist angle vs. cell gap curve. Utilizing the proposed approach, it is shown that the strong anchoring strength between a rubbed polyimide (PI) alignment layer and E7 liquid crystal is around 160 μJ/m(2) while that between a rubbed PI alignment layer and MLC-7023 liquid crystal is approximately 32 μJ/m(2). PMID:20941014

  19. Identification of a lipid-anchored heparan sulfate proteoglycan in Schwann cells

    OpenAIRE

    1990-01-01

    Schwann cells synthesize both hydrophobic and peripheral cell surface heparan sulfate proteoglycans (HSPGs). Previous analysis of the kinetics of radiolabeling suggested the peripheral HSPGs are derived from the membrane-anchored forms (Carey, D., and D. Evans. 1989. J. Cell Biol. 108:1891-1897). Peripheral cell surface HSPGs were purified from phytic acid extracts of cultured neonatal rat sciatic nerve Schwann cells by anion exchange, gel filtration, and laminin-affinity chromatography. Appr...

  20. Analysis on anchoring plate retaining wall of Shanxi Science and Technology Museum%山西省科技馆锚定板挡土墙分析

    Institute of Scientific and Technical Information of China (English)

    张波

    2015-01-01

    介绍了锚定板式挡土墙的受力原理及适用条件,结合工程实例,从参数设计、土压力计算、锚定板抗拔力计算、稳定性分析等方面,阐述了锚定板挡土墙的具体设计方法,指出锚定板挡土墙具有施工速度快、造价低等优势。%This paper introduced the force principle and applicable conditions of anchoring plate retaining wall,combining with the engineering examples,from the parameter design,soil pressure calculation,anchor plate pulling force calculation,stability analysis and other aspects,elabo-rated the specific design method of anchoring plate retaining wall,pointed out that the anchoring plate retaining wall had quick construction speed,low cost and other advantages.

  1. Cell wall remodelling enzymes modulate fungal cell wall elasticity and osmotic stress resistance

    OpenAIRE

    Ene, Iuliana; Walker, Louise; Schiavone, Marion; Lee, Keunsook K.; Dague, Etienne; Gow, Neil A.R.; Munro, Carol A

    2015-01-01

    The fungal cell wall confers cell morphology and protection against environmental insults. For fungal pathogens, the cell wall is a key immunological modulator and an ideal therapeutic target. Yeast cell walls possess an inner matrix of interlinked β-glucan and chitin that is thought to provide tensile strength and rigidity. Yeast cells remodel their walls over time in response to environmental change, a process controlled by evolutionarily conserved stress (Hog1) and cell integrity (Mkc1, Ce...

  2. Regulation of Cell Wall Biogenesis in Saccharomyces cerevisiae: The Cell Wall Integrity Signaling Pathway

    OpenAIRE

    Levin, David E.

    2011-01-01

    The yeast cell wall is a strong, but elastic, structure that is essential not only for the maintenance of cell shape and integrity, but also for progression through the cell cycle. During growth and morphogenesis, and in response to environmental challenges, the cell wall is remodeled in a highly regulated and polarized manner, a process that is principally under the control of the cell wall integrity (CWI) signaling pathway. This pathway transmits wall stress signals from the cell surface to...

  3. Architecture of dermatophyte cell Walls: Electron microscopic and biochemical analysis

    Science.gov (United States)

    Nozawa, Y.; Kitajima, Y.

    1984-01-01

    A review with 83 references on the cell wall structure of dermatophytes is presented. Topics discussed include separation and preparation of cell walls; microstructure of cell walls by electron microscopy; chemical composition of cell walls; structural model of cell walls; and morphological structure of cell walls.

  4. Chemically Modified Multi-walled Carbon Nano tubes (MWCNTs) with Anchored Acidic Groups

    International Nuclear Information System (INIS)

    Surface functionalization of multi-walled carbon nano tubes (MWCNTs) was carried out using a gas phase treatment in a Universal Temperature Program (UTP) reactor by flowing SO3 gas onto the CNTs while being heated at different temperatures. The functionalized nano tubes were characterized using X-ray Fluorescence (XRF), Fourier Transform Infrared Spectroscopy (FT-IR) and Raman spectroscopy. The amount of oxygen and sulfur containing groups was determined by acid-base titration. The titration results were in good agreement with elemental analysis using x-ray fluorescence. FTIR analysis showed the presence of oxygen and sulfur containing groups, S=O, C-S, C=O and -COOH. Raman spectroscopy confirmed that oxygen and sulfur containing acidic groups covalently attached to the sidewall of the MWCNTs. (author)

  5. Cell Wall Assembly in Saccharomyces cerevisiae

    OpenAIRE

    Lesage, Guillaume; Bussey, Howard

    2006-01-01

    An extracellular matrix composed of a layered meshwork of β-glucans, chitin, and mannoproteins encapsulates cells of the yeast Saccharomyces cerevisiae. This organelle determines cellular morphology and plays a critical role in maintaining cell integrity during cell growth and division, under stress conditions, upon cell fusion in mating, and in the durable ascospore cell wall. Here we assess recent progress in understanding the molecular biology and biochemistry of cell wall synthesis and it...

  6. A cell-surface-anchored ratiometric i-motif sensor for extracellular pH detection.

    Science.gov (United States)

    Ying, Le; Xie, Nuli; Yang, Yanjing; Yang, Xiaohai; Zhou, Qifeng; Yin, Bincheng; Huang, Jin; Wang, Kemin

    2016-06-14

    A FRET-based sensor is anchored on the cell surface through streptavidin-biotin interactions. Due to the excellent properties of the pH-sensitive i-motif structure, the sensor can detect extracellular pH with high sensitivity and excellent reversibility. PMID:27241716

  7. How do plant cell walls extend?

    Science.gov (United States)

    Cosgrove, D. J.

    1993-01-01

    This article briefly summarizes recent work that identifies the biophysical and biochemical processes that give rise to the extension of plant cell walls. I begin with the biophysical notion of stress relaxation of the wall and follow with recent studies of wall enzymes thought to catalyze wall extension and relaxation. Readers should refer to detailed reviews for more comprehensive discussion of earlier literature (Taiz, 1984; Carpita and Gibeaut, 1993; Cosgrove, 1993).

  8. Anchor-free NEMS non-volatile memory cell for harsh environment data storage

    International Nuclear Information System (INIS)

    This work demonstrates a novel anchor-free nano-electromechanical (NEMS) based non-volatile memory cell, suitable for high temperature (T ≤ 300 °C) and radiation prone harsh environment applications. The anchor-free circular metal beam is actuated by electrostatic force and is held in one of the bi-stable memory states by adhesion force between two smooth metal surfaces in contact. Smooth metal layers form strong van der Waals stiction between two surfaces in contact and memory detection (Logic-‘1’ / Logic-‘0’) is obtained by detecting the conductance between two fixed contacts. This anchor-free design offers highest density (9F2 footprint) compared to other mechanical memory devices reported to date. (paper)

  9. GPI-anchored influenza hemagglutinin induces hemifusion to both red blood cell and planar bilayer membranes

    OpenAIRE

    1995-01-01

    Under fusogenic conditions, fluorescent dye redistributed from the outer monolayer leaflet of red blood cells (RBCs) to cells expressing glycophosphatidylinositol-anchored influenza virus hemagglutinin (GPI- HA) without transfer of aqueous dye. This suggests that hemifusion, but not full fusion, occurred (Kemble, G. W., T. Danieli, and J. M. White. 1994. Cell. 76:383-391). We extended the evidence for hemifusion by labeling the inner monolayer leaflets of RBCs with FM4-64 and observing that t...

  10. Microanalysis of Plant Cell Wall Polysaccharides

    Institute of Scientific and Technical Information of China (English)

    Nicolai Obel; Veronika Erben; Tatjana Schwarz; Stefan Kühne; Andrea Fodor; Markus Pauly

    2009-01-01

    Oligosaccharide Mass Profiling (OLIMP) allows a fast and sensitive assessment of cell wall polymer structure when coupled with Matrix Assisted Laser Desorption Ionisation Time Of Flight Mass Spectrometry (MALDI-TOF MS). The short time required for sample preparation and analysis makes possible the study of a wide range of plant organs, revealing a high degree of heterogeneity in the substitution pattern of wall polymers such as the cross-linking glycan xyloglucan and the pectic polysaccharide homogalacturonan. The high sensitivity of MALDI-TOF allows the use of small amounts of samples, thus making it possible to investigate the wall structure of single cell types when material is collected by such methods as laser micro-dissection. As an example, the analysis of the xyloglucan structure in the leaf cell types outer epidermis layer, entire epidermis cell layer, palisade mesophyll cells, and vascular bundles were investigated. OLIMP is amenable to in situ wall analysis, where wall polymers are analyzed on unprepared plant tissue itself without first iso-lating cell walls. In addition, OLIMP enables analysis of wall polymers in Golgi-enriched fractions, the location of nascent matrix polysaccharide biosynthesis, enabling separation of the processes of wall biosynthesis versus post-deposition apo-plastic metabolism. These new tools will make possible a semi-quantitative analysis of the cell wall at an unprecedented level.

  11. Glycoprotein component of plant cell walls

    International Nuclear Information System (INIS)

    The primary wall surrounding most dicotyledonous plant cells contains a hydroxyproline-rich glycoprotein (HRGP) component named extensin. A small group of glycopeptides solubilized from isolated cell walls by proteolysis contained a repeated pentapeptide glycosylated by tri- and tetraarabinosides linked to hydroxyproline and, by galactose, linked to serine. Recently, two complementary approaches to this problem have provided results which greatly increase the understanding of wall extensin. In this paper the authors describe what is known about the structure of soluble extensin secreted into the walls of the carrot root cells

  12. Displaying Lipase B from Candida antarctica in Pichia pastoris Using the Yeast Surface Display Approach: Prospection of a New Anchor and Characterization of the Whole Cell Biocatalyst.

    Directory of Open Access Journals (Sweden)

    Marcelo Victor Holanda Moura

    Full Text Available Yeast Surface Display (YSD is a strategy to anchor proteins on the yeast cell wall which has been employed to increase enzyme stability thus decreasing production costs. Lipase B from Candida antarctica (LipB is one of the most studied enzymes in the context of industrial biotechnology. This study aimed to assess the biochemical features of this important biocatalyst when immobilized on the cell surface of the methylotrophic yeast Pichia pastoris using the YSD approach. For that purpose, two anchors were tested. The first (Flo9 was identified after a prospection of the P. pastoris genome being related to the family of flocculins similar to Flo1 but significantly smaller. The second is the Protein with Internal Repeats (Pir1 from P. pastoris. An immunolocalization assay showed that both anchor proteins were able to display the reporter protein EGFP in the yeast outer cell wall. LipB was expressed in P. pastoris fused either to Flo9 (FLOLIPB or Pir1 (PIRLIPB. Both constructions showed hydrolytic activity towards tributyrin (>100 U/mgdcw and >80 U/mgdcw, respectively, optimal hydrolytic activity around 45°C and pH 7.0, higher thermostability at 45°C and stability in organic solvents when compared to a free lipase.

  13. Displaying Lipase B from Candida antarctica in Pichia pastoris Using the Yeast Surface Display Approach: Prospection of a New Anchor and Characterization of the Whole Cell Biocatalyst

    Science.gov (United States)

    Moura, Marcelo Victor Holanda; da Silva, Giulia Pontes; Machado, Antônio Carlos de Oliveira; Torres, Fernando Araripe Gonçalves; Freire, Denise Maria Guimarães; Almeida, Rodrigo Volcan

    2015-01-01

    Yeast Surface Display (YSD) is a strategy to anchor proteins on the yeast cell wall which has been employed to increase enzyme stability thus decreasing production costs. Lipase B from Candida antarctica (LipB) is one of the most studied enzymes in the context of industrial biotechnology. This study aimed to assess the biochemical features of this important biocatalyst when immobilized on the cell surface of the methylotrophic yeast Pichia pastoris using the YSD approach. For that purpose, two anchors were tested. The first (Flo9) was identified after a prospection of the P. pastoris genome being related to the family of flocculins similar to Flo1 but significantly smaller. The second is the Protein with Internal Repeats (Pir1) from P. pastoris. An immunolocalization assay showed that both anchor proteins were able to display the reporter protein EGFP in the yeast outer cell wall. LipB was expressed in P. pastoris fused either to Flo9 (FLOLIPB) or Pir1 (PIRLIPB). Both constructions showed hydrolytic activity towards tributyrin (>100 U/mgdcw and >80 U/mgdcw, respectively), optimal hydrolytic activity around 45°C and pH 7.0, higher thermostability at 45°C and stability in organic solvents when compared to a free lipase. PMID:26510006

  14. Cell wall composition of chlorococcal algae

    OpenAIRE

    Blumreisinger, Maria; Meindl, Doris; Loos, Eckhard

    1983-01-01

    The cell walls of representatives of the genera Chlorella, Monoraphidium, Ankistrodesmus and Scenedesmus contained 24–74% neutral sugars, 1–24% uronic acids, 2–16% protein and 0–15% glucosamine. Two types of cell walls could be discerned containing as main sugars either rhamnose and galactose or mannose and glucose with a lack of galactose.

  15. WallProtDB, a database resource for plant cell wall proteomics

    OpenAIRE

    San Clemente, Hélène; Jamet, Elisabeth

    2015-01-01

    Background During the last fifteen years, cell wall proteomics has become a major research field with the publication of more than 50 articles describing plant cell wall proteomes. The WallProtDB database has been designed as a tool to facilitate the inventory, the interpretation of cell wall proteomics data and the comparisons between cell wall proteomes. Results WallProtDB (http://www.polebio.lrsv.ups-tlse.fr/WallProtDB/) presently contains 2170 proteins and ESTs identified experimentally i...

  16. Phospholipase D specific for the phosphatidylinositol anchor of cell-surface proteins is abundant in plasma

    International Nuclear Information System (INIS)

    An enzyme activity capable of degrading the glycosyl-phosphatidylinositol membrane anchor of cell-surface proteins has previously been reported in a number of mammalian tissues. The experiments reported here demonstrate that this anchor-degrading activity is also abundant in mammalian plasma. The activity was inhibited by EGTA or 1,10-phenanthroline. It was capable of removing the anchor from alkaline phosphatase, 5'-nucleotidase, and variant surface glycoprotein but had little or no activity toward phosphatidylinositol or phosphatidylcholine. Phosphatidic acid was the only 3H-labeled product when this enzyme hydrolyzed [3H]myristate-labeled variant surface glycoprotein. It could be distinguished from the Ca2=-dependent inositol phospholipid-specific phospholipase C activity in several rat tissues on the basis of its molecular size and its sensitivity to 1,10-phenanthroline. The data therefore suggest that this activity is due to a phospholipase D with specificity for glycosylphosphatidylinositol structures. Although the precise physiological function of this anchor-specific phospholipase D remains to be determined, these findings indicate that it could play an important role in regulating the expression and release of cell-surface proteins in vivo

  17. Safranine fluorescent staining of wood cell walls.

    Science.gov (United States)

    Bond, J; Donaldson, L; Hill, S; Hitchcock, K

    2008-06-01

    Safranine is an azo dye commonly used for plant microscopy, especially as a stain for lignified tissues such as xylem. Safranine fluorescently labels the wood cell wall, producing green/yellow fluorescence in the secondary cell wall and red/orange fluorescence in the middle lamella (ML) region. We examined the fluorescence behavior of safranine under blue light excitation using a variety of wood- and fiber-based samples of known composition to interpret the observed color differentiation of different cell wall types. We also examined the basis for the differences in fluorescence emission using spectral confocal microscopy to examine lignin-rich and cellulose-rich cell walls including reaction wood and decayed wood compared to normal wood. Our results indicate that lignin-rich cell walls, such as the ML of tracheids, the secondary wall of compression wood tracheids, and wood decayed by brown rot, tend to fluoresce red or orange, while cellulose-rich cell walls such as resin canals, wood decayed by white rot, cotton fibers and the G-layer of tension wood fibers, tend to fluoresce green/yellow. This variation in fluorescence emission seems to be due to factors including an emission shift toward red wavelengths combined with dye quenching at shorter wavelengths in regions with high lignin content. Safranine fluorescence provides a useful way to differentiate lignin-rich and cellulose-rich cell walls without counterstaining as required for bright field microscopy. PMID:18802812

  18. Cell wall proteins: a new insight through proteomics

    OpenAIRE

    Jamet, Elisabeth; Canut, Hervé; Boudart, Georges; Pont-Lezica, Rafael F

    2006-01-01

    Cell wall proteins are essential constituents of plant cell walls; they are involved in modifications of cell wall components, wall structure, signaling and interactions with plasma membrane proteins at the cell surface. The application of proteomic approaches to the cell wall compartment raises important questions: are there technical problems specific to cell wall proteomics? What kinds of proteins can be found in Arabidopsis walls? Are some of them unexpected? What sort of post-translation...

  19. Construction of an artificial cell membrane anchor using DARC as a fitting for artificial extracellular functionalities of eukaryotic cells

    Directory of Open Access Journals (Sweden)

    von Nickisch-Rosenegk Markus

    2012-01-01

    Full Text Available Abstract The need to functionalize cell membranes in a directed way for specific applications as single cell arrays or to force close cell-to-cell contact for artificial intercellular interaction and/or induction concerning stem cell manipulation or in general to have a tool for membrane and cell surface-associated processes, we envisaged a neutral inactive membrane anchor for extracellular entities to facillitate the above mentioned functionalities. The silent Duffy antigen/receptor for chemokines (DARC is a receptor-like membrane protein of erythrocytes and mediates no cell transduction not at least regarding a missing or truncated G-loop and therefore it seemed to be the candidate for our cell membrane anchor. We isolated the genetic information of DARC from human genomic DNA and cloned it in a mammalian cell line as a fusion protein via a suitable plasmid vector. In this report we demonstrate that the human plasma membrane protein DARC can be used as an artificial anchor molecule in cell surface engineering applications. We constructed the fusion protein SNAP-tag-DARC, consisting of DARC and the self-labeling protein tag SNAP-tag® (Covalys. The SNAP-tag® served as an example for a molecular-technological developed protein that is artificially attached to the extracellular side of the plasma membrane through our DARC-anchor. SnapTag should serve as an example for any extracellular entity and was easy to detect by a commercial detection system. The synthesis of SNAP-tag-DARC, its correct incorporation into the cell membrane and the functionality of the SNAP-tag® were verified by RT-PCR, Western blotting and confocal fluorescence microscopy and showed the desired functionality as an membrane anchor for an extracellular application entity.

  20. Anchored and soluble gangliosides contribute to myelosupportivity of stromal cells

    International Nuclear Information System (INIS)

    Stroma-mediated myelopoiesis depends upon growth factors and an appropriate intercellular microenvironment. Previous studies have demonstrated that gangliosides, produced by hepatic stromal cell types, are required for optimal myelosupportive function. Here, we compared the mielossuportive functions of a bone marrow stroma (S17) and skin fibroblasts (SF) regarding their ganglioside pattern of synthesis and shedding. The survival and proliferation of a myeloid precursor cell (FDC-P1) were used as reporter. Although the ganglioside synthesis of the two stromal cells was similar, their relative content and shedding were distinct. The ganglioside requirement for mielossuportive function was confirmed by the decreased proliferation of FDC-P1 cells in ganglioside synthesis-inhibited cultures and in presence of an antibody to GM3 ganglioside. The distinct mielossuportive activities of the S17 and SF stromata may be related to differences on plasma membrane ganglioside concentrations or to differences on the gangliosides shed and their subsequent uptake by myeloid cells, specially, GM3 ganglioside.

  1. Anchored and soluble gangliosides contribute to myelosupportivity of stromal cells

    Energy Technology Data Exchange (ETDEWEB)

    Ziulkoski, Ana L. [Programa de Pos-Graduacao em Ciencias Biologicas: Bioquimica, ICBS, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS (Brazil); Departamento de Bioquimica, ICBS, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS (Brazil); Instituto de Ciencias da Saude, Centro Universitario Feevale, Novo Hamburgo, RS (Brazil); Santos, Aline X.S. dos; Andrade, Claudia M.B.; Trindade, Vera M.T. [Programa de Pos-Graduacao em Ciencias Biologicas: Bioquimica, ICBS, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS (Brazil); Departamento de Bioquimica, ICBS, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS (Brazil); Daniotti, Jose Luis [Departamento de Quimica Biologica, Faculdad de Ciencias Quimicas, Universidad Nacional de Cordoba, Cordoba (Argentina); Borojevic, Radovan [Departamento de Histologia e Embriologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro (Brazil); Guma, Fatima C.R., E-mail: fatima.guma@ufrgs.br [Programa de Pos-Graduacao em Ciencias Biologicas: Bioquimica, ICBS, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS (Brazil); Departamento de Bioquimica, ICBS, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS (Brazil)

    2009-10-09

    Stroma-mediated myelopoiesis depends upon growth factors and an appropriate intercellular microenvironment. Previous studies have demonstrated that gangliosides, produced by hepatic stromal cell types, are required for optimal myelosupportive function. Here, we compared the mielossuportive functions of a bone marrow stroma (S17) and skin fibroblasts (SF) regarding their ganglioside pattern of synthesis and shedding. The survival and proliferation of a myeloid precursor cell (FDC-P1) were used as reporter. Although the ganglioside synthesis of the two stromal cells was similar, their relative content and shedding were distinct. The ganglioside requirement for mielossuportive function was confirmed by the decreased proliferation of FDC-P1 cells in ganglioside synthesis-inhibited cultures and in presence of an antibody to GM3 ganglioside. The distinct mielossuportive activities of the S17 and SF stromata may be related to differences on plasma membrane ganglioside concentrations or to differences on the gangliosides shed and their subsequent uptake by myeloid cells, specially, GM3 ganglioside.

  2. Display of GPI-anchored anti-EGFR nanobodies on extracellular vesicles promotes tumour cell targeting

    Directory of Open Access Journals (Sweden)

    Sander A. A. Kooijmans

    2016-03-01

    Full Text Available Background: Extracellular vesicles (EVs are attractive candidate drug delivery systems due to their ability to functionally transport biological cargo to recipient cells. However, the apparent lack of target cell specificity of exogenously administered EVs limits their therapeutic applicability. In this study, we propose a novel method to equip EVs with targeting properties, in order to improve their interaction with tumour cells. Methods: EV producing cells were transfected with vectors encoding for anti-epidermal growth factor receptor (EGFR nanobodies, which served as targeting ligands for tumour cells, fused to glycosylphosphatidylinositol (GPI anchor signal peptides derived from decay-accelerating factor (DAF. EVs were isolated using ultrafiltration/size-exclusion liquid chromatography and characterized using western blotting, Nanoparticle Tracking Analysis, and electron microscopy. EV–tumour cell interactions were analyzed under static conditions using flow cytometry and under flow conditions using a live-cell fluorescence microscopy-coupled perfusion system. Results: V analysis showed that GPI-linked nanobodies were successfully displayed on EV surfaces and were highly enriched in EVs compared with parent cells. Display of GPI-linked nanobodies on EVs did not alter general EV characteristics (i.e. morphology, size distribution and protein marker expression, but greatly improved EV binding to tumour cells dependent on EGFR density under static conditions. Moreover, nanobody-displaying EVs showed a significantly improved cell association to EGFR-expressing tumour cells under flow conditions. Conclusions: We show that nanobodies can be anchored on the surface of EVs via GPI, which alters their cell targeting behaviour. Furthermore, this study highlights GPI-anchoring as a new tool in the EV toolbox, which may be applied for EV display of a variety of proteins, such as antibodies, reporter proteins and signaling molecules.

  3. Molecular regulation of plant cell wall extensibility

    Science.gov (United States)

    Cosgrove, D. J.

    1998-01-01

    Gravity responses in plants often involve spatial and temporal changes in cell growth, which is regulated primarily by controlling the ability of the cell wall to extend. The wall is thought to be a cellulose-hemicellulose network embedded in a hydrated matrix of complex polysaccharides and a small amount of structural protein. The wall extends by a form of polymer creep, which is mediated by expansins, a novel group of wall-loosening proteins. Expansins were discovered during a molecular dissection of the "acid growth" behavior of cell walls. Expansin alters the rheology of plant walls in profound ways, yet its molecular mechanism of action is still uncertain. It lacks detectable hydrolytic activity against the major components of the wall, but it is able to disrupt noncovalent adhesion between wall polysaccharides. The discovery of a second family of expansins (beta-expansins) sheds light on the biological role of a major group of pollen allergens and implies that expansins have evolved for diverse developmental functions. Finally, the contribution of other processes to wall extensibility is briefly summarized.

  4. Plant cell wall proteomics: the leadership of Arabidopsis thaliana

    OpenAIRE

    Albenne, Cécile; Canut, Hervé; Jamet, Elisabeth

    2013-01-01

    Plant cell wall proteins (CWPs) progressively emerged as crucial components of cell walls although present in minor amounts. Cell wall polysaccharides such as pectins, hemicelluloses, and cellulose represent more than 90% of primary cell wall mass, whereas hemicelluloses, cellulose, and lignins are the main components of lignified secondary walls. All these polymers provide mechanical properties to cell walls, participate in cell shape and prevent water loss in aerial organs. However, cell wa...

  5. 2003 Plant Cell Walls Gordon Conference

    Energy Technology Data Exchange (ETDEWEB)

    Daniel J. Cosgrove

    2004-09-21

    This conference will address recent progress in many aspects of cell wall biology. Molecular, genetic, and genomic approaches are yielding major advances in our understanding of the composition, synthesis, and architecture of plant cell walls and their dynamics during growth, and are identifying the genes that encode the machinery needed to make their biogenesis possible. This meeting will bring together international scientists from academia, industry and government labs to share the latest breakthroughs and perspectives on polysaccharide biosynthesis, wood formation, wall modification, expansion and interaction with other organisms, and genomic & evolutionary analyses of wall-related genes, as well as to discuss recent ''nanotechnological'' advances that take wall analysis to the level of a single cell.

  6. Refractive index of plant cell walls

    Science.gov (United States)

    Gausman, H. W.; Allen, W. A.; Escobar, D. E.

    1974-01-01

    Air was replaced with media of higher refractive indices by vacuum infiltration in leaves of cucumber, blackeye pea, tomato, and string bean plants, and reflectance of noninfiltrated and infiltrated leaves was spectrophotometrically measured. Infiltrated leaves reflected less light than noninfiltrated leaves over the 500-2500-nm wavelength interval because cell wall-air interfaces were partly eliminated. Minimal reflectance should occur when the average refractive index of plant cell walls was matched by the infiltrating fluid. Although refractive indices that resulted in minimal reflectance differed among the four plant genera, an average value of 1.425 approximates the refractive index of plant cell walls for the four plant genera.

  7. Disturbance of the bacterial cell wall specifically interferes with biofilm formation.

    Science.gov (United States)

    Bucher, Tabitha; Oppenheimer-Shaanan, Yaara; Savidor, Alon; Bloom-Ackermann, Zohar; Kolodkin-Gal, Ilana

    2015-12-01

    In nature, bacteria communicate via chemical cues and establish complex communities referred to as biofilms, wherein cells are held together by an extracellular matrix. Much research is focusing on small molecules that manipulate and prevent biofilm assembly by modifying cellular signalling pathways. However, the bacterial cell envelope, presenting the interface between bacterial cells and their surroundings, is largely overlooked. In our study, we identified specific targets within the biosynthesis pathways of the different cell wall components (peptidoglycan, wall teichoic acids and teichuronic acids) hampering biofilm formation and the anchoring of the extracellular matrix with a minimal effect on planktonic growth. In addition, we provide convincing evidence that biofilm hampering by transglycosylation inhibitors and D-Leucine triggers a highly specific response without changing the overall protein levels within the biofilm cells or the overall levels of the extracellular matrix components. The presented results emphasize the central role of the Gram-positive cell wall in biofilm development, resistance and sustainment. PMID:26472159

  8. STUDYING MEMBRANE ANCHOR ORGANIZATION IN LIVING CELL MEMBRANES

    OpenAIRE

    Huang, Hector Han-Li

    2011-01-01

    The cell membrane is a complex mixture of various lipids, proteins and other biomolecules that are all organized into a fluid 2-dimensional bilayer. A rather unique trait of this organelle is the lateral mobility of the component molecules. Surprisingly, these molecules are not necessarily distributed homogeneously in the membrane. From a physical perspective, these inhomogeneities are interesting because they indicate some level of organization in the membrane. From a biological perspect...

  9. Homogenization of a viscoelastic model for plant cell wall biomechanics

    OpenAIRE

    Ptashnyk, Mariya; Seguin, Brian

    2015-01-01

    The microscopic structure of a plant cell wall is given by cellulose microfibrils embedded in a cell wall matrix. In this paper we consider a microscopic model for interactions between viscoelastic deformations of a plant cell wall and chemical processes in the cell wall matrix. We consider elastic deformations of the cell wall microfibrils and viscoelastic Kelvin--Voigt type deformations of the cell wall matrix. Using homogenization techniques (two-scale convergence and periodic unfolding me...

  10. Permanent ground anchors: Nicholson design criteria

    Science.gov (United States)

    Nicholson, P. J.; Uranowski, D. D.; Wycliffe-Jones, P. T.

    1982-09-01

    The methods used by Nicholson Construction Company in the design of permanent ground anchors specifically as related to retaining walls are discussed. Basic soil parameters, design concepts, drilling and grouting methods for ground anchors are discussed. Particular emphasis is placed on anchors founded in soil rather than rock formations. Also, soil properties necessary for the proper design of anchored retaining walls are detailed. The second chapter of the report is devoted to a general discussion of retaining wall and anchor design. In addition, a design example of an anchored retaining wall is presented in a step by step manner.

  11. Function of laccases in cell wall biosynthesis

    DEFF Research Database (Denmark)

    Larsen, Anders; Holm, Preben Bach; Andersen, Jeppe Reitan

    2011-01-01

    Laccases are multicopper oxidases capable of polymerizing monolignols. Histochemical assays have shown temporal and spatial correlation with secondary cell wall formation in both herbs and woody perennials. However, in plants laccases constitutes a relatively large group of isoenzymes with unique...... substrate specificities and expression patterns. As part of the strategic research centre Bio4Bio, the present project deals with laccase functions in relation to cell wall formation in grasses based on a study of the model species Brachypodium distachyon. Thirty-one isozymes have been retrieved from the...... hybridization. Specific isozymes that show high correlation with the process of secondary cell wall formation will be further studied in a reverse genetic study in which candidates will be knocked out using RNA interference. Phenotypes of knock-out mutants are to be described in relation to cell wall...

  12. Decay accelerating factor of complement is anchored to cells by a C-terminal glycolipid

    International Nuclear Information System (INIS)

    Membrane-associated decay accelerating factor (DAF) of human erythrocytes (E/sup hu/) was analyzed for a C-terminal glycolipid anchoring structure. Automated amino acid analysis of DAF following reductive radiomethylation revealed ethanolamine and glucosamine residues in proportions identical with those present in the E/sup hu/ acetylcholinesterase (AChE) anchor. Cleavage of radiomethylated 70-kilodalton (kDa) DAF with papain released the labeled ethanolamine and glucosamine and generated 61- and 55-kDa DAF products that retained all labeled Lys and labeled N-terminal Asp. Incubation of intact E/sup hu/ with phosphatidylinositol-specific phospholipase C (PI-PLC), which cleaves the anchors in trypanosome membrane form variant surface glycoproteins (mfVSGs) and murine thymocyte Thy-1 antigen, released 15% of the cell-associated DAF antigen. The released 67-kDa PI-PLC DAF derivative retained its ability to decay the classical C3 convertase C4b2a but was unable to membrane-incorporate and displayed physicochemical properties similar to urine DAF, a hydrophilic DAF form that can be isolated for urine. Nitrous acid deamination cleavage of E/sup hu/ DAF at glucosamine following labeling with the lipophilic photoreagent 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine ([125I]TID) released the [125I]TID label in a parallel fashion as from [125I]TID-labeled AChE. Biosynthetic labeling of HeLa cells with [3H] ethanolamine resulted in rapid 3H incorporation into both 48-kDa pro-DAF and 72-kDa mature epithelial cell DAF. The findings indicate that DAF and AChE are anchored in E/sup hu/ by the same or a similar glycolipid structure and that, like VSGs, this structure is incorporated into DAF early in DAF biosynthesis prior to processing of pro-DAF in the Golgi

  13. Cell wall remodeling under abiotic stress

    OpenAIRE

    Tenhaken, Raimund

    2015-01-01

    Plants exposed to abiotic stress respond to unfavorable conditions on multiple levels. One challenge under drought stress is to reduce shoot growth while maintaining root growth, a process requiring differential cell wall synthesis and remodeling. Key players in this process are the formation of reactive oxygen species (ROS) and peroxidases, which initially cross-link phenolic compounds and glycoproteins of the cell walls causing stiffening. The function of ROS shifts after having converted a...

  14. Cell-wall dynamics in growing bacteria

    Science.gov (United States)

    Furchtgott, Leon; Wingreen, Ned; Huang, Kerwyn Casey

    2010-03-01

    Bacterial cells come in a large variety of shapes, and cell shape plays an important role in the regulation of many biological functions. Cell shape in bacterial cells is dictated by a cell wall composed of peptidoglycan, a polymer made up of long, stiff glycan strands and flexible peptide crosslinks. Although much is understood about the structural properties of peptidoglycan, little is known about the dynamics of cell wall organization in bacterial cells. In particular, during cell growth, how does the bacterial cell wall continuously expand and reorganize while maintaining cell shape? In order to investigate this question quantitatively, we model the cell wall of the Gram-negative bacterium Escherichia coli using a simple elastic model, in which glycan and peptide subunits are treated as springs with different spring constants and relaxed lengths. We consider the peptidoglycan network as a single-layered network of these springs under tension due to an internal osmotic pressure. Within this model, we simulate possible hypotheses for cell growth as different combinations of addition of new springs and breakage of old springs.

  15. The Cell Wall Protein Ecm33 of Candida albicans is Involved in Chronological Life Span, Morphogenesis, Cell Wall Regeneration, Stress Tolerance, and Host–Cell Interaction

    Science.gov (United States)

    Gil-Bona, Ana; Reales-Calderon, Jose A.; Parra-Giraldo, Claudia M.; Martinez-Lopez, Raquel; Monteoliva, Lucia; Gil, Concha

    2016-01-01

    Ecm33 is a glycosylphosphatidylinositol-anchored protein in the human pathogen Candida albicans. This protein is known to be involved in fungal cell wall integrity (CWI) and is also critical for normal virulence in the mouse model of hematogenously disseminated candidiasis, but its function remains unknown. In this work, several phenotypic analyses of the C. albicans ecm33/ecm33 mutant (RML2U) were performed. We observed that RML2U displays the inability of protoplast to regenerate the cell wall, activation of the CWI pathway, hypersensitivity to temperature, osmotic and oxidative stresses and a shortened chronological lifespan. During the exponential and stationary culture phases, nuclear and actin staining revealed the possible arrest of the cell cycle in RML2U cells. Interestingly, a “veil growth,” never previously described in C. albicans, was serendipitously observed under static stationary cells. The cells that formed this structure were also observed in cornmeal liquid cultures. These cells are giant, round cells, without DNA, and contain large vacuoles, similar to autophagic cells observed in other fungi. Furthermore, RML2U was phagocytozed more than the wild-type strain by macrophages at earlier time points, but the damage caused to the mouse cells was less than with the wild-type strain. Additionally, the percentage of RML2U apoptotic cells after interaction with macrophages was fewer than in the wild-type strain. PMID:26870022

  16. Bone Anchor Fixation in Abdominal Wall Reconstruction: A Useful Adjunct in Suprapubic and Para-iliac Hernia Repair.

    Science.gov (United States)

    Blair, Laurel J; Cox, Tiffany C; Huntington, Ciara R; Ross, Samuel W; Kneisl, Jeffrey S; Augenstein, Vedra A; Heniford, B Todd

    2015-07-01

    Suprapubic hernias, parailiac or flank hernias, and lumbar hernias are difficult to repair and are associated with high-recurrence rates owing to difficulty in obtaining substantive overlap and especially mesh fixation due to bone being a margin of the hernia. Orthopedic suture anchors used for ligament reconstruction have been used to attach prosthetic material to bony surfaces and can be used in the repair of these hernias where suture fixation was impossible. A prospective, single institution study of ventral hernia repairs involving bone anchor mesh fixation was performed. Demographics, operative details, and outcomes data were collected. Twenty patients were identified, with a mean age 53 (range: 35-70 years) and mean body mass index 28.4 kg/m(2) (range 21-38). Ten lumbar, seven suprapubic, and three parailiac hernias were studied. The majority were recurrent hernias (n = 13), with one to seven previously failed repairs. The mean hernia defect size was very large (270 cm(2); range: 56-832 cm(2)) with average mesh size of 1090 cm(2) (range 224-3640 cm(2)). Both Mitek GII (Depuy, Raynham, MA) and JuggerKnot 2.9-mm (Biomet, Biomedical Instruments, Warsaw, IN) anchors were used, with an average of four anchors/case (range: 1-16). Mean operative time was 218 minutes (120-495). There were three minor complications, no operative mortality, and no recurrences during an average follow-up of 24 months. Pelvic bone anchors permit mesh fixation in high-recurrence areas not amenable to traditional suture fixation. The ability to safely and effectively use bone anchor fixation is an essential tool in complex open ventral hernia repair. PMID:26140889

  17. Cell wall trapping of autocrine peptides for human G-protein-coupled receptors on the yeast cell surface.

    Directory of Open Access Journals (Sweden)

    Jun Ishii

    Full Text Available G-protein-coupled receptors (GPCRs regulate a wide variety of physiological processes and are important pharmaceutical targets for drug discovery. Here, we describe a unique concept based on yeast cell-surface display technology to selectively track eligible peptides with agonistic activity for human GPCRs (Cell Wall Trapping of Autocrine Peptides (CWTrAP strategy. In our strategy, individual recombinant yeast cells are able to report autocrine-positive activity for human GPCRs by expressing a candidate peptide fused to an anchoring motif. Following expression and activation, yeast cells trap autocrine peptides onto their cell walls. Because captured peptides are incapable of diffusion, they have no impact on surrounding yeast cells that express the target human GPCR and non-signaling peptides. Therefore, individual yeast cells can assemble the autonomous signaling complex and allow single-cell screening of a yeast population. Our strategy may be applied to identify eligible peptides with agonistic activity for target human GPCRs.

  18. "Steiner trees" between cell walls of sisal

    Institute of Scientific and Technical Information of China (English)

    LI GuanShi; YIN YaJun; LI Yan; ZHONG Zheng

    2009-01-01

    Through careful analysis on the cross-section of sisal fibers,it is found that the middle lamellae between the cell walls have clear geometric characteristics:between the cell walls of three neighboring cells,the middle lamellae form a three-way junction with 120°symmetry. If the neighboring three-way junctions are connected,a network of Steiner tree with angular symmetry and topological invariability is formed. If more and more Steiner trees are connected,a network of Steiner rings is generated. In another word,idealized cell walls and the middle lamellae are dominated by the Steiner geometry. This geometry not only depicts the geometric symmetry,the topological invariability and minimal property of the middle lamellae,but also controls the mechanics of sisal fibers.

  19. Exoskeleton anchoring to tendon cells and muscles in molting isopod crustaceans

    Directory of Open Access Journals (Sweden)

    Nada Žnidaršič

    2012-03-01

    Full Text Available Specialized mechanical connection between exoskeleton and underlying muscles in arthropods is a complex network of interconnected matrix constituents, junctions and associated cytoskeletal elements, which provides prominent mechanical attachment of the epidermis to the cuticle and transmits muscle tensions to the exoskeleton. This linkage involves anchoring of the complex extracellular matrix composing the cuticle to the apical membrane of tendon cells and linking of tendon cells to muscles basally. The ultrastructural arhitecture of these attachment complexes during molting is an important issue in relation to integument integrity maintenance in the course of cuticle replacement and in relation to movement ability. The aim of this work was to determine the ultrastructural organization of exoskeleton – muscles attachment complexes in the molting terrestrial isopod crustaceans, in the stage when integumental epithelium is covered by both, the newly forming cuticle and the old detached cuticle. We show that the old exoskeleton is extensively mechanically connected to the underlying epithelium in the regions of muscle attachment sites by massive arrays of fibers in adult premolt Ligia italica and in prehatching embryos and premolt marsupial mancas of Porcellio scaber. Fibers expand from the tendon cells, traverse the new cuticle and ecdysal space and protrude into the distal layers of the detached cuticle. They likely serve as final anchoring sites before exuviation and may be involved in animal movements in this stage. Tendon cells in the prehatching embryo and in marsupial mancas display a substantial apicobasally oriented transcellular arrays of microtubules, evidently engaged in myotendinous junctions and in apical anchoring of the cuticular matrix. The structural framework of musculoskeletal linkage is basically established in described intramarsupial developmental stages, suggesting its involvement in animal motility within the marsupium.

  20. Study of two dimensional nematic liquid crystal reorientations and anchoring effects

    OpenAIRE

    S Shoarinejad; MA Shahzamanian

    2010-01-01

    In this paper, the director distribution is calculated for a nematic liquid crystal, in the cell with different surface anchoring conditions and external fields. The effects of finite and infinite surface anchoring on molecular orientations for one dimensional geometry are discussed. In these situations, the planar alignment is considered. Then, in a two dimensional geometry the planar and homotropic anchoring conditions are assumed for wall- interfaces of confined nematic, and the reorientat...

  1. Non random usage of T cell receptor alpha gene expression in atopy using anchored PCR.

    Science.gov (United States)

    Mansur, A H; Gelder, C M; Holland, D; Campell, D A; Griffin, A; Cunliffe, W; Markham, A F; Morrison, J F

    1996-01-01

    The T cell receptor (TCR) alpha beta heterodimer recognises antigenic peptide fragments presented by Class II MHC. This interaction initiates T cell activation and cytokine release with subsequent recruitment of inflammatory cells. Previous work from our group suggests a qualitative difference in variable alpha gene expression in atopy as compared to non atopic controls. In this study we examine TCR alpha repertoire using anchored PCR to provide a quantitative assessment of the V alpha and J alpha repertoire. One atopic (DRB1*0701,DRB1*15: DRB4*0101, DRB5*01: DQB1* 0303, DQB1*601/2) and one non-atopic (DRB1*0701,DRB1*03011/2: DRB4*01, DRB3*0x: DQB1* 0303, DQB1*0201/2) control were studied. Variable gene usage was markedly limited in the atopic individual. V alpha 1, 3, 8 accounted for 60% and J alpha 12, 31 30% of the gene usage. There was evidence of preferential V alpha-J alpha gene pairing and clonal expansion. We conclude that there is a marked non random TCR alpha gene distribution in atopy using both V alpha family and anchored PCR. This may be due in part to antigen driven clonal expansion. PMID:9095269

  2. Roles of membrane trafficking in plant cell wall dynamics

    OpenAIRE

    Ebine, Kazuo; Ueda, Takashi

    2015-01-01

    The cell wall is one of the characteristic components of plant cells. The cell wall composition differs among cell types and is modified in response to various environmental conditions. To properly generate and modify the cell wall, many proteins are transported to the plasma membrane or extracellular space through membrane trafficking, which is one of the key protein transport mechanisms in eukaryotic cells. Given the diverse composition and functions of the cell wall in plants, the transpor...

  3. Sialic Acid on the Glycosylphosphatidylinositol Anchor Regulates PrP-mediated Cell Signaling and Prion Formation.

    Science.gov (United States)

    Bate, Clive; Nolan, William; Williams, Alun

    2016-01-01

    The prion diseases occur following the conversion of the cellular prion protein (PrP(C)) into disease-related isoforms (PrP(Sc)). In this study, the role of the glycosylphosphatidylinositol (GPI) anchor attached to PrP(C) in prion formation was examined using a cell painting technique. PrP(Sc) formation in two prion-infected neuronal cell lines (ScGT1 and ScN2a cells) and in scrapie-infected primary cortical neurons was increased following the introduction of PrP(C). In contrast, PrP(C) containing a GPI anchor from which the sialic acid had been removed (desialylated PrP(C)) was not converted to PrP(Sc). Furthermore, the presence of desialylated PrP(C) inhibited the production of PrP(Sc) within prion-infected cortical neurons and ScGT1 and ScN2a cells. The membrane rafts surrounding desialylated PrP(C) contained greater amounts of sialylated gangliosides and cholesterol than membrane rafts surrounding PrP(C). Desialylated PrP(C) was less sensitive to cholesterol depletion than PrP(C) and was not released from cells by treatment with glimepiride. The presence of desialylated PrP(C) in neurons caused the dissociation of cytoplasmic phospholipase A2 from PrP-containing membrane rafts and reduced the activation of cytoplasmic phospholipase A2. These findings show that the sialic acid moiety of the GPI attached to PrP(C) modifies local membrane microenvironments that are important in PrP-mediated cell signaling and PrP(Sc) formation. These results suggest that pharmacological modification of GPI glycosylation might constitute a novel therapeutic approach to prion diseases. PMID:26553874

  4. The Cell Wall Teichuronic Acid Synthetase (TUAS) Is an Enzyme Complex Located in the Cytoplasmic Membrane of Micrococcus luteus

    OpenAIRE

    Anderson, John S.; Alexander, Alice A.; Lingyi Lynn Deng; Sijin Lei

    2010-01-01

    The cell wall teichuronic acid (TUA) of Micrococcus luteus is a long-chain polysaccharide composed of disaccharide repeating units [-4-β-D-ManNAcAp-(1→6)α-D-Glcp−1-]n, which is covalently anchored to the peptidoglycan on the inner cell wall and extended to the outer surface of the cell envelope. An enzyme complex responsible for the TUA chain biosynthesis was purified and characterized. The 440 kDa enzyme complex, named teichuronic acid synthetase (TUAS), is...

  5. Cell Wall Heterogeneity in Root Development of Arabidopsis.

    Science.gov (United States)

    Somssich, Marc; Khan, Ghazanfar Abbas; Persson, Staffan

    2016-01-01

    Plant cell walls provide stability and protection to plant cells. During growth and development the composition of cell walls changes, but provides enough strength to withstand the turgor of the cells. Hence, cell walls are highly flexible and diverse in nature. These characteristics are important during root growth, as plant roots consist of radial patterns of cells that have diverse functions and that are at different developmental stages along the growth axis. Young stem cell daughters undergo a series of rapid cell divisions, during which new cell walls are formed that are highly dynamic, and that support rapid anisotropic cell expansion. Once the cells have differentiated, the walls of specific cell types need to comply with and support different cell functions. For example, a newly formed root hair needs to be able to break through the surrounding soil, while endodermal cells modify their walls at distinct positions to form Casparian strips between them. Hence, the cell walls are modified and rebuilt while cells transit through different developmental stages. In addition, the cell walls of roots readjust to their environment to support growth and to maximize nutrient uptake. Many of these modifications are likely driven by different developmental and stress signaling pathways. However, our understanding of how such pathways affect cell wall modifications and what enzymes are involved remain largely unknown. In this review we aim to compile data linking cell wall content and re-modeling to developmental stages of root cells, and dissect how root cell walls respond to certain environmental changes. PMID:27582757

  6. GPI-anchored proteins do not reside in ordered domains in the live cell plasma membrane

    Science.gov (United States)

    Sevcsik, Eva; Brameshuber, Mario; Fölser, Martin; Weghuber, Julian; Honigmann, Alf; Schütz, Gerhard J.

    2015-04-01

    The organization of proteins and lipids in the plasma membrane has been the subject of a long-lasting debate. Membrane rafts of higher lipid chain order were proposed to mediate protein interactions, but have thus far not been directly observed. Here we use protein micropatterning combined with single-molecule tracking to put current models to the test: we rearranged lipid-anchored raft proteins (glycosylphosphatidylinositol(GPI)-anchored-mGFP) directly in the live cell plasma membrane and measured the effect on the local membrane environment. Intriguingly, this treatment does neither nucleate the formation of an ordered membrane phase nor result in any enrichment of nanoscopic-ordered domains within the micropatterned regions. In contrast, we find that immobilized mGFP-GPIs behave as inert obstacles to the diffusion of other membrane constituents without influencing their membrane environment over distances beyond their physical size. Our results indicate that phase partitioning is not a fundamental element of protein organization in the plasma membrane.

  7. Arrangement of peptidoglycan in the cell wall of Staphylococcus spp.

    OpenAIRE

    Amako, K; Umeda, A.; Murata, K

    1982-01-01

    The arrangement of peptidoglycan in the cell wall of Staphylococcus was observed with the newly developed freeze-fracture technique, using n-octanol instead of water as the freezing medium. The replica of the trichloroacetic acid-extracted cell wall (TCA-wall) showed two areas. One of them has a concentric circular structure, a characteristic surface structure of the staphylococcal cell wall, and the other showed an irregular and rough surface. The chemical analysis of the wall revealed that ...

  8. Fluorescent tags to explore cell wall structure and dynamics

    OpenAIRE

    Gonneau, Martine; Höfte, Herman; Vernhettes, Samantha

    2012-01-01

    Plant cell walls are highly dynamic and heterogeneous structures, which vary between cell types, growth stages but also between microdomains within a single cell wall. In this review, we summarize the imaging techniques using fluorescent tags that are currently being used and which should in the coming years revolutionize our understanding of the dynamics of cell wall architecture and the cellular processes involved in the synthesis of cell wall components.

  9. Measuring in vitro extensibility of growing plant cell walls.

    Science.gov (United States)

    Cosgrove, Daniel J

    2011-01-01

    This article summarizes the theory and practical aspects of measuring cell wall properties by four different extensometer techniques and how the results of these methods relate to the concept and ideal measurement of cell wall extensibility in the context of cell growth. These in vivo techniques are particularly useful for studies of the molecular basis of cell wall extension. Measurements of breaking strength, elastic compliance, and plastic compliance may be informative about changes in cell wall structure, whereas measurements of wall stress relaxation and creep are sensitive to both changes in wall structure and wall-loosening processes, such as those mediated by expansins and some lytic enzymes. A combination of methods is needed to obtain a broader view of cell wall behavior and properties connected with the concept of cell wall extensibility. PMID:21222092

  10. Plant Cell Wall Matrix Polysaccharide Biosynthesis

    Institute of Scientific and Technical Information of China (English)

    Ajay Pal S. Sandhu; Gursharn S. Randhawa; Kanwarpal S. Dhugga

    2009-01-01

    The wall of an expanding plant cell consists primarily of cellulose microfibrils embedded in a matrix of hemi-cellulosic and pectic polysaccharides along with small amounts of structural and enzymatic proteins. Matrix polysacchar-ides are synthesized in the Golgi and exported to the cell wall by exocytosis, where they intercalate among cellulose microfibrUs, which are made at the plasma membrane and directly deposited into the cell wall. Involvement of Golgi glucan synthesis in auxin-induced cell expansion has long been recognized; however, only recently have the genes corresponding to glucan synthases been identified. Biochemical purification was unsuccessful because of the labile nature and very low abundance of these enzymes. Mutational genetics also proved fruitless. Expression of candidate genes identified through gene expression profiling or comparative genomics in heterologous systems followed by functional characterization has been relatively successful. Several genes from the cellulose synthase-like (Cs/) family have been found to be involved in the synthesis of various hemicellulosic glycans. The usefulness of this approach, however, is limited to those enzymes that probably do not form complexes consisting of unrelated proteins. Nonconventional approaches will continue to incre-mentally unravel the mechanisms of Golgi polysaccharide biosynthesis.

  11. Alfalfa stem tissues: Cell wall deposition, composition, and degradability

    NARCIS (Netherlands)

    Jung, H.G.; Engels, F.M.

    2002-01-01

    Declining cell wall degradability of alfalfa (Medicago sativa L.) stems with maturation limits the nutritional value of alfalfa for ruminants. This study characterized changes in cell wall concentration, composition, and degradability by rumen microbes resulting from alfalfa stem tissue proliferatio

  12. Carbazole dye with phosphonic acid anchoring groups for long-term heat stability of dye-sensitized solar cells

    International Nuclear Information System (INIS)

    In dye-sensitized solar cells (DSSCs), the binding strength between the dye and TiO2 surface may affect the heat stability of the DSSCs. Carboxylic acids have generally been used as the anchoring groups for the adsorption of dyes on TiO2 surfaces. However, a phosphonic acid anchor is expected to effect more stable bonding to a TiO2 surface because of its tridentate binding capability. In this study, we developed a new carbazole dye that employed a phosphonic acid anchor, MK-94. We studied the effect of the dye anchoring group on the heat stability of DSSCs by comparing MK-94, MK-2 (a carbazole dye with a carboxylic acid anchoring group), and Z-907 (a well-known, robust Ru complex dye with a carboxylic acid anchoring group). In the initial performance assessment, the DSSC with MK-94 exhibited a lower short circuit current (jsc) but a higher open circuit voltage (ΔVoc) than MK-2. With respect to heat stability, decreases in jsc and ΔVoc were suppressed with MK-94, and the drop of the power conversion efficiency (PCE) was kept within 6% after 504 h of heating. In addition, MK-94 demonstrated the longest electron lifetime and largest charge transfer resistance than the other dyes during the stability test. In contrast, the PCE decreased 27% from the initial value in the DSSC with MK-2. Thus, the phosphonic acid anchor was effective in extending the long-term heat stability of the DSSCs

  13. SUPPRESSION OF CELL ADHESION ON POLYACRYLONITRILE-BASED MEMBRANES BY THE ANCHORING OF PHOSPHOLIPID MOIETIES

    Institute of Scientific and Technical Information of China (English)

    Xiao-jun Huang; Xiao-dan Huang; Ai-fu Che; Zhi-kang Xu; Ke Yao

    2006-01-01

    In this work, the membrane surface of poly(acrylonitrile-co-2-hydroxyethyl methacrylate) (PANCHEMA) waschemically modified by anchoring of phospholipid moieties. The process involved the reaction of hydroxyl groups on the membrane surface with 2-chloro-2-oxo-1,3,2-dioxaphospholane (COP) followed by the ring-opening reaction of COP with trimethylamine. Chemical differences between the original and the modified membranes were characterized by FT-IR and XPS. It was found that the amount of macrophage adhered on the modified membrane surface is substantially lower than that on polyacrylonitrile (PAN) and PANCHEMA membranes under the same condition. The morphological change of the adherent cell is also suppressed by the generation ofphospholipid moieties on the membrane surface.

  14. Endo-b-1,4-glucanases impact plant cell wall development by influencing cellulose crystallization

    Institute of Scientific and Technical Information of China (English)

    Magdalena Glass; Sarah Barkwill; Faride Unda; Shawn D. Mansfield

    2015-01-01

    Cell walls are vital to the normal growth and development of plants as they protect the protoplast and provide rigidity to the stem. Here, two poplar and Arabidopsis orthologous endoglucanases, which have been proposed to play a role in secondary cell wall development, were examined. The class B endoglucanases, PtGH9B5 and AtGH9B5, are secreted enzymes that have a predicted glycosylphosphatidylinositol anchor, while the class C endo-glucanases, PtGH9C2 and AtGH9C2, are also predicted to be secreted but instead contain a carbohydrate-binding module. The poplar endoglucanases were expressed in Arabidopsis using both a 35S promoter and the Arabidopsis secondary cell wall-specific CesA8 promoter. Additionally, Arabidopsis t-DNA insertion lines and an RNAi construct was created to downregulate AtGH9C2 in Arabidopsis. All of the plant lines were examined for changes in cell morphology and pattern-ing, growth and development, cell wall crystallinity, microfibril angle, and proportion of cell wall carbohydrates. Misregula-tion of PtGH9B5/AtGH9B5 resulted in changes in xylose content, while misregulation of PtGH9C2/AtGH9C2 resulted in changes in crystallinity, which was inversely correlated with changes in plant height and rosette diameter. Together, these results suggest that these endoglucanases affect secondary cell wall development by contributing to the cell wall crystallization process.

  15. Structure-property relationships in vegetable cell wall suspensions

    OpenAIRE

    Sankaran, Ashwin Karthik

    2015-01-01

    Plant cell wall suspensions are widely present in daily food, such as soups, dressings and sauces. Cell walls of edible plants are made up of an intricate biopolymer network of mainly cellulose microfibrils, pectins, and hemicelluloses. Foodsnbsp;as soups, ketchup, etc are made up of cell wall components. Modern processing methods alter the chemical and physical nature of the cell wall which in turn affect the properties of the end product. There is a need in the industry to build a fundament...

  16. Bio-based composites that mimic the plant cell wall

    OpenAIRE

    Li, Zhuo

    2009-01-01

    Nature creates high performance materials under modest conditions, i.e., neutral pH and ambient temperature and pressure. One of the most significant materials is the plant cell wall. The plant cell wall is a composite of oriented cellulose microfibrils reinforcing a lignin/hemicellulose matrix. In principle, the plant cell wall composite is designed much like a synthetic fiber-reinforced polymer composite. Unlike synthetic composites, the plant cell wall has an excellent combination of h...

  17. Fluorescent tags to explore cell wall structure and dynamics.

    OpenAIRE

    Martine eGonneau; Herman eHöfte; Samantha eVernhettes

    2012-01-01

    Plant cell walls are highly dynamic and heterogeneic structures, which vary between celltypes, growth stages but also between microdomains within a single cell wall. In this review, we summarize the imaging techniques using fluorescent tags that are currently being used and which should in the coming years revolutionize our understanding of the dynamics of cell wall architecture and the cellular processes involved in synthesis of cell wall components.

  18. Dystroglycan loss disrupts polarity and beta-casein induction inmammary epithelial cells by perturbing laminin anchoring

    Energy Technology Data Exchange (ETDEWEB)

    Weir, M. Lynn; Oppizzi, Maria Luisa; Henry, Michael D.; Onishi,Akiko; Campbell, Kevin P.; Bissell, Mina J.; Muschler, John L.

    2006-02-17

    Precise contact between epithelial cells and their underlying basement membrane is critical to the maintenance of tissue architecture and function. To understand the role that the laminin receptor dystroglycan (DG) plays in these processes, we assayed cell responses to laminin-111 following conditional ablation of DG expression in cultured mammary epithelial cells (MECs). Strikingly, DG loss disrupted laminin-111-induced polarity and {beta}-casein production, and abolished laminin assembly at the step of laminin binding to the cell surface. DG re-expression restored these deficiencies. Investigations of mechanism revealed that DG cytoplasmic sequences were not necessary for laminin assembly and signaling, and only when the entire mucin domain of extracellular DG was deleted did laminin assembly not occur. These results demonstrate that DG is essential as a laminin-111 co-receptor in MECs that functions by mediating laminin anchoring to the cell surface, a process that allows laminin polymerization, tissue polarity, and {beta}-casein induction. The observed loss of laminin-111 assembly and signaling in DG-/-MECs provides insights into the signaling changes occurring in breast carcinomas and other cancers, where DG's laminin-binding function is frequently defective.

  19. Association Mapping of Cell Wall Synthesis Regulatory Genes and Cell Wall Quality in Switchgrass

    Energy Technology Data Exchange (ETDEWEB)

    Bartley, Laura [Univ. of Oklahoma, Norman, OK (United States). Dept. of Microbiology and Plant Biology; Wu, Y. [Oklahoma State Univ., Stillwater, OK (United States); Zhu, L. [Oklahoma State Univ., Stillwater, OK (United States); Brummer, E. C. [Noble Foundation, Ardmore, OK (United States); Saha, M. [Noble Foundation, Ardmore, OK (United States)

    2016-05-31

    Inefficient conversion of biomass to biofuels is one of the main barriers for biofuel production from such materials. Approximately half of polysaccharides in biomass remain unused by typical biochemical conversion methods. Conversion efficiency is influenced by the composition and structure of cell walls of biomass. Grasses such as wheat, maize, and rice, as well as dedicated perennial bioenergy crops, like switchgrass, make up ~55% of biomass that can be produced in the United States. Grass cell walls have a different composition and patterning compared with dicotyledonous plants, including the well-studied model plant, Arabidopsis. This project identified genetic determinants of cell wall composition in grasses using both naturally occurring genetic variation of switchgrass and gene network reconstruction and functional assays in rice. In addition, the project linked functional data in rice and other species to switchgrass improvement efforts through curation of the most abundant class of regulators in the switchgrass genome. Characterizing natural diversity of switchgrass for variation in cell wall composition and properties, also known as quality, provides an unbiased avenue for identifying biologically viable diversity in switchgrass cell walls. To characterizing natural diversity, this project generated cell wall composition and enzymatic deconstruction data for ~450 genotypes of the Switchgrass Southern Association Collection (SSAC), a diverse collection composed of 36 switchgrass accessions from the southern U.S. distribution of switchgrass. Comparing these data with other measures of cell wall quality for the same samples demonstrated the complementary nature of the diverse characterization platforms now being used for biomass characterization. Association of the composition data with ~3.2K single nucleotide variant markers identified six significant single nucleotide variant markers co-associated with digestibility and another compositional trait. These

  20. Wall relaxation and the driving forces for cell expansive growth

    Science.gov (United States)

    Cosgrove, D. J.

    1987-01-01

    When water uptake by growing cells is prevented, the turgor pressure and the tensile stress in the cell wall are reduced by continued wall loosening. This process, termed in vivo stress relaxation, provides a new way to study the dynamics of wall loosening and to measure the wall yield threshold and the physiological wall extensibility. Stress relaxation experiments indicate that wall stress supplies the mechanical driving force for wall yielding. Cell expansion also requires water absorption. The driving force for water uptake during growth is created by wall relaxation, which lowers the water potential of the expanding cells. New techniques for measuring this driving force show that it is smaller than believed previously; in elongating stems it is only 0.3 to 0.5 bar. This means that the hydraulic resistance of the water transport pathway is small and that rate of cell expansion is controlled primarily by wall loosening and yielding.

  1. Plant cell wall dynamics and wall-related susceptibility in plant–pathogen interactions

    OpenAIRE

    Bellincampi, Daniela; Cervone, Felice; Lionetti, Vincenzo

    2014-01-01

    The cell wall is a dynamic structure that often determines the outcome of the interactions between plants and pathogens. It is a barrier that pathogens need to breach to colonize the plant tissue. While fungal necrotrophs extensively destroy the integrity of the cell wall through the combined action of degrading enzymes, biotrophic fungi require a more localized and controlled degradation of the cell wall in order to keep the host cells alive and utilize their feeding structures. Also bacteri...

  2. Plant cell wall dynamics and wall-related susceptibility in plant-pathogen interactions

    OpenAIRE

    Daniela eBellincampi; Felice eCervone; Vincenzo eLionetti

    2014-01-01

    The cell wall is a dynamic structure that often determines the outcome of the interactions between plants and pathogens. It is a barrier that pathogens need to breach to colonize the plant tissue. While fungal necrotrophs extensively destroy the integrity of the cell wall through the combined action of degrading enzymes, biotrophic fungi require a more localized and controlled degradation of the cell wall in order to keep the host cells alive and utilize their feeding structures. Also bacteri...

  3. Evidence that a glycolipid tail anchors antigen 117 to the plasma membrane of Dictyostelium discoideum cells

    International Nuclear Information System (INIS)

    The authors describe the biochemical features of the putative cell cohesion molecule antigen 117, indicating that it is anchored to the plasma membrane by a glycolipid tail. Antigen 117 can be radiolabeled with [3H]myristate, [3H]palmitate, and [14C]ethanolamine. The fatty acid label is removed by periodate oxidation and nitrous acid deamination, indicating that the fatty acid is attached to the protein by a structure containing carbohydrate and an unsubstituted glucosamine. As cells develop aggregation competence, the antigen is released from the cell surface in a soluble form that can still be radiolabeled with [14C]ethanolamine but not with [3H]myristate of [3H]-palmitate. The molecular weight of the released antigen is similar to that found in the plasma membrane, but it preferentially partitions in Triton X-114 as a hydrophilic, as opposed to a hydrophobic, protein. Plasma membranes contain the enzyme activity responsible for the release of the antigen in a soluble form

  4. Cell Geometry Guides the Dynamic Targeting of Apoplastic GPI-Linked Lipid Transfer Protein to Cell Wall Elements and Cell Borders in Arabidopsis thaliana

    Science.gov (United States)

    Wasteneys, Geoffrey

    2013-01-01

    During cellular morphogenesis, changes in cell shape and cell junction topology are fundamental to normal tissue and organ development. Here we show that apoplastic Glycophosphatidylinositol (GPI)-anchored Lipid Transfer Protein (LTPG) is excluded from cell junctions and flat wall regions, and passively accumulates around their borders in the epidermal cells of Arabidopsis thaliana. Beginning with intense accumulation beneath highly curved cell junction borders, this enrichment is gradually lost as cells become more bulbous during their differentiation. In fully mature epidermal cells, YFP-LTPG often shows a fibrous cellulose microfibril-like pattern within the bulging outer faces. Physical contact between a flat glass surface and bulbous cell surface induces rapid and reversible evacuation from contact sites and accumulation to the curved wall regions surrounding the contact borders. Thus, LTPG distribution is dynamic, responding to changes in cell shape and wall curvature during cell growth and differentiation. We hypothesize that this geometry-based mechanism guides wax-carrying LTPG to functional sites, where it may act to “seal” the vulnerable border surrounding cell-cell junctions and assist in cell wall fortification and cuticular wax deposition. PMID:24260561

  5. Construction Technology of Anchor Support in Silo Wall of Large Underground Coal Storage%大型地下储煤槽仓仓壁锚杆支护施工方法

    Institute of Scientific and Technical Information of China (English)

    杨圣扩

    2012-01-01

    The silo wall in large underground coal storage is supported by anchor, which can assure stability of the anchored soil. This paper introduces in detail the characteristics of anchor support, technique principle, key operating points and quality control measures. The anchor support method is used in several projects. The application effect shows that the application of anchor in slope reinforcement can reduce soil excavation and discarding soil, decrease land use area, and so as to reduce damage to surrounding environment. The temporary anchor support surface of silo wall, also as permanent silo wall structure,can effectively shorten construction period, reduce project cost.%在大型地下储煤槽仓仓壁采用锚杆进行支护,能够保证被锚固土体的稳定.主要介绍了锚杆支护特点、工艺原理、操作要点和质量控制措施.该方法在多个项目成功应用,应用效果表明预应力锚杆在边坡加固中应用,有利于减少土方开挖和弃方量,减少用地面积,从而减少对周边环境的破坏.临时性仓壁支护锚杆面层与永久性结构仓壁合二为一,有效地缩短了施工工期,大大降低了工程投资.

  6. An accessory protein required for anchoring and assembly of amyloid fibres in B. subtilis biofilms.

    Science.gov (United States)

    Romero, Diego; Vlamakis, Hera; Losick, Richard; Kolter, Roberto

    2011-06-01

    Cells within Bacillus subtilis biofilms are held in place by an extracellular matrix that contains cell-anchored amyloid fibres, composed of the amyloidogenic protein TasA. As biofilms age they disassemble because the cells release the amyloid fibres. This release appears to be the consequence of incorporation of D-tyrosine, D-leucine, D-tryptophan and D-methionine into the cell wall. Here, we characterize the in vivo roles of an accessory protein TapA (TasA anchoring/assembly protein; previously YqxM) that serves both to anchor the fibres to the cell wall and to assemble TasA into fibres. TapA is found in discrete foci in the cell envelope and these foci disappear when cells are treated with a mixture of D-amino acids. Purified cell wall sacculi retain a functional form of this anchoring protein such that purified fibres can be anchored to the sacculi in vitro. In addition, we show that TapA is essential for the proper assembly of the fibres. Its absence results in a dramatic reduction in TasA levels and what little TasA is left produces only thin fibres that are not anchored to the cell. PMID:21477127

  7. Glycosytransferases involved in arabinosylation of cell wall extensins

    DEFF Research Database (Denmark)

    Petersen, Bent L; Harholt, Jesper; Jørgensen, Bodil;

    2011-01-01

    Extensins are a group of ancient hydroxyproline rich cell wall glycoproteins that are found in some chlorophyte algae (such as Chlamydomonas), where they constitute the main wall building block, as well as in higher plant cell walls, where they constitute a relatively minor component of particular...

  8. Cell wall integrity signaling and innate immunity in plants

    OpenAIRE

    Nühse, Thomas S.

    2012-01-01

    All plant pathogens and parasites have had to develop strategies to overcome cell walls in order to access the host’s cytoplasm. As a mechanically strong, multi-layered composite exoskeleton, the cell wall not only enables plants to grow tall but also protects them from such attacks. Many plant pathogens employ an arsenal of cell wall degrading enzymes, and it has long been thought that the detection of breaches in wall integrity contributes to the induction of defense. Cell wall fragments ar...

  9. Phosphatidylinositol-anchored glycoproteins of PC12 pheochromocytoma cells and brain

    Energy Technology Data Exchange (ETDEWEB)

    Margolis, R.K.; Goossen, B.; Margolis, R.U.

    1988-05-03

    PC12 pheochromocytoma cells and cultures of early postnatal rat cerebellium were labeled with (/sup 3/H)glucosamine, (/sup 3/H)fucose, (/sup 3/H)leucine, (/sup 3/H)ethanolamine, or sodium (/sup 35/S)sulfate and treated with a phosphatidylinositol-specific phospholipase C. Enzyme treatment of (/sup 3/H) glucosamine- or (/sup 3/H)fucose-labeled PC12 cells led to a 15-fold increase in released glycoproteins. On sodium dodecyl sulfate-polyacrylamide gel ectrophoresis, most of the released material migrated as a broad band with an apparent molecular size of 32,000 daltons (Da), which was specifically immunoprecipitated by a monoclonal antibody to the Thy-l glycoprotein. A second glycoprotein, with an apparent molecular size of 158,000 Da, was also released. After treatment with endo-..beta..-galactosidase, 40-45% of the (/sup 3/H)glucosamine of (/sup 3/H)fucose radioactivity in the phospholipase-released glycoproteins was converted to products of disaccharide size, and the molecular size of the 158-kDa glycoprotein decreased to 145 kDa, demonstrating that it contains fucosylated poly-(N-acetyllactosaminyl) oligosaccharides. The phospholipase also released labeled Thy-1 and the 158-kDa glycoprotein from PC12 cells cultured in the presence of (/sup 3/H)ethanolamine, which specifically labels this component of the phosphatidylinositol membrane-anchoring sequence,while in the lipid-free protein residue of cells not treated with phospholipase, Thy-1 and a doublet at 46/48 kDa were the only labeled proteins. Sulfated glycoproteins of 155, 132/134, 61, and 21 kDa are the predominant species released by phospholipase, which does not affect a major 44-kDa protein seen in (/sup 3/H)ethanolamine-labeled brain cultures. The 44-48- and 155/158-kDa proteins may be common to both PC12 cells and brain.

  10. Phosphatidylinositol-anchored glycoproteins of PC12 pheochromocytoma cells and brain

    International Nuclear Information System (INIS)

    PC12 pheochromocytoma cells and cultures of early postnatal rat cerebellium were labeled with [3H]glucosamine, [3H]fucose, [3H]leucine, [3H]ethanolamine, or sodium [35S]sulfate and treated with a phosphatidylinositol-specific phospholipase C. Enzyme treatment of [3H] glucosamine- or [3H]fucose-labeled PC12 cells led to a 15-fold increase in released glycoproteins. On sodium dodecyl sulfate-polyacrylamide gel ectrophoresis, most of the released material migrated as a broad band with an apparent molecular size of 32,000 daltons (Da), which was specifically immunoprecipitated by a monoclonal antibody to the Thy-l glycoprotein. A second glycoprotein, with an apparent molecular size of 158,000 Da, was also released. After treatment with endo-β-galactosidase, 40-45% of the [3H]glucosamine of [3H]fucose radioactivity in the phospholipase-released glycoproteins was converted to products of disaccharide size, and the molecular size of the 158-kDa glycoprotein decreased to 145 kDa, demonstrating that it contains fucosylated poly-(N-acetyllactosaminyl) oligosaccharides. The phospholipase also released labeled Thy-1 and the 158-kDa glycoprotein from PC12 cells cultured in the presence of [3H]ethanolamine, which specifically labels this component of the phosphatidylinositol membrane-anchoring sequence,while in the lipid-free protein residue of cells not treated with phospholipase, Thy-1 and a doublet at 46/48 kDa were the only labeled proteins. Sulfated glycoproteins of 155, 132/134, 61, and 21 kDa are the predominant species released by phospholipase, which does not affect a major 44-kDa protein seen in [3H]ethanolamine-labeled brain cultures. The 44-48- and 155/158-kDa proteins may be common to both PC12 cells and brain

  11. Chemical and functional properties of cell wall polymers from two cherry varieties at two developmental stages.

    Science.gov (United States)

    Basanta, María F; de Escalada Plá, Marina F; Stortz, Carlos A; Rojas, Ana M

    2013-01-30

    The cell wall polysaccharides of Regina and Sunburst cherry varieties at two developmental stages were extracted sequentially, and their changes in monosaccharide composition and functional properties were studied. The loosely-attached pectins presented a lower d-galacturonic acid/rhamnose ratio than ionically-bound pectins, as well as lower thickening effects of their respective 2% aqueous solution: the lowest Newtonian viscosity and shear rate dependence during the pseudoplastic phase. The main constituents of the cell wall matrix were covalently bound pectins (probably through diferulate cross-linkings), with long arabinan side chains at the RG-I cores. This pectin domain was also anchored into the XG-cellulose elastic network. Ripening occurred with a decrease in the proportion of HGs, water extractable GGM and xylogalacturonan, and with a concomitant increase in neutral sugars. Ripening was also associated with higher viscosities and thickening effects, and to larger distribution of molecular weights. The highest firmness and compactness of Regina cherry may be associated with its higher proportion of calcium-bound HGs localized in the middle lamellae of cell walls, as well as to some higher molar proportion of NS (Rha and Ara) in covalently bound pectins. These pectins showed significantly better hydration properties than hemicellulose and cellulose network. Chemical composition and functional properties of cell wall polymers were dependent on cherry variety and ripening stage, and helped explain the contrasting firmness of Regina and Sunburst varieties. PMID:23218373

  12. Enzymes and other agents that enhance cell wall extensibility

    Science.gov (United States)

    Cosgrove, D. J.

    1999-01-01

    Polysaccharides and proteins are secreted to the inner surface of the growing cell wall, where they assemble into a network that is mechanically strong, yet remains extensible until the cells cease growth. This review focuses on the agents that directly or indirectly enhance the extensibility properties of growing walls. The properties of expansins, endoglucanases, and xyloglucan transglycosylases are reviewed and their postulated roles in modulating wall extensibility are evaluated. A summary model for wall extension is presented, in which expansin is a primary agent of wall extension, whereas endoglucanases, xyloglucan endotransglycosylase, and other enzymes that alter wall structure act secondarily to modulate expansin action.

  13. Anthocyanins influence tannin-cell wall interactions.

    Science.gov (United States)

    Bautista-Ortín, Ana Belén; Martínez-Hernández, Alejandro; Ruiz-García, Yolanda; Gil-Muñoz, Rocío; Gómez-Plaza, Encarna

    2016-09-01

    The rate of tannin extraction was studied in a vinification of red grapes and the results compared with another vinification made with white grapes fermented as for typical red wine, in the presence of skins and seeds. Even though the grapes presented a quite similar skin and seed tannin content, the differences in tannin concentration between both vinifications was very large, despite the fact that the only apparent difference between the phenolic composition of both wines was the anthocyanin content. This suggests that anthocyanins play an important role in tannin extractability, perhaps because they affect the extent of the tannin-cell wall interaction, a factor that largely controls the resulting quantity of tannins in wines. To confirm this observation, the effect of anthocyanins on the tannin extractability from grape seeds and skin and on the interaction between tannins and grape cell walls suspended in model solutions were studied. The results indicated that anthocyanins favored skin and seed tannin extraction and that there is a competition for the adsorption sites between anthocyanins and tannins that increases the tannin content when anthocyanins are present. PMID:27041322

  14. Disruption of hydrogen bonding between plant cell wall polymers by proteins that induce wall extension.

    Science.gov (United States)

    McQueen-Mason, S; Cosgrove, D J

    1994-07-01

    Plant cell enlargement is controlled by the ability of the constraining cell wall to expand. This ability has been postulated to be under the control of polysaccharide hydrolases or transferases that weaken or rearrange the loadbearing polymeric networks in the wall. We recently identified a family of wall proteins, called expansins, that catalyze the extension of isolated plant cell walls. Here we report that these proteins mechanically weaken pure cellulose paper in extension assays and stress relaxation assays, without detectable cellulase activity (exo- or endo- type). Because paper derives its mechanical strength from hydrogen bonding between cellulose microfibrils, we conclude that expansins can disrupt hydrogen bonding between cellulose fibers. This conclusion is further supported by experiments in which expansin-mediated wall extension (i) was increased by 2 M urea (which should weaken hydrogen bonding between wall polymers) and (ii) was decreased by replacement of water with deuterated water, which has a stronger hydrogen bond. The temperature sensitivity of expansin-mediated wall extension suggests that units of 3 or 4 hydrogen bonds are broken by the action of expansins. In the growing cell wall, expansin action is likely to catalyze slippage between cellulose microfibrils and the polysaccharide matrix, and thereby catalyze wall stress relaxation, followed by wall surface expansion and plant cell enlargement. PMID:11607483

  15. Two endogenous proteins that induce cell wall extension in plants

    Science.gov (United States)

    McQueen-Mason, S.; Durachko, D. M.; Cosgrove, D. J.

    1992-01-01

    Plant cell enlargement is regulated by wall relaxation and yielding, which is thought to be catalyzed by elusive "wall-loosening" enzymes. By employing a reconstitution approach, we found that a crude protein extract from the cell walls of growing cucumber seedlings possessed the ability to induce the extension of isolated cell walls. This activity was restricted to the growing region of the stem and could induce the extension of isolated cell walls from various dicot stems and the leaves of amaryllidaceous monocots, but was less effective on grass coleoptile walls. Endogenous and reconstituted wall extension activities showed similar sensitivities to pH, metal ions, thiol reducing agents, proteases, and boiling in methanol or water. Sequential HPLC fractionation of the active wall extract revealed two proteins with molecular masses of 29 and 30 kD associated with the activity. Each protein, by itself, could induce wall extension without detectable hydrolytic breakdown of the wall. These proteins appear to mediate "acid growth" responses of isolated walls and may catalyze plant cell wall extension by a novel biochemical mechanism.

  16. Advanced technologies for plant cell wall evolution and diversity

    DEFF Research Database (Denmark)

    Fangel, Jonatan Ulrik

    cannot really be synthesised or sequenced. The work described in this thesis is focused to a large extent on the development of a microarray-based high-throughput method for cell wall analysis known as Comprehensive microarray polymer profiling or CoMPP. The procedure uses highly specific molecular...... produced has provided new insight into cell wall evolution and biosynthesis and has contributed to the commercial development of cell wall materials. A major focus of the work has been the wide scale sampling of cell wall diversity across the plant kingdom, from unicellular algae to highly evolved......Plant cell walls consist of polysaccharides, glycoproteins and phenolic polymers interlinked together in a highly complex network. The detailed analysis of cell walls is challenging because of their inherent complexity and heterogeneity. Also, complex carbohydrates, unlike proteins and nucleotides...

  17. An asymmetric supercapacitor with ultrahigh energy density based on nickle cobalt sulfide nanocluster anchoring multi-wall carbon nanotubes hybrid

    Science.gov (United States)

    Wen, Ping; Fan, Mingjin; Yang, Desuo; Wang, Yan; Cheng, Hualei; Wang, Jinqing

    2016-07-01

    The development of novel electrode materials with high energy density and long cycling life is critical to realize electrochemical capacitive energy storage for practical applications. In this paper, the hybrids of nickle cobalt sulfide/multi-wall carbon nanotubes (NiCo2S4/MWCNTs) with different contents of MWCNTs are prepared using a facile one-pot solvothermal reaction. As novel active materials for supercapacitors, the electrochemistry tests show that the hybrid of NiCo2S4/MWCNTs-5 is able to deliver a high specific capacitance of 2080 F g-1 at the current density of 1 A g-1, even superior rate capability of 61% capacitance retention after a 20-fold increase in current densities, when the content of MWCNTs is up to 5%. More importantly, an asymmetric supercapacitor assembled by NiCo2S4/MWCNTs-5 as positive electrode and reduced graphene oxide (rGO) as negative electrode delivers a high energy density of 51.8 Wh Kg-1 at a power density of 865 W kg-1, and 85.7% of its initial capacitance is retained at the current density of 4 A g-1 after 5000 charge-discharge cycles, exhibiting potential prospect for practical applications.

  18. Characterisation of cell wall polysaccharides in bilberries and black currants

    OpenAIRE

    Hilz, H

    2007-01-01

    During berry juice production, polysaccharides are released from the cell walls and cause thickening and high viscosity when the berries are mashed. Consequences are a low juice yield and a poor colour. This can be prevented by the use of enzymes that degrade these polysaccharides. To use these enzymes most efficiently, the structure and composition of the cell walls had to be known. This thesis describes a detailed composition of the cell walls of bilberries and black currants. The obtained ...

  19. Calpain-Mediated Positional Information Directs Cell Wall Orientation to Sustain Plant Stem Cell Activity, Growth and Development.

    Science.gov (United States)

    Liang, Zhe; Brown, Roy C; Fletcher, Jennifer C; Opsahl-Sorteberg, Hilde-Gunn

    2015-09-01

    Eukaryotic development and stem cell control depend on the integration of cell positional sensing with cell cycle control and cell wall positioning, yet few factors that directly link these events are known. The DEFECTIVE KERNEL1 (DEK1) gene encoding the unique plant calpain protein is fundamental for development and growth, being essential to confer and maintain epidermal cell identity that allows development beyond the globular embryo stage. We show that DEK1 expression is highest in the actively dividing cells of seeds, meristems and vasculature. We further show that eliminating Arabidopsis DEK1 function leads to changes in developmental cues from the first zygotic division onward, altered microtubule patterns and misshapen cells, resulting in early embryo abortion. Expression of the embryonic marker genes WOX2, ATML1, PIN4, WUS and STM, related to axis organization, cell identity and meristem functions, is also altered in dek1 embryos. By monitoring cell layer-specific DEK1 down-regulation, we show that L1- and 35S-induced down-regulation mainly affects stem cell functions, causing severe shoot apical meristem phenotypes. These results are consistent with a requirement for DEK1 to direct layer-specific cellular activities and set downstream developmental cues. Our data suggest that DEK1 may anchor cell wall positions and control cell division and differentiation, thereby balancing the plant's requirement to maintain totipotent stem cell reservoirs while simultaneously directing growth and organ formation. A role for DEK1 in regulating microtubule-orchestrated cell wall orientation during cell division can explain its effects on embryonic development, and suggests a more general function for calpains in microtubule organization in eukaryotic cells. PMID:26220906

  20. Composition of lignin in outer cell-wall layers

    OpenAIRE

    Christiernin, Maria

    2006-01-01

    The composition of lignin in the outer cell-wall layers of spruce and poplar has been studied and the data obtained have been compared with those of the mature reference wood in which the secondary cell wall predominates. Materials with exclusively or predominantly outer cell-wall layers were examined. Accurate data relating to the lignin monomer composition and the number of β-O-4´ bonds were obtained from pure middle lamella/primary cell wall lignin. Firstly, a 10 000 year old white spruce ...

  1. Cosegregation of cell wall and DNA in Bacillus subtilis.

    OpenAIRE

    Schlaeppi, J M; Karamata, D

    1982-01-01

    Cosegregation of cell wall and DNA of a lysis-negative mutant of Bacillus subtilis was examined by continuously labeling (i) cell wall, (ii) DNA, and (iii) both cell wall and DNA. After four to five generations of chase in liquid media it was found by light microscope autoradiography that the numbers of wall segregation units per cell are 29 and 9 in rich and minimal medium, respectively. Under the same conditions the numbers of segregation units of DNA were almost 50% lower: 15 and 5, respec...

  2. Cell wall structure and biogenesis in Aspergillus species.

    Science.gov (United States)

    Yoshimi, Akira; Miyazawa, Ken; Abe, Keietsu

    2016-09-01

    Aspergillus species are among the most important filamentous fungi from the viewpoints of industry, pathogenesis, and mycotoxin production. Fungal cells are exposed to a variety of environmental stimuli, including changes in osmolality, temperature, and pH, which create stresses that primarily act on fungal cell walls. In addition, fungal cell walls are the first interactions with host cells in either human or plants. Thus, understanding cell wall structure and the mechanism of their biogenesis is important for the industrial, medical, and agricultural fields. Here, we provide a systematic review of fungal cell wall structure and recent findings regarding the cell wall integrity signaling pathways in aspergilli. This accumulated knowledge will be useful for understanding and improving the use of industrial aspergilli fermentation processes as well as treatments for some fungal infections. PMID:27140698

  3. Cellulose synthesis in two secondary cell wall processes in a single cell type

    OpenAIRE

    Mendu, Venugopal; Stork, Jozsef; Harris, Darby; DeBolt, Seth

    2011-01-01

    Plant cells have a rigid cell wall that constrains internal turgor pressure yet extends in a regulated and organized manner to allow the cell to acquire shape. The primary load-bearing macromolecule of a plant cell wall is cellulose, which forms crystalline microfibrils that are organized with respect to a cell's function and shape requirements. A primary cell wall is deposited during expansion whereas secondary cell wall is synthesized post expansion during differentiation. A complex form of...

  4. Measurement of streptococcal cell wall in tissues of rats resistant or susceptible to cell wall-induced chronic erosive arthritis.

    OpenAIRE

    Anderle, S K; Allen, J B; Wilder, R L; Eisenberg, R A; Cromartie, W J; Schwab, J. H.

    1985-01-01

    The quantity of streptococcal cell wall localized in the joints of rats of strains which are either susceptible (Sprague-Dawley, LEW/N, M520/N) or resistant (Buffalo, WKY/N, F344/N) to cell wall-induced chronic erosive arthritis was measured after intraperitoneal injection of group A streptococcal cell wall fragments. Susceptibility or resistance was not associated with a difference in the amount of cell wall localized in limbs or other tissues. It is concluded that although localization of c...

  5. Micropipette aspiration on the outer hair cell lateral wall.

    OpenAIRE

    Sit, P S; Spector, A A; Lue, A J; Popel, A S; Brownell, W.E.

    1997-01-01

    The mechanical properties of the lateral wall of the guinea pig cochlear outer hair cell were studied using the micropipette aspiration technique. A fire-polished micropipette with an inner diameter of approximately 4 microm was brought into contact with the lateral wall and negative pressure was applied. The resulting deformation of the lateral wall was recorded on videotape and subjected to morphometric analysis. The relation between the length of the aspirated portion of the cell and aspir...

  6. Regulation of Meristem Morphogenesis by Cell Wall Synthases in Arabidopsis

    OpenAIRE

    Yang, Weibing; Schuster, Christoph; Beahan, Cherie T.; Charoensawan, Varodom; Peaucelle, Alexis; Bacic, Antony; Doblin, Monika S.; Wightman, Raymond; Meyerowitz, Elliot M.

    2016-01-01

    The cell walls of the shoot apical meristem (SAM), containing the stem cell niche that gives rise to the above-ground tissues, are crucially involved in regulating differentiation. It is currently unknown how these walls are built and refined or their role, if any, in influencing meristem developmental dynamics. We have combined polysaccharide linkage analysis, immuno-labeling, and transcriptome profiling of the SAM to provide a spatiotemporal plan of the walls of this dynamic structure. We f...

  7. Assembly and enlargement of the primary cell wall in plants

    Science.gov (United States)

    Cosgrove, D. J.

    1997-01-01

    Growing plant cells are shaped by an extensible wall that is a complex amalgam of cellulose microfibrils bonded noncovalently to a matrix of hemicelluloses, pectins, and structural proteins. Cellulose is synthesized by complexes in the plasma membrane and is extruded as a self-assembling microfibril, whereas the matrix polymers are secreted by the Golgi apparatus and become integrated into the wall network by poorly understood mechanisms. The growing wall is under high tensile stress from cell turgor and is able to enlarge by a combination of stress relaxation and polymer creep. A pH-dependent mechanism of wall loosening, known as acid growth, is characteristic of growing walls and is mediated by a group of unusual wall proteins called expansins. Expansins appear to disrupt the noncovalent bonding of matrix hemicelluloses to the microfibril, thereby allowing the wall to yield to the mechanical forces generated by cell turgor. Other wall enzymes, such as (1-->4) beta-glucanases and pectinases, may make the wall more responsive to expansin-mediated wall creep whereas pectin methylesterases and peroxidases may alter the wall so as to make it resistant to expansin-mediated creep.

  8. Structural Studies of Complex Carbohydrates of Plant Cell Walls

    Energy Technology Data Exchange (ETDEWEB)

    Darvill, Alan [Univ. of Georgia, Athens, GA (United States); Hahn, Michael G. [Univ. of Georgia, Athens, GA (United States); O' Neill, Malcolm A. [Univ. of Georgia, Athens, GA (United States); York, William S. [Univ. of Georgia, Athens, GA (United States)

    2015-02-17

    Most of the solar energy captured by land plants is converted into the polysaccharides (cellulose, hemicellulose, and pectin) that are the predominant components of the cell wall. These walls, which account for the bulk of plant biomass, have numerous roles in the growth and development of plants. Moreover, these walls have a major impact on human life as they are a renewable source of biomass, a source of diverse commercially useful polymers, a major component of wood, and a source of nutrition for humans and livestock. Thus, understanding the molecular mechanisms that lead to wall assembly and how cell walls and their component polysaccharides contribute to plant growth and development is essential to improve and extend the productivity and value of plant materials. The proposed research will develop and apply advanced analytical and immunological techniques to study specific changes in the structures and interactions of the hemicellulosic and pectic polysaccharides that occur during differentiation and in response to genetic modification and chemical treatments that affect wall biosynthesis. These new techniques will make it possible to accurately characterize minute amounts of cell wall polysaccharides so that subtle changes in structure that occur in individual cell types can be identified and correlated to the physiological or developmental state of the plant. Successful implementation of this research will reveal fundamental relationships between polysaccharide structure, cell wall architecture, and cell wall functions.

  9. Hemicellulose biosynthesis and degradation in tobacco cell walls

    NARCIS (Netherlands)

    Compier, M.G.M.

    2005-01-01

    Natural fibres have a wide range of technological applications, such as in paper and textile industries. The basic properties and the quality of plant fibres are determined by the composition of the plant cell wall. Characteristic for fibres are thick secondary cell walls, which consist of cellulose

  10. Characterising the cellulose synthase complexes of cell walls

    NARCIS (Netherlands)

    Mansoori Zangir, N.

    2012-01-01

    One of the characteristics of the plant kingdom is the presence of a structural cell wall. Cellulose is a major component in both the primary and secondary cell walls of plants. In higher plants cellulose is synthesized by so called rosette protein complexes with cellulose synthases (CESAs) as the c

  11. Engineering the Oryza sativa cell wall with rice NAC transcription factors regulating secondary wall formation

    Directory of Open Access Journals (Sweden)

    Kouki eYoshida

    2013-10-01

    Full Text Available Plant tissues that require structural rigidity synthesize a thick, strong secondary cell wall of lignin, cellulose and hemicelluloses in a complicated bridged structure. Master regulators of secondary wall synthesis were identified in dicots, and orthologs of these regulators have been identified in monocots, but regulation of secondary cell wall formation in monocots has not been extensively studied. Here we demonstrate that the rice transcription factors SECONDARY WALL NAC DOMAIN PROTEINs (SWNs can regulate secondary wall formation in rice (Oryza sativa and are potentially useful for engineering the monocot cell wall. The OsSWN1 promoter is highly active in sclerenchymatous cells of the leaf blade and less active in xylem cells. By contrast, the OsSWN2 promoter is highly active in xylem cells and less active in sclerenchymatous cells. OsSWN2 splicing variants encode two proteins; the shorter protein (OsSWN2S has very low transcriptional activation ability, but the longer protein (OsSWN2L and OsSWN1 have strong transcriptional activation ability. In rice, expression of an OsSWN2S chimeric repressor, driven by the OsSWN2 promoter, resulted in stunted growth and para-wilting (leaf rolling and browning under normal water conditions due to impaired vascular vessels. The same OsSWN2S chimeric repressor, driven by the OsSWN1 promoter, caused a reduction of cell wall thickening in sclerenchymatous cells, a drooping leaf phenotype, reduced lignin and xylose contents and increased digestibility as forage. These data suggest that OsSWNs regulate secondary wall formation in rice and manipulation of OsSWNs may enable improvements in monocotyledonous crops for forage or biofuel applications.

  12. Engineering the Oryza sativa cell wall with rice NAC transcription factors regulating secondary wall formation.

    Science.gov (United States)

    Yoshida, Kouki; Sakamoto, Shingo; Kawai, Tetsushi; Kobayashi, Yoshinori; Sato, Kazuhito; Ichinose, Yasunori; Yaoi, Katsuro; Akiyoshi-Endo, Miho; Sato, Hiroko; Takamizo, Tadashi; Ohme-Takagi, Masaru; Mitsuda, Nobutaka

    2013-01-01

    Plant tissues that require structural rigidity synthesize a thick, strong secondary cell wall of lignin, cellulose and hemicelluloses in a complicated bridged structure. Master regulators of secondary wall synthesis were identified in dicots, and orthologs of these regulators have been identified in monocots, but regulation of secondary cell wall formation in monocots has not been extensively studied. Here we demonstrate that the rice transcription factors SECONDARY WALL NAC DOMAIN PROTEINs (SWNs) can regulate secondary wall formation in rice (Oryza sativa) and are potentially useful for engineering the monocot cell wall. The OsSWN1 promoter is highly active in sclerenchymatous cells of the leaf blade and less active in xylem cells. By contrast, the OsSWN2 promoter is highly active in xylem cells and less active in sclerenchymatous cells. OsSWN2 splicing variants encode two proteins; the shorter protein (OsSWN2S) has very low transcriptional activation ability, but the longer protein (OsSWN2L) and OsSWN1 have strong transcriptional activation ability. In rice, expression of an OsSWN2S chimeric repressor, driven by the OsSWN2 promoter, resulted in stunted growth and para-wilting (leaf rolling and browning under normal water conditions) due to impaired vascular vessels. The same OsSWN2S chimeric repressor, driven by the OsSWN1 promoter, caused a reduction of cell wall thickening in sclerenchymatous cells, a drooping leaf phenotype, reduced lignin and xylose contents and increased digestibility as forage. These data suggest that OsSWNs regulate secondary wall formation in rice and manipulation of OsSWNs may enable improvements in monocotyledonous crops for forage or biofuel applications. PMID:24098302

  13. On-Off Switches for Secondary Cell Wall Biosynthesis

    Institute of Scientific and Technical Information of China (English)

    Huan-Zhong Wang; Richard A.Dixon

    2012-01-01

    Secondary cell walls provide plants with rigidity and strength to support their body weight and ensure water and nutrient transport.They also provide textiles,timber,and potentially second-generation biofuels for human use.Genes responsible for synthesis of the different cell wall components,namely cellulose,hemicelluloses,and lignin,are coordinately expressed and under transcriptional regulation.In the past several years,cell wall-related NAC and MYB transcription factors have been intensively investigated in different species and shown to be master switches of secondary cell wall biosynthesis.Positive and negative regulators,which function upstream of NAC master switches,have also been identified in different plant tissues.Further elucidation of the regulatory mechanisms of cell wall synthesis will facilitate the engineering of plant feedstocks suitable for biofuel production.

  14. The role of wall calcium in the extension of cell walls of soybean hypocotyls

    Science.gov (United States)

    Virk, S. S.; Cleland, R. E.

    1990-01-01

    Calcium crosslinks are load-bearing bonds in soybean (Glycine max (L.) Merr.) hypocotyl cell walls, but they are not the same load-bearing bonds that are broken during acid-mediated cell elongation. This conclusion is reached by studying the relationship between wall calcium, pH and the facilitated creep of frozen-thawed soybean hypocotyl sections. Supporting data include the following observations: 1) 2-[(2-bis-[carboxymethyl]amino-5-methylphenoxy)methyl]-6-methoxy-8-bis[car boxymethyl]aminoquinoline (Quin 2) and ethylene glycol-bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) caused only limited facilitated creep as compared with acid, despite removal of comparable or larger amounts of wall calcium; 2) the pH-response curves for calcium removal and acid-facilitated creep were different; 3) reversible acid-extension occurred even after removal of almost all wall calcium with Quin 2; and 4) growth of abraded sections did not involve a proportional loss of wall calcium. Removal of wall calcium, however, increased the capacity of the walls to undergo acid-facilitated creep. These data indicate that breakage of calcium crosslinks is not a major mechanism of cell-wall loosening in soybean hypocotyl tissues.

  15. Pectin, a versatile polysaccharide present in plant cell walls

    OpenAIRE

    Voragen, A.G.J.; Coenen, G.J.; Verhoef, R.P.; Schols, H.A.

    2009-01-01

    Pectin or pectic substances are collective names for a group of closely associated polysaccharides present in plant cell walls where they contribute to complex physiological processes like cell growth and cell differentiation and so determine the integrity and rigidity of plant tissue. They also play an important role in the defence mechanisms against plant pathogens and wounding. As constituents of plant cell walls and due to their anionic nature, pectic polysaccharides are considered to be ...

  16. The state of cell wall pectin monitored by wall associated kinases: A model

    OpenAIRE

    Kohorn, Bruce D

    2015-01-01

    The Wall Associated Kinases (WAKs) bind to both cross-linked polymers of pectin in the plant cell wall, but have a higher affinity for smaller fragmented pectins that are generated upon pathogen attack or wounding. WAKs are required for cell expansion during normal seedling development and this involves pectin binding and a signal transduction pathway involving MPK3 and invertase induction. Alternatively WAKs bind pathogen generated pectin fragments to activate a distinct MPK6 dependent stres...

  17. Glucuronoarabinoxylan structure in the walls of Aechmea leaf chlorenchyma cells is related to wall strength.

    Science.gov (United States)

    Ceusters, Johan; Londers, Elsje; Brijs, Kristof; Delcour, Jan A; De Proft, Maurice P

    2008-09-01

    In CAM-plants rising levels of malic acid in the early morning cause elevated turgor pressures in leaf chlorenchyma cells. Under specific conditions this process is lethal for sensitive plants resulting in chlorenchyma cell burst while other species can cope with these high pressures and do not show cell burst under comparable conditions. The non-cellulosic polysaccharide composition of chlorenchyma cell walls was investigated and compared in three cultivars of Aechmea with high sensitivity for chlorenchyma cell burst and three cultivars with low sensitivity. Chlorenchyma layers were cut from the leaf and the non-cellulosic carbohydrate fraction of the cell wall fraction was analyzed by gas-liquid chromatography. Glucuronoarabinoxylans (GAXs) were the major non-cellulosic polysaccharides in Aechmea. The fine structure of these GAXs was strongly related to chlorenchyma wall strength. Chlorenchyma cell walls from cultivars with low sensitivity to cell burst were characterized by an A/X ratio of ca. 0.13 while those from cultivars with high sensitivity showed an A/X ratio of ca. 0.23. Xylose chains from cultivars with high cell burst sensitivity were ca. 40% more substituted with arabinose compared to cultivars with low sensitivity for cell burst. The results indicate a relationship in vivo between glucuronoarabinoxylan fine structure and chlorenchyma cell wall strength in Aechmea. The evidence obtained supports the hypothesis that GAXs with low degrees of substitution cross-link cellulose microfibrils, while GAXs with high degrees of substitution do not. A lower degree of arabinose substitution on the xylose backbone implies stronger cell walls and the possibility of withstanding higher internal turgor pressures without cell bursting. PMID:18632122

  18. Lipid II: a central component in bacterial cell wall synthesis and a target for antibiotics.

    Science.gov (United States)

    de Kruijff, Ben; van Dam, Vincent; Breukink, Eefjan

    2008-01-01

    The bacterial cell wall is mainly composed of peptidoglycan, which is a three-dimensional network of long aminosugar strands located on the exterior of the cytoplasmic membrane. These strands consist of alternating MurNAc and GlcNAc units and are interlinked to each other via peptide moieties that are attached to the MurNAc residues. Peptidoglycan subunits are assembled on the cytoplasmic side of the bacterial membrane on a polyisoprenoid anchor and one of the key components in the synthesis of peptidoglycan is Lipid II. Being essential for bacterial cell survival, it forms an attractive target for antibacterial compounds such as vancomycin and several lantibiotics. Lipid II consists of one GlcNAc-MurNAc-pentapeptide subunit linked to a polyiosoprenoid anchor 11 subunits long via a pyrophosphate linker. This review focuses on this special molecule and addresses three questions. First, why are special lipid carriers as polyprenols used in the assembly of peptidoglycan? Secondly, how is Lipid II translocated across the bacterial cytoplasmic membrane? And finally, how is Lipid II used as a receptor for lantibiotics to kill bacteria? PMID:19008088

  19. How cell wall complexity influences saccharification efficiency in Miscanthus sinensis.

    Science.gov (United States)

    De Souza, Amanda P; Alvim Kamei, Claire L; Torres, Andres F; Pattathil, Sivakumar; Hahn, Michael G; Trindade, Luisa M; Buckeridge, Marcos S

    2015-07-01

    The production of bioenergy from grasses has been developing quickly during the last decade, with Miscanthus being among the most important choices for production of bioethanol. However, one of the key barriers to producing bioethanol is the lack of information about cell wall structure. Cell walls are thought to display compositional differences that lead to emergence of a very high level of complexity, resulting in great diversity in cell wall architectures. In this work, a set of different techniques was used to access the complexity of cell walls of different genotypes of Miscanthus sinensis in order to understand how they interfere with saccharification efficiency. Three genotypes of M. sinensis displaying different patterns of correlation between lignin content and saccharification efficiency were subjected to cell wall analysis by quantitative/qualitative analytical techniques such as monosaccharide composition, oligosaccharide profiling, and glycome profiling. When saccharification efficiency was correlated negatively with lignin, the structural features of arabinoxylan and xyloglucan were found to contribute positively to hydrolysis. In the absence of such correlation, different types of pectins, and some mannans contributed to saccharification efficiency. Different genotypes of M. sinensis were shown to display distinct interactions among their cell wall components, which seem to influence cell wall hydrolysis. PMID:25908240

  20. Messenger Functions of the Bacterial Cell Wall-derived Muropeptides

    OpenAIRE

    Boudreau, Marc A.; Fisher, Jed F.; Mobashery, Shahriar

    2012-01-01

    Bacterial muropeptides are soluble peptidoglycan structures central to recycling of the bacterial cell wall, and messengers in diverse cell-signaling events. Bacteria sense muropeptides as signals that antibiotics targeting cell-wall biosynthesis are present, and eukaryotes detect muropeptides during the innate immune response to bacterial infection. This review summarizes the roles of bacterial muropeptides as messengers, with a special emphasis on bacterial muropeptide structures and the re...

  1. Cell wall remodeling in mycorrhizal symbiosis: a way towards biotrophism

    Science.gov (United States)

    Balestrini, Raffaella; Bonfante, Paola

    2014-01-01

    Cell walls are deeply involved in the molecular talk between partners during plant and microbe interactions, and their role in mycorrhizae, i.e., the widespread symbiotic associations established between plant roots and soil fungi, has been investigated extensively. All mycorrhizal interactions achieve full symbiotic functionality through the development of an extensive contact surface between the plant and fungal cells, where signals and nutrients are exchanged. The exchange of molecules between the fungal and the plant cytoplasm takes place both through their plasma membranes and their cell walls; a functional compartment, known as the symbiotic interface, is thus defined. Among all the symbiotic interfaces, the complex intracellular interface of arbuscular mycorrhizal (AM) symbiosis has received a great deal of attention since its first description. Here, in fact, the host plasma membrane invaginates and proliferates around all the developing intracellular fungal structures, and cell wall material is laid down between this membrane and the fungal cell surface. By contrast, in ectomycorrhizae (ECM), where the fungus grows outside and between the root cells, plant and fungal cell walls are always in direct contact and form the interface between the two partners. The organization and composition of cell walls within the interface compartment is a topic that has attracted widespread attention, both in ecto- and endomycorrhizae. The aim of this review is to provide a general overview of the current knowledge on this topic by integrating morphological observations, which have illustrated cell wall features during mycorrhizal interactions, with the current data produced by genomic and transcriptomic approaches. PMID:24926297

  2. A proteomic and genetic analysis of the Neurospora crassa conidia cell wall proteins identifies two glycosyl hydrolases involved in cell wall remodeling.

    Science.gov (United States)

    Ao, Jie; Aldabbous, Mash'el; Notaro, Marysa J; Lojacono, Mark; Free, Stephen J

    2016-09-01

    A proteomic analysis of the conidial cell wall identified 35 cell wall proteins. A comparison with the proteome of the vegetative hyphae showed that 16 cell wall proteins were shared, and that these shared cell wall proteins were cell wall biosynthetic proteins or cell wall structural proteins. Deletion mutants for 34 of the genes were analyzed for phenotypes indicative of conidial cell wall defects. Mutants for two cell wall glycosyl hydrolases, the CGL-1 β-1,3-glucanase (NCU07523) and the NAG-1 exochitinase (NCU10852), were found to have a conidial separation phenotype. These two enzymes function in remodeling the cell wall between adjacent conidia to facilitate conidia formation and dissemination. Using promoter::RFP and promoter::GFP constructs, we demonstrated that the promoters for 15 of the conidia-specific cell wall genes, including cgl-1 and nag-1, provided for conidia-specific gene expression or for a significant increase in their expression during conidiation. PMID:27381444

  3. Mechanical Properties of Plant Cell Walls Probed by Relaxation Spectra

    DEFF Research Database (Denmark)

    Hansen, Steen Laugesen; Ray, Peter Martin; Karlsson, Anders Ola;

    2011-01-01

    type. This may be due to the plant’s ability to compensate for the wall modification or because the biophysical method that is often employed, determination of simple elastic modulus and breakstrength, lacks the resolving power necessary for detecting subtle mechanical phenotypes. Here, we apply a...... method, determination of relaxation spectra, which probes, and can separate, the viscoelastic properties of different cell wall components (i.e. those properties that depend on the elastic behavior of load-bearing wall polymers combined with viscous interactions between them). A computer program, Bayes......Relax, that deduces relaxation spectra from appropriate rheological measurements is presented and made accessible through a Web interface. BayesRelax models the cell wall as a continuum of relaxing elements, and the ability of the method to resolve small differences in cell wall mechanical properties is...

  4. Regulation of Meristem Morphogenesis by Cell Wall Synthases in Arabidopsis.

    Science.gov (United States)

    Yang, Weibing; Schuster, Christoph; Beahan, Cherie T; Charoensawan, Varodom; Peaucelle, Alexis; Bacic, Antony; Doblin, Monika S; Wightman, Raymond; Meyerowitz, Elliot M

    2016-06-01

    The cell walls of the shoot apical meristem (SAM), containing the stem cell niche that gives rise to the above-ground tissues, are crucially involved in regulating differentiation. It is currently unknown how these walls are built and refined or their role, if any, in influencing meristem developmental dynamics. We have combined polysaccharide linkage analysis, immuno-labeling, and transcriptome profiling of the SAM to provide a spatiotemporal plan of the walls of this dynamic structure. We find that meristematic cells express only a core subset of 152 genes encoding cell wall glycosyltransferases (GTs). Systemic localization of all these GT mRNAs by in situ hybridization reveals members with either enrichment in or specificity to apical subdomains such as emerging flower primordia, and a large class with high expression in dividing cells. The highly localized and coordinated expression of GTs in the SAM suggests distinct wall properties of meristematic cells and specific differences between newly forming walls and their mature descendants. Functional analysis demonstrates that a subset of CSLD genes is essential for proper meristem maintenance, confirming the key role of walls in developmental pathways. PMID:27212401

  5. Cell Wall Composition, Biosynthesis and Remodeling during Pollen Tube Growth

    Directory of Open Access Journals (Sweden)

    Jean-Claude Mollet

    2013-03-01

    Full Text Available The pollen tube is a fast tip-growing cell carrying the two sperm cells to the ovule allowing the double fertilization process and seed setting. To succeed in this process, the spatial and temporal controls of pollen tube growth within the female organ are critical. It requires a massive cell wall deposition to promote fast pollen tube elongation and a tight control of the cell wall remodeling to modify the mechanical properties. In addition, during its journey, the pollen tube interacts with the pistil, which plays key roles in pollen tube nutrition, guidance and in the rejection of the self-incompatible pollen. This review focuses on our current knowledge in the biochemistry and localization of the main cell wall polymers including pectin, hemicellulose, cellulose and callose from several pollen tube species. Moreover, based on transcriptomic data and functional genomic studies, the possible enzymes involved in the cell wall remodeling during pollen tube growth and their impact on the cell wall mechanics are also described. Finally, mutant analyses have permitted to gain insight in the function of several genes involved in the pollen tube cell wall biosynthesis and their roles in pollen tube growth are further discussed.

  6. Modification of cell wall polysaccharides during retting of cassava roots.

    Science.gov (United States)

    Ngolong Ngea, Guillaume Legrand; Guillon, Fabienne; Essia Ngang, Jean Justin; Bonnin, Estelle; Bouchet, Brigitte; Saulnier, Luc

    2016-12-15

    Retting is an important step in traditional cassava processing that involves tissue softening of the roots to transform the cassava into flour and various food products. The tissue softening that occurs during retting was attributed to the degradation of cell wall pectins through the action of pectin-methylesterase and pectate-lyase that possibly originated from a microbial source or the cassava plant itself. Changes in cell wall composition were investigated during retting using chemical analysis, specific glycanase degradation and immuno-labelling of cell wall polysaccharides. Pectic 1,4-β-d-galactan was the main cell wall polysaccharide affected during the retting of cassava roots. This result suggested that better control of pectic galactan degradation and a better understanding of the degradation mechanism by endogenous endo-galactanase and/or exogenous microbial enzymes might contribute to improve the texture properties of cassava products. PMID:27451197

  7. Roles of tRNA in cell wall biosynthesis

    DEFF Research Database (Denmark)

    Dare, Kiley; Ibba, Michael

    2012-01-01

    Recent research into various aspects of bacterial metabolism such as cell wall and antibiotic synthesis, degradation pathways, cellular stress, and amino acid biosynthesis has elucidated roles of aminoacyl-transfer ribonucleic acid (aa-tRNA) outside of translation. Although the two enzyme families...... specificity of this diverse enzymatic family is necessary to aid current efforts in designing potential bactericidal agents. These two enzyme families are linked only by the substrate with which they modify the cell wall, aa-tRNA; their structure, cell wall modification processes and the physiological changes...... responsible for cell wall modifications, aminoacyl-phosphatidylglycerol synthases (aaPGSs) and Fem, were discovered some time ago, they have recently become of intense interest for their roles in the antimicrobial resistance of pathogenic microorganisms. The addition of positively charged amino acids to...

  8. Stress Analysis of Building Curtain Wall Post Embedment Anchor Bolt%建筑幕墙后置埋件锚栓受力分析

    Institute of Scientific and Technical Information of China (English)

    陆新鑫; 吴强

    2014-01-01

    本文根据JGJ 145—2013《混凝土结构后锚固技术规程》第5.2.2条,对后置埋件在双向受弯矩、双向受剪力、以及扭矩、拉力的共同作用下进行的锚栓受力做了分析,提出了复合力作用下后置埋件锚栓的计算公式。%This paper, according to article 5.2.2 of JGJ 145 -2013 “technical specification for concrete structure of anch-oring force”, made an analysis of the force analysis of post anchor bolt under the interaction of bi-directional torque, shear force, bending, put ing forward the calculation formula of posts anchor bolt under composite force.

  9. Patterns of expression of cell wall related genes in sugarcane

    Directory of Open Access Journals (Sweden)

    Lima D.U.

    2001-01-01

    Full Text Available Our search for genes related to cell wall metabolism in the sugarcane expressed sequence tag (SUCEST database (http://sucest.lbi.dcc.unicamp.br resulted in 3,283 reads (1% of the total reads which were grouped into 459 clusters (potential genes with an average of 7.1 reads per cluster. To more clearly display our correlation coefficients, we constructed surface maps which we used to investigate the relationship between cell wall genes and the sugarcane tissues libraries from which they came. The only significant correlations that we found between cell wall genes and/or their expression within particular libraries were neutral or synergetic. Genes related to cellulose biosynthesis were from the CesA family, and were found to be the most abundant cell wall related genes in the SUCEST database. We found that the highest number of CesA reads came from the root and stem libraries. The genes with the greatest number of reads were those involved in cell wall hydrolases (e.g. beta-1,3-glucanases, xyloglucan endo-beta-transglycosylase, beta-glucosidase and endo-beta-mannanase. Correlation analyses by surface mapping revealed that the expression of genes related to biosynthesis seems to be associated with the hydrolysis of hemicelluloses, pectin hydrolases being mainly associated with xyloglucan hydrolases. The patterns of cell wall related gene expression in sugarcane based on the number of reads per cluster reflected quite well the expected physiological characteristics of the tissues. This is the first work to provide a general view on plant cell wall metabolism through the expression of related genes in almost all the tissues of a plant at the same time. For example, developing flowers behaved similarly to both meristematic tissues and leaf-root transition zone tissues. Besides providing a basis for future research on the mechanisms of plant development which involve the cell wall, our findings will provide valuable tools for plant engineering in the

  10. Transformation of Abdominal Wall Endometriosis to Clear Cell Carcinoma

    OpenAIRE

    Maria Paula Ruiz; Darryl Lewis Wallace; Matthew Thomas Connell

    2015-01-01

    Clear cell carcinoma is the least common of the malignant transformations reported in nonpelvic sites of endometriosis. Two cases with clear cell carcinoma transformation arising from endometriosis in abdominal wall scars are presented. These patients underwent total abdominal hysterectomy with bilateral salpingo-oophorectomy, pelvic washings, and abdominal wall lesion resection. The first case had initial treatment with chemotherapy, while chemotherapy and radiation therapy were given for th...

  11. Analyzing the complex machinery of cell wall biosynthesis

    OpenAIRE

    Timmers, J.F.P.

    2009-01-01

    The plant cell wall polymers make up most of the plant biomass and provide the raw material for many economically important products including food, feed, bio-materials, chemicals, textiles, and biofuel. This broad range of functions and applications make the biosynthesis of these polysaccharides a highly interesting target of scientific research. In this thesis a protein-protein interaction strategy was used to gain insight in the cell wall biosynthesis of Arabidopsis thaliana and to identif...

  12. Biosynthetic origin of mycobacterial cell wall arabinosyl residues.

    OpenAIRE

    Scherman, M.; Weston, A; Duncan, K; Whittington, A; Upton, R; Deng, L.; Comber, R; Friedrich, J D; McNeil, M

    1995-01-01

    Designing new drugs that inhibit the biosynthesis of the D-arabinan moiety of the mycobacterial cell wall arabinogalactan is one important basic approach for treatment of mycobacterial diseases. However, the biosynthetic origin of the D-arabinosyl monosaccharide residues themselves is not known. To obtain information on this issue, mycobacteria growing in culture were fed glucose labeled with 14C or 3H in specific positions. The resulting radiolabeled cell walls were isolated and hydrolyzed, ...

  13. Pectin, a versatile polysaccharide present in plant cell walls

    NARCIS (Netherlands)

    Voragen, A.G.J.; Coenen, G.J.; Verhoef, R.P.; Schols, H.A.

    2009-01-01

    Pectin or pectic substances are collective names for a group of closely associated polysaccharides present in plant cell walls where they contribute to complex physiological processes like cell growth and cell differentiation and so determine the integrity and rigidity of plant tissue. They also pla

  14. An Accessory Protein Required for Anchoring and Assembly of Amyloid Fibers in B. subtilis Biofilms

    OpenAIRE

    Romero, Diego; Vlamakis, Hera; Losick, Richard; Kolter, Roberto

    2011-01-01

    Cells within Bacillus subtilis biofilms are held in place by an extracellular matrix that contains cell-anchored amyloid fibers, composed of the amyloidogenic protein TasA. As biofilms age they disassemble because the cells release the amyloid fibers. This release appears to be the consequence of incorporation of D-tyrosine, D-leucine, D-tryptophan and D-methionine into the cell wall. Here, we characterize the in vivo roles of an accessory protein TapA (TasA anchoring/assembly protein; previo...

  15. Evaluation of cell wall preparations for proteomics: a new procedure for purifying cell walls from Arabidopsis hypocotyls

    Directory of Open Access Journals (Sweden)

    Canut Hervé

    2006-05-01

    Full Text Available Abstract Background The ultimate goal of proteomic analysis of a cell compartment should be the exhaustive identification of resident proteins; excluding proteins from other cell compartments. Reaching such a goal closely depends on the reliability of the isolation procedure for the cell compartment of interest. Plant cell walls possess specific difficulties: (i the lack of a surrounding membrane may result in the loss of cell wall proteins (CWP during the isolation procedure, (ii polysaccharide networks of cellulose, hemicelluloses and pectins form potential traps for contaminants such as intracellular proteins. Several reported procedures to isolate cell walls for proteomic analyses led to the isolation of a high proportion (more than 50% of predicted intracellular proteins. Since isolated cell walls should hold secreted proteins, one can imagine alternative procedures to prepare cell walls containing a lower proportion of contaminant proteins. Results The rationales of several published procedures to isolate cell walls for proteomics were analyzed, with regard to the bioinformatic-predicted subcellular localization of the identified proteins. Critical steps were revealed: (i homogenization in low ionic strength acid buffer to retain CWP, (ii purification through increasing density cushions, (iii extensive washes with a low ionic strength acid buffer to retain CWP while removing as many cytosolic proteins as possible, and (iv absence of detergents. A new procedure was developed to prepare cell walls from etiolated hypocotyls of Arabidopsis thaliana. After salt extraction, a high proportion of proteins predicted to be secreted was released (73%, belonging to the same functional classes as proteins identified using previously described protocols. Finally, removal of intracellular proteins was obtained using detergents, but their amount represented less than 3% in mass of the total protein extract, based on protein quantification. Conclusion The

  16. DCB-adapted plant cells possess unique wall structure

    Energy Technology Data Exchange (ETDEWEB)

    Shedletzky, E.; Shmuel, M. (Hebrew Univ., Jerusalem (Israel)); Delmer, D. (Hebrew Univ., Jerusalem (Israel) Michigan State Univ., East Lansing (USA)); Lamport, D. (Michigan State Univ., East Lansing (USA))

    1990-05-01

    Suspension-cultured cells of tomato (Lycopersicon esculentum VF 36) haven been adapted to growth on high concentrations of 2,6-dichloro-benzonitrile (DCB), an herbicide which inhibits cellulose biosynthesis. The mechanism of adaptation appears to rest largely on the ability of thee cells to divide and expand in the virtual absence of a cellulose-xyloglucan network. Walls of adapted cells growing on DCB also differ from non-adapted cells by having reduced levels of hydroxyproline in protein, both in bound and salt-elutable form, and in having a much higher proportion of homogalacturonon and rhamnogalacturonan-like polymers. Most of these latter polymers are apparently cross-linked in the wall via phenolic-esters and/or phenolic ether linkages, and these polymers appear to represent the major load-bearing network in thee unusual cell walls. The surprising finding that plant cells can survive in the virtual absence of a major load-bearing network in their primary cell walls indicates that plants possess remarkable flexibility for tolerating changes in wall composition.

  17. Another brick in the cell wall: biosynthesis dependent growth model.

    Directory of Open Access Journals (Sweden)

    Adelin Barbacci

    Full Text Available Expansive growth of plant cell is conditioned by the cell wall ability to extend irreversibly. This process is possible if (i a tensile stress is developed in the cell wall due to the coupling effect between turgor pressure and the modulation of its mechanical properties through enzymatic and physicochemical reactions and if (ii new cell wall elements can be synthesized and assembled to the existing wall. In other words, expansive growth is the result of coupling effects between mechanical, thermal and chemical energy. To have a better understanding of this process, models must describe the interplay between physical or mechanical variable with biological events. In this paper we propose a general unified and theoretical framework to model growth in function of energy forms and their coupling. This framework is based on irreversible thermodynamics. It is then applied to model growth of the internodal cell of Chara corallina modulated by changes in pressure and temperature. The results describe accurately cell growth in term of length increment but also in term of cell pectate biosynthesis and incorporation to the expanding wall. Moreover, the classical growth model based on Lockhart's equation such as the one proposed by Ortega, appears as a particular and restrictive case of the more general growth equation developed in this paper.

  18. Involvement of MAK-1 and MAK-2 MAP kinases in cell wall integrity in Neurospora crassa.

    Science.gov (United States)

    Kamei, Masayuki; Yamashita, Kazuhiro; Takahashi, Masakazu; Fukumori, Fumiyasu; Ichiishi, Akihiko; Fujimura, Makoto

    2016-09-01

    Among three MAPK disruptants of Neurospora crassa, Δmak-1 was sensitive and Δmak-2 was hypersensitive to micafungin, a beta-1,3-glucan synthase inhibitor, than the wild-type or Δos-2 strains. We identified six micafungin-inducible genes that are involved in cell wall integrity (CWI) and found that MAK-1 regulated the transcription of non-anchored cell wall protein gene, ncw-1, and the beta-1,3-endoglucanase gene, bgt-2, whereas MAK-2 controlled the expression of the glycosylhydrolase-like protein gene, gh76-5, and the C4-dicarboxylate transporter gene, tdt-1. Western blotting analysis revealed that, in the wild-type strain, MAK-1 was constitutively phosphorylated from conidial germination to hyphal development. In contrast, the phosphorylation of MAK-2 was growth phase-dependent, and micafungin induced the phosphorylation of unphosphorylated MAK-2. It should be noted that the phosphorylation of MAK-1 was virtually abolished in the Δmak-2 strain, but was significantly induced by micafungin, suggesting functional cross talk between MAK-1 and MAK-2 signalling pathway in CWI. PMID:27268441

  19. Cell wall proteomics of the green alga Haematococcus pluvialis (Chlorophyceae).

    Science.gov (United States)

    Wang, Sheng-Bing; Hu, Qiang; Sommerfeld, Milton; Chen, Feng

    2004-03-01

    The green microalga Haematococcus pluvialis can synthesize and accumulate large amounts of the ketocarotenoid astaxanthin, and undergo profound changes in cell wall composition and architecture during the cell cycle and in response to environmental stresses. In this study, cell wall proteins (CWPs) of H. pluvialis were systematically analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) coupled with peptide mass fingerprinting (PMF) and sequence-database analysis. In total, 163 protein bands were analyzed, which resulted in positive identification of 81 protein orthologues. The highly complex and dynamic composition of CWPs is manifested by the fact that the majority of identified CWPs are differentially expressed at specific stages of the cell cycle along with a number of common wall-associated 'housekeeping' proteins. The detection of cellulose synthase orthologue in the vegetative cells suggested that the biosynthesis of cellulose occurred during primary wall formation, in contrast to earlier observations that cellulose was exclusively present in the secondary wall of the organism. A transient accumulation of a putative cytokinin oxidase at the early stage of encystment pointed to a possible role in cytokinin degradation while facilitating secondary wall formation and/or assisting in cell expansion. This work represents the first attempt to use a proteomic approach to investigate CWPs of microalgae. The reference protein map constructed and the specific protein markers obtained from this study provide a framework for future characterization of the expression and physiological functions of the proteins involved in the biogenesis and modifications in the cell wall of Haematococcus and related organisms. PMID:14997492

  20. Inhibitors targeting on cell wall biosynthesis pathway of MRSA.

    Science.gov (United States)

    Hao, Haihong; Cheng, Guyue; Dai, Menghong; Wu, Qinghua; Yuan, Zonghui

    2012-11-01

    Methicillin resistant Staphylococcus aureus (MRSA), widely known as a type of new superbug, has aroused world-wide concern. Cell wall biosynthesis pathway is an old but good target for the development of antibacterial agents. Peptidoglycan and wall teichoic acids (WTAs) biosynthesis are two main processes of the cell wall biosynthesis pathway (CWBP). Other than penicillin-binding proteins (PBPs), some key factors (Mur enzymes, lipid I or II precursor, etc.) in CWBP are becoming attractive molecule targets for the discovery of anti-MRSA compounds. A number of new compounds, with higher affinity for PBPs or with inhibitory activity on such molecule targets in CWBP of MRSA, have been in the pipeline recently. This review concludes recent research achievements and provides a complete picture of CWBP of MRSA, including the peptidoglycan and wall teichoic acids synthesis pathway. The potential inhibitors targeting on CWBP are subsequently presented to improve development of novel therapeutic strategies for MRSA. PMID:22898792

  1. Cell wall staining with Trypan blue enables quantitative analysis of morphological changes in yeast cells

    OpenAIRE

    Liesche, Johannes; Marek, Magdalena; Günther-Pomorski, Thomas

    2015-01-01

    Yeast cells are protected by a cell wall that plays an important role in the exchange of substances with the environment. The cell wall structure is dynamic and can adapt to different physiological states or environmental conditions. For the investigation of morphological changes, selective staining with fluorescent dyes is a valuable tool. Furthermore, cell wall staining is used to facilitate sub-cellular localization experiments with fluorescently-labeled proteins and the detection of yeast...

  2. Co-delivery of cell-wall-forming enzymes in the same vesicle for coordinated fungal cell wall formation.

    Science.gov (United States)

    Schuster, Martin; Martin-Urdiroz, Magdalena; Higuchi, Yujiro; Hacker, Christian; Kilaru, Sreedhar; Gurr, Sarah J; Steinberg, Gero

    2016-01-01

    Fungal cells are surrounded by an extracellular cell wall. This complex matrix of proteins and polysaccharides protects against adverse stresses and determines the shape of fungal cells. The polysaccharides of the fungal wall include 1,3-β-glucan and chitin, which are synthesized by membrane-bound synthases at the growing cell tip. A hallmark of filamentous fungi is the class V chitin synthase, which carries a myosin-motor domain. In the corn smut fungus Ustilago maydis, the myosin-chitin synthase Mcs1 moves to the plasma membrane in secretory vesicles, being delivered by kinesin-1 and myosin-5. The myosin domain of Mcs1 enhances polar secretion by tethering vesicles at the site of exocytosis. It remains elusive, however, how other cell-wall-forming enzymes are delivered and how their activity is coordinated post secretion. Here, we show that the U. maydis class VII chitin synthase and 1,3-β-glucan synthase travel in Mcs1-containing vesicles, and that their apical secretion depends on Mcs1. Once in the plasma membrane, anchorage requires enzyme activity, which suggests co-synthesis of chitin and 1,3-β-glucan polysaccharides at sites of exocytosis. Thus, delivery of cell-wall-forming enzymes in Mcs1 vesicles ensures local foci of fungal cell wall formation. PMID:27563844

  3. Primary Cell Wall Structure in the Evolution of Land Plants

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Investigation of the primary cell walls of lower plants improves our understanding of the cell biology of these organisms but also has the potential to improve our understanding of cell wall structure and function in angiosperms that evolved from lower plants. Cell walls were prepared from eight species, ranging from a moss to advanced gymnosperms, and subjected to sequential chemical extraction to separate the main polysaccharide fractions. The glycosyl compositions of these fractions were then determined by gas chromatography. The results were compared among the eight plants and among data from related studies reported in the existing published reports to identify structural features that have been either highly conserved or clearly modified during evolution. Among the highly conserved features are the presence of a cellulose framework, the presence of certain hemicelluloses such as xyloglucan, and the presence of rhamnogalacturonan Ⅱ, a domain in pectic polysaccharides. Among the modified features are the abundance of mannosyl-containing hemicelluloses and the presence of methylated sugars.

  4. Simulated microgravity inhibits cell wall regeneration of Penicillium decumbens protoplasts

    Science.gov (United States)

    Zhao, C.; Sun, Y.; Yi, Z. C.; Rong, L.; Zhuang, F. Y.; Fan, Y. B.

    2010-09-01

    This work compares cell wall regeneration from protoplasts of the fungus Penicillium decumbens under rotary culture (simulated microgravity) and stationary cultures. Using an optimized lytic enzyme mixture, protoplasts were successfully released with a yield of 5.3 × 10 5 cells/mL. Under simulated microgravity conditions, the protoplast regeneration efficiency was 33.8%, lower than 44.9% under stationary conditions. Laser scanning confocal microscopy gave direct evidence for reduced formation of polysaccharides under simulated conditions. Scanning electron microscopy showed the delayed process of cell wall regeneration by simulated microgravity. The delayed regeneration of P. decumbens cell wall under simulated microgravity was likely caused by the inhibition of polysaccharide synthesis. This research contributes to the understanding of how gravitational loads affect morphological and physiological processes of fungi.

  5. How cell wall complexity influences saccharification efficiency in Miscanthus sinensis

    NARCIS (Netherlands)

    Souza, De Amanda P.; Lessa Alvim Kamei, Claire; Torres Salvador, Andres Francisco; Pattathil, Sivakumar; Hahn, Michael G.; Trindade, Luisa M.; Buckeridge, Marcos S.

    2015-01-01

    The production of bioenergy from grasses has been developing quickly during the last decade, with Miscanthus being among the most important choices for production of bioethanol. However, one of the key barriers to producing bioethanol is the lack of information about cell wall structure. Cell wal

  6. Identification of FtsW as a transporter of lipid-linked cell wall precursors across the membrane.

    Science.gov (United States)

    Mohammadi, Tamimount; van Dam, Vincent; Sijbrandi, Robert; Vernet, Thierry; Zapun, André; Bouhss, Ahmed; Diepeveen-de Bruin, Marlies; Nguyen-Distèche, Martine; de Kruijff, Ben; Breukink, Eefjan

    2011-04-20

    Bacterial cell growth necessitates synthesis of peptidoglycan. Assembly of this major constituent of the bacterial cell wall is a multistep process starting in the cytoplasm and ending in the exterior cell surface. The intracellular part of the pathway results in the production of the membrane-anchored cell wall precursor, Lipid II. After synthesis this lipid intermediate is translocated across the cell membrane. The translocation (flipping) step of Lipid II was demonstrated to require a specific protein (flippase). Here, we show that the integral membrane protein FtsW, an essential protein of the bacterial division machinery, is a transporter of the lipid-linked peptidoglycan precursors across the cytoplasmic membrane. Using Escherichia coli membrane vesicles we found that transport of Lipid II requires the presence of FtsW, and purified FtsW induced the transbilayer movement of Lipid II in model membranes. This study provides the first biochemical evidence for the involvement of an essential protein in the transport of lipid-linked cell wall precursors across biogenic membranes. PMID:21386816

  7. Alterations in auxin homeostasis suppress defects in cell wall function.

    Directory of Open Access Journals (Sweden)

    Blaire J Steinwand

    Full Text Available The plant cell wall is a highly dynamic structure that changes in response to both environmental and developmental cues. It plays important roles throughout plant growth and development in determining the orientation and extent of cell expansion, providing structural support and acting as a barrier to pathogens. Despite the importance of the cell wall, the signaling pathways regulating its function are not well understood. Two partially redundant leucine-rich-repeat receptor-like kinases (LRR-RLKs, FEI1 and FEI2, regulate cell wall function in Arabidopsis thaliana roots; disruption of the FEIs results in short, swollen roots as a result of decreased cellulose synthesis. We screened for suppressors of this swollen root phenotype and identified two mutations in the putative mitochondrial pyruvate dehydrogenase E1α homolog, IAA-Alanine Resistant 4 (IAR4. Mutations in IAR4 were shown previously to disrupt auxin homeostasis and lead to reduced auxin function. We show that mutations in IAR4 suppress a subset of the fei1 fei2 phenotypes. Consistent with the hypothesis that the suppression of fei1 fei2 by iar4 is the result of reduced auxin function, disruption of the WEI8 and TAR2 genes, which decreases auxin biosynthesis, also suppresses fei1 fei2. In addition, iar4 suppresses the root swelling and accumulation of ectopic lignin phenotypes of other cell wall mutants, including procuste and cobra. Further, iar4 mutants display decreased sensitivity to the cellulose biosynthesis inhibitor isoxaben. These results establish a role for IAR4 in the regulation of cell wall function and provide evidence of crosstalk between the cell wall and auxin during cell expansion in the root.

  8. Anchored nanostructure materials and method of fabrication

    Science.gov (United States)

    Seals, Roland D; Menchhofer, Paul A; Howe, Jane Y; Wang, Wei

    2012-11-27

    Anchored nanostructure materials and methods for their fabrication are described. The anchored nanostructure materials may utilize nano-catalysts that include powder-based or solid-based support materials. The support material may comprise metal, such as NiAl, ceramic, a cermet, or silicon or other metalloid. Typically, nanoparticles are disposed adjacent a surface of the support material. Nanostructures may be formed as anchored to nanoparticles that are adjacent the surface of the support material by heating the nano-catalysts and then exposing the nano-catalysts to an organic vapor. The nanostructures are typically single wall or multi-wall carbon nanotubes.

  9. Changes of lipid domains in Bacillus subtilis cells with disrupted cell wall peptidoglycan

    OpenAIRE

    Muchová, Katarína; Wilkinson, Anthony J.; Barák, Imrich

    2011-01-01

    The cell wall is responsible for cell integrity and the maintenance of cell shape in bacteria. The Gram-positive bacterial cell wall consists of a thick peptidoglycan layer located on the outside of the cytoplasmic membrane. Bacterial cell membranes, like eukaryotic cell membranes, are known to contain domains of specific lipid and protein composition. Recently, using the membrane-binding fluorescent dye FM4-64, helix-like lipid structures extending along the long axis of the cell and consist...

  10. The Interplay between Cell Wall Mechanical Properties and the Cell Cycle in Staphylococcus aureus

    OpenAIRE

    Bailey, Richard G.; Turner, Robert D.; Mullin, Nic; Clarke, Nigel,; Foster, Simon J.; Hobbs, Jamie K.

    2014-01-01

    The nanoscale mechanical properties of live Staphylococcus aureus cells during different phases of growth were studied by atomic force microscopy. Indentation to different depths provided access to both local cell wall mechanical properties and whole-cell properties, including a component related to cell turgor pressure. Local cell wall properties were found to change in a characteristic manner throughout the division cycle. Splitting of the cell into two daughter cells followed a local softe...

  11. Structure, function, and biosynthesis of plant cell walls: proceedings of the seventh annual symposium in botany

    Energy Technology Data Exchange (ETDEWEB)

    Dugger, W.M.; Bartnicki-Garcia, S. (eds.)

    1984-01-01

    Papers in the following areas were included in these symposium proceedings: (1) cell wall chemistry and biosynthesis; (2) cell wall hydrolysis and associated physiology; (3) cellular events associated with cell wall biosynthesis; and (4) interactions of plant cell walls with pathogens and related responses. Papers have been individually abstracted for the data base. (ACR)

  12. Immunocytochemical characterization of the cell walls of bean cell suspensions during habituation and dehabituation to dichlobenil

    DEFF Research Database (Denmark)

    Garcia-Angulo, P.; Willats, W. G. T.; Encina, A. E.;

    2006-01-01

    analysed showed calcofluor-stained appositions. However, in habituated and dehabituated cells, appositions were not recognized by an anticallose antibody. This finding suggested the accumulation of an extracellular polysaccharide different to callose, probably a 1,4-ß-glucan in these cell lines......The effects of the cellulose inhibitor dichlobenil on the cell wall composition and structure during the habituation/dehabituation process of suspension-cultured bean cells were assessed. A range of techniques were used including cell wall fractionation, sugar analysis, immunofluorescence and...... fluorochrome labelling of resin-embedded sections, and immunodot assays (IDAs) of cell wall fractions. The cell walls from bean cell suspensions with initial levels of habituation to dichlobenil had decreased levels of cellulose, but this effect lessened with increasing numbers of subcultures. All cell walls...

  13. Ultrastructure of organic cell walls in Proterozoic microalgae

    Science.gov (United States)

    Moczydlowska-Vidal, M.

    2009-04-01

    The antiquity of life has been well appreciated since the discoveries of microfossils and confirmation of their authenticity, as well as the recognition of geochemical signs of biogenicity in the Archean successions. Resolving the biological affinities of early biota is essential for the unravelling the changes that led to modern biodiversity, but also for the detection of possible biogenic records outside of the terrestrial biosphere. Advanced techniques in microscopy, tomography and spectroscopy applied to examine individual microfossils at the highest attainable spatial resolution have provided unprecedented insights into micro- and nano-scale structure and composition of organic matter. Transmission and scanning electron microscopy studies of the wall ultrastructure of sphaeromorphic and ornamented acritarchs have revealed complex, single to multilayered walls, having a unique texture in sub-layers and an occasionally preserved trilaminar sheath structure (TLS) of the cell wall. A variety of optical characteristics, the electron density and texture of fabrics of discrete layers, and the properties of biopolymers may indicate the polyphyletic affiliations of such microfossils and/or the preservation of various stages (vegetative, resting) in their life cycle. I evaluate the morphological features of organic-walled unicellular microfossils in conjunction with their cell wall ultrastructure to infer their life cycle and to recognize various developmental stages represented among microfossils attributed to a single form-taxon. Several cases of fine wall ultrastructure in microfossils have been documented and have had a conclusive influence on understanding their affinities. Some Proterozoic and Cambrian leiosphaerids are of algal affinities. Certain specimens represent chlorophyceaens, having the multilayered composite wall with TLS structure known from vegetative and resting cells in modern genera of the Chlorococcales and Volvocales. The wall ultrastructure of

  14. Transcriptional Wiring of Cell Wall-Related Genes in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    Marek Mutwil; Colin Ruprecht; Federico M. Giorgi; Martin Bringmann; Bj(o)rn Usadel; Staffan Persson

    2009-01-01

    Transcriptional coordination, or co-expression, of genes may signify functional relatedness of the correspond-ing proteins. For example, several genes involved in secondary cell wall cellulose biosynthesis are co-expressed with genes engaged in the synthesis of xylan, which is a major component of the secondary cell wall. To extend these types of anal-yses, we investigated the co-expression relationships of all Carbohydrate-Active enZYmes (CAZy)-related genes for Arabidopsis thaliana. Thus, the intention was to transcriptionally link different cell wall-related processes to each other, and also to other biological functions. To facilitate easy manual inspection, we have displayed these interactions as networks and matrices, and created a web-based interface (http://aranet.mpimp-golm.mpg.de/corecarb) containing downloadable files for all the transcriptional associations.

  15. Fluorescent Probes for Exploring Plant Cell Wall Deconstruction: A Review

    Directory of Open Access Journals (Sweden)

    Gabriel Paës

    2014-07-01

    Full Text Available Plant biomass is a potential resource of chemicals, new materials and biofuels that could reduce our dependency on fossil carbon, thus decreasing the greenhouse effect. However, due to its chemical and structural complexity, plant biomass is recalcitrant to green biological transformation by enzymes, preventing the establishment of integrated bio-refineries. In order to gain more knowledge in the architecture of plant cell wall to facilitate their deconstruction, many fluorescent probes bearing various fluorophores have been devised and used successfully to reveal the changes in structural motifs during plant biomass deconstruction, and the molecular interactions between enzymes and plant cell wall polymers. Fluorescent probes are thus relevant tools to explore plant cell wall deconstruction.

  16. Synthesis and Application of Plant Cell Wall Oligogalactans

    DEFF Research Database (Denmark)

    Andersen, Mathias Christian Franch

    The plant cell walls represent almost 50% of the biomass found in plants and are therefore one of the main targets for biotechnological research. Major motivators are their potential as a renewable energy source for transport fuels, as functional foods, and as a source of raw materials to generate...... chemical building blocks for industrial processes. To achieve a sustainable development it is necessary to optimize plant production and utilization. This will require a better understanding of the cell wall structure and function at the molecular level. The cell wall is composed by an intricate network of...... as part of the arabinogalactans series. The fragments were applied in the characterization of a glycosyl transferase, a hydrolase and to study the important cancer biomarker galectin-3. The work done during an external stay at University of Oxford is also presented. This concerns isolation and...

  17. The role of the cell wall in plant immunity

    DEFF Research Database (Denmark)

    Malinovsky, Frederikke Gro; Fangel, Jonatan Ulrik; Willats, William George Tycho

    2014-01-01

    The battle between plants and microbes is evolutionarily ancient, highly complex, and often co-dependent. A primary challenge for microbes is to breach the physical barrier of host cell walls whilst avoiding detection by the plant's immune receptors. While some receptors sense conserved microbial...... features, others monitor physical changes caused by an infection attempt. Detection of microbes leads to activation of appropriate defense responses that then challenge the attack. Plant cell walls are formidable and dynamic barriers. They are constructed primarily of complex carbohydrates joined by...... numerous distinct connection types, and are subject to extensive post-synthetic modification to suit prevailing local requirements. Multiple changes can be triggered in cell walls in response to microbial attack. Some of these are well described, but many remain obscure. The study of the myriad of subtle...

  18. Cell wall integrity signalling in human pathogenic fungi.

    Science.gov (United States)

    Dichtl, Karl; Samantaray, Sweta; Wagener, Johannes

    2016-09-01

    Fungi are surrounded by a rigid structure, the fungal cell wall. Its plasticity and composition depend on active regulation of the underlying biosynthesis and restructuring processes. This involves specialised signalling pathways that control gene expression and activities of biosynthetic enzymes. The cell wall integrity (CWI) pathway is the central signalling cascade required for the adaptation to a wide spectrum of cell wall perturbing conditions, including heat, oxidative stress and antifungals. In the recent years, great efforts were made to analyse the CWI pathway of diverse fungi. It turned out that the CWI signalling cascade is mostly conserved in the fungal kingdom. In this review, we summarise as well as compare the current knowledge on the canonical CWI pathway in the human pathogenic fungi Candida albicans, Candida glabrata, Aspergillus fumigatus and Cryptococcus neoformans. Understanding the differences and similarities in the stress responses of these organisms could become a key to improving existing or developing new antifungal therapies. PMID:27155139

  19. Histochemical effects of γ radiation on soft fruit cell walls

    International Nuclear Information System (INIS)

    Irradiation effects in peaches, tomatoes, cherries and grapes on the composition of cell wall polysaccharides were investigated by histochemical techniques. Cell wall polysaccharides, separated by a modified Jensen's method were pectins, hemicellulose, non-cellulosic polysaccharides and cellulose. The extinction values of Periodic Acid Schiff stained tissues was measured by microscopical photometry. Irradiation induced highly significant changes in polysaccharide composition of mesocarp cell walls; these changes were found to be a function of time of irradiation after harvest and of the species tested. A general influence on polysaccharide molecules was not found. Variations produced by irradiation are postulated to be an interference with a regulatory system rather than a breakdown of a functional molecule (metabolic enzyme or polysaccharide. (author)

  20. Stem and progenitor cells in biostructure of blood vessel walls

    Directory of Open Access Journals (Sweden)

    Krzysztof Korta

    2013-09-01

    Full Text Available Development of vascular and hematopoietic systems during organogenesis occurs at the same time. During vasculogenesis, a small part of cells does not undergo complete differentiation but stays on this level, “anchored” in tissue structures described as stem cell niches. The presence of blood vessels within tissue stem cell niches is typical and led to identification of niches and ensures that they are functioning. The three-layer biostructure of vessel walls for artery and vein, tunica: intima, media and adventitia, for a long time was defined as a mechanical barrier between vessel light and the local tissue environment. Recent findings from vascular biology studies indicate that vessel walls are dynamic biostructures, which are equipped with stem and progenitor cells, described as vascular wall-resident stem cells/progenitor cells (VW-SC/PC. Distinct zones for vessel wall harbor heterogeneous subpopulations of VW-SC/PC, which are described as “subendothelial or vasculogenic zones”. Recent evidence from in vitro and in vivo studies show that prenatal activity of stem and progenitor cells is not only limited to organogenesis but also exists in postnatal life, where it is responsible for vessel wall homeostasis, remodeling and regeneration. It is believed that VW-SC/PC could be engaged in progression of vascular disorders and development of neointima. We would like to summarize current knowledge about mesenchymal and progenitor stem cell phenotype with special attention to distribution and biological properties of VW-SC/PC in biostructures of intima, media and adventitia niches. It is postulated that in the near future, niches for VW-SC/PC could be a good source of stem and progenitor cells, especially in the context of vessel tissue bioengineering as a new alternative to traditional revascularization therapies.

  1. Involvement of cell surface phosphatidylinositol-anchored glycoproteins in cell-cell adhesion of chick embryo myoblasts

    OpenAIRE

    1989-01-01

    During myogenesis myoblasts fuse to form multinucleate cells that express muscle-specific proteins. A specific cell-cell adhesion process precedes lipid bilayer union during myoblast fusion (Knudsen, K. A., and A. F. Horwitz. 1977. Dev. Biol. 58:328-338) and is mediated by cell surface glycoproteins (Knudsen, K. A., 1985. J. Cell Biol. 101:891- 897). In this paper we show that myoblast adhesion and myotube formation are inhibited by treating fusion-competent myoblasts with phosphatidylinosito...

  2. Evolution of the cell wall components during terrestrialization

    Directory of Open Access Journals (Sweden)

    Alicja Banasiak

    2014-12-01

    Full Text Available Colonization of terrestrial ecosystems by the first land plants, and their subsequent expansion and diversification, were crucial for the life on the Earth. However, our understanding of these processes is still relatively poor. Recent intensification of studies on various plant organisms have identified the plant cell walls are those structures, which played a key role in adaptive processes during the evolution of land plants. Cell wall as a structure protecting protoplasts and showing a high structural plasticity was one of the primary subjects to changes, giving plants the new properties and capabilities, which undoubtedly contributed to the evolutionary success of land plants. In this paper, the current state of knowledge about some main components of the cell walls (cellulose, hemicelluloses, pectins and lignins and their evolutionary alterations, as preadaptive features for the land colonization and the plant taxa diversification, is summarized. Some aspects related to the biosynthesis and modification of the cell wall components, with particular emphasis on the mechanism of transglycosylation, are also discussed. In addition, new surprising discoveries related to the composition of various cell walls, which change how we perceive their evolution, are presented, such as the presence of lignin in red algae or MLG (1→3,(1→4-β-D-glucan in horsetails. Currently, several new and promising projects, regarding the cell wall, have started, deciphering its structure, composition and metabolism in the evolutionary context. That additional information will allow us to better understand the processes leading to the terrestrialization and the evolution of extant land plants.

  3. Bacterial Cell Wall-Induced Arthritis: Chemical Composition and Tissue Distribution of Four Lactobacillus Strains

    OpenAIRE

    Šimelyte, Egle; Rimpiläinen, Marja; Lehtonen, Leena; Zhang, Xiang; Toivanen, Paavo

    2000-01-01

    To study what determines the arthritogenicity of bacterial cell walls, cell wall-induced arthritis in the rat was applied, using four strains of Lactobacillus. Three of the strains used proved to induce chronic arthritis in the rat; all were Lactobacillus casei. The cell wall of Lactobacillus fermentum did not induce chronic arthritis. All arthritogenic bacterial cell walls had the same peptidoglycan structure, whereas that of L. fermentum was different. Likewise, all arthritogenic cell walls...

  4. Particle Trajectories in Rotating Wall Cell Culture Devices

    Science.gov (United States)

    Ramachandran N.; Downey, J. P.

    1999-01-01

    Cell cultures are extremely important to the medical community since such cultures provide an opportunity to perform research on human tissue without the concerns inherent in experiments on individual humans. Development of cells in cultures has been found to be greatly influenced by the conditions of the culture. Much work has focused on the effect of the motions of cells in the culture relative to the solution. Recently rotating wall vessels have been used with success in achieving improved cellular cultures. Speculation and limited research have focused on the low shear environment and the ability of rotating vessels to keep cells suspended in solution rather than floating or sedimenting as the primary reasons for the improved cellular cultures using these devices. It is widely believed that the cultures obtained using a rotating wall vessel simulates to some degree the effect of microgravity on cultures. It has also been speculated that the microgravity environment may provide the ideal acceleration environment for culturing of cellular tissues due to the nearly negligible levels of sedimentation and shear possible. This work predicts particle trajectories of cells in rotating wall vessels of cylindrical and annular design consistent with the estimated properties of typical cellular cultures. Estimates of the shear encountered by cells in solution and the interactions with walls are studied. Comparisons of potential experiments in ground and microgravity environments are performed.

  5. Cell Wall Polysaccharides of Candida albicans Induce Mast Cell Degranulation in the Gut

    OpenAIRE

    Sakurai, Atsuko; Yamaguchi, Natsu; Sonoyama, Kei

    2012-01-01

    We investigated Candida albicans-induced mast cell degranulation in vitro and in vivo. Cell wall fraction but not culture supernatant and cell membrane fraction prepared from hyphally grown C. albicans induced β-hexosaminidase release in RBL-2H3 cells. Cell wall mannan and soluble β-glucan fractions also induced β-hexosaminidase release. Histological examination of mouse forestomach showed that C. albicans gut colonization induces mast cell degranulation. However, intragastric administration ...

  6. Characters of Fractal Ultrastructure in Wood Cell Wall

    Institute of Scientific and Technical Information of China (English)

    LI Beimei; ZHAO Guangjie

    2006-01-01

    Fractal theory was introduced in order to describe the ultrastructure of wood cell wall in this paper.The cellulose chain clusters around nano-scale were viewed as a fractal object that consists of many fibrillar structural units with different scales including microfibrils.On the basis of the morphological data of wood cell wall.fractal dimensions of multi-level fibrillar structural units were calculated by fractal-geometry approach,and then the morphological and structural characteristics of fibers as well as the influences on wood properties were investigated according to the dimensions.Besides,the fractal self-nesting character of the ultrastruture was also analyzed.

  7. Phagocytic properties of lung alveolar wall cells

    Directory of Open Access Journals (Sweden)

    Tanaka,Akisuke

    1974-04-01

    Full Text Available For the purpose to define the mechanism of heavy metal intoxication by inhalation, morphologic observations were made on rat lungs after nasal instillation of iron colloid particles of positive and negative electric charges. Histochemical observation was also made on the liver and spleen of these animals. The instilled iron colloid particles reach the alveolar cavity easily, as can be seen in the tissue sections stained by Prussian blue reaction. Alveolar macrophages do take up them avidly both of positive and negative charges, though much less the positive particles than negative ones. In contrast, the alveolar epithelial cells take up solely positive particles by phagocytosis but not negative ones. Electron microscope observation revealed that the positive particles are ingested by Type I epithelial cells by pinocytosis and by Type II cells by phagocytosis as well. Then the iron colloid particles are transferred into the basement membrane by exocytosis. Travelling through the basement membrane they are again taken up by capillary endothelial cells by phagocytosis. Some particles were found in the intercellular clefts of capillary endothelial cells but not any iron colloid particles in the intercellular spaces of epithelial cells and in the capillary lumen. However, the liver and spleen tissues of the animals given iron colloid showed a strong positive iron reaction. On the basis of these observations, the mechanism of acute intoxication by inhaling heavy metal dusts like lead fume is discussed from the view point of selective uptake of alveolar epithelial and capillary endothelial cells for the particles of the positive electric cha'rge.

  8. Cell wall modification in grapevine cells in response to UV stress investigated by atomic force microscopy

    International Nuclear Information System (INIS)

    Despite cell wall reinforcement being a well-known defence mechanism of plants, it remains poorly characterized from a physical point of view. The objective of this work was to further describe this mechanism. Vitis vinifera cv Gamay cells were treated with UV-light (254 nm), a well-known elicitor of defence mechanisms in grapevines, and physical cell wall modifications were observed using the atomic force microscopy (AFM) under native conditions. The grapevine cell suspensions were continuously observed in their culture medium from 30 min to 24 h after elicitation. In the beginning, cellulose fibrils covered by a matrix surrounded the control and treated cells. After 3 h, the elicited cells displayed sprouted expansions around the cell wall that correspond to pectin chains. These expansions were not observed on untreated grapevine cells. The AFM tip was used to determine the average surface elastic modulus of cell wall that account for cell wall mechanical properties. The elasticity is diminished in UV-treated cells. In a comparative study, grapevine cells showed the same decrease in cell wall elasticity when treated with a fungal biotic elicitor of defence response. These results demonstrate cell wall strengthening by UV stress

  9. The Cellulose System in the Cell Wall of Micrasterias

    Science.gov (United States)

    Kim; Herth; Vuong; Chanzy

    1996-11-01

    The cellulose system of the cell wall of Micrasterias denticulata and Micrasterias rotata was analyzed by diffraction contrast transmission electron microscopy, electron diffraction, and X-ray analysis. The studies, achieved on disencrusted cell ghosts, confirmed that the cellulose microfibrils occurred in crisscrossed bands consisting of a number of parallel ribbon-like microfibrils. The individual microfibrils had thicknesses of 5 nm for a width of around 20 nm, but in some instances, two or three microfibrils merged into one another to yield larger monocrystalline domains reaching up to 60 nm in lateral size. The orientation of the cellulose of Micrasterias is very unusual, as it was found that in the cell wall, the equatorial crystallographic planes of cellulose having a d-spacing of 0.60 nm [(11;0) in the Ibeta cellulose unit cell defined by Sugiyama et al., 1991, Macromolecules 24, 4168-4175] were oriented perpendicular to the cell wall surface. Up to now, such orientation has been found only in Spirogyra, another member of the Zygnemataceae group. The unusual structure of the secondary wall cellulose of Micrasterias may be tentatively correlated with the unique organization of the terminal complexes, which in this alga occur as hexagonal arrays of rosettes. PMID:8986649

  10. The metabolic enzyme ManA reveals a link between cell wall integrity and chromosome morphology.

    OpenAIRE

    Maya Elbaz; Sigal Ben-Yehuda

    2010-01-01

    Author Summary The bacterial cell is resistant to extremes of osmotic pressure and protected against mechanical damages by the existence of a rigid outer shell defined as the cell wall. The strength of the cell wall is achieved by the presence of long glycan strands cross-linked by peptide side bridges. The cell wall is a dynamic structure continuously being synthesized and modified to allow for cell growth and division. Damaging the cell wall leads to abnormal cellular morphologies and cell ...

  11. A model of cell wall expansion based on thermodynamics of polymer networks

    Science.gov (United States)

    Veytsman, B. A.; Cosgrove, D. J.

    1998-01-01

    A theory of cell wall extension is proposed. It is shown that macroscopic properties of cell walls can be explained through the microscopic properties of interpenetrating networks of cellulose and hemicellulose. The qualitative conclusions of the theory agree with the existing experimental data. The dependence of the cell wall yield threshold on the secretion of the wall components is discussed.

  12. Cell wall staining with Trypan blue enables quantitative analysis of morphological changes in yeast cells

    DEFF Research Database (Denmark)

    Liesche, Johannes; Marek, Magdalena; Günther-Pomorski, Thomas

    2015-01-01

    staining with fluorescent dyes is a valuable tool. Furthermore, cell wall staining is used to facilitate sub-cellular localization experiments with fluorescently-labeled proteins and the detection of yeast cells in non-fungal host tissues. Here, we report staining of Saccharomyces cerevisiae cell wall with...... Trypan Blue, which emits strong red fluorescence upon binding to chitin and yeast glucan; thereby, it facilitates cell wall analysis by confocal and super-resolution microscopy. The staining pattern of Trypan Blue was similar to that of the widely used UV-excitable, blue fluorescent cell wall stain...... Calcofluor White. Trypan Blue staining facilitated quantification of cell size and cell wall volume when utilizing the optical sectioning capacity of a confocal microscope. This enabled the quantification of morphological changes during growth under anaerobic conditions and in the presence of chemicals...

  13. Display of GPI-anchored anti-EGFR nanobodies on extracellular vesicles promotes tumour cell targeting

    OpenAIRE

    Sander A. A. Kooijmans; Gómez Aleza, Clara; Roffler, Steve R; van Solinge, Wouter W.; Vader, Pieter; Schiffelers, Raymond M.

    2016-01-01

    Background: Extracellular vesicles (EVs) are attractive candidate drug delivery systems due to their ability to functionally transport biological cargo to recipient cells. However, the apparent lack of target cell specificity of exogenously administered EVs limits their therapeutic applicability. In this study, we propose a novel method to equip EVs with targeting properties, in order to improve their interaction with tumour cells.Methods: EV producing cells were transfected with vectors enco...

  14. Scaffolding during the cell cycle by A-kinase anchoring proteins

    NARCIS (Netherlands)

    Han, B; Poppinga, W J; Schmidt, M

    2015-01-01

    Cell division relies on coordinated regulation of the cell cycle. A process including a well-defined series of strictly regulated molecular mechanisms involving cyclin-dependent kinases, retinoblastoma protein, and polo-like kinases. Dysfunctions in cell cycle regulation are associated with disease

  15. Catalysts of plant cell wall loosening [version 1; referees: 2 approved

    OpenAIRE

    Cosgrove, Daniel J.

    2016-01-01

    The growing cell wall in plants has conflicting requirements to be strong enough to withstand the high tensile forces generated by cell turgor pressure while selectively yielding to those forces to induce wall stress relaxation, leading to water uptake and polymer movements underlying cell wall expansion. In this article, I review emerging concepts of plant primary cell wall structure, the nature of wall extensibility and the action of expansins, family-9 and -12 endoglucanases, family-16 xyl...

  16. An emerging role of pectic rhamnogalacturonanII for cell wall integrity

    OpenAIRE

    Reboul, Rebecca; Tenhaken, Raimund

    2012-01-01

    The plant cell wall is a complex network of different polysaccharides and glycoproteins, showing high diversity in nature. The essential components, tethering cell wall are under debate, as novel mutants challenge established models. The mutant ugd2,3 with a reduced supply of the important wall precursor UDP-glucuronic acid reveals the critical role of the pectic compound rhamnogalacturonanII for cell wall stability. This polymer seems to be more important for cell wall integrity than the pre...

  17. Cell wall structure and function in lactic acid bacteria.

    Science.gov (United States)

    Chapot-Chartier, Marie-Pierre; Kulakauskas, Saulius

    2014-08-29

    The cell wall of Gram-positive bacteria is a complex assemblage of glycopolymers and proteins. It consists of a thick peptidoglycan sacculus that surrounds the cytoplasmic membrane and that is decorated with teichoic acids, polysaccharides, and proteins. It plays a major role in bacterial physiology since it maintains cell shape and integrity during growth and division; in addition, it acts as the interface between the bacterium and its environment. Lactic acid bacteria (LAB) are traditionally and widely used to ferment food, and they are also the subject of more and more research because of their potential health-related benefits. It is now recognized that understanding the composition, structure, and properties of LAB cell walls is a crucial part of developing technological and health applications using these bacteria. In this review, we examine the different components of the Gram-positive cell wall: peptidoglycan, teichoic acids, polysaccharides, and proteins. We present recent findings regarding the structure and function of these complex compounds, results that have emerged thanks to the tandem development of structural analysis and whole genome sequencing. Although general structures and biosynthesis pathways are conserved among Gram-positive bacteria, studies have revealed that LAB cell walls demonstrate unique properties; these studies have yielded some notable, fundamental, and novel findings. Given the potential of this research to contribute to future applied strategies, in our discussion of the role played by cell wall components in LAB physiology, we pay special attention to the mechanisms controlling bacterial autolysis, bacterial sensitivity to bacteriophages and the mechanisms underlying interactions between probiotic bacteria and their hosts. PMID:25186919

  18. Bidirectional cell-surface anchoring function of C-terminal repeat region of peptidoglycan hydrolase of Lactococcus lactis IL1403.

    Science.gov (United States)

    Tarahomjoo, Shirin; Katakura, Yoshio; Satoh, Eiichi; Shioya, Suteaki

    2008-02-01

    With the aim of constructing an efficient protein display system for lactic acid bacteria (LABs), the effect of fusion direction on the cell-surface binding activity of the C-terminal region of the peptidoglycan hydrolase (CPH) of Lactococcus lactis IL1403 was studied. CPH fused to the alpha-amylase (AMY) of Streptococcus bovis 148 either at its C-terminus (CPH-AMY) or at its N-terminus (AMY-CPH) was expressed intracellularly in Escherichia coli. This domain was able to direct binding of AMY to the surface of L. lactis ATCC 19435 in both constructs. However, the number of bound molecules per cell and the specific activity for starch digestion in the case of CPH-AMY were 3 and 14 times greater than those in the case of AMY-CPH, respectively. Of the LABs tested, L. lactis ATCC 19435 showed the highest binding capability for CPH-AMY, up to 6 x 10(4) molecules per cell, with a dissociation rate constant of 5.00 x 10(-5) s(-1). The binding of CPH-AMY to the surface of Lactobacillus delbrueckii ATCC 9649 cells was very stable with a dissociation rate constant of 6.96 x 10(-6) s(-1). The production of CPH-AMY in the soluble form increased 3-fold as a result of coexpression with a molecular chaperone, trigger factor. The results of this study suggest the usefulness of CPH as a bidirectional anchor protein for the production of cell-surface adhesive enzymes in E. coli. Furthermore, the importance of the fusion direction of CPH in determining cell-surface binding and enzymatic activities was shown. PMID:18343337

  19. Analyzing the complex machinery of cell wall biosynthesis

    NARCIS (Netherlands)

    Timmers, J.F.P.

    2009-01-01

    The plant cell wall polymers make up most of the plant biomass and provide the raw material for many economically important products including food, feed, bio-materials, chemicals, textiles, and biofuel. This broad range of functions and applications make the biosynthesis of these polysaccharides a

  20. The identification of cell wall degrading enzymes in Globodera rostochiensis

    NARCIS (Netherlands)

    Popeijus, H.E.

    2002-01-01

    This thesis describes the identification of cell wall degrading enzymes of the potato cyst nematode Globodera rostochiensis . A robust method using expressed sequence tags (ESTs) was applied to identify new parasitism related enzymes. One of the ESTs revealed the first pectate lyase from a metazoan

  1. Magnetic domain wall conduits for single cell applications

    DEFF Research Database (Denmark)

    Donolato, Marco; Torti, A.; Kostesha, Natalie;

    2011-01-01

    The ability to trap, manipulate and release single cells on a surface is important both for fundamental studies of cellular processes and for the development of novel lab-on-chip miniaturized tools for biological and medical applications. In this paper we demonstrate how magnetic domain walls...

  2. Characterisation of cell wall polysaccharides in bilberries and black currants

    NARCIS (Netherlands)

    Hilz, H.

    2007-01-01

    During berry juice production, polysaccharides are released from the cell walls and cause thickening and high viscosity when the berries are mashed. Consequences are a low juice yield and a poor colour. This can be prevented by the use of enzymes that degrade these polysaccharides. To use these enzy

  3. Characterisation of cell-wall polysaccharides from mandarin segment membranes

    NARCIS (Netherlands)

    Coll-Almela, L.; Saura-Lopez, D.; Laencina-Sanchez, J.; Schols, H.A.; Voragen, A.G.J.; Ros-García, J.M.

    2015-01-01

    In an attempt to develop a process of enzymatic peeling of mandarin segments suitable for use on an industrial scale, the cell wall fraction of the segment membrane of Satsuma mandarin fruits was extracted to obtain a chelating agent-soluble pectin fraction (ChSS), a dilute sodium hydroxide-soluble

  4. Evidence for a Melanin Cell Wall Component in Pneumocystis carinii

    OpenAIRE

    Icenhour, Crystal R.; Kottom, Theodore J.; Limper, Andrew H.

    2003-01-01

    Fluorescein isothiocyanate-labeled monoclonal antibodies specific for fungal melanin were used in this study to visualize melanin-like components of the Pneumocystis carinii cell wall. A colorimetric enzyme assay confirmed these findings. This is the first report of melanin-like pigments in Pneumocystis.

  5. Hetero-oligomeric cell wall channels (porins) of Nocardia farcinica.

    Science.gov (United States)

    Kläckta, Christian; Knörzer, Philipp; Riess, Franziska; Benz, Roland

    2011-06-01

    The cell wall of Nocardia farcinica contains a cation-selective cell wall channel, which may be responsible for the limited permeability of the cell wall of N. farcinica for negatively charged antibiotics. Based on partial sequencing of the protein responsible for channel formation derived from N. farcinica ATTC 3318 we were able to identify the corresponding genes (nfa15890 and nfa15900) within the known genome of N. farcinica IFM 10152. The corresponding genes of N. farcinica ATTC 3318 were separately expressed in the Escherichia coli BL21DE3Omp8 strain and the N-terminal His10-tagged proteins were purified to homogeneity using immobilized metal affinity chromatography. The pure proteins were designated NfpANHis and NfpBNHis, for N. farcinica porin A and N. farcinica porin B. The two proteins were checked separately for channel formation in lipid bilayers. Our results clearly indicate that the proteins NfpANHis and NfpBNHis expressed in E. coli could only together form a channel in lipid bilayer membranes. This means that the cell wall channel of N. farcinica is formed by a heterooligomer. NfpA and NfpB form together a channel that may structurally be related to MspA of Mycobacterium smegmatis based on amino acid comparison and renaturation procedure. PMID:21092733

  6. Structure of cellulose microfibrils in primary cell walls from Collenchyma

    Czech Academy of Sciences Publication Activity Database

    Thomas, L. H.; Forsyth, V. T.; Šturcová, Adriana; Kennedy, C. J.; May, R. P.; Altaner, C. M.; Apperley, D. C.; Wess, T. J.; Jarvis, M. C.

    2013-01-01

    Roč. 161, č. 1 (2013), s. 465-476. ISSN 0032-0889 R&D Projects: GA ČR GAP108/12/0703 Institutional support: RVO:61389013 Keywords : primary cell wall * cellulose microfibril structure * chain packing disorder Subject RIV: CD - Macromolecular Chemistry Impact factor: 7.394, year: 2013

  7. Polymer mobility in cell walls of cucumber hypocotyls

    Science.gov (United States)

    Fenwick, K. M.; Apperley, D. C.; Cosgrove, D. J.; Jarvis, M. C.

    1999-01-01

    Cell walls were prepared from the growing region of cucumber (Cucumis sativus) hypocotyls and examined by solid-state 13C NMR spectroscopy, in both enzymically active and inactivated states. The rigidity of individual polymer segments within the hydrated cell walls was assessed from the proton magnetic relaxation parameter, T2, and from the kinetics of cross-polarisation from 1H to 13C. The microfibrils, including most of the xyloglucan in the cell wall, as well as cellulose, behaved as very rigid solids. A minor xyloglucan fraction, which may correspond to cross-links between microfibrils, shared a lower level of rigidity with some of the pectic galacturonan. Other pectins, including most of the galactan side-chain residues of rhamnogalacturonan I, were much more mobile and behaved in a manner intermediate between the solid and liquid states. The only difference observed between the enzymically active and inactive cell walls, was the loss of a highly mobile, methyl-esterified galacturonan fraction, as the result of pectinesterase activity.

  8. Environmental stability of stem cell wall traits in alfalfa

    Science.gov (United States)

    The concentration of stem cell wall constituents in alfalfa, Medicago sativa L., herbage can affect dry matter intake and energy availability in dairy and beef production systems and impact energy conversion efficiency when alfalfa is used to produce biofuels. Stem Klason lignin, glucose, xylose, an...

  9. Plant Cell Wall Carbohydrates as Substrates for Azospirillum brasiliense†

    OpenAIRE

    Myers, Mary L.; Hubbell, David H.

    1987-01-01

    Carbohydrate components (simple sugars and polysaccharides) of cell walls of pearl millet (Pennisetum americanum L., cv. Gahi) were studied as potential substrates for the root-associated diazotroph Azospirillum brasiliense Sp. 7. Simple sugars were utilized, but no evidence was obtained to support the suggestion that the polysaccharide components tested might serve as substrates for growth following hydrolysis by the associated azospirilla.

  10. Display of active enzymes on the cell surface of Escherichia coli using PgsA anchor protein and their application to bioconversion.

    Science.gov (United States)

    Narita, Junya; Okano, Kenji; Tateno, Toshihiro; Tanino, Takanori; Sewaki, Tomomitsu; Sung, Moon-Hee; Fukuda, Hideki; Kondo, Akihiko

    2006-05-01

    We have developed a novel Escherichia coli cell surface display system by employing PgsA as an anchoring motif. In our display system, C-terminal fusion to PgsA anchor protein from Bacillus subtilis was used. The enzymes selected for display were alpha-amylase (AmyA) from Streptococcus bovis 148 and lipase B (CALB) from Candida antarctica. The molecular mass values of AmyA and CALB are approximately 77 and 34 kDa, respectively. The enzymes were displayed on the surface as a fusion protein with a FLAG peptide tag at the C terminus. Both the PgsA-AmyA-FLAG and PgsA-CALB-FLAG fusion proteins were shown to be displayed by immunofluorescence labeling using anti-FLAG antibody. The displayed enzymes were active forms, and AmyA and CALB activities reached 990 U/g (dry cell weight) and 4.6 U/g (dry cell weight), respectively. AmyA-displaying E. coli cells grew utilizing cornstarch as the sole carbon source, while CALB-displaying E. coli cells catalyzed enantioselective transesterification, indicating that they are effective whole-cell biocatalysts. Since a target enzyme with a size of 77 kDa and an industrially useful lipase have been successfully displayed on the cell surface of E. coli for the first time, PgsA protein is probably a useful anchoring motif to display various enzymes. PMID:16133338

  11. MIG-10 (lamellipodin) has netrin-independent functions and is a FOS-1A transcriptional target during anchor cell invasion in C. elegans

    OpenAIRE

    Wang, Zheng; Chi, Qiuyi; Sherwood, David R.

    2014-01-01

    To transmigrate basement membrane, cells must coordinate distinct signaling activities to breach and pass through this dense extracellular matrix barrier. Netrin expression and activity are strongly associated with invasion in developmental and pathological processes, but how netrin signaling is coordinated with other pathways during invasion is poorly understood. Using the model of anchor cell (AC) invasion in C. elegans, we have previously shown that the integrin receptor heterodimer INA-1/...

  12. Cell wall structure and function in lactic acid bacteria

    OpenAIRE

    Kulakauskas, Saulius

    2014-01-01

    The cell wall of Gram-positive bacteria is a complex assemblage of glycopolymers and proteins. It consists of a thick peptidoglycan sacculus that surrounds the cytoplasmic membrane and that is decorated with teichoic acids, polysaccharides, and proteins. It plays a major role in bacterial physiology since it maintains cell shape and integrity during growth and division; in addition, it acts as the interface between the bacterium and its environment. Lactic acid bacteria (LAB) are traditionall...

  13. Ultrastructure and biochemistry of the cell wall of Methanococcus voltae.

    OpenAIRE

    Koval, S F; Jarrell, K F

    1987-01-01

    The ultrastructure and chemical composition of the cell wall of the marine archaebacterium Methanococcus voltae were studied by negative-staining and freeze-etch electron microscopy and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. M. voltae possesses a single regularly structured (RS) protein layer external to the plasma membrane. Freeze-etch preparations of cells indicated that the protein subunits are hexagonally arranged with a center-to-center spacing of approximately 10 ...

  14. A Phosphorylated Pseudokinase Complex Controls Cell Wall Synthesis in Mycobacteria

    OpenAIRE

    Gee, Christine L.; Papavinasasundaram, Kadamba G.; Blair, Sloane R.; Baer, Christina E.; Falick, Arnold M.; King, David S.; Griffin, Jennifer E.; Venghatakrishnan, Harene; Zukauskas, Andrew; Wei, Jun-Rong; Dhiman, Rakesh K.; Crick, Dean C.; Rubin, Eric J.; Sassetti, Christopher M.; Alber, Tom

    2012-01-01

    Prokaryotic cell wall biosynthesis is coordinated with cell growth and division, but the mechanisms regulating this dynamic process remain obscure. Here, we describe a phosphorylation-dependent regulatory complex that controls peptidoglycan (PG) biosynthesis in Mycobacterium tuberculosis. We found that PknB, a PG-responsive Ser-Thr protein kinase (STPK), initiates complex assembly by phosphorylating a kinase-like domain in the essential PG biosynthetic protein, MviN. This domain was structura...

  15. Molecular Mechanisms for Vascular Development and Secondary Cell Wall Formation

    OpenAIRE

    Yang, Jung Hyun; Wang, Huanzhong

    2016-01-01

    Vascular tissues are important for transporting water and nutrients throughout the plant and as physical support of upright growth. The primary constituents of vascular tissues, xylem, and phloem, are derived from the meristematic vascular procambium and cambium. Xylem cells develop secondary cell walls (SCWs) that form the largest part of plant lignocellulosic biomass that serve as a renewable feedstock for biofuel production. For the last decade, research on vascular development and SCW bio...

  16. The Novel Tail-anchored Membrane Protein Mff Controls Mitochondrial and Peroxisomal Fission in Mammalian Cells

    OpenAIRE

    Gandre-Babbe, Shilpa; van der Bliek, Alexander M.

    2008-01-01

    Few components of the mitochondrial fission machinery are known, even though mitochondrial fission is a complex process of vital importance for cell growth and survival. Here, we describe a novel protein that controls mitochondrial fission. This protein was identified in a small interfering RNA (siRNA) screen using Drosophila cells. The human homologue of this protein was named Mitochondrial fission factor (Mff). Mitochondria of cells transfected with Mff siRNA form a closed network similar t...

  17. Scaffolding during the cell cycle by A-kinase anchoring proteins

    OpenAIRE

    Han, B.; Poppinga, W J; Schmidt, M.

    2015-01-01

    Cell division relies on coordinated regulation of the cell cycle. A process including a well-defined series of strictly regulated molecular mechanisms involving cyclin-dependent kinases, retinoblastoma protein, and polo-like kinases. Dysfunctions in cell cycle regulation are associated with disease such as cancer, diabetes, and neurodegeneration. Compartmentalization of cellular signaling is a common strategy used to ensure the accuracy and efficiency of cellular responses. Compartmentalizati...

  18. The Cell Wall Teichuronic Acid Synthetase (TUAS Is an Enzyme Complex Located in the Cytoplasmic Membrane of Micrococcus luteus

    Directory of Open Access Journals (Sweden)

    Lingyi Lynn Deng

    2010-01-01

    composed of disaccharide repeating units [-4-β-D-ManNAcAp-(1→6α-D-Glcp−1-]n, which is covalently anchored to the peptidoglycan on the inner cell wall and extended to the outer surface of the cell envelope. An enzyme complex responsible for the TUA chain biosynthesis was purified and characterized. The 440 kDa enzyme complex, named teichuronic acid synthetase (TUAS, is an octomer composed of two kinds of glycosyltransferases, Glucosyltransferase, and ManNAcA-transferase, which is capable of catalyzing the transfer of disaccharide glycosyl residues containing both glucose and the N-acetylmannosaminuronic acid residues. TUAS displays hydrophobic properties and is found primarily associated with the cytoplasmic membrane. The purified TUAS contains carotinoids and lipids. TUAS activity is diminished by phospholipase digestion. We propose that TUAS serves as a multitasking polysaccharide assembling station on the bacterial membrane.

  19. Crushing Strength of Aluminum Honeycomb with Thinning Cell Wall

    Science.gov (United States)

    Ogasawara, Nagahisa; Chiba, Norimasa; Kobayashi, Eiji; Kikuchi, Yuji

    To evaluate the crash safety of automobiles, various collision tests are performed by the auto industry. In the offset frontal collision test and the side collision test, the target is an aluminum honeycomb material which has thinning cell walls. In this study, based on the analyses of the shock absorption mechanism, a new crushing strength formula is proposed. First, load-displacement curves obtained from compression tests in quasi-static condition showed an almost linear relation between a thinning rate of cell walls and a crushing strength. Second, based on Wierzbicki's theory, a new formula was proposed, which can estimate a crushing strength of a honeycomb material with thinning wall. In addition, a correcting equation which considered an elastic deformation was also proposed. Third, parametric analyses were carried out with a FE model which can simulate a delamination between cell walls. The results obtained from the theory and FEM almost corresponded to each other for a wide range of the thinning rate. Fourth, impact tests were carried out, in which the weight was dropped freely at the speed used for the automobile tests. Those results almost agreed well with the sum of the theoretical crush strength and the inside air pressure.

  20. Nucleated assembly of Chlamydomonas and Volvox cell walls.

    Science.gov (United States)

    Adair, W S; Steinmetz, S A; Mattson, D M; Goodenough, U W; Heuser, J E

    1987-11-01

    The Chlamydomonas reinhardtii cell wall is made up of hydroxyproline-rich glycoproteins, arranged in five distinct layers. The W6 (crystalline) layer contains three major glycoproteins (GP1, GP2, GP3), selectively extractable with chaotropic agents, that self-assemble into crystals in vitro. A system to study W6 assembly in a quantitative fashion was developed that employs perchlorate-extracted Chlamydomonas cells as nucleating agents. Wall reconstitution by biotinylated W6 monomers was monitored by FITC-streptavidin fluorescence and quick-freeze/deep-etch electron microscopy. Optimal reconstitution was obtained at monomer concentrations (0.2-0.3 mg/ml) well below those required for nonnucleated assembly. Assembly occurred from multiple nucleation sites, and faithfully reflected the structure of the intact W6 layer. Specificity of nucleated assembly was demonstrated using two cell-wall mutants (cw-2 and cw-15); neither served as a substrate for assembly of wild-type monomers. In addition, W6 sublayers were assembled from purified components: GP2 and GP3 coassembled to form the inner (W6A) sublayer; this then served as a substrate for self-assembly of GP1 into the outer (W6B) sublayer. Finally, evolutionary relationships between C. reinhardtii and two additional members of the Volvocales (Chlamydomonas eugametos and Volvox carteri) were explored by performing interspecific reconstitutions. Hybrid walls were obtained between C. reinhardtii and Volvox but not with C. eugametos, confirming taxonomic assignments based on structural criteria. PMID:3680387

  1. In situ microscopic observation of chitin and fungal cells with chitinous cell walls in hydrothermal conditions

    OpenAIRE

    Shigeru Deguchi; Kaoru Tsujii; Koki Horikoshi

    2015-01-01

    Recent findings of intact chitin in fossil records suggest surprisingly high recalcitrance of this biopolymer during hydrothermal treatments. We also know in the experience of everyday life that mushroom, cells of which have chitinous cell walls, do not fall apart however long they are simmered. We used in situ optical microscopy to examine chitin and fungal cells with chitinous cell walls during hydrothermal treatments, and obtained direct evidence that they remained undegraded at temperatur...

  2. Effect of Yeast Cell Morphology, Cell Wall Physical Structure and Chemical Composition on Patulin Adsorption

    OpenAIRE

    Luo, Ying; Wang, Jianguo; Liu, Bin; Wang, Zhouli; Yuan, Yahong; Yue, Tianli

    2015-01-01

    The capability of yeast to adsorb patulin in fruit juice can aid in substantially reducing the patulin toxic effect on human health. This study aimed to investigate the capability of yeast cell morphology and cell wall internal structure and composition to adsorb patulin. To compare different yeast cell morphologies, cell wall internal structure and composition, scanning electron microscope, transmission electron microscope and ion chromatography were used. The results indicated that patulin ...

  3. Augmented delivery of gemcitabine in lung cancer cells exploring mannose anchored solid lipid nanoparticles.

    Science.gov (United States)

    Soni, Namrata; Soni, Neetu; Pandey, Himanshu; Maheshwari, Rahul; Kesharwani, Prashant; Tekade, Rakesh Kumar

    2016-11-01

    Gemcitabine (GmcH) is an effective anti-cancer agent used in the chemotherapy of lung cancer. However, the clinical applications of GmcH has been impeded primarily due to its low blood residence time, unfavorable pharmacokinetic and pharmacodynamic (PK/PD) profile, and poor penetration in the complex environment of lung cancer cells. Thus, the present study aims to formulate GmcH loaded mannosylated solid lipid nanoparticles (GmcH-SLNs) for improving its drug uptake into the lung cancer cells. GmcH-SLNs were prepared by emulsification and solvent evaporation process, and surface modification was done with mannose using ring opening technique. The cellular toxicity and cell uptake studies were performed in A549 lung adenocarcinoma cell line. The developed nanoformulation appears to be proficient in targeted delivery of GmcH with improved therapeutic effectiveness and enhanced safety. PMID:27459173

  4. Cell surface engineering of renal cell carcinoma with glycosylphosphatidylinositol-anchored TIMP-1 blocks TGF- β 1 activation and reduces regulatory ID gene expression.

    Science.gov (United States)

    Notohamiprodjo, Susan; Djafarzadeh, Roghieh; Rieth, Nicole; Hofstetter, Monika; Jaeckel, Carsten; Nelson, Peter J

    2012-12-01

    Tissue inhibitor of metalloproteinase 1 (TIMP-1) controls matrix metalloproteinase activity through 1:1stoichiometric binding. Human TIMP-1 fused to a glycosylphosphatidylinositol(GPI) anchor (TIMP-1 - GPI) shifts the activity of TIMP-1 from the extracellular matrix to the cell surface. TIMP-1 - GPI treated renal cell carcinoma cells show increased apoptosis and reduced proliferation.Transcriptomic profiling and regulatory pathway mapping were used to identify the potential mechanisms driving these effects. Significant changes in the DNA binding inhibitors, TGF- β 1/SMAD and BMP pathways resulted from TIMP-1 - GPI treatment. These events were linked to reduced TGF- β 1 signaling mediated by inhibition of proteolytic processing of latent TGF- β 1 by TIMP-1 - GPI. PMID:23667903

  5. Identification of Cell Wall Synthesis Regulatory Genes Controlling Biomass Characteristics and Yield in Rice (Oryza Sativa)

    Energy Technology Data Exchange (ETDEWEB)

    Peng, Zhaohua PEng [Mississippi State University; Ronald, Palmela [UC-Davis; Wang, Guo-Liang [The Ohio State University

    2013-04-26

    This project aims to identify the regulatory genes of rice cell wall synthesis pathways using a cell wall removal and regeneration system. We completed the gene expression profiling studies following the time course from cell wall removal to cell wall regeneration in rice suspension cells. We also completed, total proteome, nuclear subproteome and histone modification studies following the course from cell wall removal and cell wall regeneration process. A large number of differentially expressed regulatory genes and proteins were identified. Meanwhile, we generated RNAi and over-expression transgenic rice for 45 genes with at least 10 independent transgenic lines for each gene. In addition, we ordered T-DNA and transposon insertion mutants for 60 genes from Korea, Japan, and France and characterized the mutants. Overall, we have mutants and transgenic lines for over 90 genes, exceeded our proposed goal of generating mutants for 50 genes. Interesting Discoveries a) Cell wall re-synthesis in protoplasts may involve a novel cell wall synthesis mechanism. The synthesis of the primary cell wall is initiated in late cytokinesis with further modification during cell expansion. Phragmoplast plays an essential role in cell wall synthesis. It services as a scaffold for building the cell plate and formation of a new cell wall. Only one phragmoplast and one new cell wall is produced for each dividing cell. When the cell wall was removed enzymatically, we found that cell wall re-synthesis started from multiple locations simultaneously, suggesting that a novel mechanism is involved in cell wall re-synthesis. This observation raised many interesting questions, such as how the starting sites of cell wall synthesis are determined, whether phragmoplast and cell plate like structures are involved in cell wall re-synthesis, and more importantly whether the same set of enzymes and apparatus are used in cell wall re-synthesis as during cytokinesis. Given that many known cell wall

  6. Ectopic lignification in primary cellulose-deficient cell walls of maize cell suspension cultures

    Institute of Scientific and Technical Information of China (English)

    Hugo Melida; Antonio Encina; Asier Largo-Gosens; Esther Novo-Uzal; Rogelio Santiago; Federico Pomar; Pedro Garca; Penelope Garca-Angulo; Jose Luis Acebes; Jesus Alvarez

    2015-01-01

    Maize (Zea mays L.) suspension-cultured cells with up to 70% less cellulose were obtained by stepwise habituation to dichlobenil (DCB), a cellulose biosynthesis inhibitor. Cellulose deficiency was accompanied by marked changes in cell wall matrix polysaccharides and phenolics as revealed by Fourier transform infrared (FTIR) spectroscopy. Cell wall compositional analysis indicated that the cellulose-deficient cell walls showed an enhancement of highly branched and cross-linked arabinoxylans, as well as an increased content in ferulic acid, diferulates and p-coumaric acid, and the presence of a polymer that stained positive for phloroglucinol. In accordance with this, cellulose-deficient cell walls showed a fivefold increase in Klason-type lignin. Thioacidolysis/GC-MS analysis of cellulose-deficient cell walls indicated the presence of a lignin-like polymer with a Syringyl/Guaiacyl ratio of 1.45, which differed from the sensu stricto stress-related lignin that arose in response to short-term DCB-treatments. Gene expression analysis of these cells indicated an overexpression of genes specific for the biosynthesis of monolignol units of lignin. A study of stress signaling pathways revealed an overexpression of some of the jasmonate signaling pathway genes, which might trigger ectopic lignification in response to cell wall integrity disruptions. In summary, the structural plasticity of primary cell walls is proven, since a lignification process is possible in response to cellulose impoverishment.

  7. Surface Analyses and Immune Reactivities of Major Cell Wall-Associated Proteins of Group A Streptococcus

    OpenAIRE

    Cole, Jason N; Ramirez, Ruben D.; Currie, Bart J.; Cordwell, Stuart J.; Djordjevic, Steven P.; Mark J Walker

    2005-01-01

    A proteomic analysis was undertaken to identify cell wall-associated proteins of Streptococcus pyogenes. Seventy-four distinct cell wall-associated proteins were identified, 66 of which were novel. Thirty-three proteins were immunoreactive with pooled S. pyogenes-reactive human antisera. Biotinylation of the GAS cell surface identified 23 cell wall-associated proteins that are surface exposed.

  8. 基于框架预应力锚杆加固技术的重力式挡土墙的稳定性分析%Stability analysis of gravity retaining wall reinforced with framed prestressed anchors

    Institute of Scientific and Technical Information of China (English)

    朱彦鹏; 王勇; 周勇

    2012-01-01

    Based on the instability phenomenon of gravity retaining wall, the framed anchor with prestress was used to reinforce the retaining wall. On the basis of limit equilibrium theory and Coulomb earth pressure theory, the stability of retaining walls was analyzed and the solution formula of safety coefficient of stability of anti-slip and anti-overturning was derived. It was shown by illustrative computation and cont-rastive analysis, the result showed that the safety coefficient of stability of the gravity retaining wall with this kind of reinforcement techniques was significantly improved. The safety coefficient of stability would be increased by about 50% when the prestress was increased by 50 kN. However, taking the economic and security aspects into account, 100 kN prestress applied to the anchors would be the most optimaL%基于重力式挡土墙不稳定的现象,运用框架预应力锚杆加固技术对重力式挡土墙进行加固.以极限平衡理论及库仑土压力理论为基础,对挡土墙进行稳定性分析,推导出挡土墙的抗滑移稳定性和抗倾覆稳定性安全系数的求解计算公式.通过算例计算及对比分析,结果显示引入此加固技术的重力式挡墙的稳定性安全系数明显提高,在锚杆预应力每增加50 kN时,稳定性安全系数提高的幅度达到50%左右;从经济性和安全性两方面综合考虑,对锚杆施加100kN的预应力是最佳的.

  9. Reduced Wall Acetylation Proteins Play Vital and Distinct Roles in Cell Wall O-Acetylation in Arabidopsis

    DEFF Research Database (Denmark)

    Manabe, Yuzuki; Verhertbruggen, Yves; Gille, Sascha;

    2013-01-01

    The Reduced Wall Acetylation (RWA) proteins are involved in cell wall acetylation in plants. Previously, we described a single mutant, rwa2, which has about 20% lower level of O-acetylation in leaf cell walls and no obvious growth or developmental phenotype. In this study, we generated double...... quadruple rwa mutant can be completely complemented with the RWA2 protein expressed under 35S promoter, indicating the functional redundancy of the RWA proteins. Nevertheless, the degree of acetylation of xylan, (gluco) mannan, and xyloglucan as well as overall cell wall acetylation is affected differently...... in different combinations of triple mutants, suggesting their diversity in substrate preference. The overall degree of wall acetylation in the rwa quadruple mutant was reduced by 63% compared with the wild type, and histochemical analysis of the rwa quadruple mutant stem indicates defects in cell...

  10. Construction of a highly active xylanase displaying oleaginous yeast: comparison of anchoring systems.

    Directory of Open Access Journals (Sweden)

    Sophie Duquesne

    Full Text Available Three Yarrowia lipolytica cell wall proteins (YlPir, YlCWP1 and YlCBM were evaluated for their ability to display the xylanase TxXYN from Thermobacillus xylanilyticus on the cell surface of Y. lipolytica. The fusion proteins were produced in Y. lipolytica JMY1212, a strain engineered for mono-copy chromosomal insertion, and enabling accurate comparison of anchoring systems. The construction using YlPir enabled cell bound xylanase activity to be maximised (71.6 U/g. Although 48% of the activity was released in the supernatant, probably due to proteolysis at the fusion zone, this system is three times more efficient for the anchoring of TxXYN than the YlCWP1 system formerly developed for Y. lipolytica. As far as we know it represents the best displayed xylanase activity ever published. It could be an attractive alternative anchoring system to display enzymes in Y. lipolytica.

  11. Bio-inspired Construction of Advanced Fuel Cell Cathode with Pt Anchored in Ordered Hybrid Polymer Matrix

    Science.gov (United States)

    Xia, Zhangxun; Wang, Suli; Jiang, Luhua; Sun, Hai; Liu, Shuang; Fu, Xudong; Zhang, Bingsen; Sheng Su, Dang; Wang, Jianqiang; Sun, Gongquan

    2015-11-01

    The significant use of platinum for catalyzing the cathodic oxygen reduction reactions (ORRs) has hampered the widespread use of polymer electrolyte membrane fuel cells (PEMFCs). The construction of well-defined electrode architecture in nanoscale with enhanced utilization and catalytic performance of Pt might be a promising approach to address such barrier. Inspired by the highly efficient catalytic processes in enzymes with active centers embedded in charge transport pathways, here we demonstrate for the first time a design that allocates platinum nanoparticles (Pt NPs) at the boundaries with dual-functions of conducting both electrons by aid of polypyrrole and protons via Nafion® ionomer within hierarchical nanoarrays. By mimicking enzymes functionally, an impressive ORR activity and stability is achieved. Using this brand new electrode architecture as the cathode and the anode of a PEMFC, a high mass specific power density of 5.23 W mg-1Pt is achieved, with remarkable durability. These improvements are ascribed to not only the electron decoration and the anchoring effects from the Nafion® ionomer decorated PPy substrate to the supported Pt NPs, but also the fast charge and mass transport facilitated by the electron and proton pathways within the electrode architecture.

  12. Antigenicity and immunogenicity of an extract from the cell wall and cell membrane of Histoplasma capsulatum yeast cells.

    OpenAIRE

    Gómez, A M; Rhodes, J C; Deepe, G S

    1991-01-01

    In order to identify T-cell antigens from Histoplasma capsulatum yeast cells, we prepared a detergent extract of the cell wall and cell membrane of yeast-phase H. capsulatum G217B and analyzed its antigenicity and immunogenicity. Mice injected with viable H. capsulatum yeast cells or with 500 or 1,000 micrograms of the extract mounted a delayed-type hypersensitivity response to solubilized cell wall and cell membrane. Vaccination with this antigenic preparation conferred a protective immune r...

  13. Orbital wall infarction in child with sickle cell disease.

    Science.gov (United States)

    Janssens, C; Claeys, L; Maes, P; Boiy, T; Wojciechowski, M

    2015-12-01

    We present the case of a 17-year-old boy, known with homozygous sickle cell disease, who was admitted because of generalised pain. He developed bilateral periorbital oedema and proptosis, without pain or visual disturbances. In addition to hyperhydration, oxygen and analgesia IV antibiotics were started, to cover a possible osteomyelitis. Patients with sickle cell disease are at risk for vaso-occlusive crises, when the abnormally shaped red blood cells aggregate and block the capillaries. Such a crisis typically presents at a location with high bone marrow activity, as the vertebrae and long bones. At an early age, the bone marrow is still active at other sites, for example the orbital wall, and thus infarction can also occur there. Thus, in young persons with sickle cell disease, it is important to consider orbital wall infarction in the differential diagnosis, since the approach is different from osteomyelitis. If the disease is complicated by an orbital compression syndrome, corticosteroids or surgical intervention may be necessary to preserve the vision. In our patient, an MRI of the orbitae demonstrated periorbital oedema with bone anomalies in the orbital and frontal bones, confirming orbital wall infarction. Ophthalmological examination revealed no signs of pressure on the nervus opticus. The patient recovered gradually with conservative treatment. PMID:26790559

  14. Preparation and Characterization of Squaraine Dyes containing Mono- and Bis-Anchoring Groups as the Light Absorber in Dye Sensitized Solar Cells

    International Nuclear Information System (INIS)

    Graphical abstract: - Highlights: • We aim to enhance charge transfer between squaraine dyes (SQ) and TiO2. • Smaller amounts of SSQ dye is loaded on the TiO2 than the USQ dye. • SSQ shows better electronic coupling with TiO2 than USQ. • The enhanced electronic coupling leads SSQ to show better photovoltaic performance. - Abstract: Enhancing the charge transfer rate in dye sensitized solar cells (DSSCs) is one of the most important criteria determining the cell efficiency. This paper reports a novel strategy for enhancing charge transfer between squaraine dyes and TiO2 by increasing the anchoring number. Symmetrical (SSQ) and unsymmetrical squaraine (USQ) sensitizers with different numbers of anchoring groups in their chromophores are synthesized. SSQ dye containing a bis-anchoring group shows better electronic coupling with TiO2 than the USQ dye with a mono-bridge. This enhanced electronic coupling leads SSQ to exhibit better photovoltaic performance, though smaller amounts of SSQ dye is loaded on the TiO2 than the USQ dye. Under standard global AM 1.5 solar conditions, the optimized SSQ-sensitized solar cells show 7.76 mA cm−2 of short-circuit photocurrent density, 0.62 V of open-circuit voltage, and 0.65 of fill factor (FF), to give overall conversion efficiency of 3.20%, which is approximately 25% higher than the USQ-sensitized solar cells

  15. Effect of Yeast Cell Morphology, Cell Wall Physical Structure and Chemical Composition on Patulin Adsorption.

    Directory of Open Access Journals (Sweden)

    Ying Luo

    Full Text Available The capability of yeast to adsorb patulin in fruit juice can aid in substantially reducing the patulin toxic effect on human health. This study aimed to investigate the capability of yeast cell morphology and cell wall internal structure and composition to adsorb patulin. To compare different yeast cell morphologies, cell wall internal structure and composition, scanning electron microscope, transmission electron microscope and ion chromatography were used. The results indicated that patulin adsorption capability of yeast was influenced by cell surface areas, volume, and cell wall thickness, as well as 1,3-β-glucan content. Among these factors, cell wall thickness and 1,3-β-glucan content serve significant functions. The investigation revealed that patulin adsorption capability was mainly affected by the three-dimensional network structure of the cell wall composed of 1,3-β-glucan. Finally, patulin adsorption in commercial kiwi fruit juice was investigated, and the results indicated that yeast cells could adsorb patulin from commercial kiwi fruit juice efficiently. This study can potentially simulate in vitro cell walls to enhance patulin adsorption capability and successfully apply to fruit juice industry.

  16. Effects of spaceflight on polysaccharides of Saccharomyces cerevisiae cell wall.

    Science.gov (United States)

    Liu, Hong-Zhi; Wang, Qiang; Liu, Xiao-Yong; Tan, Sze-Sze

    2008-12-01

    Freeze-dried samples of four Saccharomyces cerevisiae strains, namely, FL01, FL03, 2.0016, and 2.1424, were subjected to spaceflight. After the satellite's landing on Earth, the samples were recovered and changes in yeast cell wall were analyzed. Spaceflight strains of all S. cerevisiae strains showed significant changes in cell wall thickness (P growth curve analysis showed spaceflight S. cerevisiae 2.0016 had a faster growth rate, shorter lag phase periods, higher final biomass, and higher content of beta-glucan. Genetic stability analysis showed that prolonged subculturing of spaceflight strain S. cerevisiae 2.0016 did not lead to the appearance of variants, indicating that the genetic stability of S. cerevisiae 2.0016 mutant could be sufficient for its exploitation of beta-glucan production. PMID:18797865

  17. A Communal Bacterial Adhesin Anchors Biofilm and Bystander Cells to Surfaces

    OpenAIRE

    Absalon, Cedric; Van Dellen, Katrina; Paula I. Watnick

    2011-01-01

    Author Summary The bacterial multilayer biofilm consists of matrix-enclosed cells attached to each other to form large aggregates. The base of these aggregates may be attached to a living or non-living surface. The biofilm matrix most often contains at least one exopolysaccharide component and may also contain protein and DNA. While much is known about the exopolysaccharide component of the Gram-negative biofilm matrix, little is known about the function of biofilm matrix proteins. We hypothe...

  18. Nitrate sensing and cell wall modification in Staphylococci

    OpenAIRE

    Niemann, Volker

    2015-01-01

    This thesis highlights two topics concerning the regulation of energy metabolism and the cell wall biosynthesis in Staphylococci. Most members of this genus are facultative anaerobic microorganisms able to respire on nitrate as final electron acceptor. The completely apathogenic organism Staphylococcus carnosus is used as starter culture in food industry. Dissimilatory nitrate reduction causes desired effects during the ripening process of sausages. First, the nitrate concentra...

  19. Transcriptome analysis of secondary cell wall development in Medicago truncatula

    OpenAIRE

    Wang, Huanzhong; Yang, Jung Hyun; Chen, Fang; Torres-Jerez, Ivone; Tang, Yuhong; Wang, Mingyi; Du, Qian; Cheng, Xiaofei; Wen, Jiangqi; Dixon, Richard

    2016-01-01

    Background Legumes are important to humans by providing food, feed and raw materials for industrial utilizations. Some legumes, such as alfalfa, are potential bioenergy crops due to their high biomass productivity. Global transcriptional profiling has been successfully used to identify genes and regulatory pathways in secondary cell wall thickening in Arabidopsis, but such transcriptome data is lacking in legumes. Results A systematic microarray assay and high through-put real time PCR analys...

  20. Antioxidant properties of cell wall polysaccharides of Stevia rebaudiana leaves

    OpenAIRE

    Mediesse Kengne Francine; Woguia Alice Louise; Fogue Souopgui Pythagore; Atogho-Tiedeu Barbara; Simo Gustave; Thaddée Boudjeko

    2014-01-01

    Objective: To examine the total phenolic and protein contents, and the antioxidant activities of cell wall polysaccharide fractions of Stevia rebaudiana leaves. Methods: Three different polysaccharide-enriched fractions, namely FPE (extract with 50 mmol/ L ethylene diamine tetra acetic acid), FPK (extract with 0.05 mol/L KOH) and FH (extract with 4 mol/L KOH) were extracted from Stevia rebaudiana leaves. The antioxidant activity of these fractions was evaluated based on thei...

  1. Cell-wall hemicelluloses as mobile carbon stores in plants

    OpenAIRE

    Schädel, Christina

    2009-01-01

    Hemicelluloses are the second most abundant polysaccharide in nature after cellulose. So far, the chemical heterogeneity of cell-wall hemicelluloses and the relatively large sample-volume required in existing methods represent major obstacles for large-scale, cross-species analyses of this important plant compounds. Here, we apply a new micro-extraction method to analyse hemicelluloses and the ratio of ‘cellulose and lignin’ to hemicelluloses in different tissues of 28 plant species comprisin...

  2. The Mechanisms of Plant Cell Wall Deconstruction during Enzymatic Hydrolysis

    OpenAIRE

    Thygesen, Lisbeth G; Thybring, Emil E.; Johansen, Katja S.; Claus Felby

    2014-01-01

    Mechanical agitation during enzymatic hydrolysis of insoluble plant biomass at high dry matter contents is indispensable for the initial liquefaction step in biorefining. It is known that particle size reduction is an important part of liquefaction, but the mechanisms involved are poorly understood. Here we put forward a simple model based on mechanical principles capable of capturing the result of the interaction between mechanical forces and cell wall weakening via hydrolysis of glucosidic ...

  3. Plant cell walls: New insights from ancient species

    DEFF Research Database (Denmark)

    Sørensen, Iben; Willats, William George Tycho

    2008-01-01

    ¿4)-linked ß-D-Glcp are joined by occasional (1¿3)-linkages. This mixed linkage glucan (MLG) has been the subject of extensive research because of the economic importance of several Poales species including rice, barley and wheat and because MLG has proven health benefits. The recent discovery of MLG......-D-glucan is not unique to the Poales and is an abundant component of Equisetum arvense cell walls. Plant J 2008; 54:510-21....

  4. Cell wall perturbation sensitizes fungi to the antimalarial drug chloroquine

    OpenAIRE

    Islahudin, Farida; Khozoie, Combiz; Bates, Steven; Ting, Kang-Nee; Pleass, Richard J.; Avery, Simon V.

    2013-01-01

    Chloroquine (CQ) has been a mainstay of antimalarial drug treatment for several decades. Additional therapeutic actions of CQ have been described, including some reports of fungal inhibition. Here we investigated the action of CQ in fungi, including the yeast model Saccharomyces cerevisiae. A genomewide yeast deletion strain collection was screened against CQ, revealing that bck1Δ and slt2Δ mutants of the cell wall integrity pathway are CQ hypersensitive. This phenotype was rescued with sorbi...

  5. Enzyme Amplified Detection of Microbial Cell Wall Components

    Science.gov (United States)

    Wainwright, Norman R.

    2004-01-01

    This proposal is MBL's portion of NASA's Johnson Space Center's Astrobiology Center led by Principal Investigator, Dr. David McKay, entitled: 'Institute for the Study of Biomarkers in Astromaterials.' Dr. Norman Wainwright is the principal investigator at MBL and is responsible for developing methods to detect trace quantities of microbial cell wall chemicals using the enzyme amplification system of Limulus polyphemus and other related methods.

  6. Control of lipid storage and cell size between adipocytes by vesicle-associated glycosylphosphatidylinositol-anchored proteins.

    Science.gov (United States)

    Müller, Günter

    2011-02-01

    Adipose tissue mass in mammals is expanding by increasing the average cell volume as well as the total number of the adipocytes. Up-regulation of lipid storage in fully differentiated adipocytes resulting in their enlargement is well documented and thought to be a critical mechanism for the expansion of adipose tissue depots during the growth of both lean and obese animals and human beings. A novel molecular mechanism for the regulation of lipid storage and cell size in rat adipocytes has recently been elucidated for the physiological stimuli, palmitate and hydrogen peroxide, the anti-diabetic sulfonylurea drug, glimepiride, and insulin-mimetic phosphoinositolglycans. It encompasses (i) the release of small vesicles, so-called adiposomes, harbouring the glycosylphosphatidylinositol-anchored (c)AMP-degrading phosphodiesterase Gce1 and 5'-nuceotidase CD73 from large donor adipocytes, (ii) the transfer of the adiposomes and their interaction with detergent-insoluble glycolipid-enriched microdomains of the plasma membrane of small acceptor adipocytes, (iii) the translocation of Gce1 and CD73 from the adiposomes to the intracellular lipid droplets of the acceptor adipocytes and (iv) the degradation of (c)AMP at the lipid droplet surface zone by Gce1 and CD73 in the acceptor adipocytes. In concert, this sequence of events leads to up-regulation of esterification of fatty acids into triacylglycerol and down-regulation of their release from triacylglycerol. This apparent mechanism for shifting the triacylglycerol burden from large to small adipocytes may provide novel strategies for the therapy of metabolic diseases, such as type 2 diabetes and obesity. PMID:20883086

  7. Changes in alfalfa cell wall structure during vegetation

    Directory of Open Access Journals (Sweden)

    Božičković Aleksa Đ.

    2014-01-01

    Full Text Available The investigation was done on 141 samples of one alfalfa cultivar, collected from the same location during the first three growth cycles: spring growth, the first and the second regrowth. Within each growth cycle, sampling was done during the whole growing period, commencing when plant height was below 150 mm and continuing until plants were bearing ripe seeds. On all collected samples the following cell wall characteristics were determined: neutral detergent fibre (NDF, acid detergent fibre (ADF, acid detergent lignin (ADL, neutral detergent insoluble crude protein (NDICP, acid detergent insoluble crude protein (ADICP. Cellulose and hemicellulose were detected on the base of the mentioned chemical parameters. Significantly lower (p<0.01 content of aNDF, ADF, ADL, ADICP and cellulose is found in the second regrowth, while there were no significant differences between the other two growth cycles. Except in NDICP and ADICP, the increase in all accompanying components of the cell wall was observed, and expressed in average daily changes. There was no consistent trend in NDICP and ADICP. During the spring growth from late bud to full-bloom stage the ’plateau’ was observed. The plateau was represented as almost constant content of aNDF, ADF, ADL and cellulose. The correlations between all components of the cell wall were shown. The equation aNDF = 36.713 + 1.181 × ADF is recommended for conversion of ADF into aNDF in alfalfa. [Projekat Ministarstva nauke Republike Srbije, br. III 46012

  8. Secondary cell wall polysaccharides in Bacillus anthracis and Bacillus cereus strains

    OpenAIRE

    Leoff, Christine

    2009-01-01

    This thesis presents a systematic comparison of cell wall carbohydrates, in particular the non classical secondary cell wall polysaccharides from closely related strains within the Bacillus cereus group. The results suggest that the cell wall glycosyl composition of the various Bacillus cereus group strains display differences that correlate with their phylogenetic relatedness. Comparative structural analysis of polysaccharide components that were released from the cell walls of the various s...

  9. The cell wall and endoplasmic reticulum stress responses are coordinately regulated in Saccharomyces cerevisiae

    OpenAIRE

    Krysan, Damian J.

    2009-01-01

    The unfolded protein response (UPR) is an intracellular signaling pathway that regulates the cellular response to the accumulation of misfolded proteins in eukaryotes. Our group has demonstrated that cell wall stress activates UPR in yeast through signals transmitted by the cell wall integrity (CWI) mitogen-activated protein (MAP) kinase cascade. The UPR is required to maintain cell wall integrity; mutants lacking a functional UPR have defects in cell wall biosynthesis and are hypersensitive ...

  10. Homogenization of a system of elastic and reaction-diffusion equations modelling plant cell wall biomechanics

    OpenAIRE

    Ptashnyk, Mariya; Seguin, Brian

    2014-01-01

    In this paper we present a derivation and multiscale analysis of a mathematical model for plant cell wall biomechanics that takes into account both the microscopic structure of a cell wall coming from the cellulose microfibrils and the chemical reactions between the cell wall's constituents. Particular attention is paid to the role of pectin and the impact of calcium-pectin cross-linking chemistry on the mechanical properties of the cell wall. We prove the existence and uniqueness of the stro...

  11. Plectasin, a Fungal Defensin, Targets the Bacterial Cell Wall Precursor Lipid II

    DEFF Research Database (Denmark)

    Schneider, Tanja; Kruse, Thomas; Wimmer, Reinhard;

    2010-01-01

    plectasin, a fungal defensin, acts by directly binding the bacterial cell-wall precursor Lipid II. A wide range of genetic and biochemical approaches identify cell-wall biosynthesis as the pathway targeted by plectasin. In vitro assays for cell-wall synthesis identified Lipid II as the specific cellular...

  12. Staphylococcus aureus Cell Wall Stress Stimulon Gene-lacZ Fusion Strains: Potential for Use in Screening for Cell Wall-Active Antimicrobials▿

    OpenAIRE

    Steidl, Rebecca; Pearson, Stacy; Stephenson, Robert E.; Ledala, Nagender; Sitthisak, Sutthirat; Wilkinson, Brian J; Jayaswal, Radheshyam K.

    2008-01-01

    lacZ fusion strains were constructed using the promoters of five cell wall stress stimulon genes: pbp2, tcaA, vraSR, sgtB, and lytR. All fusion strains were induced only in the presence of cell wall-active antibiotics, suggesting the potential of these strains for use in high-throughput screening for new cell wall-active agents.

  13. The Unfolded Protein Response Is Induced by the Cell Wall Integrity Mitogen-activated Protein Kinase Signaling Cascade and Is Required for Cell Wall Integrity in Saccharomyces cerevisiae

    OpenAIRE

    Scrimale, Thomas; DiDone, Louis; de Mesy Bentley, Karen L.; Krysan, Damian J.

    2009-01-01

    The yeast cell wall is an extracellular structure that is dependent on secretory and membrane proteins for its construction. We investigated the role of protein quality control mechanisms in cell wall integrity and found that the unfolded protein response (UPR) and, to a lesser extent, endoplasmic reticulum (ER)-associated degradation (ERAD) pathways are required for proper cell wall construction. Null mutation of IRE1, double mutation of ERAD components (hrd1Δ and ubc7Δ) and ire1Δ, or expres...

  14. Effect of Wall Charge on Striation in Plasma Display Cells

    Institute of Scientific and Technical Information of China (English)

    HE Feng; OUYANG Jiting; CAO Jing; FENG Shuo; MIAO Jinsong; WANG Jianqi

    2007-01-01

    Different configurations and driving voltages have been employed to investigate the effect of the wall charge on the striations in macroscopic plasma display panel (PDP) cells.The experimental results show that a discharge channel near the dielectric layer is indispensable to striation occurring in the anode area during a discharge,while the pre-accumulated charge on the dielectric layer and the surface state are not important.The origin of the striation is related only to the physical process in the cell.The dielectric layer acts as a charge collector during a PDP discharge.

  15. Stress analysis for wall structure in mobile hot cell design

    Energy Technology Data Exchange (ETDEWEB)

    Bahrin, Muhammad Hannan, E-mail: hannan@nuclearmalaysia.gov.my; Rahman, Anwar Abdul, E-mail: anwar@nuclearmalaysia.gov.my; Hamzah, Mohd Arif, E-mail: arif@nuclearmalaysia.gov.my; Mamat, Mohd Rizal; Azman, Azraf; Hasan, Hasni [Prototype and Plant Development Centre, Technical Services Division, Malaysian Nuclear Agency (Malaysia)

    2016-01-22

    Malaysian Nuclear Agency is developing a Mobile Hot Cell (MHC) in order to handle and manage Spent High Activity Radioactive Sources (SHARS) such as teletherapy heads and irradiators. At present, there are only two units of MHC in the world, in South Africa and China. Malaysian Mobile Hot cell is developed by Malaysian Nuclear Agency with the assistance of IAEA expert, based on the design of South Africa and China, but with improved features. Stress analysis has been performed on the design in order to fulfil the safety requirement in operation of MHC. This paper discusses the loading analysis effect from the sand to the MHC wall structure.

  16. A Model for Cell Wall Dissolution in Mating Yeast Cells: Polarized Secretion and Restricted Diffusion of Cell Wall Remodeling Enzymes Induces Local Dissolution

    OpenAIRE

    Huberman, Lori B.; Murray, Andrew W.

    2014-01-01

    Mating of the budding yeast, Saccharomyces cerevisiae, occurs when two haploid cells of opposite mating types signal using reciprocal pheromones and receptors, grow towards each other, and fuse to form a single diploid cell. To fuse, both cells dissolve their cell walls at the point of contact. This event must be carefully controlled because the osmotic pressure differential between the cytoplasm and extracellular environment causes cells with unprotected plasma membranes to lyse. If the cell...

  17. Pectic homogalacturonan masks abundant sets of xyloglucan epitopes in plant cell walls

    DEFF Research Database (Denmark)

    Marcus, Susan E; Verhertbruggen, Yves; Hervé, Cécile;

    2008-01-01

    BACKGROUND: Molecular probes are required to detect cell wall polymers in-situ to aid understanding of their cell biology and several studies have shown that cell wall epitopes have restricted occurrences across sections of plant organs indicating that cell wall structure is highly developmentally...... regulated. Xyloglucan is the major hemicellulose or cross-linking glycan of the primary cell walls of dicotyledons although little is known of its occurrence or functions in relation to cell development and cell wall microstructure. RESULTS: Using a neoglycoprotein approach, in which a XXXG heptasaccharide...... hapten inhibition of binding studies. The use of LM15 for the analysis of xyloglucan in the cell walls of tamarind and nasturtium seeds, in which xyloglucan occurs as a storage polysaccharide, indicated that the LM15 xyloglucan epitope occurs throughout the thickened cell walls of the tamarind seed and...

  18. Expansins are among plant cell wall modifying agents specifically expressed during development of nematode-induced syncytia

    OpenAIRE

    Fudali, Sylwia; Sobczak, Miroslaw; Janakowski, Slawomir; Griesser, Michaela; Grundler, Florian MW; Golinowski, Wladyslaw

    2008-01-01

    Cyst nematodes are economically important pests. As obligatory biotrophic endoparasites they invade host roots and induce formation of syncytia, structures that serve them as the only source of nutrients. During syncytium development, extensive cell wall modifications take place. Cell wall dissolution occurs during cell wall opening formation, cell walls expand during hypertrophy of syncytial elements and local cell wall synthesis leads to the thickening of syncytial cell wall and the formati...

  19. Impact of processing on the noncovalent interactions between procyanidin and apple cell wall.

    Science.gov (United States)

    Le Bourvellec, Carine; Watrelot, Aude A; Ginies, Christian; Imberty, Anne; Renard, Catherine M G C

    2012-09-19

    Procyanidins can bind cell wall material in raw product, and it could be supposed that the same mechanism of retention of procyanidins by apple cell walls takes place in cooked products. To evaluate the influence of cell wall composition and disassembly during cooking on the cell walls' capacity to interact with procyanidins, four cell wall materials differing in their protein contents and physical characteristics were prepared: cell wall with proteins, cell wall devoid of protein, and two processed cell walls differing by their drying method. Protein contents varied from 23 to 99 mg/g and surface areas from 1.26 to 3.16 m(2)/g. Apple procyanidins with an average polymerization degree of 8.7 were used. The adsorption of apple procyanidins on solid cell wall material was quantified using the Langmuir isotherm formulation. The protein contents in cell wall material had no effect on procyanidin/cell wall interactions, whereas modification of the cell wall material by boiling, which reduces pectin content, and drying decreased the apparent affinity and increased the apparent saturation levels when constants were expressed relative to cell wall weight. However, boiling and drying increased apparent saturation levels and had no effect on apparent affinity when the same data were expressed per surface units. Isothermal titration calorimetry indicated strong affinity (K(a) = 1.4 × 10(4) M(-1)) between pectins solubilized by boiling and procyanidins. This study higllights the impact of highly methylated pectins and drying, that is, composition and structure of cell wall in the cell wall/procyanidin interactions. PMID:22861056

  20. Seed coat mucilage cells of Arabidopsis thaliana as a model for plant cell wall research.

    Science.gov (United States)

    Arsovski, Andrej A; Haughn, George W; Western, Tamara L

    2010-07-01

    Plant cells are encased within a complex polysaccharide wall that strengthens the cell and has key roles in all aspects of plant cell growth, differentiation, and interaction with the environment. This dynamic structure is under continual modification during plant development, and its synthesis and modification require the activity of a myriad of enzymes. The mucilage secretory cells (MSCs) of the Arabidopsis thaliana seed coat provide a model for the discovery of novel genes involved in the synthesis, secretion and modification of cell wall components, particularly pectin. These cells synthesize copious amounts of pectinaceous mucilage during development and, upon hydration of the desiccated seed, the mucilage rapidly swells, bursts from the MSCs and surrounds the seed in a gelatinous capsule. Several genes affecting MSC differentiation, pectin synthesis, and mucilage release have been identified and additional genes involved in these and related processes including pectin secretion and the mechanical alteration of cell walls await to be discovered. PMID:20505351

  1. Cell wall pH and auxin transport velocity

    Science.gov (United States)

    Hasenstein, K. H.; Rayle, D.

    1984-01-01

    According to the chemiosmotic polar diffusion hypothesis, auxin pulse velocity and basal secretion should increase with decreasing cell wall pH. Experiments were designed to test this prediction. Avena coleoptile sections were preincubated in either fusicoccin (FC), cycloheximide, pH 4.0, or pH 8.0 buffer and subsequently their polar transport capacities were determined. Relative to controls, FC enhanced auxin (IAA) uptake while CHI and pH 8.0 buffer reduced IAA uptake. Nevertheless, FC reduced IAA pulse velocity while cycloheximide increased velocity. Additional experiments showed that delivery of auxin to receivers is enhanced by increased receiver pH. This phenomenon was overcome by a pretreatment of the tissue with IAA. Our data suggest that while acidic wall pH values facilitate cellular IAA uptake, they do not enhance pulse velocity or basal secretion. These findings are inconsistent with the chemiosmotic hypothesis for auxin transport.

  2. Scattering properties of microalgae: the effect of cell size and cell wall

    Science.gov (United States)

    Svensen, Øyvind; Frette, Øyvind; Rune Erga, Svein

    2007-08-01

    The main objective of this work was to investigate how the cell size and the presence of a cell wall influence the scattering properties of the green microalgae Chlamydomonas reinhardtii. The growth cycle of two strains, one with a cell wall and one without, was synchronized to be in the same growth phase. Measurements were conducted at two different phases of the growth cycle on both strains of the algae. It was found that the shape of the scattering phase function was very similar for both strains at both growth phases, but the regular strain with a cell wall scatters more strongly than the wall-less mutant. It was also found that the mutant strain has a stronger increase in scattering than the regular strain, as the algae grow, and that the scattering from the regular strain is more wavelength dependent than from the mutant strain.

  3. Immunotherapy with BCG cell wall plus irradiated tumor cells

    International Nuclear Information System (INIS)

    Two different fibrosarcomas (MCB-I, MCB-II) were induced by methylcholcholanthrene in syngeneic Balb/C mice were used. The tumor cells irradiated with 5,000 to 30,000 rads did not growth in mice on 30 days after inoculation. The viable tumor cells were challenged intradermally to mice on 7 days after inoculation of the tumor cells irradiated with 5,000 to 30,000 rads. The challenged tumor cells were all rejected at 30 days after inoculation. Mice were challenged with 5 x 105 viable tumor cells on 7 days after inoculation of 103 to 108 irradiated tumor cells. Mice pretreated with 105 or 106 irradiated tumor cells rejected the tumor cells completely. The viable tumor cells were challenged to mice on 7 days after inoculation of BCG-CW emulsion plus 106 irradiated tumor cells. 0, 50, 100, 200, and 400 mu g of BCG-CW emulsion were mixed in 106 irradiated tumor cells. Optimal dosage of BCG-CW emulsion was 50 or 100 mu g. BCG-CW emulsion plus irradiated tumor cells were injected subcutaneously to the mice after tumor cells inoculation. Three injections of the vaccine significantly suppressed the tumor outgrowth, but not one or two injections in no-treated mice. However, in the mice pretreated with BCG-CW emulsion, the tumor growth was significantly suppressed by one or two injections of the vaccine. Especially, the three injections of the vaccine significantly suppressed the tumor growth and the 25% of the mice were completely cured. The effect of the vaccine was almost the same grade by contralateral or ipsilateral treatment. The irradiated MCB-II tumor cells plus BCG-CW emulsion were not effective to the MCB-1 tumor bearing mice, suggesting the anti-tumor effect of this vaccine was immunologically specific

  4. Immunotherapy with BCG cell wall plus irradiated tumor cells

    Energy Technology Data Exchange (ETDEWEB)

    Mizukuro, Tomoyuki (Kyoto Prefectural Univ. of Medicine (Japan))

    1983-04-01

    Two different fibrosarcomas (MCB-I, MCB-II) were induced by methylcholcholanthrene in syngeneic Balb/C mice were used. The tumor cells irradiated with 5,000 to 30,000 rads did not growth in mice on 30 days after inoculation. The viable tumor cells were challenged intradermally to mice on 7 days after inoculation of the tumor cells irradiated with 5,000 to 30,000 rads. The challenged tumor cells were all rejected at 30 days after inoculation. Mice were challenged with 5 x 10/sup 5/ viable tumor cells on 7 days after inoculation of 10/sup 3/ to 10/sup 8/ irradiated tumor cells. Mice pretreated with 10/sup 5/ or 10/sup 6/ irradiated tumor cells rejected the tumor cells completely. The viable tumor cells were challenged to mice on 7 days after inoculation of BCG-CW emulsion plus 10/sup 6/ irradiated tumor cells. 0, 50, 100, 200, and 400 mu g of BCG-CW emulsion were mixed in 10/sup 6/ irradiated tumor cells. Optimal dosage of BCG-CW emulsion was 50 or 100 mu g. BCG-CW emulsion plus irradiated tumor cells were injected subcutaneously to the mice after tumor cells inoculation. Three injections of the vaccine significantly suppressed the tumor outgrowth, but not one or two injections in no-treated mice. However, in the mice pretreated with BCG-CW emulsion, the tumor growth was significantly suppressed by one or two injections of the vaccine. Especially, the three injections of the vaccine significantly suppressed the tumor growth and the 25% of the mice were completely cured. The effect of the vaccine was almost the same grade by contralateral or ipsilateral treatment. The irradiated MCB-II tumor cells plus BCG-CW emulsion were not effective to the MCB-1 tumor bearing mice, suggesting the anti-tumor effect of this vaccine was immunologically specific.

  5. Penium margaritaceum: A Unicellular Model Organism for Studying Plant Cell Wall Architecture and Dynamics

    OpenAIRE

    Domozych, David S

    2014-01-01

    Penium margaritaceum is a new and valuable unicellular model organism for studying plant cell wall structure and developmental dynamics. This charophyte has a cell wall composition remarkably similar to the primary cell wall of many higher plants and clearly-defined inclusive zones containing specific polymers. Penium has a simple cylindrical phenotype with a distinct region of focused wall synthesis. Specific polymers, particularly pectins, can be identified using monoclonal antibodies rais...

  6. [Hydroxyproline: Rich glycoproteins of the plant and cell wall

    Energy Technology Data Exchange (ETDEWEB)

    Varner, J.E.

    1993-01-01

    Since xylem tissue includes the main cell types which are lignified, we are interested in gene expression of glycine-rich proteins and proline-rich proteins, and other proteins which are involved in secondary cell wall thickening during xylogenesis. Since the main feature of xylogenesis is the deposition of additional wall components, study of the mechanism of xylogenesis will greatly advance our knowledge of the synthesis and assembly of wall macromolecules. We are using the in vitro xylogenesis system from isolated Zinnia mesophyll cells to isolate genes which are specifically expressed during xylogenesis. We have used subtractive hybridization methods to isolate a number of cDNA clones for differentially regulated genes from the cells after hormonal induction. So far, we have partially characterized 18 different cDNA clones from 239 positive clones. These differentially regulated genes can be divided into three sets according to the characteristics of gene expression in the induction medium and the control medium. The first set is induced in both the induction medium and the control medium without hormones. The second set is induced mainly in the induction medium and in the control medium with the addition of NAA alone. Two of thesegenes are exclusively induced by auxin. The third set of genes is induced mainly in the induction medium. Since these genes are not induced by either auxin or cytokinin alone, they may be directly involved in the process of xylogenesis. Our experiments on the localization of H[sub 2]O[sub 2] production reinforce the earlier ideas of others that H[sub 2]O[sub 2] is involved in normal lignification.

  7. Induction kinetics of the Staphylococcus aureus cell wall stress stimulon in response to different cell wall active antibiotics

    Directory of Open Access Journals (Sweden)

    Berger-Bächi Brigitte

    2011-01-01

    Full Text Available Abstract Background Staphylococcus aureus activates a protective cell wall stress stimulon (CWSS in response to the inhibition of cell wall synthesis or cell envelope damage caused by several structurally and functionally different antibiotics. CWSS induction is coordinated by the VraSR two-component system, which senses an unknown signal triggered by diverse cell wall active agents. Results We have constructed a highly sensitive luciferase reporter gene system, using the promoter of sas016 (S. aureus N315, which detects very subtle differences in expression as well as measuring > 4 log-fold changes in CWSS activity, to compare the concentration dependence of CWSS induction kinetics of antibiotics with different cell envelope targets. We compared the effects of subinhibitory up to suprainhibitory concentrations of fosfomycin, D-cycloserine, tunicamycin, bacitracin, flavomycin, vancomycin, teicoplanin, oxacillin, lysostaphin and daptomycin. Induction kinetics were both strongly antibiotic- and concentration-dependent. Most antibiotics triggered an immediate response with induction beginning within 10 min, except for tunicamycin, D-cycloserine and fosfomycin which showed lags of up to one generation before induction commenced. Induction characteristics, such as the rate of CWSS induction once initiated and maximal induction reached, were strongly antibiotic dependent. We observed a clear correlation between the inhibitory effects of specific antibiotic concentrations on growth and corresponding increases in CWSS induction kinetics. Inactivation of VraR increased susceptibility to the antibiotics tested from 2- to 16-fold, with the exceptions of oxacillin and D-cycloserine, where no differences were detected in the methicillin susceptible S. aureus strain background analysed. There was no apparent correlation between the induction capacity of the various antibiotics and the relative importance of the CWSS for the corresponding resistance phenotypes

  8. Chromatin and Cell Wall Staining of Schizosaccharomyces pombe.

    Science.gov (United States)

    Hagan, Iain M

    2016-01-01

    Fission yeasts grow by tip extension, maintaining a constant width until they reach a critical size threshold and divide. Division by medial fission-which gives these yeast their name-generates a new end that arises from the site of cytokinesis. The old end, which was produced during the previous cell cycle, initiates progression of the new cell cycle, and in G2, the new end is activated in a process termed new-end takeoff (NETO). In this protocol, the fluorescent stains calcofluor and 4',6-diamidino-2-phenylindole (DAPI) are used to give a rapid and informative assessment of morphogenesis and cell-cycle progression in the fission yeast Schizosaccharomyces pombe Calcofluor reveals the timing of NETO because it stains the birth scars that are generated at new ends by cytokinesis less efficiently than the rest of the cell wall. Intense calcofluor staining of the septum and measurement of cell length are also widely used to identify dividing cells and to gauge the timing of mitotic commitment. Staining nuclei with DAPI identifies mono- and binucleated cells and complements the calcofluor staining procedure to evaluate the stages of the cell cycle and identify mitotic errors. Equally simple DAPI staining procedures reveal chromatin structure in higher resolution, facilitating more accurate staging of mitotic progression and characterization of mitotic errors. PMID:27250942

  9. Profiling the Hydrolysis of Isolated Grape Berry Skin Cell Walls by Purified Enzymes.

    Science.gov (United States)

    Zietsman, Anscha J J; Moore, John P; Fangel, Jonatan U; Willats, William G T; Vivier, Melané A

    2015-09-23

    The unraveling of crushed grapes by maceration enzymes during winemaking is difficult to study because of the complex and rather undefined nature of both the substrate and the enzyme preparations. In this study we simplified both the substrate, by using isolated grape skin cell walls, and the enzyme preparations, by using purified enzymes in buffered conditions, to carefully follow the impact of the individual and combined enzymes on the grape skin cell walls. By using cell wall profiling techniques we could monitor the compositional changes in the grape cell wall polymers due to enzyme activity. Extensive enzymatic hydrolysis, achieved with a preparation of pectinases or pectinases combined with cellulase or hemicellulase enzymes, completely removed or drastically reduced levels of pectin polymers, whereas less extensive hydrolysis only opened up the cell wall structure and allowed extraction of polymers from within the cell wall layers. Synergistic enzyme activity was detectable as well as indications of specific cell wall polymer associations. PMID:26309153

  10. Auxin-induced modifications of cell wall polysaccharides in cat coleoptile segments. Effect of galactose

    International Nuclear Information System (INIS)

    Galactose inhibits auxin-induced cell elongation in oat coleoptile segments. Cell elongation induced by exogenously applied auxin is controlled by factors such as auxin uptake, cell wall loosening, osmotic concentration of sap and hydraulic conductivity. However, galactose does not have any effect on these factors. The results discussed in this paper led to the conclusion that galactose does not affect cell wall loosening which controls rapid growth, but inhibits cell wall synthesis which is required to maintain long-term growth

  11. Members of the Hsp70 family of proteins in the cell wall of Saccharomyces cerevisiae.

    OpenAIRE

    López-Ribot, J L; Chaffin, W L

    1996-01-01

    Western blot (immunoblot) analysis of cell wall and cytosolic extracts obtained from parental and ssa1 and ssa2 single- and double-mutant strains of Saccharomyces cerevisiae showed that the heat shock protein 70 (Hsp70) products of these genes, previously thought to be restricted to the cell interior, are also present in the cell wall. A cell wall location was further confirmed by indirect immunofluorescence with intact cells and biotinylation of extracellular Hsp70. Hsp70s have been implicat...

  12. Cellulose-hemicellulose interaction in wood secondary cell-wall

    Science.gov (United States)

    Zhang, Ning; Li, Shi; Xiong, Liming; Hong, Yu; Chen, Youping

    2015-12-01

    The wood cell wall features a tough and relatively rigid fiber reinforced composite structure. It acts as a pressure vessel, offering protection against mechanical stress. Cellulose microfibrils, hemicellulose and amorphous lignin are the three major components of wood. The structure of secondary cell wall could be imagined as the same as reinforced concrete, in which cellulose microfibrils acts as reinforcing steel bar and hemicellulose-lignin matrices act as the concrete. Therefore, the interface between cellulose and hemicellulose/lignin plays a significant role in determine the mechanical behavior of wood secondary cell wall. To this end, we present a molecular dynamics (MD) simulation study attempting to quantify the strength of the interface between cellulose microfibrils and hemicellulose. Since hemicellulose binds with adjacent cellulose microfibrils in various patterns, the atomistic models of hemicellulose-cellulose composites with three typical binding modes, i.e. bridge, loop and random binding modes are constructed. The effect of the shape of hemicellulose chain on the strength of hemicellulose-cellulose composites under shear loadings is investigated. The contact area as well as hydrogen bonds between cellulose and hemicellulose, together with the covalent bonds in backbone of hemicellulose chain are found to be the controlling parameters which determine the strength of the interfaces in the composite system. For the bridge binding model, the effect of shear loading direction on the strength of the cellulose material is also studied. The obtained results suggest that the shear strength of wood-inspired engineering composites can be optimized through maximizing the formations of the contributing hydrogen bonds between cellulose and hemicellulose.

  13. Nutrient depletion modifies cell wall adsorption activity of wine yeast.

    Science.gov (United States)

    Sidari, R; Caridi, A

    2016-06-01

    Yeast cell wall is a structure that helps yeasts to manage and respond to many environmental stresses. The mannosylphosphorylation is a modification in response to stress that provides the cell wall with negative charges able to bind compounds present in the environment. Phenotypes related to the cell wall modification such as the filamentous growth in Saccharomyces cerevisiae are affected by nutrient depletion. The present work aimed at describing the effect of carbon and/or nitrogen limitation on the aptitude of S. cerevisiae strains to bind coloured polyphenols. Carbon- and nitrogen-rich or deficient media supplemented with grape polyphenols were used to simulate different grape juice conditions-early, mid, 'adjusted' for nitrogen, and late fermentations. In early fermentation condition, the R+G+B values range from 106 (high adsorption, strain Sc1128) to 192 (low adsorption, strain Σ1278b), in mid-fermentation the values range from 111 (high adsorption, strain Sc1321) to 258 (low adsorption, strain Sc2306), in 'adjusted' for nitrogen conditions the values range from 105 (high adsorption, strain Sc1321) to 194 (low adsorption, strain Sc2306) while in late fermentation conditions the values range from 101 (high adsorption, strain Sc384) to 293 (low adsorption, strain Sc2306). The effect of nutrient availability is not univocal for all the strains and the different media tested modified the strains behaviour. In all the media the strains show significant differences. Results demonstrate that wine yeasts decrease/increase their parietal adsorption activity according to the nutrient availability. The wide range of strain variability observed could be useful in selecting wine starters. PMID:27116955

  14. A radioimmunoassay for lignin in plant cell walls

    International Nuclear Information System (INIS)

    Lignin detection and determination in herbaceous tissue requires selective, specific assays which are not currently available. A radioimmunoassay (RIA) was developed to study lignin metabolism in these tissues. A β-aryl ether lignin model compound was synthesized, linked to keyhole limpet hemocyanin using a water-soluble carbodiimide, and injected into rabbits. The highest titer of the antiserum obtained was 34 ηg/mL of model derivatized BSA. An in vitro system was developed to characterize the RIA. The model compound was linked to amino activated polyacrylamide beads to mimic lignin in the cell walls. 125I Radiolabelled protein A was used to detect IgG antibody binding. The RIA was shown in the in vitro system to exhibit saturable binding. The amount of antibody bound decreased when the serum was diluted. Immunoelectrophoresis and competitive binding experiments confirmed that both aromatic rings of the lignin model compound had been antigenic. Chlorogenic acid, a phenolic known to be present in plant cells, did not compete for antibody binding. The RIA was used to measure lignin in milled plant samples and barley seedlings. Antiserum binding to wheat cell walls and stressed barley segments was higher than preimmune serum binding. Antibody binding to stressed barley tissue decreased following NaClO2 delignification. The RIA was found to be less sensitive than expected, so several avenues for improving the method are discussed

  15. Lignification in poplar tension wood lignified cell wall layers.

    Science.gov (United States)

    Yoshinaga, Arata; Kusumoto, Hiroshi; Laurans, Françoise; Pilate, Gilles; Takabe, Keiji

    2012-09-01

    The lignification process in poplar tension wood lignified cell wall layers, specifically the S(1) and S(2) layers and the compound middle lamella (CML), was analysed using ultraviolet (UV) and transmission electron microscopy (TEM). Variations in the thickness of the gelatinous layer (G-layer) were also measured to clarify whether the lignified cell wall layers had completed their lignification before the deposition of G-layers, or, on the contrary, if lignification of these layers was still active during G-layer formation. Observations using UV microscopy and TEM indicated that both UV absorbance and the degree of potassium permanganate staining increased in the CML and S(1) and S(2) layers during G-layer formation, suggesting that the lignification of these lignified layers is still in progress during G-layer formation. In the context of the cell-autonomous monolignol synthesis hypothesis, our observations suggest that monolignols must go through the developing G-layer during the lignification of CML and the S(1) and S(2) layers. The alternative hypothesis of external synthesis (in the rays) does not require that monolignols go through the G-layer before being deposited in the CML, or the S(1) and S(2) layers. Interestingly, the previous observation of lignin in the poplar G-layer was not confirmed with the microscopy techniques used in the present study. PMID:22933655

  16. Absence of correlation between rates of cell wall turnover and autolysis shown by Bacillus subtilis mutants.

    OpenAIRE

    Vitković, L; Cheung, H. Y.; Freese, E

    1984-01-01

    Bacillus subtilis mutants with reduced rates of cell wall autolysis reached a constant rate of wall turnover after a longer lag than the standard strain but eventually showed the same turnover rate. In reverse, a turnover-deficient mutant autolysed at a slightly higher rate than the standard strain. Consequently, there is no correlation between the rates of cell wall turnover and autolysis.

  17. Cell wall loosening proteins of the stigma exudate

    OpenAIRE

    Nieuwland, J.H.

    2004-01-01

    Outline of the thesis: The idea, formulated by Cosgrove, that cell wall loosening of the maternal tissue facilitates pollen tube growth is a central theme of this thesis. This idea was originally proposed for beta-expansins released by maize pollen. Since the pollen coat of dry stigma type plants, like maize, bears a functional similarity with the exudate of wet stigma type plants (e.g. tobacco and petunia), this research was started with the analysis of the putative function of the pistil-sp...

  18. Clinical analysis of lateral oropharyngeal-wall squamous cell carcinoma

    International Nuclear Information System (INIS)

    We retrospectively reviewed 98 cases of lateral-oropharyngeal wall squamous cell carcinoma seen from January 1999 to March 2011. The majority-75 cases-involeved advanced cancer. For these, we conducted concurrent chemoradiotherapy (CCRT) with cisplatin, docetaxel, and 5-FU from 2007. Five-year overall survival was 64.4%. In advanced cases, three-year overall survival was 77.8% in surgery, 71.2% in radiation therapy, and 84.6% in CCRT. While no statistically significant difference was seen, CCRT, appeared to provide more curative effectiveness. (author)

  19. Mass spectrometry for characterizing plant cell wall polysaccharides

    Directory of Open Access Journals (Sweden)

    Stefan eBauer

    2012-03-01

    Full Text Available Mass spectrometry is a selective and powerful technique to obtain identification and structural information on compounds present in complex mixtures. Since it requires only small sample amount it is an excellent tool for researchers interested in detecting changes in composition of complex carbohydrates of plants. This mini-review gives an overview of common mass spectrometry techniques applied to the analysis of plant cell wall carbohydrates. It presents examples in which mass spectrometry has been used to elucidate the structure of oligosaccharides derived from hemicelluloses and pectins and illustrates how information on sequence, linkages, branching and modifications are obtained from characteristic fragmentation patterns.

  20. The Mechanisms of Plant Cell Wall Deconstruction during Enzymatic Hydrolysis

    DEFF Research Database (Denmark)

    Thygesen, Lisbeth Garbrecht; E. Thybring, Emil; Johansen, Katja Salomon;

    2014-01-01

    Mechanical agitation during enzymatic hydrolysis of insoluble plant biomass at high dry matter contents is indispensable for the initial liquefaction step in biorefining. It is known that particle size reduction is an important part of liquefaction, but the mechanisms involved are poorly understood....... Here we put forward a simple model based on mechanical principles capable of capturing the result of the interaction between mechanical forces and cell wall weakening via hydrolysis of glucosidic bonds. This study illustrates that basic material science insights are relevant also within biochemistry...

  1. Chitosan Obtained from Cell Wall of Aspergillus Niger Mycelium

    Institute of Scientific and Technical Information of China (English)

    HUANG Hui-li; LIN Wen-luan; LIN Jian-ming

    2004-01-01

    Chitin from cell walls of Aspergillus Niger mycelium was prepared. A new method for the preparation of high deacetylation degree chitosan was studied in a dilute sodium hydroxide solution at a high pressure. The experimental results indicate that the deacetylation degree of the chitosan can reach 80% under the condition of a 5.00 mol/L sodium hydroxide solution at 0.1 MPa of pressure for 1 h. This method shows the advantages of the applications in the industry production and environment protection.

  2. Cellulose synthesis inhibition, cell expansion, and patterns of cell wall deposition in Nitella internodes

    International Nuclear Information System (INIS)

    The authors have investigated the pattern of wall deposition and maturation and correlated it with cell expansion and cellulose biosynthesis. The herbicide 2,6-dichlorobenzonitrile (DCB) was found to be a potent inhibitor of cellulose synthesis, but not of cell expansion in Nitella internodal cells. Although cellulose synthesis is inhibited during DCB treatment, matrix substances continue to be synthesized and deposited. The inhibition of cellulose microfibril deposition can be demonstrated by various techniques. These results demonstrate that matrix deposition is by apposition, not by intussusception, and that the previously deposited wall moves progressively outward while stretching and thinning as a result of cell expansion

  3. Determination of the pore size of cell walls of living plant cells

    Energy Technology Data Exchange (ETDEWEB)

    Carpita, N.; Sabularse, D.; Montezinos, D.; Delmer, D.P.

    1979-09-14

    The limiting diameter of pores in the walls of living plant cells through which molecules can freely pass has been determined by a solute exclusion technique to be 35 to 38 angstroms for hair cells of Raphanus sativus roots and fibers of Gossypium hirsutum, 38 to 40 angstroms for cultured cells of Acer pseudoplatanus, and 45 to 52 angstroms for isolated palisade parenchyma cells of the leaves of Xanthium strumarium and Commelina communis. These results indicate that molecules with diameters larger than these pores would be restricted in their ability to penetrate such a cell wall, and that such a wall may represent a more significant barrier to cellular communication than has been previously assumed.

  4. Assembly and activation of alternative complement components on endothelial cell-anchored ultra-large von Willebrand factor links complement and hemostasis-thrombosis.

    Directory of Open Access Journals (Sweden)

    Nancy A Turner

    Full Text Available BACKGROUND: Vascular endothelial cells (ECs express and release protein components of the complement pathways, as well as secreting and anchoring ultra-large von Willebrand factor (ULVWF multimers in long string-like structures that initiate platelet adhesion during hemostasis and thrombosis. The alternative complement pathway (AP is an important non-antibody-requiring host defense system. Thrombotic microangiopathies can be associated with defective regulation of the AP (atypical hemolytic-uremic syndrome or with inadequate cleavage by ADAMTS-13 of ULVWF multimeric strings secreted by/anchored to ECs (thrombotic thrombocytopenic purpura. Our goal was to determine if EC-anchored ULVWF strings caused the assembly and activation of AP components, thereby linking two essential defense mechanisms. METHODOLOGY/PRINCIPAL FINDINGS: We quantified gene expression of these complement components in cultured human umbilical vein endothelial cells (HUVECs by real-time PCR: C3 and C5; complement factor (CF B, CFD, CFP, CFH and CFI of the AP; and C4 of the classical and lectin (but not alternative complement pathways. We used fluorescent microscopy, monospecific antibodies against complement components, fluorescent secondary antibodies, and the analysis of >150 images to quantify the attachment of HUVEC-released complement proteins to ULVWF strings secreted by, and anchored to, the HUVECs (under conditions of ADAMTS-13 inhibition. We found that HUVEC-released C4 did not attach to ULVWF strings, ruling out activation of the classical and lectin pathways by the strings. In contrast, C3, FB, FD, FP and C5, FH and FI attached to ULVWF strings in quantitative patterns consistent with assembly of the AP components into active complexes. This was verified when non-functional FB blocked the formation of AP C3 convertase complexes (C3bBb on ULVWF strings. CONCLUSIONS/SIGNIFICANCE: AP components are assembled and activated on EC-secreted/anchored ULVWF multimeric

  5. Type II PKAs are anchored to mature insulin secretory granules in INS-1 β-cells and required for cAMP-dependent potentiation of exocytosis.

    Science.gov (United States)

    Villalpando, Sabrina; Cazevieille, Chantal; Fernandez, Anne; Lamb, Ned J; Hani, El-Habib

    2016-06-01

    Specificity of the cAMP-dependent protein kinase (PKA) pathway relies on an extremely sophisticated compartmentalization mechanism of the kinase within a given cell, based on high-affinity binding of PKA tetramer pools to different A-Kinase Anchoring Proteins (AKAPs). We and others have previously shown that AKAPs-dependent PKA subcellular targeting is a requisite for optimal cAMP-dependent potentiation of insulin exocytosis. We thus hypothesized that a PKA pool may directly anchor to the secretory compartment to potentiate insulin exocytosis. Here, using immunofluorescence analyses combined to subcellular fractionations and purification of insulin secretory granules (ISGs), we identified discrete subpools of type II PKAs, RIIα and RIIβ PKAs, along with the catalytic subunit, physically associated with ISGs within pancreatic insulin-secreting β-cells. Ultrastructural analysis of native rodent β-cells confirmed in vivo the occurrence of PKA on dense-core ISGs. Isoform-selective disruption of binding of PKAs to AKAPs reinforced the requirement of type II PKA isoforms for cAMP potentiation of insulin exocytosis. This granular localization of PKA was of critical importance since siRNA-mediated depletion of either RIIα or RIIβ PKAs resulted in a significant reduction of cAMP-dependent potentiation of insulin release. The present work provides evidence for a previously unrecognized pool of type II PKAs physically anchored to the β-cell ISGs compartment and supports a non-redundant function for type II PKAs during cAMP potentiation of exocytosis. PMID:26898328

  6. wall

    Directory of Open Access Journals (Sweden)

    Irshad Kashif

    2016-01-01

    Full Text Available Maintaining indoor climatic conditions of buildings compatible with the occupant comfort by consuming minimum energy, especially in a tropical climate becomes a challenging problem for researchers. This paper aims to investigate this problem by evaluating the effect of different kind of Photovoltaic Trombe wall system (PV-TW on thermal comfort, energy consumption and CO2 emission. A detailed simulation model of a single room building integrated with PV-TW was modelled using TRNSYS software. Results show that 14-35% PMV index and 26-38% PPD index reduces as system shifted from SPV-TW to DGPV-TW as compared to normal buildings. Thermal comfort indexes (PMV and PPD lie in the recommended range of ASHARE for both DPV-TW and DGPV-TW except for the few months when RH%, solar radiation intensity and ambient temperature were high. Moreover PVTW system significantly reduces energy consumption and CO2 emission of the building and also 2-4.8 °C of temperature differences between indoor and outdoor climate of building was examined.

  7. The cell-wall glycoproteins of the green alga Scenedesmus obliquus. The predominant cell-wall polypeptide of Scenedesmus obliquus is related to the cell-wall glycoprotein gp3 of Chlamydomonas reinhardtii.

    Science.gov (United States)

    Voigt, Jürgen; Stolarczyk, Adam; Zych, Maria; Malec, Przemysław; Burczyk, Jan

    2014-02-01

    The green alga Scenedesmus obliquus contains a multilayered cell wall, ultrastructurally similar to that of Chlamydomonas reinhardtii, although its proportion of hydroxyproline is considerably lower. Therefore, we have investigated the polypeptide composition of the insoluble and the chaotrope-soluble wall fractions of S. obliquus. The polypeptide pattern of the chaotrope-soluble wall fraction was strongly modified by chemical deglycosylation with anhydrous hydrogen fluoride (HF) in pyridine indicating that most of these polypeptides are glycosylated. Polypeptide constituents of the chaotrope-soluble cell-wall fraction with apparent molecular masses of 240, 270, 265, and 135 kDa cross-reacted with a polyclonal antibody raised against the 100 kDa deglycosylation product of the C. reinhardtii cell-wall glycoprotein GP3B. Chemical deglycosylation of the chaotrope-soluble wall fraction resulted in a 135 kDa major polypeptide and a 106 kDa minor component reacting with the same antibody. This antibody recognized specific peptide epitopes of GP3B. When the insoluble wall fraction of S. obliquus was treated with anhydrous HF/pyridine, three polypeptides with apparent molecular masses of 144, 135, and 65 kDa were solubilized, which also occured in the deglycosylated chaotrope-soluble wall fraction. These findings indicate that theses glycoproteins are cross-linked to the insoluble wall fraction via HF-sensitive bonds. PMID:24388513

  8. Rhizobium sp. Degradation of Legume Root Hair Cell Wall at the Site of Infection Thread Origin

    OpenAIRE

    Ridge, Robert W.; Rolfe, Barry G.

    1985-01-01

    Using a new microinoculation technique, we demonstrated that penetration of Rhizobium sp. into the host root hair cell occurs at 20 to 22 h after inoculation. It did this by dissolving the cell wall maxtrix, leaving a layer of depolymerized wall microfibrils. Colony growth pressure “stretched” the weakened wall, forming a bulge into an interfacial zone between the wall and plasmalemma. At the same time vesicular bodies, similar to plasmalemmasomes, accumulated at the penetration site in a man...

  9. Transcriptomic and molecular genetic analysis of the cell wall salvage response of Aspergillus niger to the absence of galactofuranose synthesis.

    Science.gov (United States)

    Park, Joohae; Hulsman, Mark; Arentshorst, Mark; Breeman, Matthijs; Alazi, Ebru; Lagendijk, Ellen L; Rocha, Marina C; Malavazi, Iran; Nitsche, Benjamin M; van den Hondel, Cees A M J J; Meyer, Vera; Ram, Arthur F J

    2016-09-01

    The biosynthesis of cell surface-located galactofuranose (Galf)-containing glycostructures such as galactomannan, N-glycans and O-glycans in filamentous fungi is important to secure the integrity of the cell wall. UgmA encodes an UDP-galactopyranose mutase, which is essential for the formation of Galf. Consequently, the ΔugmA mutant lacks Galf-containing molecules. Our previous work in Aspergillus niger work suggested that loss of function of ugmA results in activation of the cell wall integrity (CWI) pathway which is characterized by increased expression of the agsA gene, encoding an α-glucan synthase. In this study, the transcriptional response of the ΔugmA mutant was further linked to the CWI pathway by showing the induced and constitutive phosphorylation of the CWI-MAP kinase in the ΔugmA mutant. To identify genes involved in cell wall remodelling in response to the absence of galactofuranose biosynthesis, a genome-wide expression analysis was performed using RNAseq. Over 400 genes were higher expressed in the ΔugmA mutant compared to the wild-type. These include genes that encode enzymes involved in chitin (gfaB, gnsA, chsA) and α-glucan synthesis (agsA), and in β-glucan remodelling (bgxA, gelF and dfgC), and also include several glycosylphosphatidylinositol (GPI)-anchored cell wall protein-encoding genes. In silico analysis of the 1-kb promoter regions of the up-regulated genes in the ΔugmA mutant indicated overrepresentation of genes with RlmA, MsnA, PacC and SteA-binding sites. The importance of these transcription factors for survival of the ΔugmA mutant was analysed by constructing the respective double mutants. The ΔugmA/ΔrlmA and ΔugmA/ΔmsnA double mutants showed strong synthetic growth defects, indicating the importance of these transcription factors to maintain cell wall integrity in the absence of Galf biosynthesis. PMID:27264789

  10. Hypersensitivity to Suture Anchors

    Directory of Open Access Journals (Sweden)

    Masafumi Goto

    2013-01-01

    Full Text Available Hypersensitivity to suture anchor is extremely rare. Herein, we present a case in which hypersensitivity to suture anchor was strongly suspected. The right rotator cuff of a 50-year-old woman was repaired with a metal suture anchor. Three weeks after the surgery, she developed erythema around her face, trunk, and hands, accompanied by itching. Infection was unlikely because no abnormalities were detected by blood testing or by medical examination. Suspicious of a metallic allergy, a dermatologist performed a patch testing 6 months after the first surgery. The patient had negative reactions to tests for titanium, aluminum, and vanadium, which were the principal components of the suture anchor. The anchor was removed 7 months after the first surgery, and the erythema disappeared immediately. When allergic symptoms occur and persist after the use of a metal anchor, removal should be considered as a treatment option even if the patch test result is negative.

  11. Proteomic Analysis to Identify Tightly-Bound Cell Wall Protein in Rice Calli

    OpenAIRE

    Cho, Won Kyong; Hyun, Tae Kyung; Kumar, Dhinesh; Rim, Yeonggil; Chen, Xiong Yan; Jo, Yeonhwa; Kim, Suwha; Lee, Keun Woo; Park, Zee-Yong; Lucas, William J.; Kim, Jae-Yean

    2015-01-01

    Rice is a model plant widely used for basic and applied research programs. Plant cell wall proteins play key roles in a broad range of biological processes. However, presently, knowledge on the rice cell wall proteome is rudimentary in nature. In the present study, the tightly-bound cell wall proteome of rice callus cultured cells using sequential extraction protocols was developed using mass spectrometry and bioinformatics methods, leading to the identification of 1568 candidate proteins. Ba...

  12. Seed coat mucilage cells of Arabidopsis thaliana as a model for plant cell wall research

    OpenAIRE

    Arsovski, Andrej A; Haughn, George W; Western, Tamara L.

    2010-01-01

    Plant cells are encased within a complex polysaccharide wall that strengthens the cell and has key roles in all aspects of plant cell growth, differentiation and interaction with the environment. This dynamic structure is under continual modification during plant development, and its synthesis and modification require the activity of a myriad of enzymes. The mucilage secretory cells (MSCs) of the Arabidopsis thaliana seed coat provide a model for the discovery of novel genes involved in the s...

  13. Soya beans and Maize : The effect of chemical and physical structure of cell wall polysaccharides on fermentation kinetics

    OpenAIRE

    Laar, van, J.A.

    2000-01-01

    The analysis of the relationship between cell wall composition and fermentation of endosperm cell walls of soya beans and maize was approached from three different angles. Firstly, the fermentation (rate and extent of fermentation, the sugar degradation pattern, and volatile fatty acid production) of soya bean and maize cell walls was analysed, both in situ and in vitro. This analysis revealed that the physical structure of the cell wall (particle size and cell wall thickness) influences cell...

  14. The connection of cytoskeletal network with plasma membrane and the cell wall

    Institute of Scientific and Technical Information of China (English)

    Zengyu Liu; Staffan Persson; Yi Zhang

    2015-01-01

    The cell wall provides external support of the plant cells, while the cytoskeletons including the microtubules and the actin filaments constitute an internal framework. The cytoskeletons contribute to the cell wall biosynthesis by spatially and temporarily regulating the transportation and deposition of cell wall components. This tight control is achieved by the dynamic behavior of the cytoskeletons, but also through the tethering of these structures to the plasma membrane. This tethering may also extend beyond the plasma membrane and impact on the cell wall, possibly in the form of a feedback loop. In this review, we discuss the linking components between the cytoskeletons and the plasma membrane, and/or the cell wall. We also discuss the prospective roles of these components in cell wall biosyn-thesis and modifications, and aim to provide a platform for further studies in this field.

  15. A temperature-sensitive allele of a putative mRNA splicing helicase down-regulates many cell wall genes and causes radial swelling in Arabidopsis thaliana.

    Science.gov (United States)

    Howles, Paul A; Gebbie, Leigh K; Collings, David A; Varsani, Arvind; Broad, Ronan C; Ohms, Stephen; Birch, Rosemary J; Cork, Ann H; Arioli, Tony; Williamson, Richard E

    2016-05-01

    The putative RNA helicase encoded by the Arabidopsis gene At1g32490 is a homolog of the yeast splicing RNA helicases Prp2 and Prp22. We isolated a temperature-sensitive allele (rsw12) of the gene in a screen for root radial swelling mutants. Plants containing this allele grown at the restrictive temperature showed weak radial swelling, were stunted with reduced root elongation, and contained reduced levels of cellulose. The role of the protein was further explored by microarray analysis. By using both fold change cutoffs and a weighted gene coexpression network analysis (WGCNA) to investigate coexpression of genes, we found that the radial swelling phenotype was not linked to genes usually associated with primary cell wall biosynthesis. Instead, the mutation has strong effects on expression of secondary cell wall related genes. Many genes potentially associated with secondary walls were present in the most significant WGCNA module, as were genes coding for arabinogalactans and proteins with GPI anchors. The proportion of up-regulated genes that possess introns in rsw12 was above that expected if splicing was unrelated to the activity of the RNA helicase, suggesting that the helicase does indeed play a role in splicing in Arabidopsis. The phenotype may be due to a change in the expression of one or more genes coding for cell wall proteins. PMID:27008640

  16. Cell Wall Microstructure Analysis Implicates Hemicellulose Polysaccharides in Cell Adhesion in Tomato Fruit Pericarp Parenchyma

    Institute of Scientific and Technical Information of China (English)

    Jose J. Ordaz-Ortiz; Susan E. Marcus; J. Paul Knox

    2009-01-01

    Methods developed to isolate intact cells from both unripe and ripe tomato fruit pericarp parenchyma have allowed the cell biological analysis of polysaccharide epitopes at the surface of separated cells. The LM7 pectic homoga-lacturonan epitope is a marker of the junctions of adhesion planes and intercellular spaces in parenchyma systems. The LM7 epitope persistently marked the former edge of adhesion planes at the surface of cells separated from unripe and ripened tomato fruit and also from fruits with the Cnr mutation. The LM 11 xylan epitope was associated, in sections, with cell walls lining intercellular space but the epitope was not detected at the surface of isolated cells, being lost during cell isolation. The LM15 xyloglucan epitope was present at the surface of cells isolated from unripe fruit in a pattern reflecting the former edge of cell adhesion planes/intercellular space but with gaps and apparent breaks, An equivalent pattern ofLM15 epitope occurrence was revealed at the surface of cells isolated by pectate lyase action but was not present in cells isolated from ripe fruit or from Cnr fruit. In contrast to wild-type cells, the LM5 galactan and LM21 mannan epitopes oc-curred predominantly in positions reflecting intercellular space in Cnr, suggesting a concerted alteration in cell wall mi-crostructure in response to this mutation. Galactanase and mannanase, along with pectic homogalacturonan-degrading enzymes, were capable of releasing cells from unripe fruit parenchyma. These observations indicate that hemicellulose polymers are present in architectural contexts reflecting cell adhesion and that several cell wall polysaccharide classes are likely to contribute to cell adhesion/cell separation in tomato fruit pericarp parenchyma.

  17. Area Expansivity Moduli of Regenerating Plant Protoplast Cell Walls Exposed to Shear Flows

    Science.gov (United States)

    Fujimura, Yuu; Iino, Masaaki; Watanabe, Ugai

    2005-05-01

    To control the elasticity of the plant cell wall, protoplasts isolated from cultured Catharanthus roseus cells were regenerated in shear flows of 115 s-1 (high shear) and 19.2 s-1 (low shear, as a control). The surface area expansivity modulus and the surface breaking strength of these regenerating protoplasts were measured by a micropipette aspiration technique. Cell wall synthesis was also measured using a cell wall-specific fluorescent dye. High shear exposure for 3 h doubled both the surface area modulus and breaking strength observed under low shear, significantly decreased cell wall synthesis, and roughly quadrupled the moduli of the cell wall. Based on the cell wall synthesis data, we estimated the three-dimensional modulus of the cell wall to be 4.1± 1.2 GPa for the high shear, and 0.35± 0.2 GPa for the low shear condition, using the surface area expansivity modulus divided by the cell wall thickness, which is identical with the Young’s modulus divided by 2(1-σ), where σ is Poisson's ratio. We concluded that high shear exposure considerably strengthens the newly synthesized cell wall.

  18. Starting to Gel: How Arabidopsis Seed Coat Epidermal Cells Produce Specialized Secondary Cell Walls

    Directory of Open Access Journals (Sweden)

    Cătălin Voiniciuc

    2015-02-01

    Full Text Available For more than a decade, the Arabidopsis seed coat epidermis (SCE has been used as a model system to study the synthesis, secretion and modification of cell wall polysaccharides, particularly pectin. Our detailed re-evaluation of available biochemical data highlights that Arabidopsis seed mucilage is more than just pectin. Typical secondary wall polymers such as xylans and heteromannans are also present in mucilage. Despite their low abundance, these components appear to play essential roles in controlling mucilage properties, and should be further investigated. We also provide a comprehensive community resource by re-assessing the mucilage phenotypes of almost 20 mutants using the same conditions. We conduct an in-depth functional evaluation of all the SCE genes described in the literature and propose a revised model for mucilage production. Further investigation of SCE cells will improve our understanding of plant cell walls.

  19. Elucidation of the chemical fine structure of polysaccharides from soybean and maize kernel cell walls

    OpenAIRE

    Huisman, M.M.H.

    2000-01-01

    The subject of this thesis was the elucidation of the chemical fine structure of polysaccharides from cell walls of soybean and maize kernel. The two species investigated represent different taxonomic groups, soybean belonging to the dicotyledonous and maize to the monocotyledonous plants. Besides representing the most important structures present in cell wall material, these raw materials are of great importance in food and feed industry.The characterisation of the soybean cell wall polysacc...

  20. In vivo cell wall loosening by hydroxyl radicals during cress seed germination and elongation growth

    OpenAIRE

    Müller, Kerstin; Linkies, Ada; Vreeburg, Robert A. M.; Fry, Stephen C; Krieger-Liszkay, Anja; Leubner-Metzger, Gerhard

    2009-01-01

    Loosening of cell walls is an important developmental process in key stages of plant life cycles, including seed germination, elongation growth and fruit ripening. Here we report direct in vivo evidence for hydroxyl radical (•OH)-mediated cell wall loosening during plant seed germination and seedling growth. We used electron paramagnetic resonance (EPR)-spectroscopy to show that •OH is generated in the cell wall during radicle elongation and weakening of the endosperm of cress (Lepidium sativ...

  1. Cell wall synthesis and initiation of deoxyribonucleic acid replication in Bacillus subtilis.

    OpenAIRE

    Sandler, N.; Keynan, A

    1981-01-01

    We have observed a connection between cell wall synthesis and the initiation of chromosome replication in Bacillus subtilis. Initiation of chromosome replication was prevented in synchronous cultures in the presence of the cell wall synthesis inhibitor vancomycin. When vancomycin was added to the cultures after initiation of chromosome replication, one round of replication was completed but no reinitiation occurred. Similar results were obtained when cell wall synthesis was inhibited by risto...

  2. Diversity of beetle genes encoding novel plant cell wall degrading enzymes

    OpenAIRE

    Pauchet, Y.; Wilkinson, P.; Chauhan, R.; Ffrench-Constant, R.

    2010-01-01

    Plant cell walls are a heterogeneous mixture of polysaccharides and proteins that require a range of different enzymes to degrade them. Plant cell walls are also the primary source of cellulose, the most abundant and useful biopolymer on the planet. Plant cell wall degrading enzymes (PCWDEs) are therefore important in a wide range of biotechnological processes from the production of biofuels and food to waste processing. However, despite the fact that the last common ancestor of all deuterost...

  3. Ultrastructure of Fibre and Parenchyma Cell Walls During Early Stages of Culm Development in Dendrocalamus asper

    OpenAIRE

    Gritsch, Cristina Sanchis; Murphy, Richard J.

    2005-01-01

    • Background and Aims The anatomy of bamboo culms and the multilayered structure of fibre cell walls are known to be the main determinant factors for its physical and mechanical properties. Studies on the bamboo cell wall have focussed mainly on fully elongated and mature fibres. The main aim of this study was to describe the ultrastructure of primary and secondary cell walls in culm tissues of Dendrocalamus asper at different stages of development.

  4. Two cationic peroxidases from cell walls of Araucaria araucana seeds.

    Science.gov (United States)

    Riquelme, A; Cardemil, L

    1995-05-01

    We have previously reported the purification and partial characterization of two cationic peroxidases from the cell walls of seeds and seedlings of the South American conifer, Araucaria araucana. In this work, we have studied the amino acid composition and NH2-terminal sequences of both enzymes. We also compare the data obtained from these analyses with those reported for other plant peroxidases. The two peroxidases are similar in their amino acid compositions. Both are particularly rich in glycine, which comprises more than 30% of the amino acid residues. The content of serine is also high, ca 17%. The two enzymes are different in their content of arginine, alanine, valine, phenylalanine and threonine. Both peroxidases have identical NH2-terminal sequences, indicating that the two proteins are genetically related and probably are isoforms of the same kind of peroxidase. The amino acid composition and NH2-terminal sequence analyses showed marked differences from the cationic peroxidases from turnip and horseradish. PMID:7786490

  5. Single Wall Carbon Nanotube-polymer Solar Cells

    Science.gov (United States)

    Bailey, Sheila G.; Castro, Stephanie L.; Landi, Brian J.; Gennett, Thomas; Raffaelle, Ryne P.

    2005-01-01

    Investigation of single wall carbon nanotube (SWNT)-polymer solar cells has been conducted towards developing alternative lightweight, flexible devices for space power applications. Photovoltaic devices were constructed with regioregular poly(3-octylthiophene)-(P3OT) and purified, >95% w/w, laser-generated SWNTs. The P3OT composites were deposited on ITO-coated polyethylene terapthalate (PET) and I-V characterization was performed under simulated AM0 illumination. Fabricated devices for the 1.0% w/w SWNT-P3OT composites showed a photoresponse with an open-circuit voltage (V(sub oc)) of 0.98 V and a short-circuit current density (I(sub sc)) of 0.12 mA/sq cm. Optimization of carrier transport within these novel photovoltaic systems is proposed, specifically development of nanostructure-SWNT complexes to enhance exciton dissociation.

  6. Pectinous cell wall thickenings formation - A common defense strategy of plants to cope with Pb.

    Science.gov (United States)

    Krzesłowska, Magdalena; Rabęda, Irena; Basińska, Aneta; Lewandowski, Michał; Mellerowicz, Ewa J; Napieralska, Anna; Samardakiewicz, Sławomir; Woźny, Adam

    2016-07-01

    Lead, one of the most abundant and hazardous trace metals affecting living organisms, has been commonly detected in plant cell walls including some tolerant plants, mining ecotypes and hyperaccumulators. We have previously shown that in tip growing Funaria sp. protonemata cell wall is remodeled in response to lead by formation of thickenings rich in low-methylesterified pectins (pectin epitope JIM5 - JIM5-P) able to bind metal ions, which accumulate large amounts of Pb. Hence, it leads to the increase of cell wall capacity for Pb compartmentalization. Here we show that diverse plant species belonging to different phyla (Arabidopsis, hybrid aspen, star duckweed), form similar cell wall thickenings in response to Pb. These thickenings are formed in tip growing cells such as the root hairs, and in diffuse growing cells such as meristematic and root cap columella cells of root apices in hybrid aspen and Arabidopsis and in mesophyll cells in star duckweed fronds. Notably, all analyzed cell wall thickenings were abundant in JIM5-P and accumulated high amounts of Pb. In addition, the co-localization of JIM5-P and Pb commonly occurred in these cells. Hence, cell wall thickenings formed the extra compartment for Pb accumulation. In this way plant cells increased cell wall capacity for compartmentalization of this toxic metal, protecting protoplast from its toxicity. As cell wall thickenings occurred in diverse plant species and cell types differing in the type of growth we may conclude that pectinous cell wall thickenings formation is a widespread defense strategy of plants to cope with Pb. Moreover, detection of natural defense strategy, increasing plant cell walls capacity for metal accumulation, reveals a promising direction for enhancing plant efficiency in phytoremediation. PMID:27107260

  7. In-vitro fermentability of cell walls as influenced by lignin composition and cross-linking.

    Science.gov (United States)

    We assessed how diverse modifications in lignin composition and reductions in ferulate-lignin cross-linking influence the degradability of cell walls. Cell walls from nonlignified maize cell suspensions were artificially lignified with varying ratios of normal monolignols (coniferyl and sinapyl alco...

  8. Composition and architecture of the cell walls of grasses and the mechanisms of synthesis of cell wall polysaccharides. Final report for period September 1, 1988 - April 30, 2001

    Energy Technology Data Exchange (ETDEWEB)

    Carpita, Nicholas C.

    2001-10-18

    This program was devoted toward complete understanding of the polysaccharide structure and architecture of the primary cell walls grasses and cereals, and the biosynthesis of the mixed-linkage beta-glucane, a cellulose interacting polymer that is synthesized uniquely by grass species and close relatives. With these studies as focal point, the support from DOE was instrumental in the development of new analytical means that enabled us to characterize carbohydrate structure, to reveal new features of cell wall dynamics during cell growth, and to apply these techniques in other model organisms. The support by DOE in these basic studies was acknowledged on numerous occasions in review articles covering current knowledge of cell wall structure, architecture, dynamics, biosynthesis, and in all genes related to cell wall biogenesis.

  9. Principles of bacterial cell-size determination revealed by cell wall synthesis perturbations

    OpenAIRE

    Carolina Tropini; Timothy K. Lee; Jen Hsin; Samantha M. Desmarais; Tristan Ursell; Russell D. Monds; Kerwyn Casey Huang

    2014-01-01

    Although bacterial cell morphology is tightly controlled, the principles of size regulation remain elusive. In Escherichia coli, perturbation of cell-wall synthesis often results in similar morphologies, making it difficult to deconvolve the complex genotype-phenotype relationships underlying morphogenesis. Here we modulated cell width through heterologous expression of sequences encoding the essential enzyme PBP2 and through sublethal treatments with drugs that inhibit PBP2 and the MreB cyto...

  10. Antioxidant properties of cell wall polysaccharides of Stevia rebaudiana leaves

    Directory of Open Access Journals (Sweden)

    Mediesse Kengne Francine

    2014-12-01

    Full Text Available Objective: To examine the total phenolic and protein contents, and the antioxidant activities of cell wall polysaccharide fractions of Stevia rebaudiana leaves. Methods: Three different polysaccharide-enriched fractions, namely FPE (extract with 50 mmol/ L ethylene diamine tetra acetic acid, FPK (extract with 0.05 mol/L KOH and FH (extract with 4 mol/L KOH were extracted from Stevia rebaudiana leaves. The antioxidant activity of these fractions was evaluated based on their ability to scavenge DPPH (1, 1-diphenyl-2-picryl hydrazyl free radical, to reduce ferric power, to chelate ferrous ion and to protect human DNA. Results: The results indicated that protein content was found to be higher in FPK polysaccharide enriched fraction (47.48 µg per mg of FPK. Furthermore, the phenolic compound analysis according to the Folin-Ciocalteu method was higher in FPK (17.71 µg ferulic acid. The DPPH maximal inhibition percentage of the three polysaccharide-enriched fractions at 400 µg/mL was 27.66%, 59.90% and 23.21% respectively for FPE, FPK and FH. All the polysaccharide fractions exhibited a ferric reducing power except the FH one. The three fractions also exhibited lipid peroxidation inhibition, and they completely reverted the DNA damage induced by H2O2/FeCl2. FPK showed the strongest scavenging activity against the DPPH radical, the best chelating ability and lipid peroxidation inhibition. Conclusions: Stevia cell wall polysaccharide fractions are potent protective agents against oxidative stress. The analysis revealed major differences in the antioxidant activity in the three polysaccharides fractions. However, the 0.05 mol/L KOH pectin fraction (FPK showed better antioxidant activity.

  11. Antioxidant properties of cell wall polysaccharides of Stevia rebaudiana leaves

    Institute of Scientific and Technical Information of China (English)

    Mediesse Kengne Francine; Woguia Alice Louise; Fogue Souopgui Pythagore; Atogho-Tiedeu Barbara; Simo Gustave; Thadde Boudjeko

    2014-01-01

    Objective: To examine the total phenolic and protein contents, and the antioxidant activities of cell wall polysaccharide fractions of Stevia rebaudiana leaves.Methods:L ethylene diamine tetra acetic acid), FPK (extract with 0.05 mol/L KOH) and FH (extract with 4 mol/L KOH) were extracted from Stevia rebaudiana leaves. The antioxidant activity of these fractions was evaluated based on their ability to scavenge DPPH (1, 1-diphenyl-2-picryl hydrazyl) free radical, to reduce ferric power, to chelate ferrous ion and to protect human DNA. Three different polysaccharide-enriched fractions, namely FPE (extract with 50 mmol/Results: The results indicated that protein content was found to be higher in FPK polysaccharide enriched fraction (47.48 µg per mg of FPK). Furthermore, the phenolic compound analysis according to the Folin-Ciocalteu method was higher in FPK (17.71 µg ferulic acid). The DPPH maximal inhibition percentage of the three polysaccharide-enriched fractions at 400 µg/mL was 27.66%, 59.90% and 23.21% respectively for FPE, FPK and FH. All the polysaccharide fractions exhibited a ferric reducing power except the FH one. The three fractions also exhibited lipid peroxidation inhibition, and they completely reverted the DNA damage induced by H2O2/FeCl2. FPK showed the strongest scavenging activity against the DPPH radical, the best chelating ability and lipid peroxidation inhibition.Conclusions: Stevia cell wall polysaccharide fractions are potent protective agents against oxidative stress. The analysis revealed major differences in the antioxidant activity in the three polysaccharides fractions. However, the 0.05 mol/L KOH pectin fraction (FPK) showed better antioxidant activity.

  12. Susceptibility to anchoring effects

    Directory of Open Access Journals (Sweden)

    Todd McElroy

    2007-02-01

    Full Text Available Previous research on anchoring has shown this heuristic to be a very robust psychological phenomenon ubiquitous across many domains of human judgment and decision-making. Despite the prevalence of anchoring effects, researchers have only recently begun to investigate the underlying factors responsible for how and in what ways a person is susceptible to them. This paper examines how one such factor, the Big-Five personality trait of openness-to-experience, influences the effect of previously presented anchors on participants' judgments. Our findings indicate that participants high in openness-to-experience were significantly more influenced by anchoring cues relative to participants low in this trait. These findings were consistent across two different types of anchoring tasks providing convergent evidence for our hypothesis.

  13. Plant cell walls throughout evolution: towards a molecular understanding of their design principles

    Energy Technology Data Exchange (ETDEWEB)

    Sarkar, Purbasha; Bosneaga, Elena; Auer, Manfred

    2009-02-16

    Throughout their life, plants typically remain in one location utilizing sunlight for the synthesis of carbohydrates, which serve as their sole source of energy as well as building blocks of a protective extracellular matrix, called the cell wall. During the course of evolution, plants have repeatedly adapted to their respective niche,which is reflected in the changes of their body plan and the specific design of cell walls. Cell walls not only changed throughout evolution but also are constantly remodelled and reconstructed during the development of an individual plant, and in response to environmental stress or pathogen attacks. Carbohydrate-rich cell walls display complex designs, which together with the presence of phenolic polymers constitutes a barrier for microbes, fungi, and animals. Throughout evolution microbes have co-evolved strategies for efficient breakdown of cell walls. Our current understanding of cell walls and their evolutionary changes are limited as our knowledge is mainly derived from biochemical and genetic studies, complemented by a few targeted yet very informative imaging studies. Comprehensive plant cell wall models will aid in the re-design of plant cell walls for the purpose of commercially viable lignocellulosic biofuel production as well as for the timber, textile, and paper industries. Such knowledge will also be of great interest in the context of agriculture and to plant biologists in general. It is expected that detailed plant cell wall models will require integrated correlative multimodal, multiscale imaging and modelling approaches, which are currently underway.

  14. A 3-D Model of a Perennial Ryegrass Primary Cell Wall and Its Enzymatic Degradation

    OpenAIRE

    Indrakumar Vetharaniam; Kelly, William J.; Graeme T. Attwood; Harris, Philip J.

    2014-01-01

    We have developed a novel 3-D, agent-based model of cell-wall digestion to improve our understanding of ruminal cell-wall digestion. It offers a capability to study cell walls and their enzymatic modification, by providing a representation of cellulose microfibrils and non-cellulosic polysaccharides and by simulating their spatial and catalytic interactions with enzymes. One can vary cell-wall composition and the types and numbers of enzyme molecules, allowing the model to be applied to a ran...

  15. A phosphorylated pseudokinase complex controls cell wall synthesis in mycobacteria.

    Science.gov (United States)

    Gee, Christine L; Papavinasasundaram, Kadamba G; Blair, Sloane R; Baer, Christina E; Falick, Arnold M; King, David S; Griffin, Jennifer E; Venghatakrishnan, Harene; Zukauskas, Andrew; Wei, Jun-Rong; Dhiman, Rakesh K; Crick, Dean C; Rubin, Eric J; Sassetti, Christopher M; Alber, Tom

    2012-01-24

    Prokaryotic cell wall biosynthesis is coordinated with cell growth and division, but the mechanisms regulating this dynamic process remain obscure. Here, we describe a phosphorylation-dependent regulatory complex that controls peptidoglycan (PG) biosynthesis in Mycobacterium tuberculosis. We found that PknB, a PG-responsive Ser-Thr protein kinase (STPK), initiates complex assembly by phosphorylating a kinase-like domain in the essential PG biosynthetic protein, MviN. This domain was structurally diverged from active kinases and did not mediate phosphotransfer. Threonine phosphorylation of the pseudokinase domain recruited the FhaA protein through its forkhead-associated (FHA) domain. The crystal structure of this phosphorylated pseudokinase-FHA domain complex revealed the basis of FHA domain recognition, which included unexpected contacts distal to the phosphorylated threonine. Conditional degradation of these proteins in mycobacteria demonstrated that MviN was essential for growth and PG biosynthesis and that FhaA regulated these processes at the cell poles and septum. Controlling this spatially localized PG regulatory complex is only one of several cellular roles ascribed to PknB, suggesting that the capacity to coordinate signaling across multiple processes is an important feature conserved between eukaryotic and prokaryotic STPK networks. PMID:22275220

  16. CELL-WALL GROWTH AND PROTEIN SECRETION IN FUNGI

    NARCIS (Netherlands)

    SIETSMA, JH; WOSTEN, HAB; WESSELS, JGH

    1995-01-01

    Secretion of proteins is a vital process in fungi. Because hyphal walls form a diffusion barrier for proteins, a mechanism different from diffusion probably exist to transport proteins across the wall. In Schizophyllum commune, evidence has been obtained for synthesis at the hyphal apex of wall comp

  17. Wall extensibility: its nature, measurement and relationship to plant cell growth

    Science.gov (United States)

    Cosgrove, D. J.

    1993-01-01

    Expansive growth of plant cells is controlled principally by processes that loosen the wall and enable it to expand irreversibly. The central role of wall relaxation for cell expansion is reviewed. The most common methods for assessing the extension properties of plant cell walls ( wall extensibility') are described, categorized and assessed critically. What emerges are three fundamentally different approaches which test growing cells for their ability (a) to enlarge at different values of turgor, (b) to induce wall relaxation, and (c) to deform elastically or plastically in response to an applied tensile force. Analogous methods with isolated walls are similarly reviewed. The results of these different assays are related to the nature of plant cell growth and pertinent biophysical theory. I argue that the extensibilities' measured by these assays are fundamentally different from one another and that some are more pertinent to growth than others.

  18. Two novel, putatively cell wall-associated and glycosylphosphatidylinositol-anchored alpha-glucanotransferase enzymes of aspergillus niger

    NARCIS (Netherlands)

    van der Kaaij, R. A.; Yuan, X.-L.; Franken, A.; Ram, A. F. J.; Punt, P. J.; Dijkhuizen, L.; Maarel, M.J.E.C. van der

    2007-01-01

    In the genome sequence of Aspergillus niger CBS 513.88, three genes were identified with high similarity to fungal alpha-amylases. The protein sequences derived from these genes were different in two ways from all described fungal alpha-amylases: they were predicted to be glycosylphosphatidyllinosit

  19. Structural and Functional Analysis of Cell Wall-anchored Polypeptide Adhesin BspA in Streptococcus agalactiae.

    Science.gov (United States)

    Rego, Sara; Heal, Timothy J; Pidwill, Grace R; Till, Marisa; Robson, Alice; Lamont, Richard J; Sessions, Richard B; Jenkinson, Howard F; Race, Paul R; Nobbs, Angela H

    2016-07-29

    Streptococcus agalactiae (group B Streptococcus, GBS) is the predominant cause of early-onset infectious disease in neonates and is responsible for life-threatening infections in elderly and immunocompromised individuals. Clinical manifestations of GBS infection include sepsis, pneumonia, and meningitis. Here, we describe BspA, a deviant antigen I/II family polypeptide that confers adhesive properties linked to pathogenesis in GBS. Heterologous expression of BspA on the surface of the non-adherent bacterium Lactococcus lactis confers adherence to scavenger receptor gp340, human vaginal epithelium, and to the fungus Candida albicans Complementary crystallographic and biophysical characterization of BspA reveal a novel β-sandwich adhesion domain and unique asparagine-dependent super-helical stalk. Collectively, these findings establish a new bacterial adhesin structure that has in effect been hijacked by a pathogenic Streptococcus species to provide competitive advantage in human mucosal infections. PMID:27311712

  20. Anchoring of FRET Sensors-A Requirement for Spatiotemporal Resolution.

    Science.gov (United States)

    Ivanova, Elena V; Figueroa, Ricardo A; Gatsinzi, Tom; Hallberg, Einar; Iverfeldt, Kerstin

    2016-01-01

    FRET biosensors have become a routine tool for investigating mechanisms and components of cell signaling. Strategies for improving them for particular applications are continuously sought. One important aspect to consider when designing FRET probes is the dynamic distribution and propagation of signals within living cells. We have addressed this issue by directly comparing an anchored (taFS) to a non-anchored (naFS) cleavable FRET sensor. We chose a microtubule-associated protein tau as an anchor, as microtubules are abundant throughout the cytosol of cells. We show that tau-anchored FRET sensors are concentrated at the cytoskeleton and enriched in the neurite-like processes of cells, providing high intensity of the total signal. In addition, anchoring limits the diffusion of the sensor, enabling spatiotemporally resolved monitoring of subcellular variations in enzyme activity. Thus, anchoring is an important aspect to consider when designing FRET sensors for deeper understanding of cell signaling. PMID:27196902

  1. Cell wall dynamics modulate acetic acid-induced apoptotic cell death of Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    António Rego

    2014-08-01

    Full Text Available Acetic acid triggers apoptotic cell death in Saccharomyces cerevisiae, similar to mammalian apoptosis. To uncover novel regulators of this process, we analyzed whether impairing MAPK signaling affected acetic acid-induced apoptosis and found the mating-pheromone response and, especially, the cell wall integrity pathways were the major mediators, especially the latter, which we characterized further. Screening downstream effectors of this pathway, namely targets of the transcription factor Rlm1p, highlighted decreased cell wall remodeling as particularly important for acetic acid resistance. Modulation of cell surface dynamics therefore emerges as a powerful strategy to increase acetic acid resistance, with potential application in industrial fermentations using yeast, and in biomedicine to exploit the higher sensitivity of colorectal carcinoma cells to apoptosis induced by acetate produced by intestinal propionibacteria.

  2. The bacterial tubulin FtsZ requires its intrinsically disordered linker to direct robust cell wall construction.

    Science.gov (United States)

    Sundararajan, Kousik; Miguel, Amanda; Desmarais, Samantha M; Meier, Elizabeth L; Casey Huang, Kerwyn; Goley, Erin D

    2015-01-01

    The bacterial GTPase FtsZ forms a cytokinetic ring at midcell, recruits the division machinery and orchestrates membrane and peptidoglycan cell wall invagination. However, the mechanism for FtsZ regulation of peptidoglycan metabolism is unknown. The FtsZ GTPase domain is separated from its membrane-anchoring C-terminal conserved (CTC) peptide by a disordered C-terminal linker (CTL). Here we investigate CTL function in Caulobacter crescentus. Strikingly, production of FtsZ lacking the CTL (ΔCTL) is lethal: cells become filamentous, form envelope bulges and lyse, resembling treatment with β-lactam antibiotics. This phenotype is produced by FtsZ polymers bearing the CTC and a CTL shorter than 14 residues. Peptidoglycan synthesis still occurs downstream of ΔCTL; however, cells expressing ΔCTL exhibit reduced peptidoglycan crosslinking and longer glycan strands than wild type. Importantly, midcell proteins are still recruited to sites of ΔCTL assembly. We propose that FtsZ regulates peptidoglycan metabolism through a CTL-dependent mechanism that extends beyond simple protein recruitment. PMID:26099469

  3. Glycosylation of Candida albicans cell wall proteins is critical for induction of innate immune responses and apoptosis of epithelial cells.

    OpenAIRE

    Wagener, Jeanette; Weindl, Günther; de Groot, Piet W. J.; de Boer, Albert D.; Kaesler, Susanne; Thavaraj, Selvam; Bader, Oliver; Mailänder-Sanchez, Daniela; Borelli, Claudia; Weig, Michael; Biedermann, Tilo; Naglik, Julian R.; Korting, Hans Christian; Schaller, Martin

    2012-01-01

    C. albicans is one of the most common fungal pathogen of humans, causing local and superficial mucosal infections in immunocompromised individuals. Given that the key structure mediating host-C. albicans interactions is the fungal cell wall, we aimed to identify features of the cell wall inducing epithelial responses and be associated with fungal pathogenesis. We demonstrate here the importance of cell wall protein glycosylation in epithelial immune activation with a predominant role for the ...

  4. NAC-MYB-based transcriptional regulation of secondary cell wall biosynthesis in land plants

    OpenAIRE

    Nakano, Yoshimi; Yamaguchi, Masatoshi; Endo, Hitoshi; Rejab, Nur Ardiyana; Ohtani, Misato

    2015-01-01

    Plant cells biosynthesize primary cell walls (PCW) in all cells and produce secondary cell walls (SCWs) in specific cell types that conduct water and/or provide mechanical support, such as xylem vessels and fibers. The characteristic mechanical stiffness, chemical recalcitrance, and hydrophobic nature of SCWs result from the organization of SCW-specific biopolymers, i.e., highly ordered cellulose, hemicellulose, and lignin. Synthesis of these SCW-specific biopolymers requires SCW-specific enz...

  5. Primary abdominal wall clear cell carcinoma arising from incisional endometriosis

    Institute of Scientific and Technical Information of China (English)

    Burcu Gundogdu; Isin Ureyen; Gunsu Kimyon; Hakan Turan; Nurettin Boran; Gokhan Tulunay; Dilek Bulbul; Taner Turan; M Faruk Kose

    2013-01-01

    A 49 year-old patient with the complaint of a mass located in the caesarean scar was admitted. There was a fixed mass 30í30 mm in diameter with regular contour located at the right corner of the pfannenstiel incision. Computed tomography revealed a (40í50í50) mm solid mass lesion with margins that cannot be distinguished from the uterus, bladder and small intestines and a heterogeneous mass lesion (50í45í55) mm in diameter, located in the right side of the anterior abdominal wall. Cytoreductive surgery including total abdominal hysterectomy and bilateral salpingo-oophorectomy was performed. Final pathology was clear cell carcinoma. Clear cell carcinoma arising from an extraovarian endometriotic focus was diagnosed and the patient received 6 cycles paclitaxel-carboplatin chemotherapy as adjuvant treatment. The patient who was lost to follow-up applied to our clinic 2 years after surgery with a recurrent mass in the left inguinal region. After 3 cycles of chemotherapy, the patient's tumoral mass in the left inguinal region was excised. The result of the pathology was carcinoma metastasis. It is decided that the following treatment of the patient should be palliative radiation therapy. The patient who underwent palliative radiation therapy died of disease after 4 months of the second operation.

  6. Germ tube-specific antigens of Candida albicans cell walls

    International Nuclear Information System (INIS)

    Studies were performed to characterize the surface differences between blastospores and germ tubes of the pathogenic, dimorphic yeast, Candida albicans, and to identify components of yeast cells responsible for these differences. Investigation of surfaces differences of the two growth forms was facilitated by the production of rabbit antiserum prepared against Formalin-treated yeast possessing germ tubes. To prepare antiserum specific for germ tubes, this serum was adsorbed with stationary phase blastospores. Whereas the unadsorbed antiserum reacted with both blastospore and germ tube forms by immunofluorescence and Enzyme-Linked Immunosorbent Assay, the adsorbed antiserum did not react with blastospores but detected germ tube-specific antigens in hyphal forms. The differences between blastospores and germ tubes of Candida albicans, were further studied by comparing enzymatic digests of cell walls of both growth forms in radiolabeled organisms. Organisms were labeled either on the surface with 125I, or metabolically with [35S] methionine or [3H] mannose. Three-surface-located components (as shown by antibody adsorption and elution experiments) were precipitated from Zymolase digests. All three components were mannoproteins as shown by their ability to bind Concanavalin A, and to be labeled in protein labeling procedures, and two of these (200,000 and 155,000 molecular weight) were germ tube specific, as shown by their ability to be precipitated by germ tube-specific antiserum. Monoclonal antibodies were prepared to C. albicans, using blastospores bearing germ tubes as immunogen

  7. FINITE ELEMENT ANALYSIS OF THE FATIGUE BEHAVIOR OF WOOD FIBER CELL WALLS

    Directory of Open Access Journals (Sweden)

    Phichit Somboon

    2008-11-01

    Full Text Available The fatigue behavior of the wood fiber cell wall under mechanical treatment in refining was simulated dynamically using a finite element method. The effect of the amplitude and frequency of impacts on the mechanical breakdown of the fiber wall structure was examined. The proposed model of the fiber cell wall was constructed from elementary microfibrils in various orientations embedded in isotropic lignin. The fatigue of the cell wall was simulated under normal refiner mechanical pulping conditions. A cyclic load was applied on the model fiber through a hemispherical grit proposed to be applied on the surface on refiner segments. Changes in the elastic modulus of the cell wall were analyzed to determine the potential for cell wall breakdown. An increase in the amplitude of applied forces and frequency of impacts was found to have a significant influence on the reduction of the elastic modulus of the wall structure. A high frequency of impacts increased the stiffness of the cell wall, but resulted in faster reduction of the elastic modulus. At a lower amplitude of impacts, efficient breakdown of the cell wall using grits was achieved with a high frequency of impacts or a high rotational speed of refiners.

  8. Building and degradation of secondary cell walls: are there common patterns of lamellar assembly of cellulose microfibrils and cell wall delamination?

    Science.gov (United States)

    De Micco, Veronica; Ruel, Katia; Joseleau, Jean-Paul; Aronne, Giovanna

    2010-08-01

    During cell wall formation and degradation, it is possible to detect cellulose microfibrils assembled into thicker and thinner lamellar structures, respectively, following inverse parallel patterns. The aim of this study was to analyse such patterns of microfibril aggregation and cell wall delamination. The thickness of microfibrils and lamellae was measured on digital images of both growing and degrading cell walls viewed by means of transmission electron microscopy. To objectively detect, measure and classify microfibrils and lamellae into thickness classes, a method based on the application of computerized image analysis combined with graphical and statistical methods was developed. The method allowed common classes of microfibrils and lamellae in cell walls to be identified from different origins. During both the formation and degradation of cell walls, a preferential formation of structures with specific thickness was evidenced. The results obtained with the developed method allowed objective analysis of patterns of microfibril aggregation and evidenced a trend of doubling/halving lamellar structures, during cell wall formation/degradation in materials from different origin and which have undergone different treatments. PMID:20532796

  9. Anchoring visions in organizations

    DEFF Research Database (Denmark)

    Simonsen, Jesper

    1999-01-01

    This paper introduces the term 'anchoring' within systems development: Visions, developed through early systems design within an organization, need to be deeply rooted in the organization. A vision's rationale needs to be understood by those who decide if the vision should be implemented as well as...... anchoring is considered to be a main activity, is outlined. The task of anchoring visions is described, and techniques and activities are suggested with respect to those actors that have to act on the visions and the recommendations from a design proposal. The paper concludes that obtaining appropriate...

  10. Tensile test of plant cell wall analogs thin films using image stereocorrelation

    OpenAIRE

    Assor, Carole; Sabatier, Laurent; Cathala, Bernard; Aguié Béghin, Véronique; Arnould, Olivier

    2013-01-01

    The plant cell wall can be the optimal scale of investigation for understanding the properties and the variability of vegetal organs structure. At the molecular scale, the wall constituents’ organisation might have a strong influence on theses properties. Primary cell walls are separated into monocotyledons (cereals) and dicotyledons (fleshy fruits) depending on the type of molecules involved (cellulose, hemicelluloses, pectin, etc.). These polymers structures and concentration within the cel...

  11. Impact of Cell Wall Composition on Maize Resistance to Pests and Diseases

    OpenAIRE

    Malvar, Rosa A.; Rogelio Santiago; Jaime Barros-Rios

    2013-01-01

    In cereals, the primary cell wall is built of a skeleton of cellulosic microfibrils embedded in a matrix of hemicelluloses and smaller amounts of pectins, glycoproteins and hydroxycinnamates. Later, during secondary wall development, p-coumaryl, coniferyl and sinapyl alcohols are copolymerized to form mixed lignins. Several of these cell wall components show a determinative role in maize resistance to pest and diseases. However, defense mechanisms are very complex and vary among t...

  12. Photoactive curcumin-derived dyes with surface anchoring moieties used in ZnO nanoparticle-based dye-sensitized solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Ganesh, T.; Kim, Jong Hoon; Yoon, Seog Joon; Kil, Byung-Ho; Maldar, N.N. [Inorganic Nano-Materials Laboratory, Department of Chemistry, Hanyang University, Sung-Dong-Ku, Haengdang-dong 17, Seoul (Korea, Republic of); Han, Jin Wook, E-mail: jwhan@hanyang.ac.kr [Inorganic Nano-Materials Laboratory, Department of Chemistry, Hanyang University, Sung-Dong-Ku, Haengdang-dong 17, Seoul (Korea, Republic of); Han, Sung-Hwan, E-mail: shhan@hanyang.ac.kr [Inorganic Nano-Materials Laboratory, Department of Chemistry, Hanyang University, Sung-Dong-Ku, Haengdang-dong 17, Seoul (Korea, Republic of)

    2010-09-01

    Photoactive, eco-friendly and high molar extinction coefficient, curcumin-derived dyes (BCMoxo and BCtCM) have been explored in ZnO nanoparticles (NPs)-based dye-sensitized solar cells (DSSCs). The boron complex curcumin dyes modified with di-carboxylic anchor groups (BCtCM) provided surface attachment with a strong UV-vis region absorption than the dye molecule without anchor groups (BCMoxo). Photoanodes primed with poly-dispersive ZnO NPs ({approx}80-50 nm) specifically devised for these dyes and optimized for the critical thickness, sensitization time and concentration using a solvent-free ionic electrolyte so as to get current density as high as 1.66 mA/cm{sup 2} under 80 mW/cm{sup 2} irradiation. Therefore, a successful conversion of visible light into electricity by using these curcumin-derived dyes (natural derived photoactive molecules) as photosensitizer in DSSCs would be a great interest in future studies for enhancing further conversion efficiencies.

  13. Photoactive curcumin-derived dyes with surface anchoring moieties used in ZnO nanoparticle-based dye-sensitized solar cells

    International Nuclear Information System (INIS)

    Photoactive, eco-friendly and high molar extinction coefficient, curcumin-derived dyes (BCMoxo and BCtCM) have been explored in ZnO nanoparticles (NPs)-based dye-sensitized solar cells (DSSCs). The boron complex curcumin dyes modified with di-carboxylic anchor groups (BCtCM) provided surface attachment with a strong UV-vis region absorption than the dye molecule without anchor groups (BCMoxo). Photoanodes primed with poly-dispersive ZnO NPs (∼80-50 nm) specifically devised for these dyes and optimized for the critical thickness, sensitization time and concentration using a solvent-free ionic electrolyte so as to get current density as high as 1.66 mA/cm2 under 80 mW/cm2 irradiation. Therefore, a successful conversion of visible light into electricity by using these curcumin-derived dyes (natural derived photoactive molecules) as photosensitizer in DSSCs would be a great interest in future studies for enhancing further conversion efficiencies.

  14. Immuno and Affinity Cytochemical Analysis of Cell Wall Composition in the Moss Physcomitrella patens.

    Science.gov (United States)

    Berry, Elizabeth A; Tran, Mai L; Dimos, Christos S; Budziszek, Michael J; Scavuzzo-Duggan, Tess R; Roberts, Alison W

    2016-01-01

    In contrast to homeohydric vascular plants, mosses employ a poikilohydric strategy for surviving in the dry aerial environment. A detailed understanding of the structure, composition, and development of moss cell walls can contribute to our understanding of not only the evolution of overall cell wall complexity, but also the differences that have evolved in response to selection for different survival strategies. The model moss species Physcomitrella patens has a predominantly haploid lifecycle consisting of protonemal filaments that regenerate from protoplasts and enlarge by tip growth, and leafy gametophores composed of cells that enlarge by diffuse growth and differentiate into several different types. Advantages for genetic studies include methods for efficient targeted gene modification and extensive genomic resources. Immuno and affinity cytochemical labeling were used to examine the distribution of polysaccharides and proteins in regenerated protoplasts, protonemal filaments, rhizoids, and sectioned gametophores of P. patens. The cell wall composition of regenerated protoplasts was also characterized by flow cytometry. Crystalline cellulose was abundant in the cell walls of regenerating protoplasts and protonemal cells that developed on media of high osmolarity, whereas homogalactuonan was detected in the walls of protonemal cells that developed on low osmolarity media and not in regenerating protoplasts. Mannan was the major hemicellulose detected in all tissues tested. Arabinogalactan proteins were detected in different cell types by different probes, consistent with structural heterogneity. The results reveal developmental and cell type specific differences in cell wall composition and provide a basis for analyzing cell wall phenotypes in knockout mutants. PMID:27014284

  15. DNB test results for R grid thimble cold wall cells

    International Nuclear Information System (INIS)

    Two sets of type ''R'' mixing vane grid DNB data from a 4 x 4 rod bundle, where one of the central rods is a simulated control rod thimble, are presented. The range of parameters for this thimble cold wall data is given. The results show that the W-3 correlation with cold wall factor and the Modified Spacer Factor is applicable to ''R'' grid thimble cold wall rod bundle data. (7 references) (auth)

  16. Genome-Wide Association Mapping for Cell Wall Composition and Properties in Temperate Grasses

    DEFF Research Database (Denmark)

    Bellucci, Andrea

    -glucans. Plant cell wall biosynthesis is regulated by a large number of genes and regulatory factors but very few of these are known and characterized. This PhD project aimed to the identification of putative candidate genes involved in plant cell wall composition and properties using a genome wide (GWAS...

  17. Consolidated pretreatment and hydrolysis of plant biomass expressing cell wall degrading enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Raab, R. Michael; Zhang, Dongcheng; Bougri, Oleg

    2016-02-02

    Methods for consolidated pretreatment and hydrolysis of genetically engineered plants expressing cell wall degrading enzymes are provided. Expression cassettes and vectors for making transgenic plants are described. Plants engineered to express one or more cell wall degrading enzymes using expression cassettes and vectors of the invention are also provided.

  18. CONSTITUTIVE MELANIN IN THE CELL WALL OF THE ETIOLOGIC AGENT OF LOBO'S DISEASE

    Directory of Open Access Journals (Sweden)

    TABORDA Valeria B.A.

    1999-01-01

    Full Text Available Lobo's disease is a chronic granulomatous disease caused by the obligate pathogenic fungus, whose cell walls contain constitutive melanin. In contrast, melanin does not occur in the cell walls of Paracoccidioides brasiliensis when stained by the Fontana-Masson stain.

  19. Trans-Golgi Network-An Intersection of Trafficking Cell Wall Components

    Institute of Scientific and Technical Information of China (English)

    Natasha Worden; Eunsook Park; Georgia Drakakaki

    2012-01-01

    The cell wall,a crucial cell compartment,is composed of a network of polysaccharides and proteins,providing structural support and protection from external stimuli.While the cell wall structure and biosynthesis have been extensively studied,very little is known about the transport of polysaccharides and other components into the developing cell wall.This review focuses on endomembrane trafficking pathways involved in cell wall deposition.Cellulose synthase complexes are assembled in the Golgi,and are transported in vesicles to the plasma membrane.Non-cellulosic polysaccharides are synthesized in the Golgi apparatus,whereas cellulose is produced by enzyme complexes at the plasma membrane.Polvsaccharides and enzymes that are involved in cell wall modification and assembly are transported by distinct vesicle types to their destinations; however,the precise mechanisms involved in selection,sorting and delivery remain to be identified.The endomembrane system orchestrates the delivery of Golgi-derived and possibly endocytic vesicles carrying cell wall and cell membrane components to the newly-formed cell plate.However,the nature of these vesicles,their membrane compositions,and the timing of their delivery are largely unknown.Emerging technologies such as chemical genomics and proteomics are promising avenues to gain insight into the trafficking of cell wall components.

  20. Structure of the cell wall of mango after application of ionizing radiation

    International Nuclear Information System (INIS)

    Cells of the mesocarp of mango cultivar Tommy Atkins were analyzed by Transmission Electron Microscope—TEM to evaluate the effects of doses of 0.5 and 1.0 kGy applied immediately after the fruit and after storage for twenty days at a temperature of 12 °C followed by 5 days of simulated marketing at a temperature of 21 °C. No alteration was found in the structure of the cell wall, middle lamella, and plasma membrane of fruits when analyzed immediately after application of doses. The mesocarp cell structure of the cell wall, middle lamella, and the plasma membrane did however undergo changes after storage. Fruits that received a dose of 0.5 kGy displayed slight changes in cell wall structure and slight disintegration of the middle lamella. Fruits that received a dose of 1.0 kGy displayed more severe changes in the structure of the cell wall, greater middle lamella degradation, and displacement of the plasma membrane. - Highlights: ► Mesocarp cells were analyzed by Transmission Electron Microscope—TEM. ► No change in cell wall structure, middle lamella and plasma membrane was found in fruits immediately after irradiation. ► Changes in cell wall structure, middle lamella and plasma membrane happened after storage. ► Fruits subjected to 0.5 kGy showed smaller cell wall change.

  1. Clinostation influence on regeneration of cell wall in Solanum Tuberosum L. protoplasts

    Science.gov (United States)

    Nedukha, Elena M.; Sidorov, V. A.; Samoylov, V. M.

    1994-08-01

    Regeneration of cell walls in protoplasts was investigated using light- and electronmicroscopic methods. The protoplasts were isolated from mesophyll of Solanum tuberosum leaves and were cultivated on the horizontal low rotating clinostat (2 rpm) and in control for 10 days. Using a fluorescent method (with Calcofluor white) it was demonstrated that changes in vector gravity results in an regeneration inhibition of cell wall. With electron-microscopical and electro-cytochemical methods (staining with alcianum blue) dynamics of the regeneration of cell walls in protoplasts was studied; carbohydrate matrix of cell walls is deposited at the earliest stages of this process. The influence of microgravity on the cell wall regeneration is discussed in higher plants.

  2. Non-invasive imaging of cellulose microfibril orientation within plant cell walls by polarized Raman microspectroscopy.

    Science.gov (United States)

    Sun, Lan; Singh, Seema; Joo, Michael; Vega-Sanchez, Miguel; Ronald, Pamela; Simmons, Blake A; Adams, Paul; Auer, Manfred

    2016-01-01

    Cellulose microfibrils represent the major scaffold of plant cell walls. Different packing and orientation of the microfibrils at the microscopic scale determines the macroscopic properties of cell walls and thus affect their functions with a profound effect on plant survival. We developed a polarized Raman microspectroscopic method to determine cellulose microfibril orientation within rice plant cell walls. Employing an array of point measurements as well as area imaging and subsequent Matlab-assisted data processing, we were able to characterize the distribution of cellulose microfibril orientation in terms of director angle and anisotropy magnitude. Using this approach we detected differences between wild type rice plants and the rice brittle culm mutant, which shows a more disordered cellulose microfibril arrangement, and differences between different tissues of a wild type rice plant. This novel non-invasive Raman imaging approach allows for quantitative assessment of cellulose fiber orientation in cell walls of herbaceous plants, an important advancement in cell wall characterization. PMID:26137889

  3. The three-dimensional structure of the cell wall glycoprotein of Chlorogonium elongatum.

    Science.gov (United States)

    Shaw, P J; Hills, G J

    1984-06-01

    The green alga Chlorogonium elongatum, a member of the Volvocales, possesses a crystalline cell wall composed of hydroxyproline-rich glycoprotein similar to the primary cell wall glycoproteins of higher plants. Electron microscopy and computer image processing have been used to determine the crystal structure of the Chlorogonium cell wall in three dimensions to a resolution of 2.0 nm. The structure is composed of heterologous dimers. Each subunit of the dimer comprises a long, thin spacer domain and a large globular domain, which is the site of the intra- and inter-dimer interactions. There are also sites of intersubunit interactions at the opposite ends of the rod domains. We suggest that the rods are composed predominantly of glycosylated polyproline helix, as has been suggested for higher plant cell wall glycoproteins and has been shown for the cell wall glycoprotein of Chlamydomonas reinhardtii, which is closely related to Chlorogonium. PMID:6490737

  4. Understanding the relationship between cotton fiber properties and non-cellulosic cell wall polysaccharides

    DEFF Research Database (Denmark)

    Rajasundaram, Dhivyaa; Runavot, Jean-Luc; Guo, Xiaoyuan;

    2014-01-01

    A detailed knowledge of cell wall heterogeneity and complexity is crucial for understanding plant growth and development. One key challenge is to establish links between polysaccharide-rich cell walls and their phenotypic characteristics. It is of particular interest for some plant material, like...... different cotton species were studied. The glycan array was generated by sequential extraction of cell wall polysaccharides from mature cotton fibers and screening samples against eleven extensively characterized cell wall probes. Also, phenotypic characteristics of cotton fibers such as length, strength...... probes. Moreover, homogalacturonan and callose were shown to be significant predictors for fiber length. The role of these polysaccharides was already pointed out in previous cell wall elongation studies. Additional relationships were predicted for fiber strength and elongation which will need further...

  5. Susceptibility to anchoring effects

    OpenAIRE

    Todd McElroy; Keith Dowd

    2007-01-01

    Previous research on anchoring has shown this heuristic to be a very robust psychological phenomenon ubiquitous across many domains of human judgment and decision-making. Despite the prevalence of anchoring effects, researchers have only recently begun to investigate the underlying factors responsible for how and in what ways a person is susceptible to them. This paper examines how one such factor, the Big-Five personality trait of openness-to-experience, influences the effect of previously p...

  6. A new picture of cell wall protein dynamics in elongating cells of Arabidopsis thaliana: Confirmed actors and newcomers

    OpenAIRE

    Jamet Elisabeth; Pont-Lezica Rafael; Borderies Gisèle; Canut Hervé; Irshad Muhammad

    2008-01-01

    Abstract Background Cell elongation in plants requires addition and re-arrangements of cell wall components. Even if some protein families have been shown to play roles in these events, a global picture of proteins present in cell walls of elongating cells is still missing. A proteomic study was performed on etiolated hypocotyls of Arabidopsis used as model of cells undergoing elongation followed by growth arrest within a short time. Results Two developmental stages (active growth and after g...

  7. Intracellular vesicles as reproduction elements in cell wall-deficient L-form bacteria

    OpenAIRE

    Briers, Yves; Staubli, Titu; Schmid, Markus C; Wagner, Michael; Schuppler, Markus; Loessner, Martin J.

    2012-01-01

    Cell wall-deficient bacteria, or L-forms, represent an extreme example of bacterial plasticity. Stable L-forms can multiply and propagate indefinitely in the absence of a cell wall. Data presented here are consistent with the model that intracellular vesicles in Listeria monocytogenes L-form cells represent the actual viable reproductive elements. First, small intracellular vesicles are formed along the mother cell cytoplasmic membrane, originating from local phospholipid accumulation. During...

  8. Decreased Polysaccharide Feruloylation Compromises Plant Cell Wall Integrity and Increases Susceptibility to Necrotrophic Fungal Pathogens

    Directory of Open Access Journals (Sweden)

    Nathan T Reem

    2016-05-01

    Full Text Available The complexity of cell wall composition and structure determines the strength, flexibility, and function of the primary cell wall in plants. However, the contribution of the various components to cell wall integrity and function remains unclear. Modifications of cell wall composition can induce plant responses known as Cell Wall Integrity control. In this study, we used transgenic expression of the fungal feruloyl esterase AnFAE to examine the effect of post-synthetic modification of Arabidopsis and Brachypodium cell walls. Transgenic Arabidopsis plants expressing AnFAE showed a significant reduction of monomeric ferulic acid, increased amounts of wall-associated extensins, and increased susceptibility to Botrytis cinerea, compared with wild type. Transgenic Brachypodium showed reductions in monomeric and dimeric ferulic acids and increased susceptibility to Bipolaris sorokiniana. Upon infection, transgenic Arabidopsis and Brachypodium plants also showed increased expression of several defense-related genes compared with wild type. These results demonstrate a role, in both monocot and dicot plants, of polysaccharide feruloylation in plant cell wall integrity, which contributes to plant resistance to necrotrophic pathogens.

  9. Single-Wall Carbon Nanotube Anodes for Lithium Cells

    Science.gov (United States)

    Hepp, Aloysius F.; Raffaelle, Ryne; Gennett, Tom; Kumta, Prashant; Maranchi, Jeff; Heben, Mike

    2006-01-01

    In recent experiments, highly purified batches of single-wall carbon nanotubes (SWCNTs) have shown promise as superior alternatives to the graphitic carbon-black anode materials heretofore used in rechargeable thin-film lithium power cells. The basic idea underlying the experiments is that relative to a given mass of graphitic carbon-black anode material, an equal mass of SWCNTs can be expected to have greater lithium-storage and charge/discharge capacities. The reason for this expectation is that whereas the microstructure and nanostructure of a graphitic carbon black is such as to make most of the interior of the material inaccessible for intercalation of lithium, a batch of SWCNTs can be made to have a much more open microstructure and nanostructure, such that most of the interior of the material is accessible for intercalation of lithium. Moreover, the greater accessibility of SWCNT structures can be expected to translate to greater mobilities for ion-exchange processes and, hence, an ability to sustain greater charge and discharge current densities.

  10. Lysine methyltransferase G9a is not required for DNMT3A/3B anchoring to methylated nucleosomes and maintenance of DNA methylation in somatic cells

    Directory of Open Access Journals (Sweden)

    Sharma Shikhar

    2012-01-01

    Full Text Available Abstract Background DNA methylation, histone modifications and nucleosome occupancy act in concert for regulation of gene expression patterns in mammalian cells. Recently, G9a, a H3K9 methyltransferase, has been shown to play a role in establishment of DNA methylation at embryonic gene targets in ES cells through recruitment of de novo DNMT3A/3B enzymes. However, whether G9a plays a similar role in maintenance of DNA methylation in somatic cells is still unclear. Results Here we show that G9a is not essential for maintenance of DNA methylation in somatic cells. Knockdown of G9a has no measurable effect on DNA methylation levels at G9a-target loci. DNMT3A/3B remain stably anchored to nucleosomes containing methylated DNA even in the absence of G9a, ensuring faithful propagation of methylated states in cooperation with DNMT1 through somatic divisions. Moreover, G9a also associates with nucleosomes in a DNMT3A/3B and DNA methylation-independent manner. However, G9a knockdown synergizes with pharmacologic inhibition of DNMTs resulting in increased hypomethylation and inhibition of cell proliferation. Conclusions Taken together, these data suggest that G9a is not involved in maintenance of DNA methylation in somatic cells but might play a role in re-initiation of de novo methylation after treatment with hypomethylating drugs, thus serving as a potential target for combinatorial treatments strategies involving DNMTs inhibitors.

  11. Identification of the cell wall receptor for Candida nodaensis Killer toxin

    OpenAIRE

    Silva, Sónia Carina; Aguiar, Cristina; Veríssimo, P.; Pires, E.; Lucas, Cândida

    2004-01-01

    Comunicação efectuada no XIV Congresso Nacional de Bioquímica em Vilamoura (Portugal), 2004. The biological action of the K toxins involves a first step in the killing process, which correspond to the adsorption the toxin to the cell wall of sensitive cells. Here we describe the work performed towards the identification of the cell wall receptor for the zymocin under this study. For this purpose, the main cell wall components of the sensitive yeast Pichia guilliermondii were extracted. Th...

  12. Atomic Force Microscopy Measurements of the Mechanical Properties of Cell Walls on Living Bacterial Cells

    Science.gov (United States)

    Bailey, Richard; Mullin, Nic; Turner, Robert; Foster, Simon; Hobbs, Jamie

    2014-03-01

    Staphylococcus aureus is a major cause of infection in humans, including the Methicillin resistant strain, MRSA. However, very little is known about the mechanical properties of these cells. Our investigations use AFM to examine live S. aureus cells to quantify mechanical properties. These were explored using force spectroscopy with different trigger forces, allowing the properties to be extracted at different indentation depths. A value for the cell wall stiffness has been extracted, along with a second, higher value which is found upon indenting at higher forces. This higher value drops as the cells are exposed to high salt, sugar and detergent concentrations, implying that this measurement contains a contribution from the internal turgor pressure. We have monitored these properties as the cells progress through the cell cycle. Force maps were taken over the cells at different stages of the growth process to identify changes in the mechanics throughout the progression of growth and division. The effect of Oxacillin has also been studied, to better understand its mechanism of action. Finally mutant strains of S. aureus and a second species Bacillus subtilis have been used to link the mechanical properties of the cell walls with the chain lengths and substructures involved.

  13. Genetic modification of plant cell walls to enhance biomass yield and biofuel production in bioenergy crops.

    Science.gov (United States)

    Wang, Yanting; Fan, Chunfen; Hu, Huizhen; Li, Ying; Sun, Dan; Wang, Youmei; Peng, Liangcai

    2016-01-01

    Plant cell walls represent an enormous biomass resource for the generation of biofuels and chemicals. As lignocellulose property principally determines biomass recalcitrance, the genetic modification of plant cell walls has been posed as a powerful solution. Here, we review recent progress in understanding the effects of distinct cell wall polymers (cellulose, hemicelluloses, lignin, pectin, wall proteins) on the enzymatic digestibility of biomass under various physical and chemical pretreatments in herbaceous grasses, major agronomic crops and fast-growing trees. We also compare the main factors of wall polymer features, including cellulose crystallinity (CrI), hemicellulosic Xyl/Ara ratio, monolignol proportion and uronic acid level. Furthermore, the review presents the main gene candidates, such as CesA, GH9, GH10, GT61, GT43 etc., for potential genetic cell wall modification towards enhancing both biomass yield and enzymatic saccharification in genetic mutants and transgenic plants. Regarding cell wall modification, it proposes a novel groove-like cell wall model that highlights to increase amorphous regions (density and depth) of the native cellulose microfibrils, providing a general strategy for bioenergy crop breeding and biofuel processing technology. PMID:27269671

  14. A new picture of cell wall protein dynamics in elongating cells of Arabidopsis thaliana: Confirmed actors and newcomers

    Directory of Open Access Journals (Sweden)

    Jamet Elisabeth

    2008-09-01

    Full Text Available Abstract Background Cell elongation in plants requires addition and re-arrangements of cell wall components. Even if some protein families have been shown to play roles in these events, a global picture of proteins present in cell walls of elongating cells is still missing. A proteomic study was performed on etiolated hypocotyls of Arabidopsis used as model of cells undergoing elongation followed by growth arrest within a short time. Results Two developmental stages (active growth and after growth arrest were compared. A new strategy consisting of high performance cation exchange chromatography and mono-dimensional electrophoresis was established for separation of cell wall proteins. This work allowed identification of 137 predicted secreted proteins, among which 51 had not been identified previously. Apart from expected proteins known to be involved in cell wall extension such as xyloglucan endotransglucosylase-hydrolases, expansins, polygalacturonases, pectin methylesterases and peroxidases, new proteins were identified such as proteases, proteins related to lipid metabolism and proteins of unknown function. Conclusion This work highlights the CWP dynamics that takes place between the two developmental stages. The presence of proteins known to be related to cell wall extension after growth arrest showed that these proteins may play other roles in cell walls. Finally, putative regulatory mechanisms of protein biological activity are discussed from this global view of cell wall proteins.

  15. Trifluoromethanesulfonic acid-based proteomic analysis of cell wall and secreted proteins of the ascomycetous fungi Neurospora crassa and Candida albicans.

    Science.gov (United States)

    Maddi, Abhiram; Bowman, Shaun M; Free, Stephen J

    2009-10-01

    Cell wall proteins from purified Candida albicans and Neurospora crassa cell walls were released using trifluoromethanesulfonic acid (TFMS) which cleaves the cell wall glucan/chitin matrix and deglycosylates the proteins. The cell wall proteins were then characterized by SDS-PAGE and identified by proteomic analysis. The analyses for C. albicans identified 15 cell wall proteins and six secreted proteins. For N. crassa, the analyses identified 26 cell wall proteins and nine secreted proteins. Most of the C. albicans cell wall proteins are found in the cell walls of both yeast and hyphae cells, but some cell type-specific cell wall proteins were observed. The analyses showed that the pattern of cell wall proteins present in N. crassa vegetative hyphae and conidia (asexual spores) are quite different. Almost all of the cell wall proteins identified in N. crassa have close homologs in the sequenced fungal genomes, suggesting that these proteins have important conserved functions within the cell wall. PMID:19555771

  16. Cell Wall Nonlinear Elasticity and Growth Dynamics: How Do Bacterial Cells Regulate Pressure and Growth?

    Science.gov (United States)

    Deng, Yi

    In my thesis, I study intact and bulging Escherichia coli cells using atomic force microscopy to separate the contributions of the cell wall and turgor pressure to the overall cell stiffness. I find strong evidence of power--law stress--stiffening in the E. coli cell wall, with an exponent of 1.22±0.12, such that the wall is significantly stiffer in intact cells (E = 23±8 MPa and 49±20 MPa in the axial and circumferential directions) than in unpressurized sacculi. These measurements also indicate that the turgor pressure in living cells E. coli is 29±3 kPa. The nonlinearity in cell elasticity serves as a plausible mechanism to balance the mechanical protection and tension measurement sensitivity of the cell envelope. I also study the growth dynamics of the Bacillus subtilis cell wall to help understand the mechanism of the spatiotemporal order of inserting new cell wall material. High density fluorescent markers are used to label the entire cell surface to capture the morphological changes of the cell surface at sub-cellular to diffraction-limited spatial resolution and sub-minute temporal resolution. This approach reveals that rod-shaped chaining B. subtilis cells grow and twist in a highly heterogeneous fashion both spatially and temporally. Regions of high growth and twisting activity have a typical length scale of 5 μm, and last for 10-40 minutes. Motivated by the quantification of the cell wall growth dynamics, two microscopy and image analysis techniques are developed and applied to broader applications beyond resolving bacterial growth. To resolve densely distributed quantum dots, we present a fast and efficient image analysis algorithm, namely Spatial Covariance Reconstruction (SCORE) microscopy that takes into account the blinking statistics of the fluorescence emitters. We achieve sub-diffraction lateral resolution of 100 nm from 5 to 7 seconds of imaging, which is at least an order of magnitude faster than single-particle localization based methods

  17. Navigating the transcriptional roadmap regulating plant secondary cell wall deposition

    Directory of Open Access Journals (Sweden)

    Steven Grant Hussey

    2013-08-01

    Full Text Available The current status of lignocellulosic biomass as an invaluable resource in industry, agriculture and health has spurred increased interest in understanding the transcriptional regulation of secondary cell wall (SCW biosynthesis. The last decade of research has revealed an extensive network of NAC, MYB and other families of transcription factors regulating Arabidopsis SCW biosynthesis, and numerous studies have explored SCW-related transcription factors in other dicots and monocots. Whilst the general structure of the Arabidopsis network has been a topic of several reviews, they have not comprehensively represented the detailed protein-DNA and protein-protein interactions described in the literature, and an understanding of network dynamics and functionality has not yet been achieved for SCW formation. Furthermore the methodologies employed in studies of SCW transcriptional regulation have not received much attention, especially in the case of non-model organisms. In this review, we have reconstructed the most exhaustive literature-based network representations to date of SCW transcriptional regulation in Arabidopsis. We include a manipulable Cytoscape representation of the Arabidopsis SCW transcriptional network to aid in future studies, along with a list of supporting literature for each documented interaction. Amongst other topics, we discuss the various components of the network, its evolutionary conservation in plants, putative modules and dynamic mechanisms that may influence network function, and the approaches that have been employed in network inference. Future research should aim to better understand network function and its response to dynamic perturbations, whilst the development and application of genome-wide approaches such as ChIP-seq and systems genetics are in progress for the study of SCW transcriptional regulation in non-model organisms.

  18. Xyloglucans from flaxseed kernel cell wall: Structural and conformational characterisation.

    Science.gov (United States)

    Ding, Huihuang H; Cui, Steve W; Goff, H Douglas; Chen, Jie; Guo, Qingbin; Wang, Qi

    2016-10-20

    The structure of ethanol precipitated fraction from 1M KOH extracted flaxseed kernel polysaccharides (KPI-EPF) was studied for better understanding the molecular structures of flaxseed kernel cell wall polysaccharides. Based on methylation/GC-MS, NMR spectroscopy, and MALDI-TOF-MS analysis, the dominate sugar residues of KPI-EPF fraction comprised of (1,4,6)-linked-β-d-glucopyranose (24.1mol%), terminal α-d-xylopyranose (16.2mol%), (1,2)-α-d-linked-xylopyranose (10.7mol%), (1,4)-β-d-linked-glucopyranose (10.7mol%), and terminal β-d-galactopyranose (8.5mol%). KPI-EPF was proposed as xyloglucans: The substitution rate of the backbone is 69.3%; R1 could be T-α-d-Xylp-(1→, or none; R2 could be T-α-d-Xylp-(1→, T-β-d-Galp-(1→2)-α-d-Xylp-(1→, or T-α-l-Araf-(1→2)-α-d-Xylp-(1→; R3 could be T-α-d-Xylp-(1→, T-β-d-Galp-(1→2)-α-d-Xylp-(1→, T-α-l-Fucp-(1→2)-β-d-Galp-(1→2)-α-d-Xylp-(1→, or none. The Mw of KPI-EPF was calculated to be 1506kDa by static light scattering (SLS). The structure-sensitive parameter (ρ) of KPI-EPF was calculated as 1.44, which confirmed the highly branched structure of extracted xyloglucans. This new findings on flaxseed kernel xyloglucans will be helpful for understanding its fermentation properties and potential applications. PMID:27474598

  19. Characterisation of cell-wall polysaccharides from mandarin segment membranes.

    Science.gov (United States)

    Coll-Almela, Luis; Saura-López, Domingo; Laencina-Sánchez, José; Schols, Henk A; Voragen, Alfons G J; Ros-García, José María

    2015-05-15

    In an attempt to develop a process of enzymatic peeling of mandarin segments suitable for use on an industrial scale, the cell wall fraction of the segment membrane of Satsuma mandarin fruits was extracted to obtain a chelating agent-soluble pectin fraction (ChSS), a dilute sodium hydroxide-soluble pectin fraction (DASS), a 1M sodium hydroxide-soluble hemicellulose fraction (1MASS), a 4M sodium hydroxide-soluble hemicellulose fraction (4MASS) and a cellulose-rich residue (3.1, 0.9, 0.4, 0.7 and 1.6%w/w of fresh membrane, respectively). The ChSS pectin consisted mainly of galacturonic acid followed by arabinose and galactose. The DASS fraction contained less galacturonic acid and more neutral sugars than ChSS. Eighty-nine percent of the galacturonic acid present in the segment membranes was recovered in the above two pectin fractions. The two hemicellulosic fractions consisted of two different molecular weight populations, which also differed in their sugar composition. Arabinose, xylose, mannose, galactose and glucose were the main sugar constituents of these hemicellulose fractions. In addition to an (arabino)xylan and a xyloglucan, the presence of an arabinogalactan is suggested by the sugar composition of both hemicelluloses. The pectin fractions were also characterised by their degradability by the pectic enzymes polygalacturonase, pectinmethylesterase and rhamnogalacturonan hydrolase. However the degree of degradation of the pectin fractions by enzymes differed, and the amount of the polymeric materials resistant to further degradation and the oligomeric products also differed. Using pectic enzymes it is possible to obtain peeled mandarin segments ready to eat or for canning. PMID:25577048

  20. Investigating the role of pectin in carrot cell wall changes during thermal processing: A microscopic approach

    OpenAIRE

    Ribas, Albert; Van Buggenhout, Sandy; Palmero, Paola; Hendrickx, Marc; Van Loey, Ann

    2014-01-01

    Changes in cell wall integrity upon thermal treatment were assessed in carrot cells using novel microscopic approaches using Congo red and different cell wall polysaccharide specific probes (JIM7, LM10, LM11, LM15, LM21, LM22 and CBM3a). Strong thermal processing induced an increased accessibility of cellulose and hemicelluloses by Congo red and the specific probes, except galactomannan, which detection was not affected by the thermal processing. Detection of pectin by JIM7 disappeared upon t...

  1. Evaluation of Chlorella (Chlorophyta) as Source of Fermentable Sugars via Cell Wall Enzymatic Hydrolysis

    OpenAIRE

    Marcoaurélio Almenara Rodrigues; Elba Pinto da Silva Bon

    2011-01-01

    The cell wall of Chlorella is composed of up to 80% carbohydrates including cellulose. In this study, Chlorella homosphaera and Chlorella zofingiensis were evaluated as source of fermentable sugars via their cell wall enzymatic degradation. The algae were cultivated in inorganic medium, collected at the stationary growth phase and centrifuged. The cell pellet was suspended in citrate buffer, pH 4.8 and subjected to 24 hours hydrolysis at 50°C using a cellulases, xylanases, and amylases ble...

  2. Malignant transformation of ectopic pancreatic cells in the duodenal wall

    Institute of Scientific and Technical Information of China (English)

    Roberto; Bini; Paolo; Voghera; Alberto; Tapparo; Raffaele; Nunziata; Andrea; Demarchi; Matteo; Capocefalo; Renzo; Leli

    2010-01-01

    Ectopic pancreas (EP) is the relatively uncommon presence of pancreatic tissue outside the normal location of the pancreas. This condition is usually asymptomatic and rarely complicated by pancreatitis and malignant transformation. A few cases of neoplastic phenomena that developed from EP into the duodenal wall are described in the literature. Herein we report a case of gastric outlet obstruction due to adenocarcinoma arising from EP of the duodenal wall. The patient underwent a Whipple's procedure and had...

  3. Investigating the effect of carbon nanotube diameter and wall number in carbon nanotube/silicon heterojunction solar cells

    OpenAIRE

    Tom Grace; LePing Yu; Christopher Gibson; Daniel Tune; Huda Alturaif; Zeid Al Othman; Joseph Shapter

    2016-01-01

    Suspensions of single-walled, double-walled and multi-walled carbon nanotubes (CNTs) were generated in the same solvent at similar concentrations. Films were fabricated from these suspensions and used in carbon nanotube/silicon heterojunction solar cells and their properties were compared with reference to the number of walls in the nanotube samples. It was found that single-walled nanotubes generally produced more favorable results; however, the double and multi-walled nanotube films used in...

  4. 'Strengthening the fungal cell wall through chitin-glucan cross-links: effects on morphogenesis and cell integrity'.

    Science.gov (United States)

    Arroyo, Javier; Farkaš, Vladimír; Sanz, Ana Belén; Cabib, Enrico

    2016-09-01

    The cross-linking of polysaccharides to assemble new cell wall in fungi requires transglycosylation mechanisms by which preexisting glycosidic linkages are broken and new linkages are created between the polysaccharides. The molecular mechanisms for these processes, which are essential for fungal cell biology, are only now beginning to be elucidated. Recent development of in vivo and in vitro biochemical approaches has allowed characterization of important aspects about the formation of chitin-glucan covalent cell wall cross-links by cell wall transglycosylases of the CRH family and their biological function. Covalent linkages between chitin and glucan mediated by Crh proteins control morphogenesis and also play important roles in the remodeling of the fungal cell wall as part of the compensatory responses necessary to counterbalance cell wall stress. These enzymes are encoded by multigene families of redundant proteins very well conserved in fungal genomes but absent in mammalian cells. Understanding the molecular basis of fungal adaptation to cell wall stress through these and other cell wall remodeling enzymatic activities offers an opportunity to explore novel antifungal treatments and to identify potential fungal virulence factors. PMID:27185288

  5. Identification of a Streptococcus salivarius Cell Wall Component Mediating Coaggregation with Veillonella alcalescens VI

    Science.gov (United States)

    Weerkamp, Anton H.; McBride, Barry C.

    1981-01-01

    Cell walls of Streptococcus salivarius HB aggregated Veillonella alcalescens V1, but cell walls of the mutant S. salivarius HB-V5 did not. We found no correlation between the presence of fimbriae on streptococcal walls and the ability to aggregate Veillonella strains. Treatment of the walls with lysozyme solubilized a fraction which possessed Veillonella-aggregating activity. Solubilized cell wall preparations of strain HB contained three major (glyco)proteins as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and at least four antigens as determined by immunoelectrophoresis with antiserum prepared against strain HB walls. A specific antiserum, which was obtained by adsorption of anti-HB serum on strain HB-V5 cells, contained monospecific antibody that reacted with the solubilized strain HB wall preparation. Similar fractions prepared from strain HB-V5 cell walls did not possess aggregating activity and lacked one protein band (protein I) after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and one antigen (antigen b) after immunoelectrophoresis. The same antigen was absent when lysozyme-solubilized wall preparations of strain HB were reacted with anti-HB-V5 serum. Crossed-immunoisoelectric focusing indicated that this specific (glyco)protein and this antigen were identical and had an isoelectric point of 4.60. Protein I and antigen b were specifically adsorbed when solubilized strain HB cell walls were incubated with V. alcalescens V1 but were not adsorbed by nonaggregating Veillonella parvula ATCC 10790 cells. Culture supernatants of strain HB contained V. alcalescens V1-aggregating activity. Antigen b was present in the culture supernatant, but was not found in cultures of strain HB-V5. A total of 18 S. salivarius isolates possessing the streptococcal group K antigen released aggregating activity and antigen b into the culture medium, but 11 strains which lacked the K-antigen did not. Images PMID:7251145

  6. Display of alpha-amylase on the surface of Lactobacillus casei cells by use of the PgsA anchor protein, and production of lactic acid from starch.

    Science.gov (United States)

    Narita, Junya; Okano, Kenji; Kitao, Tomoe; Ishida, Saori; Sewaki, Tomomitsu; Sung, Moon-Hee; Fukuda, Hideki; Kondo, Akihiko

    2006-01-01

    We developed a new cell surface engineering system based on the PgsA anchor protein from Bacillus subtilis. In this system, the N terminus of the target protein was fused to the PgsA protein and the resulting fusion protein was expressed on the cell surface. Using this new system, we constructed a novel starch-degrading strain of Lactobacillus casei by genetically displaying alpha-amylase from the Streptococcus bovis strain 148 with a FLAG peptide tag (AmyAF). Localization of the PgsA-AmyA-FLAG fusion protein on the cell surface was confirmed by immunofluorescence microscopy and flow cytometric analysis. The lactic acid bacteria which displayed AmyAF showed significantly elevated hydrolytic activity toward soluble starch. By fermentation using AmyAF-displaying L. casei cells, 50 g/liter of soluble starch was reduced to 13.7 g/liter, and 21.8 g/liter of lactic acid was produced within about 24 h. The yield in terms of grams of lactic acid produced per gram of carbohydrate utilized was 0.60 g per g of carbohydrate consumed at 24 h. Since AmyA was immobilized on the cells, cells were recovered after fermentation and used repeatedly. During repeated utilization of cells, the lactic acid yield was improved to 0.81 g per g of carbohydrate consumed at 72 h. These results indicate that efficient simultaneous saccharification and fermentation from soluble starch to lactic acid were carried out by recombinant L. casei cells with cell surface display of AmyA. PMID:16391053

  7. Proteomics of loosely bound cell wall proteins of Arabidopsis thaliana cell suspension cultures: a critical analysis.

    OpenAIRE

    Borderies, Gisèle; Jamet, Elisabeth; Lafitte, Claude; Rossignol, Michel; Jauneau, Alain; Boudart, Georges; Monsarrat, Bernard; Esquerré-Tugayé, Marie-Thérèse; Boudet, Alain; Pont-Lezica, Rafael

    2003-01-01

    The complete sequencing of the Arabidopsis thaliana genome allows the use of the recently developed mass spectrometry techniques to identify the cell wall proteins (CWPs). Most proteomic approaches depend on the quality of sample preparation. Extraction of CWPs is particularly complex since the proteins may be free in the apoplast or are embedded in a polysaccharide matrix where they are retained by Van der Waals interactions, hydrogen bonds, hydrophobic or ionic interactions, or cross-linked...

  8. Penium margaritaceum as a model organism for cell wall analysis of expanding plant cells

    DEFF Research Database (Denmark)

    Rydahl, Maja Gro; Fangel, Jonatan Ulrik; Mikkelsen, Maria Dalgaard;

    2015-01-01

    organization of the polymeric networks of the cell wall around the protoplast also contributes to the direction of growth, the shape of the cell, and the proper positioning of the cell in a tissue. In essence, plant cell expansion represents the foundation of development. Most studies of plant cell expansion...

  9. Growth arrest of lung carcinoma cells (A549) by polyacrylate-anchored peroxovanadate by activating Rac1-NADPH oxidase signalling axis.

    Science.gov (United States)

    Chatterjee, Nirupama; Anwar, Tarique; Islam, Nashreen S; Ramasarma, T; Ramakrishna, Gayatri

    2016-09-01

    Hydrogen peroxide is often required in sublethal, millimolar concentrations to show its oxidant effects on cells in culture as it is easily destroyed by cellular catalase. Previously, we had shown that diperoxovanadate, a physiologically stable peroxovanadium compound, can substitute H2O2 effectively in peroxidation reactions. We report here that peroxovanadate when anchored to polyacrylic acid (PAPV) becomes a highly potent inhibitor of growth of lung carcinoma cells (A549). The early events associated with PAPV treatment included cytoskeletal modifications, increase in GTPase activity of Rac1, accumulation of the reactive oxygen species, and also increase in phosphorylation of H2AX (γH2AX), a marker of DNA damage. These effects persisted even at 24 h after removal of the compound and culminated in increased levels of p53 and p21 together with growth arrest. The PAPV-mediated growth arrest was significantly abrogated in cells pre-treated with the N-acetylcysteine, Rac1 knocked down by siRNA and DPI an inhibitor of NADPH oxidase. In conclusion, our results show that polyacrylate derivative of peroxovanadate efficiently arrests growth of A549 cancerous cells by activating the axis of Rac1-NADPH oxidase leading to oxidative stress and DNA damage. PMID:27435854

  10. Recent advances on the posttranslational modifications of EXTs and their roles in plant cell walls

    DEFF Research Database (Denmark)

    Velasquez, Melina; Salter, Juan Salgado; Dorosz, Javier Gloazzo;

    2012-01-01

    The genetic set up and the enzymes that define the O-glycosylation sites and transfer the activated sugars to cell wall glycoprotein Extensins (EXTs) have remained unknown for a long time. We are now beginning to see the emerging components of the molecular machinery that assembles these complex O......-glycoproteins on the plant cell wall. Genes conferring the posttranslational modifications, i.e., proline hydroxylation and subsequent O-glycosylation, of the EXTs have been recently identified. In this review we summarize the enzymes that define the O-glycosylation sites on the O-glycoproteins, i.e., the prolyl 4...... and function of extensins in plant cell walls....

  11. Temperature Gradients on the Cell Wall in the Critical Viscosity Experiment

    Science.gov (United States)

    Berg, Robert F.; Moldover, Michael R.

    1993-01-01

    Because of the diverging susceptibility delta rho/delta Tau near the liquid-vapor critical point, temperature gradients must be kept small to maintain adequate sample homogeneity. In our Science Requirements Document we paid particular attention to radial density gradients caused by equilibration of the xenon sample. Axial density gradients were addressed through the requirement that the cell's copper wall have a gradient less than 22 microK/m. This report re-examines the cell wall's temperature distribution in more detail by estimating all known significant contributions to temperature differences on the cell's wall.

  12. Evidence for land plant cell wall biosynthetic mechanisms in charophyte green algae

    DEFF Research Database (Denmark)

    Mikkelsen, Maria Dalgaard; Harholt, Jesper; Ulvskov, Peter;

    2014-01-01

    BACKGROUND AND AIMS: The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. The cell wall has been suggested to be a defining structure that enabled the green algal ancestor to...... colonize land. These cell walls provide support and protection, are a source of signalling molecules, and provide developmental cues for cell differentiation and elongation. The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non......-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. The number of GTs in...

  13. Role of (1,3)(1,4)-β-glucan in cell walls: interaction with cellulose.

    Science.gov (United States)

    Kiemle, Sarah N; Zhang, Xiao; Esker, Alan R; Toriz, Guillermo; Gatenholm, Paul; Cosgrove, Daniel J

    2014-05-12

    (1,3)(1,4)-β-D-Glucan (mixed-linkage glucan or MLG), a characteristic hemicellulose in primary cell walls of grasses, was investigated to determine both its role in cell walls and its interaction with cellulose and other cell wall polysaccharides in vitro. Binding isotherms showed that MLG adsorption onto microcrystalline cellulose is slow, irreversible, and temperature-dependent. Measurements using quartz crystal microbalance with dissipation monitoring showed that MLG adsorbed irreversibly onto amorphous regenerated cellulose, forming a thick hydrogel. Oligosaccharide profiling using endo-(1,3)(1,4)-β-glucanase indicated that there was no difference in the frequency and distribution of (1,3) and (1,4) links in bound and unbound MLG. The binding of MLG to cellulose was reduced if the cellulose samples were first treated with certain cell wall polysaccharides, such as xyloglucan and glucuronoarabinoxylan. The tethering function of MLG in cell walls was tested by applying endo-(1,3)(1,4)-β-glucanase to wall samples in a constant force extensometer. Cell wall extension was not induced, which indicates that enzyme-accessible MLG does not tether cellulose fibrils into a load-bearing network. PMID:24678830

  14. The role of the secondary cell wall in plant resistance to pathogens.

    Science.gov (United States)

    Miedes, Eva; Vanholme, Ruben; Boerjan, Wout; Molina, Antonio

    2014-01-01

    Plant resistance to pathogens relies on a complex network of constitutive and inducible defensive barriers. The plant cell wall is one of the barriers that pathogens need to overcome to successfully colonize plant tissues. The traditional view of the plant cell wall as a passive barrier has evolved to a concept that considers the wall as a dynamic structure that regulates both constitutive and inducible defense mechanisms, and as a source of signaling molecules that trigger immune responses. The secondary cell walls of plants also represent a carbon-neutral feedstock (lignocellulosic biomass) for the production of biofuels and biomaterials. Therefore, engineering plants with improved secondary cell wall characteristics is an interesting strategy to ease the processing of lignocellulosic biomass in the biorefinery. However, modification of the integrity of the cell wall by impairment of proteins required for its biosynthesis or remodeling may impact the plants resistance to pathogens. This review summarizes our understanding of the role of the plant cell wall in pathogen resistance with a focus on the contribution of lignin to this biological process. PMID:25161657

  15. Rice Brittleness Mutants: A Way to Open the 'Black Box' of Monocot Cell Wall Biosynthesis

    Institute of Scientific and Technical Information of China (English)

    Baocai Zhang; Yihua Zhou

    2011-01-01

    Rice is a model organism for studying the mechanism of cell wall biosynthesis and remolding in Gramineae.Mechanical strength is an important agronomy trait of rice(Oryza sativa L.)plants that affects crop lodging and grain yield.As a prominent physical property of cell walls,mechanical strength reflects upon the structure of different wall polymers and how they interact.Studies on the mechanisms that regulate the mechanical strength therefore consequently results in uncovering the genes functioning in cell wall biosynthesis and remodeling.Our group focuses on the study of isolation of brittle culm(bc)mutants and characterization of their corresponding genes.To date,several bc mutants have been reported.The identified genes have covered several pathways of cell wall biosynthesis,revealing many secrets of monocot cell wall biosynthesis.Here,we review the progress achieved in this research field and also highlight the perspectives in expectancy.All of those lend new insights into mechanisms of cell wall formation and are helpful for harnessing the waste rice straws for biofuel production.

  16. Changes in cell wall architecture of wheat coleoptiles grown under continuous hypergravity conditions

    Science.gov (United States)

    Wakabayashi, K.; Soga, K.; Kamisaka, S.; Hoson, T.

    Modifications of cell wall structure of wheat coleoptiles in response to continuous hypergravity (300 g) treatment were investigated. Length of coleoptiles exposed to hypergravity for 2-4 days from germination stage was 60-70% of that of 1 g control. The net amounts of cell wall polysaccharides, such as hemicellulose and cellulose, of hypergravity-treated coleoptiles increased as much as those of 1 g control coleoptiles during the incubation period. As a result, the levels of cell wall polysaccharides per unit length of coleoptile, which mean the thickness of cell walls, largely increased under hypergravity conditions. Particularly, the amounts of hemicellulosic polymers with middle molecular mass (0.2-1 MDa) largely increased from day 2 to 3 under hypergravity conditions. The major sugar components of the hemicellulose fraction are arabinose, xylose and glucose. The ratios of arabinose and xylose to glucose were higher in hypergravity-treated coleoptiles than in control coleoptiles. The fractionation of hemicellulosic polymers into the neutral and acidic polymers by the anion-exchange column showed that the levels of acidic polymers (mainly composed of arabinoxylans) in cell walls of hypergravity-treated coleoptiles were higher than those of control coleoptiles. In addition to wall polysaccharides, the amounts of cell wall-bound phenolics, such as ferulic acid and diferulic acid, substantially increased during the incubation period both in 1 g control and hypergravity-treated coleoptiles. Especially, the levels of diferulic acid which cross-links hemicellulosic polymers were higher in hypergravity-treated coleoptiles than in control coleoptiles during the incubation period. These results suggest that hypergravity stimuli from the germination stage bias the type of synthesized hemicellulosic polysaccharides, although they do not restrict the net synthesis of cell wall constituents in wheat coleoptiles. The stimulation of the synthesis of arabinoxylans and of the

  17. Specific labeling of peptidoglycan precursors as a tool for bacterial cell wall studies

    NARCIS (Netherlands)

    van Dam, V.; Olrichs, N.K.; Breukink, E.J.

    2009-01-01

    Wall chart: The predominant component of the bacterial cell wall, peptidoglycan, consists of long alternating stretches of aminosugar subunits interlinked in a large three-dimensional network and is formed from precursors through several cytosolic and membrane-bound steps. The high tolerance of the

  18. Expression of S-adenosylmethionine Hydrolase in Tissues Synthesizing Secondary Cell Walls Alters Specific Methylated Cell Wall Fractions and Improves Biomass Digestibility

    Science.gov (United States)

    Eudes, Aymerick; Zhao, Nanxia; Sathitsuksanoh, Noppadon; Baidoo, Edward E. K.; Lao, Jeemeng; Wang, George; Yogiswara, Sasha; Lee, Taek Soon; Singh, Seema; Mortimer, Jenny C.; Keasling, Jay D.; Simmons, Blake A.; Loqué, Dominique

    2016-01-01

    Plant biomass is a large source of fermentable sugars for the synthesis of bioproducts using engineered microbes. These sugars are stored as cell wall polymers, mainly cellulose and hemicellulose, and are embedded with lignin, which makes their enzymatic hydrolysis challenging. One of the strategies to reduce cell wall recalcitrance is the modification of lignin content and composition. Lignin is a phenolic polymer of methylated aromatic alcohols and its synthesis in tissues developing secondary cell walls is a significant sink for the consumption of the methyl donor S-adenosylmethionine (AdoMet). In this study, we demonstrate in Arabidopsis stems that targeted expression of AdoMet hydrolase (AdoMetase, E.C. 3.3.1.2) in secondary cell wall synthesizing tissues reduces the AdoMet pool and impacts lignin content and composition. In particular, both NMR analysis and pyrolysis gas chromatography mass spectrometry of lignin in engineered biomass showed relative enrichment of non-methylated p-hydroxycinnamyl (H) units and a reduction of dimethylated syringyl (S) units. This indicates a lower degree of methylation compared to that in wild-type lignin. Quantification of cell wall-bound hydroxycinnamates revealed a reduction of ferulate in AdoMetase transgenic lines. Biomass from transgenic lines, in contrast to that in control plants, exhibits an enrichment of glucose content and a reduction in the degree of hemicellulose glucuronoxylan methylation. We also show that these modifications resulted in a reduction of cell wall recalcitrance, because sugar yield generated by enzymatic biomass saccharification was greater than that of wild-type plants. Considering that transgenic plants show no important diminution of biomass yields, and that heterologous expression of AdoMetase protein can be spatiotemporally optimized, this novel approach provides a valuable option for the improvement of lignocellulosic biomass feedstock. PMID:27486577

  19. Expression of S-adenosylmethionine Hydrolase in Tissues Synthesizing Secondary Cell Walls Alters Specific Methylated Cell Wall Fractions and Improves Biomass Digestibility.

    Science.gov (United States)

    Eudes, Aymerick; Zhao, Nanxia; Sathitsuksanoh, Noppadon; Baidoo, Edward E K; Lao, Jeemeng; Wang, George; Yogiswara, Sasha; Lee, Taek Soon; Singh, Seema; Mortimer, Jenny C; Keasling, Jay D; Simmons, Blake A; Loqué, Dominique

    2016-01-01

    Plant biomass is a large source of fermentable sugars for the synthesis of bioproducts using engineered microbes. These sugars are stored as cell wall polymers, mainly cellulose and hemicellulose, and are embedded with lignin, which makes their enzymatic hydrolysis challenging. One of the strategies to reduce cell wall recalcitrance is the modification of lignin content and composition. Lignin is a phenolic polymer of methylated aromatic alcohols and its synthesis in tissues developing secondary cell walls is a significant sink for the consumption of the methyl donor S-adenosylmethionine (AdoMet). In this study, we demonstrate in Arabidopsis stems that targeted expression of AdoMet hydrolase (AdoMetase, E.C. 3.3.1.2) in secondary cell wall synthesizing tissues reduces the AdoMet pool and impacts lignin content and composition. In particular, both NMR analysis and pyrolysis gas chromatography mass spectrometry of lignin in engineered biomass showed relative enrichment of non-methylated p-hydroxycinnamyl (H) units and a reduction of dimethylated syringyl (S) units. This indicates a lower degree of methylation compared to that in wild-type lignin. Quantification of cell wall-bound hydroxycinnamates revealed a reduction of ferulate in AdoMetase transgenic lines. Biomass from transgenic lines, in contrast to that in control plants, exhibits an enrichment of glucose content and a reduction in the degree of hemicellulose glucuronoxylan methylation. We also show that these modifications resulted in a reduction of cell wall recalcitrance, because sugar yield generated by enzymatic biomass saccharification was greater than that of wild-type plants. Considering that transgenic plants show no important diminution of biomass yields, and that heterologous expression of AdoMetase protein can be spatiotemporally optimized, this novel approach provides a valuable option for the improvement of lignocellulosic biomass feedstock. PMID:27486577

  20. Investigation on Adsorption of Lithospermum erythrorhizon onto Fungal Cell Wall Polysaccharides

    Institute of Scientific and Technical Information of China (English)

    孟琴; 薛莲

    2003-01-01

    A culture of Lithosperrnum erythrorhizon adsorbed on fungal cell wall polysaccharides, a novel bioadsorbent made from fungal cell wall, has been established in this paper. Three steps were involved in this immobilization. The first step was preparation of suspended plant cells from tightly aggregated plant cell clumps. The disassembled ratio of 0.715g·g-1 (the disassembled cells over total cells) was obtained under optimum condition for the enzymatic reaction. Then, the adsorption of plant cells onto fungal cell wall polysaccharides was conducted and the saturated capacity of 12g cell per gram of carrier was obtained in adsorption immobilization. Finally, the culture of cells adsorbed on fungal cell wall polysaccharides was compared with that of cells entrapped in alginate or suspension cell culture. While exposed to in situ liquid paraffin extraction coupled with cell culture, the shikonin productivity of immobilized cells by adsorption was 10.67g·L-1, which was 1.8 times of that in suspension culture and 1.5 times of that entrapped in alginate.

  1. Liquid crystal director fluctuations and surface anchoring by molecular simulation

    OpenAIRE

    Andrienko, D.; Germano, G; Allen, M. P.

    2002-01-01

    We propose a simple and reliable method to measure the liquid crystal surface anchoring strength by molecular simulation. The method is based on the measurement of the long-range fluctuation modes of the director in confined geometry. As an example, molecular simulations of a liquid crystal in slab geometry between parallel walls with homeotropic anchoring have been carried out using the Monte Carlo technique. By studying different slab thicknesses, we are able to calculate separately the pos...

  2. Cell Wall Growth and Modulation Dynamics in a Model Unicellular Green Alga—Penium margaritaceum: Live Cell Labeling with Monoclonal Antibodies

    OpenAIRE

    Domozych, David S; Hannah Brechka; Alicia Britton; Marc Toso

    2011-01-01

    Penium margaritaceum is a unicellular charophycean green alga that possesses cell wall polymers similar to those of land plants. Several wall macromolecules of this alga are recognized by monoclonal antibodies specific for wall polymer epitopes of land plants. Immunofluorescence protocols using these antibodies may be employed to label specific cell wall constituents of live cells. Fluorescent labeling persists for several days, and this attribute allows for tracing of wall epitopes in both l...

  3. Identification and characterization of the BCG cell wall-stimulated suppressor cells in inbred rats

    International Nuclear Information System (INIS)

    In vitro experiments were undertaken to investigate the effect of intraperitoneal (i.p.) inoculation of BCG cell walls attached to oil droplets (BCGcw) on mitogenic and alloantigenic responses of spleen and bone marrow cells. These in vitro studies demonstrated that: (1) spleen cells from BCGcw-immunized ACI rats had decreased responsiveness to Concanavalin A (Con A) and to alloantigenic stimulation, (2) depressed Con A reactivity could also be induced in Buffalo rat spleen cells by the i.p. inoculation of BCGcw, (3) normal ACI rats had suppressor cells in their bone marrow but not in their spleens, (4) BCGcw-immunized ACI rats demonstrated an increase in the suppressive activity of their bone marrow as early as 1 day after inoculation of BCGcw, while suppressor activity was found in the spleen as early as 2 days after BCGcw inoculation, (5) characterization of the BCGcw-induced splenic suppressor cell demonstrated it to be adherent to plastic or nylon wool, radiation-resistant, and removed by treatment with carbonyl iron. These properties were consistent with the identification of the suppressor cell as a macrophage, (6) the Con A and mixed lymphocyte reactivities of normal spleen cells could be suppressed by the addition of the adherent spleen cell population from BCGcw-immunized ACI rats, and (7) the adherent suppressor cell from BCGcw-immunized rats suppressed Con A reactivity across a major histocompatability barrier. (Auth.)

  4. Trifluoromethanesulfonic acid-based proteomic analysis of cell wall and secreted proteins of the ascomycetous fungi Neurospora crassa and Candida albicans

    OpenAIRE

    Maddi, Abhiram; Bowman, Shaun M.; Free, Stephen J.

    2009-01-01

    Cell wall proteins from purified Candida albicans and Neurospora crassa cell walls were released using trifluoromethanesulfonic acid (TFMS) which cleaves the cell wall glucan/chitin matrix and deglycosylates the proteins. The cell wall proteins were then characterized by SDS PAGE and identified by proteomic analysis. The analyses for C. albicans identified 15 cell wall proteins and 6 secreted proteins. For N. crassa, the analyses identified 26 cell wall proteins and 9 secreted proteins. Most ...

  5. Turnover of galactans and other cell wall polysaccharides during development of flax plants

    International Nuclear Information System (INIS)

    We investigated the synthesis and turnover of cell wall polysaccharides of the flax (Linum usitatissimum L.) plant during development of the phloem fibers. One-month-old flax plants were exposed to a 40-min pulse with 14CO2 followed by 8-h, 24-h, and 1-month periods of chase with ambient CO2, and radioactivity in cell wall sugars was determined in various plant parts. The relative radioactivity of glucose in noncellulosic polysaccharides was the highest compared with all other cell wall sugars immediately after the pulse and decreased substantially during the subsequent chase. The relative radioactivities of the other cell wall sugars changed with differing rates, indicating turnover of specific polysaccharides. Notably, after 1 month of chase there was a marked decrease in the proportional mass and total radioactivity in cell wall galactose, indicating a long-term turnover of the galactans enriched in the fiber-containing tissues. The ratio of radiolabeled xylose to arabinose also increased during the chase, indicating a turnover of arabinose-containing polymers and interconversion to xylose. The pattern of label redistribution differed between organs, indicating that the cell wall turnover processes are tissue- and cell-specific

  6. Ultrastructural changes of cell walls under intense mechanical treatment of selective plant raw material

    International Nuclear Information System (INIS)

    Structural changes of cell walls under intense mechanical treatment of corn straw and oil-palm fibers were studied by electron and light microscopy. Differences in the character of destruction of plant biomass were revealed, and the dependence of destruction mechanisms on the structure of cell walls and lignin content was demonstrated. We suggest that the high reactivity of the particles of corn straw (about 18% of lignin) after intense mechanical treatment is related to disordering of cell walls and an increase of the surface area, while in the case of oil palm (10% of lignin) the major contribution into an increase in the reactivity is made by an increase of surface area. -- Highlights: ► Structure of cell walls determines the processes of plant materials' destruction. ► Ultrastructure of highly lignified materials strongly disordering by mechanical action. ► Ultrastructure of low-lignified materials is not disordering by mechanical action.

  7. Change in cell wall lectin activity and some phytohormones in tobacco leaves during photoperiodic induction

    International Nuclear Information System (INIS)

    The change in cell wall lectin activity and some phytohormones in the Silvestris and Mamont tobacco leaves during photoperiodic induction is studied. The ratio between two types of tobaccos changes alike

  8. The Paracoccidioides cell wall: past and present layers towards understanding interaction with the host

    Directory of Open Access Journals (Sweden)

    RosanaPuccia

    2011-12-01

    Full Text Available The cell wall of pathogenic fungi plays import roles in interaction with the host, so that its composition and structure may determine the course of infection. Here we present an overview of the current and past knowledge on the cell wall constituents of Paracoccidioides brasiliensis and P. lutzii. These are temperature-dependent dimorphic fungi that cause paracoccidioidomycosis, a systemic granulomatous and debilitating disease. Focus is given on cell wall carbohydrate and protein contents, their immune-stimulatory features, adhesion properties, drug target characteristics, and morphological phase specificity. We offer a journey towards the future understanding of the dynamic life that takes place in the cell wall and of the changes that it may suffer when living in the human host.

  9. In Situ Chemical Imaging of Plant Cell Walls Using CARS/SRS Microscopy (Poster)

    Energy Technology Data Exchange (ETDEWEB)

    Zeng, Y.; Liu, Y. S.; Saar, B. G.; Xie, X. S.; Chen, F.; Dixon, R. A.; Himmel, M. E.; Ding S. Y.

    2009-06-01

    This poster demonstrates coherent anti-Stokes Raman scattering and stimulated Raman scattering of plant cell walls. It includes simultaneous chemical imaging of lignin and cellulose (corn stover) during acidic pretreatment.

  10. The relation of apple texture with cell wall nanostructure studied using an atomic force microscope.

    Science.gov (United States)

    Cybulska, Justyna; Zdunek, Artur; Psonka-Antonczyk, Katarzyna M; Stokke, Bjørn T

    2013-01-30

    In this study, the relation of the nanostructure of cell walls with their texture was investigated for six different apple cultivars. Cell wall material (CWM) and cellulose microfibrils were imaged by atomic force microscope (AFM). The mean diameter of cellulose microfibrils for each cultivar was estimated based on the AFM height topographs obtained using the tapping mode of dried specimens. Additionally, crystallinity of cellulose microfibrils and pectin content was determined. Texture of apple cultivars was evaluated by sensory and instrumental analysis. Differences in cellulose diameter as determined from the AFM height topographs of the nanostructure of cell walls of the apple cultivars are found to relate to the degree of crystallinity and pectin content. Cultivars with thicker cellulose microfibrils also revealed crisper, harder and juicier texture, and greater acoustic emission. The data suggest that microfibril thickness affects the mechanical strength of cell walls which has consequences for sensory and instrumental texture. PMID:23218275

  11. Cell wall degrading enzymes in Trichoderma asperellum grown on wheat bran

    DEFF Research Database (Denmark)

    Bech, Lasse; Busk, Peter Kamp; Lange, Lene

    2015-01-01

    Trichoderma asperellum is a filamentous fungus that is able to produce and secrete a wide range of extracellular hydrolytic enzymes used for plant cell wall degradation. The Trichoderma genus has attracted considerable attention from the biorefinery industry due to the production of cell wall...... degrading enzymes and strong secretion ability of this genus. Here we report extensive transcriptome analysis of plant cell wall degrading enzymes in T. asperellum. The production of cell wall degrading enzymes by T. asperellum was tested on a range of cellulosic materials under various conditions. When T...... the theory that the glycoside hydrolases have evolved from a common ancestor, followed by a specialization in which saprotrophic fungi such as T. reesei and T. longibrachiatum lost a significant number of genes including several glycoside hydrolases....

  12. Study of the cell wall of Staphylococcus aureus and its sensitivity to enzybiotics

    OpenAIRE

    Čmelík, R. (Richard); Melková, K.; Kobzová, Š.; Janda, L

    2015-01-01

    The endolysin resistant and sensitive strains of Staphylococcus aureus were compared by means of LC-MS based structural analysis of peptidoglycan isolated from their cell walls. The structural explanation of the resistance was suggested.

  13. 2012 PLANT CELL WALLS GORDON RESEARCH CONFERENCE AND GORDON RESEARCH SEMINAR, AUGUST 4-10, 2012

    Energy Technology Data Exchange (ETDEWEB)

    Rose, Jocelyn

    2012-08-10

    The sub-theme of this year’s meeting, ‘Cell Wall Research in a Post-Genome World’, will be a consideration of the dramatic technological changes that have occurred in the three years since the previous cell wall Gordon Conference in the area of DNA sequencing. New technologies are providing additional perspectives of plant cell wall biology across a rapidly growing number of species, highlighting a myriad of architectures, compositions, and functions in both "conventional" and specialized cell walls. This meeting will focus on addressing the knowledge gaps and technical challenges raised by such diversity, as well as our need to understand the underlying processes for critical applications such as crop improvement and bioenergy resource development.

  14. Coniferyl Ferulate Incorporation into Lignin Enhances the Alkaline Delignification and Enzymatic Degradation of Cell Walls

    Science.gov (United States)

    Incorporating ester interunit linkages into lignin could facilitate fiber delignification and utilization. In model studies with maize cell walls, we examined how partial substitution of coniferyl alcohol (a normal monolignol) with coniferyl ferulate (an ester conjugate from lignan biosynthesis) alt...

  15. Size, Shape, and Arrangement of Cellulose Microfibril in Higher Plant Cell Walls

    Energy Technology Data Exchange (ETDEWEB)

    Ding, S. Y.

    2013-01-01

    Plant cell walls from maize (Zea mays L.) are imaged using atomic force microscopy (AFM) at the sub-nanometer resolution. We found that the size and shape of fundamental cellulose elementary fibril (CEF) is essentially identical in different cell wall types, i.e., primary wall (PW), parenchyma secondary wall (pSW), and sclerenchyma secondary wall (sSW), which is consistent with previously proposed 36-chain model (Ding et al., 2006, J. Agric. Food Chem.). The arrangement of individual CEFs in these wall types exhibits two orientations. In PW, CEFs are horizontally associated through their hydrophilic faces, and the planar faces are exposed, forming ribbon-like macrofibrils. In pSW and sSW, CEFs are vertically oriented, forming layers, in which hemicelluloses are interacted with the hydrophobic faces of the CEF and serve as spacers between CEFs. Lignification occurs between CEF-hemicelluloses layers in secondary walls. Furthermore, we demonstrated quantitative analysis of plant cell wall accessibility to and digestibility by different cellulase systems at real-time using chemical imaging (e.g., stimulated Raman scattering) and fluorescence microscopy of labeled cellulases (Ding et al., 2012, Science, in press).

  16. Anchoring the Deficit of the Anchor Deficit: Dyslexia or Attention?

    Science.gov (United States)

    Willburger, Edith; Landerl, Karin

    2010-01-01

    In the anchoring deficit hypothesis of dyslexia ("Trends Cogn. Sci.", 2007; 11: 458-465), it is proposed that perceptual problems arise from the lack of forming a perceptual anchor for repeatedly presented stimuli. A study designed to explicitly test the specificity of the anchoring deficit for dyslexia is presented. Four groups, representing all…

  17. Solid-State NMR on bacterial cells: selective cell wall signal enhancement and resolution improvement using dynamic nuclear polarization

    International Nuclear Information System (INIS)

    Dynamic nuclear polarization (DNP) enhanced solid-state nuclear magnetic resonance (NMR) has recently emerged as a powerful technique for the study of material surfaces. In this study, we demonstrate its potential to investigate cell surface in intact cells. Using Bacillus subtilis bacterial cells as an example, it is shown that the polarizing agent 1-(TEMPO-4-oxy)-3-(TEMPO-4-amino)propan-2-ol (TOTAPOL) has a strong binding affinity to cell wall polymers (peptidoglycan). This particular interaction is thoroughly investigated with a systematic study on extracted cell wall materials, disrupted cells, and entire cells, which proved that TOTAPOL is mainly accumulating in the cell wall. This property is used on one hand to selectively enhance or suppress cell wall signals by controlling radical concentrations and on the other hand to improve spectral resolution by means of a difference spectrum. Comparing DNP-enhanced and conventional solid-state NMR, an absolute sensitivity ratio of 24 was obtained on the entire cell sample. This important increase in sensitivity together with the possibility of enhancing specifically cell wall signals and improving resolution really opens new avenues for the use of DNP-enhanced solid-state NMR as an on-cell investigation tool. (authors)

  18. Evaluation of the damage of cell wall and cell membrane for various extracellular polymeric substance extractions of activated sludge.

    Science.gov (United States)

    Guo, Xuesong; Liu, Junxin; Xiao, Benyi

    2014-10-20

    Extracellular polymeric substances (EPS) are susceptible to contamination by intracellular substances released during the extraction of EPS owing to the damage caused to microbial cell structures. The damage to cell walls and cell membranes in nine EPS extraction processes of activated sludge was evaluated in this study. The extraction of EPS (including proteins, carbohydrates and DNA) was the highest using the NaOH extraction method and the lowest using formaldehyde extraction. All nine EPS extraction methods in this study resulted in cell wall and membrane damage. The damage to cell walls, evaluated by 2-keto-3-deoxyoctonate (KDO) and N-acetylglucosamine content changes in extracted EPS, was the most significant in the NaOH extraction process. Formaldehyde extraction showed a similar extent of damage to cell walls to those detected in the control method (centrifugation), while those in the formaldehyde-NaOH and cation exchange resin extractions were slightly higher than those detected in the control. N-acetylglucosamine was more suitable than KDO for the evaluation of cell wall damage in the EPS extraction of activated sludge. The damage to cell membranes was characterized by two fluorochromes (propidium iodide and FITC Annexin V) with flow cytometry (FCM) measurement. The highest proportion of membrane-damaged cells was detected in NaOH extraction (26.54% of total cells) while membrane-damaged cells comprised 8.19% of total cells in the control. PMID:25173614

  19. Effects of X-irradiation on artificial blood vessel wall degradation by invasive tumor cells

    International Nuclear Information System (INIS)

    Artificial vessel wall cultures, constructed by growing arterial endothelial cells on preformed layers of rat smooth muscle cells, were used to evaluate the effects of X-irradiation on tumor cell-induced tissue degradation. Bovine endothelial cells had radiation sensitivities similar to those of rat smooth muscle cells. Preirradiation of smooth muscle cells, before the addition of human fibrosarcoma (HT 1080) cells, did not increase the rate of degradation and destruction by the invasive cells. However, the degradation rate was decreased if the cultures were irradiated after the addition of HT 1080 cells. The presence of bovine endothelial cells markedly inhibited the destructive abilities of fibrosarcoma cells, but preirradiation of artificial vessel walls substantially decreased their capabilities to resist HT 1080-induced lysis. These findings suggest that the abilities of blood vessels to limit extravasation may be compromised by ionizing radiation

  20. Sucrose synthase affects carbon partitioning to increase cellulose production and altered cell wall ultrastructure

    OpenAIRE

    Coleman, Heather D.; Yan, Jimmy; Mansfield, Shawn D

    2009-01-01

    Overexpression of the Gossypium hirsutum sucrose synthase (SuSy) gene under the control of 2 promoters was examined in hybrid poplar (Populus alba × grandidentata). Analysis of RNA transcript abundance, enzyme activity, cell wall composition, and soluble carbohydrates revealed significant changes in the transgenic lines. All lines showed significantly increased SuSy enzyme activity in developing xylem. This activity manifested in altered secondary cell wall cellulose content per dry weight in...

  1. Activity of quinones from teak (Tectona grandis) on fungal cell wall stress.

    Science.gov (United States)

    Sumthong, Pattarawadee; Damveld, Robbert A; Choi, Young H; Arentshorst, Mark; Ram, Arthur F; van den Hondel, Cees A; Verpoorte, Rob

    2006-08-01

    Teak ( Tectona grandis L.f., Verbenaceae) sawdust extract inhibited the growth of Aspergillus niger. Centrifugal partition chromatography was used to isolate the active compounds. By (1)H-NMR the active compounds were identified as deoxylapachol and tectoquinone. Two A. niger transgenic strains which show induction of 1,3 -alpha-D-glucan synthase were used as a cell wall damage model. The result showed that deoxylapachol from T. grandis extract induced fungal cell wall stress. PMID:16972200

  2. Fungal Cell Wall Dynamics and Infection Site Microenvironments: Signal Integration and Infection Outcome

    OpenAIRE

    Shepardson, Kelly M.; Cramer, Robert A.

    2013-01-01

    Upon entrance into the host, fungi encounter a myriad of host effector products and microenvironments that they sense and adapt to for survival. Alterations of the structure and composition of the cell wall is a major fungal adaptation mechanism to evade these environments. Here we discuss recent findings of host-microenvironmental induced fungal cell wall changes, including structure, composition, and protein content, and their effects on host immune responses. A take home message from these...

  3. The measurement technique of radiation shielding performance for hot cell walls in IMEF

    International Nuclear Information System (INIS)

    Hot cell is the facility to test irradiated materials. The capability of radiation shielding through wall should be conformed to protect the workers from expose. In this report, the measurement techniques of radiation shielding performance through hot cell walls are described. Detailed contents are as following; 1. The theory of test 2. The measuring equipment of radiations capability 3. The choice of measuring points 4. Test procedures and data analysis method 5. The reinforcement of shielding lack area. (author). 13 tabs., 19 figs

  4. Plant Cell Wall Proteins: A Large Body of Data, but What about Runaways?

    OpenAIRE

    Cécile Albenne; Hervé Canut; Laurent Hoffmann; Elisabeth Jamet

    2014-01-01

    Plant cell wall proteomics has been a very dynamic field of research for about fifteen years. A full range of strategies has been proposed to increase the number of identified proteins and to characterize their post-translational modifications. The protocols are still improving to enlarge the coverage of cell wall proteomes. Comparisons between these proteomes have been done based on various working strategies or different physiological stages. In this review, two points are highlighted. The ...

  5. Golgi-localized UDP-glucose transporter is required for cell wall integrity in rice

    OpenAIRE

    Song, Xueqin; Zhang, Baocai; Zhou, Yihua

    2011-01-01

    Cell wall-related nucleotide sugar transporters (NSTs) theoretically supply the cytosolic nucleotide sugars for glycosyltransferases (GTs) to carry out ploysaccharide synthesis and modification in the Golgi apparatus. However, the regulation of cell wall synthesis by NSTs remains undescribed. Recently, we have reported the functional characterization of Oryza sativa nucleotide sugar transport (Osnst1) mutant and its corresponding gene. OsNST1/BC14 is localized in the Golgi apparatus and trans...

  6. Changes in inositol phosphates in wild carrot cells upon initiation of cell wall digestion

    International Nuclear Information System (INIS)

    Previous studies have shown that inositol trisphosphate (IP3) stimulated 45Ca+2 efflux from fusogenic carrot protoplasts and it was suggested that IP3 may serve as a second messenger for the mobilization of intracellular Ca+2 in higher plant cells. To determine whether or not inositol phosphate metabolism changes in response to external stimuli, the cells were labeled with myo-[2-3H] inositol for 18 h and exposed to cell wall digestion enzymes, Driselase. The inositol phosphates were extracted with ice cold 10% TCA and separated by anion exchange chromatography. The radioactivity of the fraction that contained IP3 increased 2-3.8 fold and that which contained inositol bisphosphate increased 1.9-2.6 fold within 1.5 min of exposure to Driselase. After 6 min, the radioactivity of both fractions increased 6-7.7 fold and an increase in inositol monophosphate was observed. These data indicate that inositol phosphate metabolism is stimulated by Driselase and suggest polyphosphoinositide hydrolysis occurs upon initiation of cell wall digestion

  7. Arabinose-rich polymers as an evolutionary strategy to plasticize resurrection plant cell walls against desiccation.

    Science.gov (United States)

    Moore, John P; Nguema-Ona, Eric E; Vicré-Gibouin, Mäite; Sørensen, Iben; Willats, William G T; Driouich, Azeddine; Farrant, Jill M

    2013-03-01

    A variety of Southern African resurrection plants were surveyed using high-throughput cell wall profiling tools. Species evaluated were the dicotyledons, Myrothamnus flabellifolia and Craterostigma plantagineum; the monocotyledons, Xerophyta viscosa, Xerophyta schlecterii, Xerophyta humilis and the resurrection grass Eragrostis nindensis, as well as a pteridophyte, the resurrection fern, Mohria caffrorum. Comparisons were made between hydrated and desiccated leaf and frond material, with respect to cell wall composition and polymer abundance, using monosaccharide composition analysis, FT-IR spectroscopy and comprehensive microarray polymer profiling in combination with multivariate data analysis. The data obtained suggest that three main functional strategies appear to have evolved to prepare plant cell walls for desiccation. Arabinan-rich pectin and arabinogalactan proteins are found in the resurrection fern M. caffrorum and the basal angiosperm M. flabellifolia where they appear to act as 'pectic plasticizers'. Dicotyledons with pectin-rich walls, such as C. plantagineum, seem to use inducible mechanisms which consist of up-regulating wall proteins and osmoprotectants. The hemicellulose-rich walls of the grass-like Xerophyta spp. and the resurrection grass E. nindensis were found to contain highly arabinosylated xylans and arabinogalactan proteins. These data support a general mechanism of 'plasticising' the cell walls of resurrection plants to desiccation and implicate arabinose-rich polymers (pectin-arabinans, arabinogalactan proteins and arabinoxylans) as the major contributors in ensuring flexibility is maintained and rehydration is facilitated in these plants. PMID:23117392

  8. Structural Insight into Cell Wall Architecture of Micanthus sinensis cv. using Correlative Microscopy Approaches.

    Science.gov (United States)

    Ma, Jianfeng; Lv, Xunli; Yang, Shumin; Tian, Genlin; Liu, Xing'e

    2015-10-01

    Structural organization of the plant cell wall is a key parameter for understanding anisotropic plant growth and mechanical behavior. Four imaging platforms were used to investigate the cell wall architecture of Miscanthus sinensis cv. internode tissue. Using transmission electron microscopy with potassium permanganate, we found a great degree of inhomogeneity in the layering structure (4-9 layers) of the sclerenchymatic fiber (Sf). However, the xylem vessel showed a single layer. Atomic force microscopy images revealed that the cellulose microfibrils (Mfs) deposited in the primary wall of the protoxylem vessel (Pxv) were disordered, while the secondary wall was composed of Mfs oriented in parallel in the cross and longitudinal section. Furthermore, Raman spectroscopy images indicated no variation in the Mf orientation of Pxv and the Mfs in Pxv were oriented more perpendicular to the cell axis than that of Sfs. Based on the integrated results, we have proposed an architectural model of Pxv composed of two layers: an outermost primary wall composed of a meshwork of Mfs and inner secondary wall containing parallel Mfs. This proposed model will support future ultrastructural analysis of plant cell walls in heterogeneous tissues, an area of increasing scientific interest particularly for liquid biofuel processing. PMID:26358178

  9. Evidence against the involvement of ionically bound cell wall proteins in pea epicotyl growth

    Science.gov (United States)

    Melan, M. A.; Cosgrove, D. J.

    1988-01-01

    Ionically bound cell wall proteins were extracted from 7 day old etiolated pea (Pisum sativum L. cv Alaska) epicotyls with 3 molar LiCl. Polyclonal antiserum was raised in rabbits against the cell wall proteins. Growth assays showed that treatment of growing region segments (5-7 millimeters) of peas with either dialyzed serum, serum globulin fraction, affinity purified immunoglobulin, or papain-cleaved antibody fragments had no effect on growth. Immunofluorescence microscopy confirmed antibody binding to cell walls and penetration of the antibodies into the tissues. Western blot analysis, immunoassay results, and affinity chromatography utilizing Sepharose-bound antibodies confirmed recognition of the protein preparation by the antibodies. Experiments employing in vitro extension as a screening measure indicated no effect upon extension by antibodies, by 50 millimolar LiCl perfusion of the apoplast or by 3 molar LiCl extraction. Addition of cell wall protein to protease pretreated segments did not restore extension nor did addition of cell wall protein to untreated segments increase extension. It is concluded that, although evidence suggests that protein is responsible for the process of extension, the class(es) of proteins which are extracted from pea cell walls with 3 molar LiCl are probably not involved in this process.

  10. Ethanolic rhizome extract from Kaempferia parviflora Wall. ex. Baker induces apoptosis in HL-60 cells.

    Science.gov (United States)

    Banjerdpongchai, Ratana; Suwannachot, Kittiphan; Rattanapanone, Viboon; Sripanidkulchai, Bungorn

    2008-01-01

    Kaempferia parviflora Wall. ex. Baker is a Thai herb containing many flavonoids that have anti-inflammatory, anti-allergic and antioxidant activities. The objective of this study was to demonstrate apoptotic effects of Kaempferia parviflora Wall. ex. Baker rhizome ethanolic extract on HL-60 cells in vitro. The extract suppressed HL-60 cell growth and decreased cell viability in a dose- and time-dependent manner. Apoptotic cell death was demonstrated by changes in cell morphology, externalization of phosphatidylserine on the cell surface, loss in mitochondrial transmembrane potential and activation of caspase 3. Apoptosis induced by K. parviflora Wall. ex. Baker rhizome ethanolic extract was enhanced by treatment with paclitaxel or doxorubicin, and inhibitors of Akt, PI3-K and MEK. PMID:19256745

  11. Cell wall metabolism in ripening fruit. IX. Synthesis of pectic and hemicellulosic cell wall polymers in the outer pericarp of mature green tomatoes (cv XMT-22)

    International Nuclear Information System (INIS)

    Discs of outer pericarp were excised from mature green tomato (Lycopersicon esculentum Mill.) fruit and kept in sterile tissue culture plates for 4 d, including 2 d of incubation with D-[U-13C]glucose. Cell walls were prepared and the water-soluble, pectic, and hemicellulosic polymers were extracted. Cell wall synthetic capacity was determined by gas chromatography-mass spectrometry analysis of incorporation of the heavy isotope label. The 'outer' 2-mm pericarp region, which included the cuticle, had a lower cell wall synthetic capacity than the 'inner' 2-mm region immediately below it (closer to the locules), based on the percentage of labeling of the neutral sugars. There were no significant differences in relative abundance of glycosidic linkages in the two tissue regions. Label was incorporated into neutral sugars and linkages typical for each polysaccharide class were identified in the cell wall preparations. Galacturonic acid and glucuronic acid were labeled to an extent similar to that of the neutral sugars in each tissue region

  12. On-line wall-free cell for laser-induced fluorescence detection in capillary electrophoresis.

    Science.gov (United States)

    Yu, Chang-Zhu; He, You-Zhao; Xie, Hai-Yang; Gao, Yong; Gan, Wu-Er; Li, Jun

    2009-05-15

    A wall-free detection method based on liquid junction in a capillary gap was proposed for laser-induced fluorescence (LIF) of capillary electrophoresis (CE). The capillary gap of the wall-free cell was fabricated by etching a 10-mm x 50-microm I.D. fused-silica capillary to obtain a polyimide coating sleeve, decoating about 6mm at one end of both 50 microm I.D. separation and liquid junction capillary, inserting the treated capillary ends into the coating sleeve oppositely, fixing the capillaries with a gap distance of 140 microm by epoxy glue and removing the coating sleeve by burning. The theoretical model, experimental results and wall-free cell images indicated that the gap distance and applied voltage were main influence factors on the wall-free detection. Since the wall-free cell increased the absorption light path and avoided the stray light from the capillary wall, it improved the ratio of signal to noise and limit of detection (LOD) of CE-LIF. Three flavin compounds of riboflavin (RF), flavin mononucleotide sodium (FMN) and flavin adenine dinucleotide disodium (FAD) were used to evaluate the wall-free detection method. Compared with on-column cell, the LODs of the wall-free cell were improved 15-, 6- and 9-fold for RF, FMN and FAD, respectively. The linear calibration concentrations of the flavins ranged from 0.005 to 5.0 micromol/L. The column efficiency was in the range from 1.0 x 10(5) to 2.5 x 10(5) plates. The wall-free detection of CE-LIF was applied to the analysis of the flavins in spinach and lettuce leaves. PMID:19329123

  13. Bidimensional distortion in ferroelectric liquid crystals with strong anchoring in bookshelf geometry

    Indian Academy of Sciences (India)

    S Ponti; M Becchi; A Strigazzi; S Torgova

    2003-08-01

    In the last decade, it has been experimentally found that in certain conditions a periodic domain pattern arises in ferroelectric liquid crystals (FLC) in bookshelf geometry. Such a periodic texture appears after switching-off an external electric field, even in strong anchoring conditions. It has a static character and is bidimensional, being dependent on directions normal to both the smectic planes and the cell plates. Here a new model explaining this phenomenon is proposed, valid in the case of FLC with strong anchoring. The model is based on the coupling between the spontaneous polarization field in the first semi-period of its modulation along the cell plates and the same field in the second semi-period. This coupling competes with the FLC medium elasticity and is trapped by the anchoring. According to our model, in the ferroelectric state the biperiodic texture is favored by increasing the values of spontaneous polarization. The critical value of the spontaneous polarization for which the undeformed state becomes unstable is found. It is shown to be proportional to the square root of the ratio between the FLC elastic constant and the cell thickness. Moreover, it is inversely proportional to the sinus of the pre-tilt angle with respect to the cell walls.

  14. Functional Analysis of Cellulose and Xyloglucan in the Walls of Stomatal Guard Cells of Arabidopsis.

    Science.gov (United States)

    Rui, Yue; Anderson, Charles T

    2016-03-01

    Stomatal guard cells are pairs of specialized epidermal cells that control water and CO2 exchange between the plant and the environment. To fulfill the functions of stomatal opening and closure that are driven by changes in turgor pressure, guard cell walls must be both strong and flexible, but how the structure and dynamics of guard cell walls enable stomatal function remains poorly understood. To address this question, we applied cell biological and genetic analyses to investigate guard cell walls and their relationship to stomatal function in Arabidopsis (Arabidopsis thaliana). Using live-cell spinning disk confocal microscopy, we measured the motility of cellulose synthase (CESA)-containing complexes labeled by green fluorescent protein (GFP)-CESA3 and observed a reduced proportion of GFP-CESA3 particles colocalizing with microtubules upon stomatal closure. Imaging cellulose organization in guard cells revealed a relatively uniform distribution of cellulose in the open state and a more fibrillar pattern in the closed state, indicating that cellulose microfibrils undergo dynamic reorganization during stomatal movements. In cesa3(je5) mutants defective in cellulose synthesis and xxt1 xxt2 mutants lacking the hemicellulose xyloglucan, stomatal apertures, changes in guard cell length, and cellulose reorganization were aberrant during fusicoccin-induced stomatal opening or abscisic acid-induced stomatal closure, indicating that sufficient cellulose and xyloglucan are required for normal guard cell dynamics. Together, these results provide new insights into how guard cell walls allow stomata to function as responsive mediators of gas exchange at the plant surface. PMID:26729799

  15. Bacterial Wall Components such as Lipothecoid Acid, Peptidoglycan, Liposaccharide and Lipid A Stimulate Cell Proliferation in Intestinal Epithelial Cells

    OpenAIRE

    Olaya, Jaime H.; Neopikhanov, Vadim; Söderman, Charlotte; Uribe, Andrés

    2011-01-01

    Earlier studies indicate that the microflora contains mitogens to intestinal epithelial cells. Our aim is to examine whether cell wall components of both Gram-negative and positive bacteria influence cell proliferation in small intestinal and colonic epithelial cells. A human colonic epithelial cell line from adenocarcinoma (IEC-6) and a nontransformed small intestinal cell line from germ-free rats (LS-123) were incubated with (a) lipothecoid acid from Streptococcus faecalis at 1.56–50 ...

  16. Monovalent cations enable cell wall turnover of the turnover-deficient lyt-15 mutant of Bacillus subtilis.

    OpenAIRE

    Cheung, H. Y.; Freese, E

    1985-01-01

    A lyt-15 mutant reported to be unable to turn over the cell wall exhibited the same rate of wall turnover as the standard strain if the medium contained 0.2 M NaCl, which did not affect growth. Cell wall autolysis was also optimal at 0.2 M NaCl.

  17. Decreased Polysaccharide Feruloylation Compromises Plant Cell Wall Integrity and Increases Susceptibility to Necrotrophic Fungal Pathogens

    Science.gov (United States)

    Reem, Nathan T.; Pogorelko, Gennady; Lionetti, Vincenzo; Chambers, Lauran; Held, Michael A.; Bellincampi, Daniela; Zabotina, Olga A.

    2016-01-01

    The complexity of cell wall composition and structure determines the strength, flexibility, and function of the primary cell wall in plants. However, the contribution of the various components to cell wall integrity (CWI) and function remains unclear. Modifications of cell wall composition can induce plant responses known as CWI control. In this study, we used transgenic expression of the fungal feruloyl esterase AnFAE to examine the effect of post-synthetic modification of Arabidopsis and Brachypodium cell walls. Transgenic Arabidopsis plants expressing AnFAE showed a significant reduction of monomeric ferulic acid, decreased amounts of wall-associated extensins, and increased susceptibility to Botrytis cinerea, compared with wild type. Transgenic Brachypodium showed reductions in monomeric and dimeric ferulic acids and increased susceptibility to Bipolaris sorokiniana. Upon infection, transgenic Arabidopsis and Brachypodium plants also showed increased expression of several defense-related genes compared with wild type. These results demonstrate a role, in both monocot and dicot plants, of polysaccharide feruloylation in plant CWI, which contributes to plant resistance to necrotrophic pathogens. PMID:27242834

  18. Decreased Polysaccharide Feruloylation Compromises Plant Cell Wall Integrity and Increases Susceptibility to Necrotrophic Fungal Pathogens.

    Science.gov (United States)

    Reem, Nathan T; Pogorelko, Gennady; Lionetti, Vincenzo; Chambers, Lauran; Held, Michael A; Bellincampi, Daniela; Zabotina, Olga A

    2016-01-01

    The complexity of cell wall composition and structure determines the strength, flexibility, and function of the primary cell wall in plants. However, the contribution of the various components to cell wall integrity (CWI) and function remains unclear. Modifications of cell wall composition can induce plant responses known as CWI control. In this study, we used transgenic expression of the fungal feruloyl esterase AnFAE to examine the effect of post-synthetic modification of Arabidopsis and Brachypodium cell walls. Transgenic Arabidopsis plants expressing AnFAE showed a significant reduction of monomeric ferulic acid, decreased amounts of wall-associated extensins, and increased susceptibility to Botrytis cinerea, compared with wild type. Transgenic Brachypodium showed reductions in monomeric and dimeric ferulic acids and increased susceptibility to Bipolaris sorokiniana. Upon infection, transgenic Arabidopsis and Brachypodium plants also showed increased expression of several defense-related genes compared with wild type. These results demonstrate a role, in both monocot and dicot plants, of polysaccharide feruloylation in plant CWI, which contributes to plant resistance to necrotrophic pathogens. PMID:27242834

  19. Sorption of volatile phenols by yeast cell walls

    OpenAIRE

    Nerea Jiménez-Moreno; Carmen Ancín-Azpilicueta

    2009-01-01

    Nerea Jiménez-Moreno, Carmen Ancín-AzpilicuetaDepartment of Applied Chemistry, Universidad Pública de Navarra, Pamplona, SpainAbstract: Yeast walls can retain different wine compounds and so its use is interesting in order to eliminate harmful substances from the must which affect alcoholic fermentation (medium chain fatty acids) or which affect wine quality in a negative way (ethyl phenols, ochratoxin A). The aim of this study was to examine the capacity of c...

  20. The best time of cytotoxicity for extracted cell wall from Lactobacillus casei and paracasei in K562 cell line

    Directory of Open Access Journals (Sweden)

    Riki M

    2013-02-01

    Full Text Available Background: The aim of this study was to evaluate the effect of extracted cell walls from Lactobacillus casei and Lactobacillus paracasei as probiotic bacteria (isolated from common carp intestine on K562 and the role of cell concentration on the results of MTT [3-(4,5-Dimethylthiazol-2-yl2,5- Diphenyl tetrazolium Bromide] test.Methods: For this purpose, bacteria were cultured in specific medium (MRS broth at anaerobic condition for 24-48 hour. After incubation period culture medium was centri-fuged, then the cells were washed twice with PBS buffer to remove additional medium. Finally, collected bacterial cell disrupted by Sonication and cell walls were separated from other components by centrifugation. After that, different concentrations of cell walls (500, 1000, 2000 and 4000 µg/ml were prepared in RPMI medium for each bacteria, separately. Then anticancer properties of the cell walls were determined in vitro at 12, 24, 48 and 72 h, also the effect of K562 concentration was assayed with MTT technique.Results: The results showed extracted cell wall from both probiotic statistically (P=0.098 have anti turmeric properties in K562 and their properties will arise in relation with concentration. As well as, we found that the number of cell had not any affect on the result of MTT assay.Conclusion: We conclude that the cytotoxicity property of extracted cell wall is related in the type of bacteria, but this anticancer property would warrant further study on the clinical application of extracted cell wall.

  1. Inorganic polyphosphate occurs in the cell wall of Chlamydomonas reinhardtii and accumulates during cytokinesis

    Directory of Open Access Journals (Sweden)

    Freimoser Florian M

    2007-09-01

    Full Text Available Abstract Background Inorganic polyphosphate (poly P, linear chains of phosphate residues linked by energy rich phosphoanhydride bonds, is found in every cell and organelle and is abundant in algae. Depending on its localization and concentration, poly P is involved in various biological functions. It serves, for example, as a phosphate store and buffer against alkali, is involved in energy metabolism and regulates the activity of enzymes. Bacteria defective in poly P synthesis are impaired in biofilm development, motility and pathogenicity. PolyP has also been found in fungal cell walls and bacterial envelopes, but has so far not been measured directly or stained specifically in the cell wall of any plant or alga. Results Here, we demonstrate the presence of poly P in the cell wall of Chlamydomonas reinhardtii by staining with specific poly P binding proteins. The specificity of the poly P signal was verified by various competition experiments, by staining with different poly P binding proteins and by correlation with biochemical quantification. Microscopical investigation at different time-points during growth revealed fluctuations of the poly P signal synchronous with the cell cycle: The poly P staining peaked during late cytokinesis and was independent of the high intracellular poly P content, which fluctuated only slightly during the cell cycle. Conclusion The presented staining method provides a specific and sensitive tool for the study of poly P in the extracellular matrices of algae and could be used to describe the dynamic behaviour of cell wall poly P during the cell cycle. We assume that cell wall poly P and intracellular poly P are regulated by distinct mechanisms and it is suggested that cell wall bound poly P might have important protective functions against toxic compounds or pathogens during cytokinesis, when cells are more vulnerable.

  2. Robotic Ankle for Omnidirectional Rock Anchors

    Science.gov (United States)

    Parness, Aaron; Frost, Matthew; Thatte, Nitish

    2013-01-01

    Future robotic exploration of near-Earth asteroids and the vertical and inverted rock walls of lava caves and cliff faces on Mars and other planetary bodies would require a method of gripping their rocky surfaces to allow mobility without gravitational assistance. In order to successfully navigate this terrain and drill for samples, the grippers must be able to produce anchoring forces in excess of 100 N. Additionally, the grippers must be able to support the inertial forces of a moving robot, as well gravitational forces for demonstrations on Earth. One possible solution would be to use microspine arrays to anchor to rock surfaces and provide the necessary load-bearing abilities for robotic exploration of asteroids. Microspine arrays comprise dozens of small steel hooks supported on individual suspensions. When these arrays are dragged along a rock surface, the steel hooks engage with asperities and holes on the surface. The suspensions allow for individual hooks to engage with asperities while the remaining hooks continue to drag along the surface. This ensures that the maximum possible number of hooks engage with the surface, thereby increasing the load-bearing abilities of the gripper. Using the microspine array grippers described above as the end-effectors of a robot would allow it to traverse terrain previously unreachable by traditional wheeled robots. Furthermore, microspine-gripping robots that can perch on cliffs or rocky walls could enable a new class of persistent surveillance devices for military applications. In order to interface these microspine grippers with a legged robot, an ankle is needed that can robotically actuate the gripper, as well as allow it to conform to the large-scale irregularities in the rock. The anchor serves three main purposes: deploy and release the anchor, conform to roughness or misalignment with the surface, and cancel out any moments about the anchor that could cause unintentional detachment. The ankle design contains a

  3. Interactions of the cell-wall glycopolymers of lactic acid bacteria with their bacteriophages

    Directory of Open Access Journals (Sweden)

    Marie-Pierre eChapot-Chartier

    2014-05-01

    Full Text Available Lactic acid bacteria (LAB are Gram positive bacteria widely used in the production of fermented food in particular cheese and yoghurts. Bacteriophage infections during fermentation processes have been for many years a major industrial concern and have stimulated numerous research efforts. Better understanding of the molecular mechanisms of bacteriophage interactions with their host bacteria is required for the development of efficient strategies to fight against infections. The bacterial cell wall plays key roles in these interactions. First, bacteriophages must adsorb at the bacterial surface through specific interactions with receptors that are cell wall components. At next step, phages must overcome the barrier constituted by cell wall peptidoglycan to inject DNA inside bacterial cell. Also at the end of the infection cycle, phages synthesize endolysins able to hydrolyze peptidoglycan and lyse bacterial cells to release phage progeny. In the last decade, concomitant development of genomics and structural analysis of cell wall components allowed considerable advances in the knowledge of their structure and function in several model LAB. Here, we describe the present knowledge on the structure of the cell wall glycopolymers of the best characterized LAB emphasizing their structural variations and we present the available data regarding their role in bacteria-phage specific interactions at the different steps of the infection cycle.

  4. Targeting and Regulation of Cell Wall Synthesis During Tip Growth in Plants

    Institute of Scientific and Technical Information of China (English)

    Fangwei Gu; Erik Nielsen

    2013-01-01

    Root hairs and pollen tubes are formed through tip growth, a process requiring synthesis of new cell wall material and the precise targeting and integration of these components to a selected apical plasma membrane domain in the growing tips of these cells. Presence of a tip-focused calcium gradient, control of actin cytoskeleton dynamics, and formation and targeting of secretory vesicles are essential to tip growth. Similar to cells undergoing diffuse growth, cellulose, hemi-celluloses, and pectins are also deposited in the growing apices of tip-growing cells. However, differences in the manner in which these cell wall components are targeted and inserted in the expanding portion of tip-growing cells is reflected by the identification of elements of the plant cell wall synthesis machinery which have been shown to play unique roles in tip-growing cells. In this review, we summarize our current understanding of the tip growth process, with a particular focus on the subcellular targeting of newly synthesized cell wall components, and their roles in this form of plant cell expansion.

  5. Measuring plant cell wall extension (creep) induced by acidic pH and by alpha-expansin.

    Science.gov (United States)

    Durachko, Daniel M; Cosgrove, Daniel J

    2009-01-01

    Growing plant cell walls characteristically exhibit a property known as 'acid growth', by which we mean they are more extensible at low pH (extensometer measurements. To measure acid-induced extension, the walls are initially buffered at neutral pH, resulting in low activity of expansins that are components of the native cell walls. Upon buffer exchange to acidic pH, expansins are activated and the cell walls extend rapidly. We also demonstrate expansin activity in a reconstitution assay. For this part, we use a brief heat treatment to denature the native expansins in the cell wall samples. These inactivated cell walls do not extend even in acidic buffer, but addition of expansins to the cell walls rapidly restores their ability to extend. PMID:19279553

  6. Cell-cell adhesion mediated by binding of membrane-anchored transforming growth factor α to epidermal growth factor receptors promotes cell proliferation

    International Nuclear Information System (INIS)

    The precursor for transforming growth factor α, pro-TGF-α, is a cell surface glycoprotein that can establish contact with epidermal growth factor (EGF) receptors on adjacent cells. To examine whether the pro-TGF-α/EGF receptor pair can simultaneously mediate cell adhesion and promote cell proliferation, the authors have expressed pro-TGF-α in a bone marrow stromal cell line labeled with [35S] cysteine. Expression of pro-TGF-α allows these cells to support long-term attachment of an EGF/interleukin-3-dependent hematopoietic progenitor cell line that expresses EGF receptors but is unable to adhere to normal stroma. This interaction is inhibited by soluble EGF receptor ligands. Further, the hematopoietic progenitor cells replicate their DNA while they are attached to the stromal cell layer and become foci of sustained cell proliferation. Thus, pro-TGF-α and the EGF receptor can function as mediators of intercellular adhesion and this interaction may promote a mitogenic response. They propose the term juxtacrine to designate this form of stimulation between adjacent cells

  7. Extracellular Vesicle-Associated Transitory Cell Wall Components and Their Impact on the Interaction of Fungi with Host Cells.

    Science.gov (United States)

    Nimrichter, Leonardo; de Souza, Marcio M; Del Poeta, Maurizio; Nosanchuk, Joshua D; Joffe, Luna; Tavares, Patricia de M; Rodrigues, Marcio L

    2016-01-01

    Classic cell wall components of fungi comprise the polysaccharides glucans and chitin, in association with glycoproteins and pigments. During the last decade, however, system biology approaches clearly demonstrated that the composition of fungal cell walls include atypical molecules historically associated with intracellular or membrane locations. Elucidation of mechanisms by which many fungal molecules are exported to the extracellular space suggested that these atypical components are transitorily located to the cell wall. The presence of extracellular vesicles (EVs) at the fungal cell wall and in culture supernatants of distinct pathogenic species suggested a highly functional mechanism of molecular export in these organisms. Thus, the passage of EVs through fungal cell walls suggests remarkable molecular diversity and, consequently, a potentially variable influence on the host antifungal response. On the basis of information derived from the proteomic characterization of fungal EVs from the yeasts Cryptoccocus neoformans and Candida albicans and the dimorphic fungi Histoplasma capsulatum and Paracoccidioides brasiliensis, our manuscript is focused on the clear view that the fungal cell wall is much more complex than previously thought. PMID:27458437

  8. Extracellular Vesicle-Associated Transitory Cell Wall Components and Their Impact on the Interaction of Fungi with Host Cells

    Science.gov (United States)

    Nimrichter, Leonardo; de Souza, Marcio M.; Del Poeta, Maurizio; Nosanchuk, Joshua D.; Joffe, Luna; Tavares, Patricia de M.; Rodrigues, Marcio L.

    2016-01-01

    Classic cell wall components of fungi comprise the polysaccharides glucans and chitin, in association with glycoproteins and pigments. During the last decade, however, system biology approaches clearly demonstrated that the composition of fungal cell walls include atypical molecules historically associated with intracellular or membrane locations. Elucidation of mechanisms by which many fungal molecules are exported to the extracellular space suggested that these atypical components are transitorily located to the cell wall. The presence of extracellular vesicles (EVs) at the fungal cell wall and in culture supernatants of distinct pathogenic species suggested a highly functional mechanism of molecular export in these organisms. Thus, the passage of EVs through fungal cell walls suggests remarkable molecular diversity and, consequently, a potentially variable influence on the host antifungal response. On the basis of information derived from the proteomic characterization of fungal EVs from the yeasts Cryptoccocus neoformans and Candida albicans and the dimorphic fungi Histoplasma capsulatum and Paracoccidioides brasiliensis, our manuscript is focused on the clear view that the fungal cell wall is much more complex than previously thought. PMID:27458437

  9. 2009 Plant Cell Walls Gordon Research Conference-August 2-7,2009

    Energy Technology Data Exchange (ETDEWEB)

    Debra Mohnen

    2009-08-07

    Plant cell walls are a complex cellular compartment essential for plant growth, development and response to biotic and abiotic stress and a major biological resource for meeting our future bioenergy and natural product needs. The goal of the 2009 Plant Cell Walls Gordon Research Conference is to summarize and critically evaluate the current level of understanding of the structure, synthesis and function of the whole plant extracellular matrix, including the polysaccharides, proteins, lignin and waxes that comprise the wall, and the enzymes and regulatory proteins that drive wall synthesis and modification. Innovative techniques to study how both primary and secondary wall polymers are formed and modified throughout plant growth will be emphasized, including rapid advances taking place in the use of anti-wall antibodies and carbohydrate binding proteins, comparative and evolutionary wall genomics, and the use of mutants and natural variants to understand and identify wall structure-function relationships. Discussions of essential research advances needed to push the field forward toward a systems biology approach will be highlighted. The meeting will include a commemorative lecture in honor of the career and accomplishments of the late Emeritus Professor Bruce A. Stone, a pioneer in wall research who contributed over 40 years of outstanding studies on plant cell wall structure, function, synthesis and remodeling including emphasis on plant cell wall beta-glucans and arabinogalactans. The dwindling supply of fossil fuels will not suffice to meet our future energy and industrial product needs. Plant biomass is the renewable resource that will fill a large part of the void left by vanishing fossil fuels. It is therefore critical that basic research scientists interact closely with industrial researchers to critically evaluate the current state of knowledge regarding how plant biomass, which is largely plant cell walls, is synthesized and utilized by the plant. A final

  10. Single-molecule imaging of the H-ras membrane-anchor reveals domains in the cytoplasmic leaflet of the cell membrane

    CERN Document Server

    Lommerse, Piet H M; Cognet, Laurent; Harms, Gregory S; Snaar-Jagalska, B Ewa; Spaink, Herman P; Schmidt, Thomas

    2004-01-01

    In the last decade evidence has accumulated that small domains of 50-700 nm in diameter are located in the exoplasmic leaflet of the plasma membrane. Most of these domains supposedly consist of specific sets of lipids and proteins, and are believed to coordinate signal transduction cascades. Whether similar domains are also present in the cytoplasmic leaflet of the plasma membrane is unclear so far. To investigate the presence of cytoplasmic leaflet domains, the H-Ras membrane-targeting sequence was fused to the C-terminus of the enhanced yellow fluorescent protein. Using single-molecule fluorescence microscopy, trajectories of individual molecules diffusing in the cytoplasmic leaflet of the plasma membrane were recorded. From these trajectories, the diffusion of individual membrane-anchored enhanced yellow fluorescent protein molecules was studied in live cells on timescales from 5 to 200 ms. The results show that the diffusion of 30-40% of the molecules is constrained in domains with a typical size of 200 n...

  11. Dissipative Particle Dynamics Simulation of Polymer- and Cell-Wall Depletion in Micro-Channels

    Science.gov (United States)

    Fedosov, Dmitry A.; Caswell, Bruce; Em Karniadakis, George

    2008-07-01

    A rising interest in physics of biological systems stimulates a great number of experiments and numerical simulations involving a variety of biological entities. These include bio-polymers and bio-molecules, real organism vesicles and capsules, artificial vesicles used in drug delivery and cells. Macromolecules, vesicles and cells are subject to wall depletion layers observed near solid-fluid interfaces. In the case of red blood cells depletion is often called the cell-free layer and is observed near blood vessel walls. We employ Dissipative Particle Dynamics (DPD) to model depletion layers in biological systems. In case of bio-polymers the simulated depletion layers compare well with the asymptotic lattice theory solution of depletion near a repulsive wall. Vesicles and cells are modeled as coarse-grained cell membranes described by in-plane viscoelastic energy, bending energy, area and volume constraints. We investigate cell-wall depletion for cells having vesicle-like shape and red blood cells, and we correlate our results with membrane coarse-graining and with material properties such as membrane stretching and bending stiffness.

  12. Aleurone Cell Walls of Wheat Grain: High Spatial Resolution Investigation Using Synchrotron Infrared Microspectroscopy

    International Nuclear Information System (INIS)

    Infrared microspectroscopy and immunolabeling techniques were employed in order to obtain deeper insight into the biochemical nature of aleurone cell walls of wheat grain. The use of a synchrotron source, thanks to its intrinsic brightness, has provided unprecedented information at the level of a few micrometers and has allowed the discrimination of various polysaccharides in cell walls. The high spectral quality obtained in the small analyzed domain has been beneficial in estimating the relative proportions of Β-glucan and arabinoxylan, through the use of principal component analysis (PCA). The highest amount of Β-glucan is found in periclinal cell walls close to the starchy endosperm. The junction regions between aleurone cells are enriched in arabinoxylan. At the early stage of wheat grain development (271 degrees D), the chemical composition along the cell walls is more heterogeneous than at the mature stage. Both synchrotron infrared microspectroscopy and immunolabeling experiments made it possible to reveal the spatial heterogeneity of the various chemical compositions of aleurone cell walls.

  13. Photoinhibition of stem elongation by blue and red light: effects on hydraulic and cell wall properties

    Science.gov (United States)

    Kigel, J.; Cosgrove, D. J.

    1991-01-01

    The underlying mechanism of photoinhibition of stem elongation by blue (BL) and red light (RL) was studied in etiolated seedlings of pea (Pisum sativum L. cv Alaska). Brief BL irradiations resulted in fast transient inhibition of elongation, while a delayed (lag approximately 60 minutes) but prolonged inhibition was observed after brief RL. Possible changes in the hydraulic and wall properties of the growing cells during photoinhibition were examined. Cell sap osmotic pressure was unaffected by BL and RL, but both irradiations increased turgor pressure by approximately 0.05 megapascal (pressure-probe technique). Cell wall yielding was analyzed by in vivo stress relaxation (pressure-block technique). BL and RL reduced the initial rate of relaxation by 38 and 54%, while the final amount of relaxation was decreased by 48 and 10%, respectively. These results indicate that RL inhibits elongation mainly by lowering the wall yield coefficient, while most of the inhibitory effect of BL was due to an increase of the yield threshold. Mechanical extensibility of cell walls (Instron technique) was decreased by BL and RL, mainly due to a reduction in the plastic component of extensibility. Thus, photoinhibitions of elongation by both BL and RL are achieved through changes in cell wall properties, and are not due to effects on the hydraulic properties of the cell.

  14. Strong adhesion of Saos-2 cells to multi-walled carbon nanotubes

    Energy Technology Data Exchange (ETDEWEB)

    Matsuoka, Makoto, E-mail: matsuoka@den.hokudai.ac.jp [Department of Oral and Maxillofacial Surgery, Graduate School of Dental Medicine, Hokkaido University, Kita 13 Nishi 7, Kita-ku, Sapporo 060-8586 (Japan); Akasaka, Tsukasa [Department of Dental Materials and Engineering, Graduate School of Dental Medicine, Hokkaido University, Kita 13 Nishi 7, Kita-ku, Sapporo 060-8586 (Japan); Totsuka, Yasunori [Department of Oral and Maxillofacial Surgery, Graduate School of Dental Medicine, Hokkaido University, Kita 13 Nishi 7, Kita-ku, Sapporo 060-8586 (Japan); Watari, Fumio [Department of Dental Materials and Engineering, Graduate School of Dental Medicine, Hokkaido University, Kita 13 Nishi 7, Kita-ku, Sapporo 060-8586 (Japan)

    2010-10-15

    In recent years, carbon nanotubes (CNTs) have been considered potential biomedical materials because of their unique character. The aim of this study was to investigate the response of a human osteoblast-like cell line - Saos-2 - on single-walled CNTs (SWCNTs) and multi-walled CNTs (MWCNTs). The surface of a culture dish was coated with CNTs, and Saos-2 cells were cultured for three days. Cell morphology, viability, alkaline phosphatase (ALP) activity, adhesion, and vinculin expression were evaluated. The result showed high cell viability and strong adhesion to MWCNTs. Saos-2 cultured on MWCNTs exhibited vinculin expression throughout the cell body, while the cells attached to SWCNTs and glass were mostly limited to their periphery. Our results suggest that CNT coatings promote cell activity and adhesiveness. These findings indicate that MWCNTs could be used as surface coating materials to promote cell adhesion.

  15. Isolated abdominal wall metastasis from renal cell carcinoma: Unusual presentation

    OpenAIRE

    Suresh Kumar; Pramod Kumar Sharma; Malay Kumar Bera

    2015-01-01

    Fifty-seven-year-old gentleman, who was a known victim of left-sided clear cell renal cell carcinoma (RCC), presented with isolated huge parietal swelling in left anterolateral aspect of abdomen. He had undergone open left radical nephrectomy 2 years back. Parietal swelling was widely excised and histopathology revealed clear cell RCC, nuclear Fuhrman grade 2.

  16. Chalcone Synthase (CHS) Gene Suppression in Flax Leads to Changes in Wall Synthesis and Sensing Genes, Cell Wall Chemistry and Stem Morphology Parameters.

    Science.gov (United States)

    Zuk, Magdalena; Działo, Magdalena; Richter, Dorota; Dymińska, Lucyna; Matuła, Jan; Kotecki, Andrzej; Hanuza, Jerzy; Szopa, Jan

    2016-01-01

    The chalcone synthase (CHS) gene controls the first step in the flavonoid biosynthesis. In flax, CHS down-regulation resulted in tannin accumulation and reduction in lignin synthesis, but plant growth was not affected. This suggests that lignin content and thus cell wall characteristics might be modulated through CHS activity. This study investigated the possibility that CHS affects cell wall sensing as well as polymer content and arrangement. CHS-suppressed and thus lignin-reduced plants showed significant changes in expression of genes involved in both synthesis of components and cell wall sensing. This was accompanied by increased levels of cellulose and hemicellulose. CHS-reduced flax also showed significant changes in morphology and arrangement of the cell wall. The stem tissue layers were enlarged averagely twofold compared to the control, and the number of fiber cells more than doubled. The stem morphology changes were accompanied by reduction of the crystallinity index of the cell wall. CHS silencing induces a signal transduction cascade that leads to modification of plant metabolism in a wide range and thus cell wall structure. PMID:27446124

  17. Chalcone Synthase (CHS) Gene Suppression in Flax Leads to Changes in Wall Synthesis and Sensing Genes, Cell Wall Chemistry and Stem Morphology Parameters

    Science.gov (United States)

    Zuk, Magdalena; Działo, Magdalena; Richter, Dorota; Dymińska, Lucyna; Matuła, Jan; Kotecki, Andrzej; Hanuza, Jerzy; Szopa, Jan

    2016-01-01

    The chalcone synthase (CHS) gene controls the first step in the flavonoid biosynthesis. In flax, CHS down-regulation resulted in tannin accumulation and reduction in lignin synthesis, but plant growth was not affected. This suggests that lignin content and thus cell wall characteristics might be modulated through CHS activity. This study investigated the possibility that CHS affects cell wall sensing as well as polymer content and arrangement. CHS-suppressed and thus lignin-reduced plants showed significant changes in expression of genes involved in both synthesis of components and cell wall sensing. This was accompanied by increased levels of cellulose and hemicellulose. CHS-reduced flax also showed significant changes in morphology and arrangement of the cell wall. The stem tissue layers were enlarged averagely twofold compared to the control, and the number of fiber cells more than doubled. The stem morphology changes were accompanied by reduction of the crystallinity index of the cell wall. CHS silencing induces a signal transduction cascade that leads to modification of plant metabolism in a wide range and thus cell wall structure. PMID:27446124

  18. The Plant Cell Wall: A Complex and Dynamic Structure As Revealed by the Responses of Genes under Stress Conditions

    Science.gov (United States)

    Houston, Kelly; Tucker, Matthew R.; Chowdhury, Jamil; Shirley, Neil; Little, Alan

    2016-01-01

    The plant cell wall has a diversity of functions. It provides a structural framework to support plant growth and acts as the first line of defense when the plant encounters pathogens. The cell wall must also retain some flexibility, such that when subjected to developmental, biotic, or abiotic stimuli it can be rapidly remodeled in response. Genes encoding enzymes capable of synthesizing or hydrolyzing components of the plant cell wall show differential expression when subjected to different stresses, suggesting they may facilitate stress tolerance through changes in cell wall composition. In this review we summarize recent genetic and transcriptomic data from the literature supporting a role for specific cell wall-related genes in stress responses, in both dicot and monocot systems. These studies highlight that the molecular signatures of cell wall modification are often complex and dynamic, with multiple genes appearing to respond to a given stimulus. Despite this, comparisons between publically available datasets indicate that in many instances cell wall-related genes respond similarly to different pathogens and abiotic stresses, even across the monocot-dicot boundary. We propose that the emerging picture of cell wall remodeling during stress is one that utilizes a common toolkit of cell wall-related genes, multiple modifications to cell wall structure, and a defined set of stress-responsive transcription factors that regulate them. PMID:27559336

  19. Pectic arabinan side chains are essential for pollen cell wall integrity during pollen development

    DEFF Research Database (Denmark)

    Cankar, Katarina; Kortstee, Anne; Toonen, Marcel A.J.;

    2014-01-01

    Pectin is a complex polysaccharide and an integral part of the primary plant cell wall and middle lamella, contributing to cell wall mechanical strength and cell adhesion. To understand the structure-function relationships of pectin in the cell wall, a set of transgenic potato lines with altered...... pectin composition was analysed. The expression of genes encoding enzymes involved in pectin acetylation, degradation of the rhamnogalacturonan backbone and type and length of neutral side chains, arabinan and galactan in particular, has been altered. Upon crossing of different transgenic lines, some...... transgenes were not transmitted to the next generation when these lines were used as a pollen donor, suggesting male sterility. Viability of mature pollen was severely decreased in potato lines with reduced pectic arabinan, but not in lines with altered galactan side chains. Anthers and pollen of different...

  20. Enhanced lipid recovery from Nannochloropsis microalgae by treatment with optimized cell wall degrading enzyme mixtures.

    Science.gov (United States)

    Zuorro, Antonio; Miglietta, Selenia; Familiari, Giuseppe; Lavecchia, Roberto

    2016-07-01

    A statistical mixture design approach was used to investigate the effects of cell wall degrading enzymes on the recovery of lipids from Nannochloropsis sp. A preliminary screening of potentially suitable enzyme preparations, including lysozyme, cellulase and different types of hemicellulases, was carried out. The most effective preparations were then taken as basic components for the formulation of enzyme mixtures. Optimized ternary mixtures consisting of cellulase and two hemicellulases were obtained which allowed the recovery of up to 37.2g of lipids per 100g of dry biomass. SEM and TEM images of the enzymatically treated microalga revealed extensive cell damage, with degradation of the cell wall and release of intracellular material. Overall, the results obtained demonstrate that the mixture design method can be used to prepare cell wall degrading enzyme cocktails that can significantly improve the recovery of lipids or other valuable components from microalgae. PMID:27078205

  1. Chemical origin of the space-charge layer in cuprous oxide front-wall solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Herion, J.

    1979-05-01

    Using Auger electron spectroscopy, the region near the front contact of cuprous oxide front-wall solar cells was investigated. In cells showing large photovoltages, a maximum of the copper concentration being by about 4 at.% percent higher than the bulk concentration was observed at a distance of 70 A from the metal-semiconductor interface. If the photovoltage was low a copper maximum adjacent to the interfacial layer was found, instead. It is concluded that changes of the stoichiometry of cuprous oxide must be taken into account in order to explain the origin of the space-charge layer in Cu/sub 2/O front-wall cells.

  2. Identification and characterization of genes involved in Arabidopsis thaliana cell wall acetylation

    OpenAIRE

    de Souza, Amancio Jose

    2014-01-01

    Most non-cellulosic plant cell wall polysaccharides including the hemicellulose xyloglucan and the pectic polysaccharides can be O-acetylated. This feature has direct significance in the use of these polymers in the food and biofuel industry. For example, increased pectin acetylation can reduce its gelling abilities and is hence detrimental in its application as a food thickener or emulsifier. In general, plant biomass with wall polymers with high acetate content can negatively influence biom...

  3. Single Walled Carbon Nanotubes Exhibit Dual-Phase Regulation to Exposed Arabidopsis Mesophyll Cells

    Science.gov (United States)

    Yuan, Hengguang; Hu, Shanglian; Huang, Peng; Song, Hua; Wang, Kan; Ruan, Jing; He, Rong; Cui, Daxiang

    2011-12-01

    Herein we are the first to report that single-walled carbon nanotubes (SWCNTs) exhibit dual-phase regulation to Arabidopsis mesophyll cells exposed to different concentration of SWCNTs. The mesophyll protoplasts were prepared by enzyme digestion, and incubated with 15, 25, 50, 100 μg/ml SWCNTs for 48 h, and then were observed by optical microscopy and transmission electron microscopy, the reactive oxygen species (ROS) generation was measured. Partial protoplasts were stained with propidium iodide and 4'-6- diamidino-2-phenylindole, partial protoplasts were incubated with fluorescein isothiocyanate-labeled SWCNTs, and observed by fluorescence microscopy. Results showed that SWCNTs could traverse both the plant cell wall and cell membrane, with less than or equal to 50 μg/ml in the culture medium, SWCNTs stimulated plant cells to grow out trichome clusters on their surface, with more than 50 μg/ml SWCNTs in the culture medium, SWCNTs exhibited obvious toxic effects to the protoplasts such as increasing generation of ROS, inducing changes of protoplast morphology, changing green leaves into yellow, and inducing protoplast cells' necrosis and apoptosis. In conclusion, single walled carbon nanotubes can get through Arabidopsis mesophyll cell wall and membrane, and exhibit dose-dependent dual-phase regulation to Arabidopsis mesophyll protoplasts such as low dose stimulating cell growth, and high dose inducing cells' ROS generation, necrosis or apoptosis.

  4. Single Walled Carbon Nanotubes Exhibit Dual-Phase Regulation to Exposed Arabidopsis Mesophyll Cells

    Directory of Open Access Journals (Sweden)

    Huang Peng

    2011-01-01

    Full Text Available Abstract Herein we are the first to report that single-walled carbon nanotubes (SWCNTs exhibit dual-phase regulation to Arabidopsis mesophyll cells exposed to different concentration of SWCNTs. The mesophyll protoplasts were prepared by enzyme digestion, and incubated with 15, 25, 50, 100 μg/ml SWCNTs for 48 h, and then were observed by optical microscopy and transmission electron microscopy, the reactive oxygen species (ROS generation was measured. Partial protoplasts were stained with propidium iodide and 4'-6- diamidino-2-phenylindole, partial protoplasts were incubated with fluorescein isothiocyanate-labeled SWCNTs, and observed by fluorescence microscopy. Results showed that SWCNTs could traverse both the plant cell wall and cell membrane, with less than or equal to 50 μg/ml in the culture medium, SWCNTs stimulated plant cells to grow out trichome clusters on their surface, with more than 50 μg/ml SWCNTs in the culture medium, SWCNTs exhibited obvious toxic effects to the protoplasts such as increasing generation of ROS, inducing changes of protoplast morphology, changing green leaves into yellow, and inducing protoplast cells' necrosis and apoptosis. In conclusion, single walled carbon nanotubes can get through Arabidopsis mesophyll cell wall and membrane, and exhibit dose-dependent dual-phase regulation to Arabidopsis mesophyll protoplasts such as low dose stimulating cell growth, and high dose inducing cells' ROS generation, necrosis or apoptosis.

  5. Hydroxycinnamate Conjugates as Potential Monolignol Replacements: In vitro Lignification and Cell Wall Studies with Rosmarinic Acid

    Energy Technology Data Exchange (ETDEWEB)

    Yuki, Tobimatsu; Sasikumar, Elumalai; Grabber, John H.; Davidson, Christy L.; Xuejun, Pan; John, Ralph

    2012-04-01

    The plasticity of lignin biosynthesis should permit the inclusion of new compatible phenolic monomers, such as rosmarinic acid (RA) and analogous catechol derivatives, into cell-wall lignins that are consequently less recalcitrant to biomass processing. In vitro lignin polymerization experiments revealed that RA readily underwent peroxidase-catalyzed copolymerization with monolignols and lignin oligomers to form polymers with new benzodioxane inter-unit linkages. Incorporation of RA permitted extensive depolymerization of synthetic lignins by mild alkaline hydrolysis, presumably by cleavage of ester intra-unit linkages within RA. Copolymerization of RA with monolignols into maize cell walls by in situ peroxidases significantly enhanced alkaline lignin extractability and promoted subsequent cell wall saccharification by fungal enzymes. Incorporating RA also improved cell wall saccharification by fungal enzymes and by rumen microflora even without alkaline pretreatments, possibly by modulating lignin hydrophobicity and/or limiting cell wall cross-linking. Consequently, we anticipate that bioengineering approaches for partial monolignol substitution with RA and analogous plant hydroxycinnamates would permit more efficient utilization of plant fiber for biofuels or livestock production.

  6. Detection of Cell Wall Chemical Variation in Zea Mays Mutants Using Near-Infrared Spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Buyck, N.; Thomas, S.

    2001-01-01

    Corn stover is regarded as the prime candidate feedstock material for commercial biomass conversion in the United States. Variations in chemical composition of Zea mays cell walls can affect biomass conversion process yields and economics. Mutant lines were constructed by activating a Mu transposon system. The cell wall chemical composition of 48 mutant families was characterized using near-infrared (NIR) spectroscopy. NIR data were analyzed using a multivariate statistical analysis technique called Principal Component Analysis (PCA). PCA of the NIR data from 349 maize leaf samples reveals 57 individuals as outliers on one or more of six Principal Components (PCs) at the 95% confidence interval. Of these, 19 individuals from 16 families are outliers on either PC3 (9% of the variation) or PC6 (1% of the variation), the two PCs that contain information about cell wall polymers. Those individuals for which altered cell wall chemistry is confirmed with wet chemical analysis will then be subjected to fermentation analysis to determine whether or not biomass conversion process kinetics, yields and/or economics are significantly affected. Those mutants that provide indications for a decrease in process cost will be pursued further to identify the gene(s) responsible for the observed changes in cell wall composition and associated changes in process economics. These genes will eventually be incorporated into maize breeding programs directed at the development of a truly dual use crop.

  7. WD40-repeat proteins in plant cell wall formation: current evidence and research prospects

    Directory of Open Access Journals (Sweden)

    Gea eGuerriero

    2015-12-01

    Full Text Available The metabolic complexity of living organisms relies on supramolecular protein structures which ensure vital processes, such as signal transduction, transcription, translation and cell wall synthesis. In eukaryotes WD40-repeat (WDR proteins often function as molecular hubs mediating supramolecular interactions. WDR proteins may display a variety of interacting partners and participate in the assembly of complexes involved in distinct cellular functions. In plants, the formation of lignocellulosic biomass involves extensive synthesis of cell wall polysaccharides, a process that requires the assembly of large transmembrane enzyme complexes, intensive vesicle trafficking, interactions with the cytoskeleton, and coordinated gene expression. Because of their function as supramolecular hubs, WDR proteins could participate in each or any of these steps, although to date only few WDR proteins have been linked to the cell wall by experimental evidence. Nevertheless, several potential cell wall-related WDR proteins were recently identified using in silico aproaches, such as analyses of co-expression, interactome and conserved gene neighbourhood. Notably, some WDR genes are frequently genomic neighbours of genes coding for GT2-family polysaccharide synthases in eukaryotes, and this WDR-GT2 collinear microsynteny is detected in diverse taxa. In angiosperms, two WDR genes are collinear to cellulose synthase genes, CESAs, whereas in ascomycetous fungi several WDR genes are adjacent to chitin synthase genes, chs. In this Perspective we summarize and discuss experimental and in silico studies on the possible involvement of WDR proteins in plant cell wall formation. The prospects of biotechnological engineering for enhanced biomass production are discussed.

  8. Altered cell wall disassembly during ripening of Cnr tomato fruit: implications for cell adhesion and fruit softening

    DEFF Research Database (Denmark)

    Orfila, C.; Huisman, M.M.H.; Willats, William George Tycho;

    2002-01-01

    The Cnr (Colourless non-ripening) tomato (Lycopersicon esculentum Mill.) mutant has an aberrant fruit-ripening phenotype in which fruit do not soften and have reduced cell adhesion between pericarp cells. Cell walls from Cnr fruit were analysed in order to assess the possible contribution of pectic...... found in the solubility and composition of the pectic polysaccharides extracted from the CWM at both stages of development. In comparison with the wild type, the ripening-associated solubilisation of homogalacturonan-rich pectic polysaccharides was reduced in Cnr. The proportion of carbohydrate that was...... larger amounts of galactosyl- and arabinosyl-containing polysaccharides that were tightly bound in the cell wall and could only be extracted with 4 M KOH, or remained in the insoluble residue. The complexity of the cell wall alterations that occur during fruit ripening and the significance of different...

  9. Transcriptome and biochemical analysis reveals that suppression of GPI-anchor synthesis leads to autophagy and possible necroptosis in Aspergillus fumigatus.

    Directory of Open Access Journals (Sweden)

    Jianghong Yan

    Full Text Available Previously, it has been shown that GPI proteins are required for cell wall synthesis and organization in Aspergillus fumigatus, a human opportunistic pathogen causing life-threatening invasive aspergillosis (IA in immunocompromised patients. Blocking GPI anchor synthesis leads to severe phenotypes such as cell wall defects, increased cell death, and attenuated virulence. However, the mechanism by which these phenotypes are induced is unclear. To gain insight into global effects of GPI anchoring in A. fumigatus, in this study a conditional expression mutant was constructed and a genome wide transcriptome analysis was carried out. Our results suggested that suppression of GPI anchor synthesis mainly led to activation of phosphatidylinositol (PtdIns signaling and ER stress. Biochemical and morphological evidence showed that autophagy was induced in response to suppression of the GPI anchor synthesis, and also an increased necroptosis was observed. Based on our results, we propose that activation of PtdIns3K and increased cytosolic Ca(2+, which was induced by both ER stress and PtdIns signaling, acted as the main effectors to induce autophagy and possible necroptosis.

  10. Temporal sequence of cell wall disassembly events in developing fruits. 2. Analysis of blueberry (Vaccinium species).

    Science.gov (United States)

    Vicente, Ariel R; Ortugno, Claudia; Rosli, Hernan; Powell, Ann L T; Greve, L Carl; Labavitch, John M

    2007-05-16

    Softening and pathogen susceptibility are the major factors limiting the marketing of blueberries as fresh fruits, and these traits are associated with fruit cell wall structure. However, few studies that characterize wall modifications occurring during development and ripening have been reported for this fruit. In this study the ripening-associated modifications of blueberry fruit cell walls (composition, pectin and hemicellulose solubilization, and depolymerization) at five stages of ripeness have been analyzed. Xylose was found to be the most abundant noncellulosic neutral sugar associated with fruit walls, and the observed high Xyl/Glc ratio suggested that xylans, which are usually a minor hemicellulosic fruit wall component, are abundant in blueberry. The pectic matrix showed increased solubilization at early and intermediate stages of ripening, but no changes were detected in late ripening. Furthermore, little reduction in pectin polymer size occurred during blueberry ripening. In contrast, hemicellulose levels decreased as ripening progressed, and a clear depolymerization of these components was observed. A model for cell wall degradation in this fruit is discussed. PMID:17428068

  11. Evaluation of Tritium Behavior in the Epoxy Painted Concrete Wall of ITER Hot Cell

    International Nuclear Information System (INIS)

    Tritium behavior released in the ITER hot cell has been investigated numerically using a combined analytical methods of a tritium transport analysis in the multi-layer wall (concrete and epoxy paint) with the one dimensional diffusion model and a tritium concentration analysis in the hot cell with the complete mixing model by the ventilation. As the results, it is revealed that tritium concentration decay and permeation issues are not serious problem in a viewpoint of safety, since it is expected that tritium concentration in the hot cell decrease rapidly within several days just after removing the tritium release source, and tritium permeation through the epoxy painted concrete wall will be negligible as long as the averaged realistic diffusion coefficient is ensured in the concrete wall. It is also revealed that the epoxy paint on the concrete wall prevents the tritium inventory increase in the concrete wall greatly (two orders of magnitudes), but still, the inventory in the wall is estimated to reach about 0.1 PBq for 20 years operation

  12. Procedure for load cell calibration at ELSA Reaction Wall

    OpenAIRE

    MOLINA RUIZ Francisco Javier; Pegon, Pierre; PERONI MARCO; Viaccoz, Bernard; PETIT Patrick

    2013-01-01

    This report describes the procedure currently applied for the calibration of the load cells used for the mechanical experiments in the ELSA laboratory. The procedure is based on the international norm ISO 7500-1 and the definitions there proposed. The calibration experiment consists of applying a number of load cycles simultaneously on the object load cell in series with a traceable measuring proving instrument externally calibrated (reference load cell). The accuracy of the measures of the o...

  13. How endogenous plant cell-wall degradation mechanisms can help achieve higher efficiency in saccharification of biomass.

    Science.gov (United States)

    Tavares, Eveline Q P; De Souza, Amanda P; Buckeridge, Marcos S

    2015-07-01

    Cell-wall recalcitrance to hydrolysis still represents one of the major bottlenecks for second-generation bioethanol production. This occurs despite the development of pre-treatments, the prospect of new enzymes, and the production of transgenic plants with less-recalcitrant cell walls. Recalcitrance, which is the intrinsic resistance to breakdown imposed by polymer assembly, is the result of inherent limitations in its three domains. These consist of: (i) porosity, associated with a pectin matrix impairing trafficking through the wall; (ii) the glycomic code, which refers to the fine-structural emergent complexity of cell-wall polymers that are unique to cells, tissues, and species; and (iii) cellulose crystallinity, which refers to the organization in micro- and/or macrofibrils. One way to circumvent recalcitrance could be by following cell-wall hydrolysis strategies underlying plant endogenous mechanisms that are optimized to precisely modify cell walls in planta. Thus, the cell-wall degradation that occurs during fruit ripening, abscission, storage cell-wall mobilization, and aerenchyma formation are reviewed in order to highlight how plants deal with recalcitrance and which are the routes to couple prospective enzymes and cocktail designs with cell-wall features. The manipulation of key enzyme levels in planta can help achieving biologically pre-treated walls (i.e. less recalcitrant) before plants are harvested for bioethanol production. This may be helpful in decreasing the costs associated with producing bioethanol from biomass. PMID:25922489

  14. Changes of myoid and endothelial cells in the peritubular wall during contraction of the seminiferous tubule.

    Science.gov (United States)

    Losinno, Antonella D; Sorrivas, Viviana; Ezquer, Marcelo; Ezquer, Fernando; López, Luis A; Morales, Alfonsina

    2016-08-01

    The wall of the seminiferous tubule in rodents consists of an inner layer of myoid cells covered by an outer layer of endothelial cells. Myoid cells are a type of smooth muscle cell containing α-actin filaments arranged in two independent layers that contract when stimulated by endothelin-1. The irregular surface relief of the tubular wall is often considered a hallmark of contraction induced by a variety of stimuli. We examine morphological changes of the rat seminiferous tubule wall during contraction by a combination of light, confocal, transmission and scanning electron microscopy. During ET-1-induced contraction, myoid cells changed from a flat to a conical shape, but their actin filaments remained in independent layers. As a consequence of myoid cell contraction, the basement membrane became wavy, orientation of collagen fibers in the extracellular matrix was altered and the endothelial cell layer became folded. To observe the basement of the myoid cell cone, the endothelial cell monolayer was removed by collagenase digestion prior to SEM study. In contracted tubules, it is possible to distinguish cell relief: myoid cells have large folds on the external surface oriented parallel to the tubular axis, whereas endothelial cells have numerous cytoplasmic projections facing the interstitium. The myoid cell cytoskeleton is unusual in that the actin filaments are arranged in two orthogonal layers, which adopt differing shapes during contraction with myoid cells becoming cone-shaped. This arrangement impacts on other components of the seminiferous tubule wall and affects the propulsion of the tubular contents to the rete testis. PMID:26987820

  15. The Natural Product Citral Can Cause Significant Damage to the Hyphal Cell Walls of Magnaporthe grisea

    Directory of Open Access Journals (Sweden)

    Rong-Yu Li

    2014-07-01

    Full Text Available In order to find a natural alternative to the synthetic fungicides currently used against the devastating rice blast fungus, Magnaporthe grisea, this study explored the antifungal potential of citral and its mechanism of action. It was found that citral not only inhibited hyphal growth of M. grisea, but also caused a series of marked hyphal morphological and structural alterations. Specifically, citral was tested for antifungal activity against M. grisea in vitro and was found to significantly inhibit colony development and mycelial growth with IC50 and IC90 values of 40.71 and 203.75 μg/mL, respectively. Furthermore, citral reduced spore germination and germ tube length in a concentration-dependent manner. Following exposure to citral, the hyphal cell surface became wrinkled with folds and cell breakage that were observed under scanning electron microscopy (SEM. There was damage to hyphal cell walls and membrane structures, loss of villous-like material outside of the cell wall, thinning of the cell wall, and discontinuities formed in the cell membrane following treatment based on transmission electron microscopy (TEM. This increase in chitinase activity both supports the morphological changes seen in the hyphae, and also suggests a mechanism of action. In conclusion, citral has strong antifungal properties, and treatment with this compound is capable of causing significant damage to the hyphal cell walls of M. grisea.

  16. α-1,6-Mannosylation of N-Linked Oligosaccharide Present on Cell Wall Proteins Is Required for Their Incorporation into the Cell Wall in the Filamentous Fungus Neurospora crassa▿†

    OpenAIRE

    Maddi, Abhiram; Free, Stephen J.

    2010-01-01

    The enzyme α-1,6-mannosyltransferase (OCH-1) is required for the synthesis of galactomannans attached to the N-linked oligosaccharides of Neurospora crassa cell wall proteins. The Neurospora crassa och-1 mutant has a tight colonial phenotype and a defective cell wall. A carbohydrate analysis of the och-1 mutant cell wall revealed a 10-fold reduction in the levels of mannose and galactose and a total lack of 1,6-linked mannose residues. Analysis of the integral cell wall protein from wild-type...

  17. Brown algae overproduce cell wall polysaccharides as a protection mechanism against the heavy metal toxicity

    International Nuclear Information System (INIS)

    Brown algae are often used as heavy metal biomonitors and biosorbents because they can accumulate high concentrations of metals. Cation-exchange performed by cell wall polysaccharides is pointed out as the main chemical mechanism for the metal sequestration. Here, we biochemically investigated if the brown alga Padina gymnospora living in a heavy metal contaminated area would modify their polysaccharidic content. We exposed non-living biomass to Cd and Pb and studied the metals adsorption and localization. We found that raw dried polysaccharides, sulfate groups, uronic acids, fucose, mannose, and galactose were significantly higher in contaminated algae compared with the control ones. Metal concentrations adsorbed by non-living biomass were rising comparatively to the tested concentrations. Electron microscopy showed numerous granules in the cell walls and X-ray microanalysis revealed Cd as the main element. We concluded that P. gymnospora overproduces cell wall polysaccharides when exposed to high metal concentrations as a defense mechanism.

  18. Abiotic and enzymatic degradation of wheat straw cell wall: a biochemical and ultrastructural investigation.

    Science.gov (United States)

    Lequart, C; Ruel, K; Lapierre, C; Pollet, B; Kurek, B

    2000-07-14

    The action of an abiotic lignin oxidant and a diffusible xylanase on wheat straw was studied and characterized at the levels of the molecular structures by chemical analysis and of the cell wall ultrastructure by transmission electron microscopy. While distinct chemical changes in the target polymers were observed when each system was used separately, a combination of the two types of catalysts did not significantly increase either lignin oxidation or hemicellulose hydrolysis. Microscopic observations however revealed that the supramolecular organization of the cell wall polymers was significantly altered. This suggests that the abiotic Mn-oxalate complex and the xylanase cooperate in modifying the cell wall architecture, without noticeably enhancing the degradation of the constitutive polymers. PMID:10949315

  19. Chemical characterisation and analysis of the cell wall polysaccharides of duckweed (Lemna minor).

    Science.gov (United States)

    Zhao, X; Moates, G K; Wellner, N; Collins, S R A; Coleman, M J; Waldron, K W

    2014-10-13

    Duckweed is potentially an ideal biofuel feedstock due to its high proportion of cellulose and starch and low lignin content. However, there is little detailed information on the composition and structure of duckweed cell walls relevant to optimising the conversion of duckweed biomass to ethanol and other biorefinery products. This study reports that, for the variety and batch evaluated, carbohydrates constitute 51.2% (w/w) of dry matter while starch accounts for 19.9%. This study, for the first time, analyses duckweed cell wall composition through a detailed sequential extraction. The cell wall is rich in cellulose and also contains 20.3% pectin comprising galacturonan, xylogalacturonan, rhamnogalacturonan; 3.5% hemicellulose comprising xyloglucan and xylan, and 0.03% phenolics. In addition, essential fatty acids (0.6%, α-linolenic and linoleic/linoelaidic acid) and p-coumaric acid (0.015%) respectively are the most abundant fatty acids and phenolics in whole duckweed. PMID:25037369

  20. Cell-wall disruption and lipid/astaxanthin extraction from microalgae: Chlorella and Haematococcus.

    Science.gov (United States)

    Kim, Dong-Yeon; Vijayan, Durairaj; Praveenkumar, Ramasamy; Han, Jong-In; Lee, Kyubock; Park, Ji-Yeon; Chang, Won-Seok; Lee, Jin-Suk; Oh, You-Kwan

    2016-01-01

    Recently, biofuels and nutraceuticals produced from microalgae have emerged as major interests, resulting in intensive research of the microalgal biorefinery process. In this paper, recent developments in cell-wall disruption and extraction methods are reviewed, focusing on lipid and astaxanthin production from the biotechnologically important microalgae Chlorella and Haematococcus, respectively. As a common, critical bottleneck for recovery of intracellular components such as lipid and astaxanthin from these microalgae, the composition and structure of rigid, thick cell-walls were analyzed. Various chemical, physical, physico-chemical, and biological methods applied for cell-wall breakage and lipid/astaxanthin extraction from Chlorella and Haematococcus are discussed in detail and compared based on efficiency, energy consumption, type and dosage of solvent, biomass concentration and status (wet/dried), toxicity, scalability, and synergistic combinations. This report could serve as a useful guide to the implementation of practical downstream processes for recovery of valuable products from microalgae including Chlorella and Haematococcus. PMID:26342788

  1. Saccharomyces Cerevisiae Cell Wall Components as Tools for Ochratoxin A Decontamination

    Directory of Open Access Journals (Sweden)

    Małgorzata Piotrowska

    2015-04-01

    Full Text Available The aim of this study was to evaluate the usefulness of Saccharomyces cerevisiae cell wall preparations in the adsorption of ochratoxin A (OTA. The study involved the use of a brewer’s yeast cell wall devoid of protein substances, glucans obtained by water and alkaline extraction, a glucan commercially available as a dietary supplement for animals and, additionally, dried brewer’s yeast for comparison. Fourier Transform Infrared (FTIR analysis of the obtained preparations showed bands characteristic for glucans in the resulting spectra. The yeast cell wall preparation, water-extracted glucan and the commercial glucan bound the highest amount of ochratoxin A, above 55% of the initial concentration, and the alkaline-extracted glucan adsorbed the lowest amount of this toxin. It has been shown that adsorption is most effective at a close-to-neutral pH, while being considerably limited in alkaline conditions.

  2. Arabinose-rich polymers as an evolutionary strategy to plasticize resurrection plant cell walls against desiccation

    DEFF Research Database (Denmark)

    Moore, John P.; Nguema-Ona, Eric E.; Vicré-Gibouin, Mäite;

    2013-01-01

    A variety of Southern African resurrection plants were surveyed using high-throughput cell wall profiling tools. Species evaluated were the dicotyledons, Myrothamnus flabellifolia and Craterostigma plantagineum; the monocotyledons, Xerophyta viscosa, Xerophyta schlecterii, Xerophyta humilis and the...... resurrection grass Eragrostis nindensis, as well as a pteridophyte, the resurrection fern, Mohria caffrorum. Comparisons were made between hydrated and desiccated leaf and frond material, with respect to cell wall composition and polymer abundance, using monosaccharide composition analysis, FT-IR spectroscopy...... and comprehensive microarray polymer profiling in combination with multivariate data analysis. The data obtained suggest that three main functional strategies appear to have evolved to prepare plant cell walls for desiccation. Arabinan-rich pectin and arabinogalactan proteins are found in the...

  3. Plant metabolism and cell wall formation in space (microgravity) and on Earth

    Science.gov (United States)

    Lewis, Norman G.

    1994-01-01

    Variations in cell wall chemistry provide vascular plants with the ability to withstand gravitational forces, as well as providing facile mechanisms for correctional responses to various gravitational stimuli, e.g., in reaction wood formation. A principal focus of our current research is to precisely and systematically dissect the essentially unknown mechanism(s) of vascular plant cell wall assembly, particularly with respect to formation of its phenolic constituents, i.e., lignins and suberins, and how gravity impacts upon these processes. Formation of these phenolic polymers is of particular interest, since it appears that elaboration of their biochemical pathways was essential for successful land adaptation. By extrapolation, we are also greatly intrigued as to how the microgravity environment impacts upon 'normal' cell wall assembly mechanisms/metabolism.

  4. pH within pores in plant fiber cell walls assessed by Fluorescence Ratio Imaging

    DEFF Research Database (Denmark)

    Hidayat, Budi Juliman; Thygesen, Lisbeth Garbrecht; Johansen, Katja S.

    2013-01-01

    The pH within cell wall pores of filter paper fibers and hemp fibers was assessed by Fluorescence Ratio Imaging (FRIM). It was found that the Donnan effect affected the pH measured within the fibers. When the conductivity of the added liquid was low (0. 7 mS), pH values were lower within the cell...... wall than in the bulk solution. This was not the case at high conductivity (22 mS). The occurrence of the Donnan effect allowed the pH values within pores in normal regions of the cell wall to be compared to the pH in regions with misaligned microfibrils (dislocations) when FRIM was carried out in a...

  5. Influence of α sex factor on the biosynthesis of the cell wall from Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Cells of Saccharomyces cerevisiae produce peptide hormones (a and α) which dramatically affect the physiology, structure, and behavior of cells from the opposite mating type, presumably in preparation for conjugation. Some cell division cycle mutants mimick several of the changes induced by α factor. Accordingly, conditional mutants cdc 28, cdc 36, cdc 37, and cdc 39 undergo arrest in G1, exhibit shmoo morphology and are able to mate when they are transferred to the restrictive temperature. Formation of shmoo cells would require increased synthesis of glycosyl transferases involved in the biosynthesis of cell wall polysaccharides. Accordingly, the authors investigated the effect of G1 arrest on the chemical composition of the cell wall and on the levels of glycosyl transferases. Arrest in G1 was obtained by two methods: addition of α factor, and transfer of a cdc 28 mutant to the restrictive temperature

  6. Understanding CrRLK1L Function: Cell Walls and Growth Control.

    Science.gov (United States)

    Nissen, Karen S; Willats, William G T; Malinovsky, Frederikke G

    2016-06-01

    To develop successfully in an ever-changing environment, it is essential for plants to monitor and control their growth. Therefore, cell expansion is carefully regulated to establish correct cell shape and size. In this review, we explore the role of the Catharanthus roseus receptor-like kinase (CrRLK1L) subfamily as regulators of cell expansion. Recently, the downstream signalling events of individual CrRLK1L pathways were discovered, implicating known modulators of cell expansion, such as reactive oxygen species (ROS) production, Ca(2+) dynamics, and exocytosis of cell wall material. Based on these intriguing new insights, we propose a model for a common pathway of CrRLK1L signalling that enables spatial and temporal control of cell wall extensibility throughout the plant. PMID:26778775

  7. Osmotic Pressure, Bacterial Cell Walls, and Penicillin: A Demonstration.

    Science.gov (United States)

    Lennox, John E.

    1984-01-01

    An easily constructed apparatus that models the effect of penicillin on the structure of bacterial cells is described. Background information and procedures for using the apparatus during a classroom demonstration are included. (JN)

  8. The Anchoring Concept

    DEFF Research Database (Denmark)

    Simonsen, Jesper

    1998-01-01

    This paper introduces the term 'anchoring' within systems development: Visions, developed through early systems design within an organization, need to be deeply rooted in the organization. A vision's rationale needs to be understood by those who decide if the vision should be implemented as well as...... by those involved in the actual implementation. A model depicting a recent trend within systems development is presented: Organizations rely on purchasing generic software products and/or software development outsourced to external contractors. A contemporary method for participatory design, where...

  9. Calcium bridges are not load-bearing cell-wall bonds in Avena coleoptiles

    Science.gov (United States)

    Rayle, D. L.

    1989-01-01

    I examined the ability of frozen-thawed Avena sativa L. coleoptile sections under applied load to extend in response to the calcium chelators ethyleneglycol-bis-(beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) and 2-[(2-bis-[carboxymethyl]amino-5-methylphenoxy)methyl]-6-methoxy-8-bis[car boxymethyl]aminoquinoline (Quin II). Addition of 5 mM EGTA to weakly buffered (0.1 mM, pH 6.2) solutions of 2(N-morpholino) ethanesulfonic acid (Mes) initiated rapid extension and wall acidification. When the buffer strength was increased (e.g. from 20 to 100 mM Mes, pH 6.2) EGTA did not initiate extension nor did it cause wall acidification. At 5 mM Quin II failed to stimulate cell extension or wall acidification at all buffer molarities tested (0.1 to 100 mM Mes). Both chelators rapidly and effectively removed Ca2+ from Avena sections. These data indicate that Ca2+ chelation per se does not result in loosening of Avena cells walls. Rather, EGTA promotes wall extension indirectly via wall acidification.

  10. Granulated metrial gland cells in the uterine wall and placenta of the pregnant woodmouse, Apodemus sylvaticus.

    OpenAIRE

    Stewart, I J; Clarke, J R

    1995-01-01

    The differentiation of the decidua and the fetal placenta in the woodmouse, Apodemus sylvaticus, was studied with particular attention to the differentiation and migration of granulated metrial gland cells. Numerous granulated metrial gland cells, characterised by their glycoprotein-containing cytoplasmic granules, were found to differentiate in the decidua basalis and metrial gland of the uterine wall during the first half of pregnancy. Many granulated metrial gland cells appeared to pass in...

  11. Differential Actions of Chlorhexidine on the Cell Wall of Bacillus subtilis and Escherichia coli

    OpenAIRE

    Cheung, Hon-Yeung; Wong, Matthew Man-Kin; Cheung, Sau-Ha; Liang, Longman Yimin; Lam, Yun-Wah; Chiu, Sung-Kay

    2012-01-01

    Chlorhexidine is a chlorinated phenolic disinfectant used commonly in mouthwash for its action against bacteria. However, a comparative study of the action of chlorhexidine on the cell morphology of Gram-positive and Gram-negative bacteria is lacking. In this study, the actions of chlorhexidine on the cell morphology were identified with the aids of electron microscopy. After exposure to chlorhexidine, numerous spots of indentation on the cell wall were found in both Bacillus subtilis and Esc...

  12. Role of Protein Glycosylation in Candida parapsilosis Cell Wall Integrity and Host Interaction

    Science.gov (United States)

    Pérez-García, Luis A.; Csonka, Katalin; Flores-Carreón, Arturo; Estrada-Mata, Eine; Mellado-Mojica, Erika; Németh, Tibor; López-Ramírez, Luz A.; Toth, Renata; López, Mercedes G.; Vizler, Csaba; Marton, Annamaria; Tóth, Adél; Nosanchuk, Joshua D.; Gácser, Attila; Mora-Montes, Héctor M.

    2016-01-01

    Candida parapsilosis is an important, emerging opportunistic fungal pathogen. Highly mannosylated fungal cell wall proteins are initial contact points with host immune systems. In Candida albicans, Och1 is a Golgi α1,6-mannosyltransferase that plays a key role in the elaboration of the N-linked mannan outer chain. Here, we disrupted C. parapsilosis OCH1 to gain insights into the contribution of N-linked mannosylation to cell fitness and to interactions with immune cells. Loss of Och1 in C. parapsilosis resulted in cellular aggregation, failure of morphogenesis, enhanced susceptibility to cell wall perturbing agents and defects in wall composition. We removed the cell wall O-linked mannans by β-elimination, and assessed the relevance of mannans during interaction with human monocytes. Results indicated that O-linked mannans are important for IL-1β stimulation in a dectin-1 and TLR4-dependent pathway; whereas both, N- and O-linked mannans are equally important ligands for TNFα and IL-6 stimulation, but neither is involved in IL-10 production. Furthermore, mice infected with C. parapsilosis och1Δ null mutant cells had significantly lower fungal burdens compared to wild-type (WT)-challenged counterparts. Therefore, our data are the first to demonstrate that C. parapsilosis N- and O-linked mannans have different roles in host interactions than those reported for C. albicans. PMID:27014229

  13. Structure of the cell wall of mango after application of ionizing radiation

    Science.gov (United States)

    Silva, Josenilda M.; Villar, Heldio P.; Pimentel, Rejane M. M.

    2012-11-01

    Cells of the mesocarp of mango cultivar Tommy Atkins were analyzed by Transmission Electron Microscope—TEM to evaluate the effects of doses of 0.5 and 1.0 kGy applied immediately after the fruit and after storage for twenty days at a temperature of 12 °C followed by 5 days of simulated marketing at a temperature of 21 °C. No alteration was found in the structure of the cell wall, middle lamella, and plasma membrane of fruits when analyzed immediately after application of doses. The mesocarp cell structure of the cell wall, middle lamella, and the plasma membrane did however undergo changes after storage. Fruits that received a dose of 0.5 kGy displayed slight changes in cell wall structure and slight disintegration of the middle lamella. Fruits that received a dose of 1.0 kGy displayed more severe changes in the structure of the cell wall, greater middle lamella degradation, and displacement of the plasma membrane.

  14. A novel in vivo cell-wall labeling approach sheds new light on peptidoglycan synthesis in Escherichia coli

    NARCIS (Netherlands)

    N.K. Olrichs; M.E.G. Aarsman; J. Verheul; C.J. Arnusch; N.I. Martin; M. Hervé; W. Vollmer; B. de Kruijff; E. Breukink; T. den Blaauwen

    2011-01-01

    Peptidoglycan synthesis and turnover in relation to cell growth and division has been studied by using a new labeling method. This method involves the incorporation of fluorescently labeled peptidoglycan precursors into the cell wall by means of the cell-wall recycling pathway. We show that Escheric

  15. Apoptosis of Alveolar Wall Cells in Chronic Obstructive Pulmonary Disease Patients with Pulmonary Emphysema Is Involved in Emphysematous Changes

    Institute of Scientific and Technical Information of China (English)

    Hongmei LIU; Lijun MA; Jizhen WU; Kai WANG; Xianliang CHEN

    2009-01-01

    s of alveolar wall cells, espe-cially apoptosis of type-Ⅱ cells, may take part in the pathogenesis of emphysema. Up-regulation of Bax expression may be responsible for the apoptosis of alveolar wall cells in the COPD patients with pulmonary emphysema.

  16. Copolymerization of Sinapyl p-coumarate With Sinapyl alcohol: Impact on Syringyl Lignin Formation and Fermentability of Maize Cell Walls

    Science.gov (United States)

    Lignins in grass biomass and forage crops are highly acylated with p-coumarate, but the function of p-coumarate and its impact on cell wall degradability are poorly understood. In this study, cell walls from maize cell suspensions were artificially lignified with coniferyl alcohol and increasing pro...

  17. Inhibitory activity of 1-farnesylpyridinium on the spatial control over the assembly of cell wall polysaccharides in Schizosaccharomyces pombe.

    Science.gov (United States)

    Hamada, Masahiro; Ohata, Ikumi; Fujita, Ken-ichi; Usuki, Yoshinosuke; Ogita, Akira; Ishiguro, Junpei; Tanaka, Toshio

    2006-12-01

    The modes of actions of 1-farnesylpyridinium (FPy) on yeast cell growth were investigated on the basis of its effects on cell cycle progression, morphogenesis and the related events for construction of cell wall architecture in Schizosacchromyces pombe. FPy predominantly inhibited the growth of the yeast cells after various cycles of cell division so that cells were arrested at the phase of separation into daughter cells accompanying morphological changes to swollen spherical cells at 24 h of incubation. FPy-treated cells were osmotically stable but were susceptible to the lytic action of (1, 3) beta-D-glucanases, and characterized by serious damages to the cell wall architecture as represented by a rough and irregular surface outlook. The isolated cell wall fraction gave a similar hexose composition with or without FPy treatment, suggesting that FPy did not inhibit the synthesis of each cell wall polysaccharide. FPy was permissive for the extracellular accumulation of amorphous cell wall materials and septum development in protoplasts, but absolutely interfered with the following morphogenetic process for construction of the rod-shaped cell wall architecture. Our results suggest the inhibitory activity of FPy on the spatial control over the assembly of cell wall polysaccharides. PMID:17092950

  18. Cell wall degrading enzymes in Trichoderma asperellum grown on wheat bran

    DEFF Research Database (Denmark)

    Bech, Lasse; Busk, Peter Kamp; Lange, Lene

    2015-01-01

    Trichoderma asperellum is a filamentous fungus that is able to produce and secrete a wide range of extracellular hydrolytic enzymes used for plant cell wall degradation. The Trichoderma genus has attracted considerable attention from the biorefinery industry due to the production of cell wall...... Pattern Recognition enabling an efficient enzyme discovery. This was furthermore used to re-annotate CAZYmes present in five publically available Trichoderma species, hereby elucidating differences in CAZYmes on a functional level in contrary to glycoside hydrolase family level. This comparison supports...

  19. High-throughput mapping of cell-wall polymers within and between plants using novel microarrays

    DEFF Research Database (Denmark)

    Moller, Isabel Eva; Sørensen, Iben; Bernal Giraldo, Adriana Jimena;

    2007-01-01

    We describe here a methodology that enables the occurrence of cell-wall glycans to be systematically mapped throughout plants in a semi-quantitative high-throughput fashion. The technique (comprehensive microarray polymer profiling, or CoMPP) integrates the sequential extraction of glycans from...... analysis of mutant and wild-type plants, as demonstrated here for the Arabidopsis thaliana mutants fra8, mur1 and mur3. CoMPP was also applied to Physcomitrella patens cell walls and was validated by carbohydrate linkage analysis. These data provide new insights into the structure and functions of plant...

  20. In vivo determination of fibril orientation in plant cell walls with polarization CSLM

    International Nuclear Information System (INIS)

    Congo Red fluorescence is used to detect cellulose in the wall of plant cells. The orientation of the cellulose fibrils is determined by using polarized light for excitation. The absorption characteristics of Congo Red make this approach a method of choice for applications with any standard confocal scanning laser microscope (CSLM). The semiquantitative character of CSLM observations combined with the non-toxicity of the stain allow a very fast and reliable assessment of cellulose orientation in the wall of living plant cells. (author)