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Sample records for interstrand dna cross-links

  1. Nitric oxide-induced interstrand cross-links in DNA.

    Science.gov (United States)

    Caulfield, Jennifer L; Wishnok, John S; Tannenbaum, Steven R

    2003-05-01

    The DNA damaging effects of nitrous acid have been extensively studied, and the formation of interstrand cross-links have been observed. The potential for this cross-linking to occur through a common nitrosating intermediate derived from nitric oxide is investigated here. Using a HPLC laser-induced fluorescence (LIF) system, the amount of interstrand cross-link formed on nitric oxide treatment of the 5'-fluorescein-labeled oligomer ATATCGATCGATAT was determined. This self-complimentary sequence contains two 5'-CG sequences, which is the preferred site for nitrous acid-induced cross-linking. Nitric oxide was delivered to an 0.5 mM oligomer solution at 15 nmol/mL/min to give a final nitrite concentration of 652 microM. The resulting concentration of the deamination product, xanthine, in this sample was found to be 211 +/- 39 nM, using GC/MS, and the amount of interstrand cross-link was determined to be 13 +/- 2.5 nM. Therefore, upon nitric oxide treatment, the cross-link is found at approximately 6% of the amount of the deamination product. Using this system, detection of the cross-link is also possible for significantly lower doses of nitric oxide, as demonstrated by treatment of the same oligomer with NO at a rate of 18 nmol/mL/min resulting in a final nitrite concentration of 126 microM. The concentration of interstrand cross-link was determined to be 3.6 +/- 0.1 nM in this sample. Therefore, using the same dose rate, when the total nitric oxide concentration delivered drops by a factor of approximately 5, the concentration of cross-link drops by a factor of about 4-indicating a qausi-linear response. It may now be possible to predict the number of cross-links in a small genome based on the number of CpG sequences and the yield of xanthine derived from nitrosative deamination.

  2. Characterization of Interstrand DNA-DNA Cross-Links Using the α-Hemolysin Protein Nanopore.

    Science.gov (United States)

    Zhang, Xinyue; Price, Nathan E; Fang, Xi; Yang, Zhiyu; Gu, Li-Qun; Gates, Kent S

    2015-12-22

    Nanopore-based sensors have been studied extensively as potential tools for DNA sequencing, characterization of epigenetic modifications such as 5-methylcytosine, and detection of microRNA biomarkers. In the studies described here, the α-hemolysin protein nanopore embedded in a lipid bilayer was used for the detection and characterization of interstrand cross-links in duplex DNA. Interstrand cross-links are important lesions in medicinal chemistry and toxicology because they prevent the strand separation that is required for read-out of genetic information from DNA in cells. In addition, interstrand cross-links are used for the stabilization of duplex DNA in structural biology and materials science. Cross-linked DNA fragments produced unmistakable current signatures in the nanopore experiment. Some cross-linked substrates gave irreversible current blocks of >10 min, while others produced long current blocks (10-100 s) before the double-stranded DNA cross-link translocated through the α-hemolysin channel in a voltage-driven manner. The duration of the current block for the different cross-linked substrates examined here may be dictated by the stability of the duplex region left in the vestibule of the nanopore following partial unzipping of the cross-linked DNA. Construction of calibration curves measuring the frequency of cross-link blocking events (1/τon) as a function of cross-link concentration enabled quantitative determination of the amounts of cross-linked DNA present in samples. The unique current signatures generated by cross-linked DNA in the α-HL nanopore may enable the detection and characterization of DNA cross-links that are important in toxicology, medicine, and materials science.

  3. Site specificity of psoralen-DNA interstrand cross-linking determined by nuclease Bal31 digestion

    DEFF Research Database (Denmark)

    Zhen, W P; Buchardt, O; Nielsen, Henrik

    1986-01-01

    A novel method for determination of psoralen photo-cross-linking sites in double-stranded DNA is described, which is based on a pronounced inhibition of Bal31 exonuclease activity by psoralen-DNA interstrand cross-links. The results using a 51 base pair fragment of plasmid pUC19 and a 346 base pair...

  4. Structure of a DNA glycosylase that unhooks interstrand cross-links

    Energy Technology Data Exchange (ETDEWEB)

    Mullins, Elwood A.; Warren, Garrett M.; Bradley, Noah P.; Eichman, Brandt F. (Vanderbilt)

    2017-04-10

    DNA glycosylases are important editing enzymes that protect genomic stability by excising chemically modified nucleobases that alter normal DNA metabolism. These enzymes have been known only to initiate base excision repair of small adducts by extrusion from the DNA helix. However, recent reports have described both vertebrate and microbial DNA glycosylases capable of unhooking highly toxic interstrand cross-links (ICLs) and bulky minor groove adducts normally recognized by Fanconi anemia and nucleotide excision repair machinery, although the mechanisms of these activities are unknown. Here we report the crystal structure of Streptomyces sahachiroi AlkZ (previously Orf1), a bacterial DNA glycosylase that protects its host by excising ICLs derived from azinomycin B (AZB), a potent antimicrobial and antitumor genotoxin. AlkZ adopts a unique fold in which three tandem winged helix-turn-helix motifs scaffold a positively charged concave surface perfectly shaped for duplex DNA. Through mutational analysis, we identified two glutamine residues and a β-hairpin within this putative DNA-binding cleft that are essential for catalytic activity. Additionally, we present a molecular docking model for how this active site can unhook either or both sides of an AZB ICL, providing a basis for understanding the mechanisms of base excision repair of ICLs. Given the prevalence of this protein fold in pathogenic bacteria, this work also lays the foundation for an emerging role of DNA repair in bacteria-host pathogenesis.

  5. Chemosensitivity of primary human fibroblasts with defective unhooking of DNA interstrand cross-links

    International Nuclear Information System (INIS)

    Clingen, Peter H.; Arlett, Colin F.; Hartley, John A.; Parris, Christopher N.

    2007-01-01

    Xeroderma pigmentosum (XP) is characterised by defects in nucleotide excision repair, ultraviolet (UV) radiation sensitivity and increased skin carcinoma. Compared to other complementation groups, XP-F patients show relatively mild cutaneous symptoms. DNA interstrand cross-linking agents are a highly cytotoxic class of DNA damage induced by common cancer chemotherapeutics such as cisplatin and nitrogen mustards. Although the XPF-ERCC1 structure-specific endonuclease is required for the repair of ICLs cellular sensitivity of primary human XP-F cells has not been established. In clonogenic survival assays, primary fibroblasts from XP-F patients were moderately sensitive to both UVC and HN2 compared to normal cells (2- to 3-fold and 3- to 5-fold, respectively). XP-A fibroblasts were considerably more sensitive to UVC (10- to 12-fold) but not sensitive to HN2. The sensitivity of XP-F fibroblasts to HN2 correlated with the defective incision or 'unhooking' step of ICL repair. Using the comet assay, XP-F cells exhibited only 20% residual unhooking activity over 24 h. Over the same time, normal and XP-A cells unhooked greater than 95% and 62% of ICLs, respectively. After HN2 treatment, ICL-associated DNA double-strand breaks (DSBs) are detected by pulse field gel electrophoresis in dividing cells. Induction and repair of DNA DSBs was normal in XP-F fibroblasts. These findings demonstrate that in primary human fibroblasts, XPF is required for the unhooking of ICLs and not for the induction or repair of ICL-associated DNA DSBs induced by HN2. In terms of cancer chemotherapy, people with mild DNA repair defects affecting ICL repair may be more prevalent in the general population than expected. Since cellular sensitivity of primary human fibroblasts usually reflects clinical sensitivity such patients with cancer would be at risk of increased toxicity

  6. 8-Methoxypsoralen DNA interstrand cross-linking of the ribosomal RNA genes in Tetrahymena thermophila. Distribution, repair and effect on rRNA synthesis

    DEFF Research Database (Denmark)

    Fengquin, X; Nielsen, Henrik; Zhen, W

    1993-01-01

    The distribution and repair of 8-methoxypsoralen-DNA interstrand cross-links in the ribosomal RNA genes (rDNA) in Tetrahymena thermophila have been studied in vivo by Southern blot analysis. It is found that the cross-links at a density of

  7. Rapid detection of DNA-interstrand and DNA-protein cross-links in mammalian cells by gravity-flow alkaline elution

    International Nuclear Information System (INIS)

    Hincks, J.R.; Coulombe, R.A. Jr.

    1989-01-01

    Alkaline elution is a sensitive and commonly used technique to detect cellular DNA damage in the form of DNA strand breaks and DNA cross-links. Conventional alkaline elution procedures have extensive equipment requirements and are tedious to perform. Our laboratory recently presented a rapid, simplified, and sensitive modification of the alkaline elution technique to detect carcinogen-induced DNA strand breaks. In the present study, we have further modified this technique to enable the rapid characterization of chemically induced DNA-interstrand and DNA-protein associated cross-links in cultured epithelial cells. Cells were exposed to three known DNA cross-linking agents, nitrogen mustard (HN 2 ), mitomycin C (MMC), or ultraviolet irradiation (UV). One hour exposures of HN 2 at 0.25, 1.0, and 4.0 microM or of MMC at 20, 40, and 60 microM produced a dose-dependent induction of total DNA cross-links by these agents. Digestion with proteinase K revealed that HN 2 and MMC induced both DNA-protein cross-links and DNA-interstrand cross-links. Ultraviolet irradiation induced both DNA cross-links and DNA strand breaks, the latter of which were either protein and nonprotein associated. The results demonstrate that gravity-flow alkaline elution is a sensitive and accurate method to characterize the molecular events of DNA cross-linking. Using this procedure, elution of DNA from treated cells is completed in 1 hr, and only three fractions per sample are analyzed. This method may be useful as a rapid screening assay for genotoxicity and/or as an adjunct to other predictive assays for potential mutagenic or carcinogenic agents

  8. 8-Methoxypsoralen DNA interstrand cross-linking of the ribosomal RNA genes in Tetrahymena thermophila. Distribution, repair and effect on rRNA synthesis

    DEFF Research Database (Denmark)

    Fengquin, X; Nielsen, Henrik; Zhen, W

    1993-01-01

    The distribution and repair of 8-methoxypsoralen-DNA interstrand cross-links in the ribosomal RNA genes (rDNA) in Tetrahymena thermophila have been studied in vivo by Southern blot analysis. It is found that the cross-links at a density of ... between three domains (terminal spacer, transcribed region and central spacer) as defined by restriction enzyme analysis (BamHI and ClaI). It is furthermore shown that a dosage resulting in approximately one cross-link per rDNA molecule (21 kbp, two genes) is sufficient to block RNA synthesis. Finally...

  9. 8-Methoxypsoralen DNA interstrand cross-linking of the ribosomal RNA genes in Tetrahymena thermophila. Distribution, repair and effect on rRNA synthesis

    DEFF Research Database (Denmark)

    Fengquin, X; Nielsen, Henrik; Zhen, W

    1993-01-01

    The distribution and repair of 8-methoxypsoralen-DNA interstrand cross-links in the ribosomal RNA genes (rDNA) in Tetrahymena thermophila have been studied in vivo by Southern blot analysis. It is found that the cross-links at a density of ... between three domains (terminal spacer, transcribed region and central spacer) as defined by restriction enzyme analysis (BamHI and ClaI). It is furthermore shown that a dosage resulting in approximately one cross-link per rDNA molecule (21 kbp, two genes) is sufficient to block RNA synthesis. Finally......, it is shown that the cross-links in the rDNA molecules are repaired at equal rate in all three domains within 24 h and that RNA synthesis is partly restored during this repair period. The majority of the cells also go through one to two cell divisions in this period but do not survive....

  10. Ouabain, a cardiac glycoside, inhibits the Fanconi anemia/BRCA pathway activated by DNA interstrand cross-linking agents.

    Directory of Open Access Journals (Sweden)

    Dong Wha Jun

    Full Text Available Modulation of the DNA repair pathway is an emerging target for the development of anticancer drugs. DNA interstrand cross-links (ICLs, one of the most severe forms of DNA damage caused by anticancer drugs such as cisplatin and mitomycin C (MMC, activates the Fanconi anemia (FA/BRCA DNA repair pathway. Inhibition of the FA/BRCA pathway can enhance the cytotoxic effects of ICL-inducing anticancer drugs and can reduce anticancer drug resistance. To find FA/BRCA pathway inhibitory small molecules, we established a cell-based high-content screening method for quantitating the activation of the FA/BRCA pathway by measuring FANCD2 foci on DNA lesions and then applied our method to chemical screening. Using commercial LOPAC1280 chemical library screening, ouabain was identified as a competent FA/BRCA pathway inhibitory compound. Ouabain, a member of the cardiac glycoside family, binds to and inhibits Na(+/K(+-ATPase and has been used to treat heart disease for many years. We observed that ouabain, as well as other cardiac glycoside family members--digitoxin and digoxin--down-regulated FANCD2 and FANCI mRNA levels, reduced monoubiquitination of FANCD2, inhibited FANCD2 foci formation on DNA lesions, and abrogated cell cycle arrest induced by MMC treatment. These inhibitory activities of ouabain required p38 MAPK and were independent of cellular Ca(2+ ion increase or the drug uptake-inhibition effect of ouabain. Furthermore, we found that ouabain potentiated the cytotoxic effects of MMC in tumor cells. Taken together, we identified an additional effect of ouabain as a FA/BRCA pathway-inhibiting chemosensitization compound. The results of this study suggest that ouabain may serve as a chemosensitizer to ICL-inducing anticancer drugs.

  11. Interstrand cross-linking implies contrasting structural consequences for DNA: insights from molecular dynamics

    Czech Academy of Sciences Publication Activity Database

    Bignon, E.; Dršata, Tomáš; Morell, C.; Lankaš, Filip; Dumont, E.

    2017-01-01

    Roč. 45, č. 4 (2017), s. 2188-2195 ISSN 0305-1048 R&D Projects: GA ČR(CZ) GA14-21893S Institutional support: RVO:61388963 Keywords : abasic sites * duplex DNA * mechanical properties Subject RIV: BO - Biophysics OBOR OECD: Biophysics Impact factor: 10.162, year: 2016 https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/gkw1253

  12. Bacillus subtilis SbcC protein plays an important role in DNA inter-strand cross-link repair

    Directory of Open Access Journals (Sweden)

    Krisnamurthy Mahalakshmi

    2006-06-01

    Full Text Available Abstract Background Several distinct pathways for the repair of damaged DNA exist in all cells. DNA modifications are repaired by base excision or nucleotide excision repair, while DNA double strand breaks (DSBs can be repaired through direct joining of broken ends (non homologous end joining, NHEJ or through recombination with the non broken sister chromosome (homologous recombination, HR. Rad50 protein plays an important role in repair of DNA damage in eukaryotic cells, and forms a complex with the Mre11 nuclease. The prokaryotic ortholog of Rad50, SbcC, also forms a complex with a nuclease, SbcD, in Escherichia coli, and has been implicated in the removal of hairpin structures that can arise during DNA replication. Ku protein is a component of the NHEJ pathway in pro- and eukaryotic cells. Results A deletion of the sbcC gene rendered Bacillus subtilis cells sensitive to DNA damage caused by Mitomycin C (MMC or by gamma irradiation. The deletion of the sbcC gene in a recN mutant background increased the sensitivity of the single recN mutant strain. SbcC was also non-epistatic with AddAB (analog of Escherichia coli RecBCD, but epistatic with RecA. A deletion of the ykoV gene encoding the B. subtilis Ku protein in a sbcC mutant strain did not resulted in an increase in sensitivity towards MMC and gamma irradiation, but exacerbated the phenotype of a recN or a recA mutant strain. In exponentially growing cells, SbcC-GFP was present throughout the cells, or as a central focus in rare cases. Upon induction of DNA damage, SbcC formed 1, rarely 2, foci on the nucleoids. Different to RecN protein, which forms repair centers at any location on the nucleoids, SbcC foci mostly co-localized with the DNA polymerase complex. In contrast to this, AddA-GFP or AddB-GFP did not form detectable foci upon addition of MMC. Conclusion Our experiments show that SbcC plays an important role in the repair of DNA inter-strand cross-links (induced by MMC, most likely

  13. Mutagenic Bypass of an Oxidized Abasic Lesion-Induced DNA Interstrand Cross-Link Analogue by Human Translesion Synthesis DNA Polymerases.

    Science.gov (United States)

    Xu, Wenyan; Ouellette, Adam; Ghosh, Souradyuti; O'Neill, Tylor C; Greenberg, Marc M; Zhao, Linlin

    2015-12-22

    5'-(2-Phosphoryl-1,4-dioxobutane) (DOB) is an oxidized abasic site that is produced by several antitumor agents and γ-radiolysis. DOB reacts reversibly with a dA opposite the 3'-adjacent nucleotide to form DNA interstrand cross-links (ICLs), genotoxic DNA lesions that can block DNA replication and transcription. Translesion synthesis (TLS) is an important step in several ICL repair pathways to bypass unhooked intermediates generated by endonucleolytic incision. The instability of DOB-ICLs has made it difficult to learn about their TLS-mediated repair capability and mutagenic potential. We recently developed a method for chemically synthesizing oligonucleotides containing a modified DOB-ICL analogue. Herein, we examined the capabilities of several highly relevant eukaryotic TLS DNA polymerases (pols), including human pol η, pol κ, pol ι, pol ν, REV1, and yeast pol ζ, to bypass this DOB-ICL analogue. The prelesion, translesion, and postlesion replication efficiency and fidelity were examined. Pol η showed moderate bypass activity when encountering the DOB-ICL, giving major products one or two nucleotides beyond the cross-linked template nucleotide. In contrast, DNA synthesis by the other pols was stalled at the position before the cross-linked nucleotide. Steady-state kinetic data and liquid chromatography-mass spectrometry sequencing of primer extension products by pol η unambiguously revealed that pol η-mediated bypass is highly error-prone. Together, our study provides the first set of in vitro evidence that the DOB-ICL is a replication-blocking and highly miscoding lesion. Compared to several other TLS pols examined, pol η is likely to contribute to the TLS-mediated repair of the DOB-ICL in vivo.

  14. O6-2'-Deoxyguanosine-butylene-O6-2'-deoxyguanosine DNA Interstrand Cross-Links Are Replication-Blocking and Mutagenic DNA Lesions.

    Science.gov (United States)

    Xu, Wenyan; Kool, Daniel; O'Flaherty, Derek K; Keating, Ashley M; Sacre, Lauralicia; Egli, Martin; Noronha, Anne; Wilds, Christopher J; Zhao, Linlin

    2016-11-21

    DNA interstrand cross-links (ICLs) are cytotoxic DNA lesions derived from reactions of DNA with a number of anti-cancer reagents as well as endogenous bifunctional electrophiles. Deciphering the DNA repair mechanisms of ICLs is important for understanding the toxicity of DNA cross-linking agents and for developing effective chemotherapies. Previous research has focused on ICLs cross-linked with the N7 and N2 atoms of guanine as well as those formed at the N6 atom of adenine; however, little is known about the mutagenicity of O 6 -dG-derived ICLs. Although less abundant, O 6 -alkylated guanine DNA lesions are chemically stable and highly mutagenic. Here, O 6 -2'-deoxyguanosine-butylene-O 6 -2'-deoxyguanosine (O 6 -dG-C4-O 6 -dG) is designed as a chemically stable ICL, which can be induced by the action of bifunctional alkylating agents. We investigate the DNA replication-blocking and mutagenic properties of O 6 -dG-C4-O 6 -dG ICLs during an important step in ICL repair, translesion DNA synthesis (TLS). The model replicative DNA polymerase (pol) Sulfolobus solfataricus P2 DNA polymerase B1 (Dpo1) is able to incorporate a correct nucleotide opposite the cross-linked template guanine of ICLs with low efficiency and fidelity but cannot extend beyond the ICLs. Translesion synthesis by human pol κ is completely inhibited by O 6 -dG-C4-O 6 -dG ICLs. Moderate bypass activities are observed for human pol η and S. solfataricus P2 DNA polymerase IV (Dpo4). Among the pols tested, pol η exhibits the highest bypass activity; however, 70% of the bypass products are mutagenic containing substitutions or deletions. The increase in the size of unhooked repair intermediates elevates the frequency of deletion mutation. Lastly, the importance of pol η in O 6 -dG-derived ICL bypass is demonstrated using whole cell extracts of Xeroderma pigmentosum variant patient cells and those complemented with pol η. Together, this study provides the first set of biochemical evidence for the

  15. The Fanconi anemia/BRCA pathway is involved in DNA interstrand cross-link repair of adriamycin-resistant leukemia cells.

    Science.gov (United States)

    Yao, Chenjiao; Du, Wei; Chen, Haibing; Xiao, Sheng; Huang, Lihua; Chen, Fangping

    2015-03-01

    The Fanconi anemia/BRCA (FA/BRCA) pathway plays a vital role in DNA damage repair induced by DNA cross-linking agents and is closely related to drug response in cancer treatment. Here we demonstrate that the FA/BRCA pathway contributes to acquired drug resistance in adriamycin (ADR)-resistant leukemia cell lines, and disruption of this pathway partially reverses the drug resistance. We observed that ADR-resistant cells have reduced DNA interstrand cross-links (ICL) compared with ADR-sensitive cells. Western blot studies demonstrated enhanced FA protein expression in ADR-resistant cells. Using siRNA to knock down FANCF in K562/R drug-resistant cells showed increases in sensitivity to ADR and ADR-induced DNA damage, and demonstrated a direct relationship between the FA/BRCA pathway and drug sensitivity. Overexpression of FANCF in K562 drug-sensitive cells partially reproduced the drug-resistant phenotype. These results show that the FA/BRCA pathway is involved in acquired ADR resistance of leukemia cells. The FA/BRCA pathway may be a new target to reverse ADR resistance in leukemia treatment.

  16. Crystal structure of a Fanconi anemia-associated nuclease homolog bound to 5' flap DNA: basis of interstrand cross-link repair by FAN1

    Energy Technology Data Exchange (ETDEWEB)

    Gwon, Gwang Hyeon; Kim, Youngran; Liu, Yaqi; Watson, Adam T.; Jo, Aera; Etheridge, Thomas J.; Yuan, Fenghua; Zhang, Yanbin; Kim, YoungChang; Carr, Anthony M.; Cho, Yunje

    2014-10-15

    Fanconi anemia (FA) is an autosomal recessive genetic disorder caused by defects in any of 15 FA genes responsible for processing DNA interstrand cross-links (ICLs). The ultimate outcome of the FA pathway is resolution of cross-links, which requires structure-selective nucleases. FA-associated nuclease 1 (FAN1) is believed to be recruited to lesions by a monoubiquitinated FANCI–FANCD2 (ID) complex and participates in ICL repair. Here, we determined the crystal structure of Pseudomonas aeruginosa FAN1 (PaFAN1) lacking the UBZ (ubiquitin-binding zinc) domain in complex with 5' flap DNA. All four domains of the right-hand-shaped PaFAN1 are involved in DNA recognition, with each domain playing a specific role in bending DNA at the nick. The six-helix bundle that binds the junction connects to the catalytic viral replication and repair (VRR) nuclease (VRR nuc) domain, enabling FAN1 to incise the scissile phosphate a few bases distant from the junction. The six-helix bundle also inhibits the cleavage of intact Holliday junctions. PaFAN1 shares several conserved features with other flap structure-selective nucleases despite structural differences. A clamping motion of the domains around the wedge helix, which acts as a pivot, facilitates nucleolytic cleavage. The PaFAN1 structure provides insights into how archaeal Holliday junction resolvases evolved to incise 5' flap substrates and how FAN1 integrates with the FA complex to participate in ICL repair.

  17. Evidence for different mechanisms of ‘unhooking’ for melphalan and cisplatin-induced DNA interstrand cross-links in vitro and in clinical acquired resistant tumour samples

    International Nuclear Information System (INIS)

    Spanswick, Victoria J; Hartley, John A; Lowe, Helen L; Newton, Claire; Bingham, John P; Bagnobianchi, Alessia; Kiakos, Konstantinos; Craddock, Charles; Ledermann, Jonathan A; Hochhauser, Daniel

    2012-01-01

    DNA interstrand cross-links (ICLs) are critical lesions produced by several cancer chemotherapy agents including platinum drugs and nitrogen mustards. We have previously shown in haematological (multiple myeloma) and solid tumours (ovarian cancer) that clinical sensitivity to such agents can result from a defect in DNA ICL processing leading to their persistence. Conversely, enhanced repair can result in clinical acquired resistance following chemotherapy. The repair of ICLs is complex but it is assumed that the ‘unhooking’ step is common to all ICLs. Using a modification of the single cell gel electrophoresis (Comet) assay we measured the formation and unhooking of melphalan and cisplatin-induced ICLs in cell lines and clinical samples. DNA damage response in the form of γ-H2AX foci formation and the formation of RAD51 foci as a marker of homologous recombination were also determined. Real-time PCR of 84 genes involved in DNA damage signalling pathways was also examined pre- and post-treatment. Plasma cells from multiple myeloma patients known to be clinically resistant to melphalan showed significant unhooking of melphalan-induced ICLs at 48 hours, but did not unhook cisplatin-induced ICLs. In ovarian cancer cells obtained from patients following platinum-based chemotherapy, unhooking of cisplatin-induced ICLs was observed at 48 hours, but no unhooking of melphalan-induced ICLs. In vitro, A549 cells were proficient at unhooking both melphalan and cisplatin-induced ICLs. γ-H2AX foci formation closely followed the formation of ICLs for both drugs, and rapidly declined following the peak of formation. RPMI8226 cells unhooked melphalan, but not cisplatin-induced ICLs. In these cells, although cross-links form with cisplatin, the γ-H2AX response is weak. In A549 cells, addition of 3nM gemcitabine resulted in complete inhibition of cisplatin-induced ICL unhooking but no effect on repair of melphalan ICLs. The RAD51 foci response was both drug and cell line

  18. Unique structural properties of DNA interstrand cross-links formed by a new antitumor dinuclear Pt(II) complex

    Czech Academy of Sciences Publication Activity Database

    Hrabina, O.; Kašpárková, J.; Suchánková, Tereza; Novohradský, Vojtěch; Guo, Z.; Brabec, Viktor

    2017-01-01

    Roč. 9, č. 5 (2017), s. 494-500 ISSN 1756-5901 Institutional support: RVO:68081707 Keywords : cisplatin-modified dna * nucleotide excision-repair * hmg domain proteins Subject RIV: CE - Biochemistry OBOR OECD: Biochemistry and molecular biology Impact factor: 3.975, year: 2016

  19. Induction and removal of DNA interstrand cross-links in V-79 Chinese hamster cells measured by hydroxylapatite chromatography after treatments with bifunctional furocoumarins

    International Nuclear Information System (INIS)

    Dardalhon, M.; Averbeck, D.

    1988-01-01

    DNA interstrand crosslinks (CL) photoinduced by bifunctional furocoumarins in V-79 Chinese hamster cells were measured by alkaline denaturation and hydroxylapatite chromatography. Treatments with 5-methoxypsoralen (5-MOP), 8-methoxypsoralen (8-MOP) and 4,5',8-trimethylpsoralen (4,5',8-TMP) and 365 nm irradiation (UVA) confer a dose-dependent linear increase in the amount of double-stranded DNA indicating the induction of CL. Determination in alkaline sucrose gradients of the molecular weight of the DNA and estimation of drug-induced strand breakage allowed quantification of the CL induced. 5-MOP was found to be slightly more effective than 8-MOP whereas 4,5',8-TMP was 9 times more effective for the induction of CL. The fate of CL during post-treatment incubation was also followed. Cells in exponential growth phase were found to be efficient in the removal of CL. (Author)

  20. Ultrafast reversible photo-cross-linking reaction: toward in situ DNA manipulation.

    Science.gov (United States)

    Yoshimura, Yoshinaga; Fujimoto, Kenzo

    2008-08-07

    We describe a novel ultrafast reversible DNA interstrand photo-cross-linking reaction via 3-cyanovinylcarbazole nucleoside ( (CNV)K). Oligodeoxynucleotide (ODN) containing (CNV)K can be photo-cross-linked by irradiation at 366 nm for 1 s, and the photo-cross-linked ODN can be split by irradiation at 312 nm for 60 s.

  1. Preparation and characterization of platinum(II) and (IV) complexes of 1,3-diaminepropane and 1,4-diaminebutane: circumvention of cisplatin resistance and DNA interstrand cross-link formation in CH1cisR ovarian tumor cells.

    Science.gov (United States)

    Alvarez-Valdés, Amparo; Pérez, José Manuel; López-Solera, Isabel; Lannegrand, Raúl; Continente, José Manuel; Amo-Ochoa, Pilar; Camazón, María José; Solans, Xavier; Font-Bardía, Mercè; Navarro-Ranninger, Carmen

    2002-04-25

    The reaction of Pt(dimethyl sulfoxide)(2)CBDCA (CBDCA = 1,1-cyclobutanedicarboxylate) with 1,4-diaminebutane and 1,3-diaminepropane ligands yields, under certain conditions, new [Pt(diamine)(2)]CBDCA complexes (1a,b), where the CBDCA ligand has been removed from the coordination sphere of the platinum atom by the diamine ligand, instead of forming the expected [Pt(diamine)CBDCA] complexes (1'a,b). The structure of complexes 1a and 1'b was solved by X-ray diffraction. Complex 1a crystallizes in the orthorhombic system, in the noncentrosymmetric C222 space group, with unit cell parameters: a = 20.053(2) A; b = 8.655(2) A, c = 5.711(3) A; V = 991.2(6) A(3); delta (calcd) = 1.627 mg/m(3); and R = 0.050. The Pt atom displays an unexpected distorted tetrahedral coordination with a N-Pt-N inner bond angle equal to 81(2) degrees for N atoms of the same 1,3-propanediamine ligand and a N-Pt-N bond angle for different ligands equal to 135.4(9) degrees. Complex 1'b crystallizes in the monoclinic system, in the centrosymmetric P2(1)/c space group, with unit cell parameters: a = 6.007(2) A; b = 15.336(4) A, c = 13.232(5) A; beta = 101.90(3) degrees; V = 1192.8(7) A(3); delta (calcd) = 2.369 mg/m(3); and R = 0.067. Cytotoxicity data show that of all the synthesized compounds, only complexes 1'a and 1'b exhibit remarkable cytotoxic properties. Thus, in contrast with carboplatin (cis-diammine-1,1-cyclobutane dicarboxilatoplatinum(II)), compounds 1'a and 1'b, which also contain the CBDCA ligand, are able to circumvent cisplatin (cis-diamminedichloroplatinum(II)) resistance in several tumor cells. Moreover, after 24 h of incubation of CH1cisR ovarian tumor cells with 10 microM of compounds 1'a and 1'b, the level of DNA interstrand cross-links (ICLs) induced by compounds 1'a and 1'b is 3.3 and 3.8 times higher, respectively, than that of carboplatin and 3.5 and 4.0 times higher, respectively, than that of cisplatin. Interestingly, under the same conditions, the intracellular

  2. Thermal stability and energetics of 15-mer DNA duplex interstrand crosslinked by trans-diamminedichloroplatinum(II)

    Czech Academy of Sciences Publication Activity Database

    Hofr, Ctirad; Brabec, Viktor

    2005-01-01

    Roč. 77, č. 4 (2005), s. 222-229 ISSN 0006-3525 R&D Projects: GA ČR(CZ) GP202/01/D110; GA ČR(CZ) GA305/02/1552; GA AV ČR(CZ) LZ1K03010 Institutional research plan: CEZ:AV0Z50040507 Keywords : DNA * interstrand cross-link * transplatin Subject RIV: BO - Biophysics Impact factor: 2.545, year: 2005

  3. Combined theoretical and computational study of interstrand DNA guanine-guanine cross-linking by trans-[Pt(pyridine)2] derived from the photoactivated prodrug trans,trans,trans-[Pt(N3)2(OH)2(pyridine)2

    Czech Academy of Sciences Publication Activity Database

    Tai, H.-Ch.; Brodbeck, R.; Kašpárková, Jana; Farrer, N.J.; Brabec, Viktor; Sadler, P.J.; Deeth, R.J.

    2012-01-01

    Roč. 51, č. 12 (2012), s. 6830-6841 ISSN 0020-1669 R&D Projects: GA ČR(CZ) GAP301/10/0598 Institutional research plan: CEZ:AV0Z50040702 Keywords : platinum * photoactivation * DNA Subject RIV: BO - Biophysics Impact factor: 4.593, year: 2012

  4. Interstrand DNA crosslinks due to AP (apurinic/apyrimidinic) sites

    International Nuclear Information System (INIS)

    Goffin, C.; Verly, W.G.

    1983-01-01

    Storage of a solution of DNA containing apurinic sites, even at 4 0 C leads to the appearance of interstrand crosslinks. Possible consequences of these crosslinks, when they appear in cell DNA, are briefly discussed. Formation of interstrand crosslinks in DNA containing tritium-labelled thymine and kept in an aqueous solution might be due, at least partly, to the loss of bases by the autoirradiated DNA. (Auth.)

  5. An electron microscopic study of the photochemical cross-linking of DNA in guinea pig epidermis by psoralen derivatives

    International Nuclear Information System (INIS)

    Cech, T.; Pathak, M.A.; Biswas, R.K.

    1979-01-01

    Albino guinea pigs were treated with psoralen derivatives plus 320-400 nm ultraviolet radiation, and DNA was extracted from their epidermis. The DNA was assayed for the presence of interstrand cross-links by standard denaturation-renaturation assays and by a new technique, electron microscopy of the DNA under totally denaturing conditions. The latter method allows individual cross-links to be directly observed and counted. When either 4,5',8-trimethylpsoralen or 8-methoxypsoralen was applied topically to the skin (8-20 μg/cm 2 ) or administered orally (10-12 mg/kg body weight), followed by exposure to 320-400 nm ultraviolet radiation, most of the epidermal DNA was found to contain a high frequency of cross-links. For example, oral or topical trimethylpsoralen treatment gave an average of one cross-link per 250 nucleotide pairs or about 3 . 10 5 cross-links per guinea pig chromosome. When the dose of either drug was decreased 20-fold to the level used in the clinical treatment of psoriasis, however, no cross-links could be detected in the epidermal DNA. The electron microscopic assay is sensitive enough that one can put an upper limit of 1 cross-link per 10 6 nucleotide pairs (80 cross-links per chromosome) for the low dose studies. The significance of these findings to the understanding of the effectiveness of psoralens in psoriasis therapy is discussed. (Auth.)

  6. X-ray-mediated cross linking of protein and DNA

    International Nuclear Information System (INIS)

    Minsky, B.D.; Braun, A.

    1977-01-01

    Using a simple filter assay for the binding of BSA or lysozyme to DNA, two mechanisms of x-ray-mediated cross linking are shown to occur. One, a fast reaction, appears to involve a radical intermediate, is inhibited by high pH and salt, and seems to be enhanced by deoxygenation. The second mechanism, a slow time-dependent component, differs from the fast reaction in its stimulation by histidine, its inhibition by catalase, and the lack of an oxygen effect. Separate irradiation of DNA or water does not lead to cross linking. However, separate irradiation of protein leads to cross linking which proceeds with slow-component kinetics

  7. Cross-linking and relaxation of supercoiled DNA by psoralen and light

    International Nuclear Information System (INIS)

    Yoakum, G.H.; Cole, R.S.

    1978-01-01

    Photoreaction of 4,5',8-trimethylpsoralen with superhelical ColE1 and ColE1amp DNA was studied. Changes in mobilities in agarose gels, formation of interstrand cross-links, and DNA strand breaks were determined. Psoralen and light treatment removed negative superhelical turns, and extensive treatments failed to produce positive superhelical turns in covalently closed plasmid DNA. The rate of relaxation of superhelical turns by psoralen photobinding appeared to be directly proportional to the number of superhelical turns remaining. A unique reaction mechanism is presented to explain these results. By this interpretation the initial rate of psoralen photobinding to superhelical DNA was estimated to be 3 times that for linear DNA, and the ratio of cross-linking to monofunctional adducts appears to be dependent on the superhelical conformation of the DNA. The estimated ratio of psoralen molecules bound to DNA strand breaks was 1.7 . 10 4 :1, and 70% of this breakage is caused by the light alone. (Auth.)

  8. Generation of Guanine – Thymidine Cross-links in DNA by Peroxynitrite/Carbon Dioxide

    Science.gov (United States)

    Yun, Byeong Hwa; Geacintov, Nicholas E.; Shafirovich, Vladimir

    2011-01-01

    Nitrosoperoxycarbonate derived from the combination of carbon dioxide and peroxynitrite, is an important chemical mediator of inflammation. In aqueous solutions, it rapidly decomposes to the reactive species CO3•− and •NO2 radicals that are known to initiate the selective oxidation and nitration of guanine in DNA. We have previously demonstrated that the reactions of carbonate radical anions with guanine in 2′-deoxyoligoribonucleotides generate a previously unknown intrastrand cross-linked guanine-thymine product G*-T* with a covalent bond between the C8 (G*) and thymine N3 (T*) atoms (Crean et al., Nucleic Acids Res., 2008, 36, 742–755). In this work we demonstrate that G*-T* cross-linked products are also formed when peroxynitrite (0.1 mM) reacts with native DNA in aqueous solutions (pH 7.5–7.7) containing 25 mM carbon dioxide/bicarbonate, in addition to the well known nitration/oxidation products of guanine such as 8-nitroguanine (8-nitroG), 5-guanidino-4-nitroimidazole (NIm), 8-oxo-7,8-dehydroguanine (8-oxoG) and spiroiminodihydantoin (Sp). The yields of these products, after enzymatic digestion with P1 nuclease and alkaline phosphatase to the nucleotide level, and reversed phase HPLC separation, were compared with those obtained with the uniformly, isotopically labeled 15N,13C-labeled 2′-deoxy oligoribonucleotides 5′-dGpT and 5′-dGpCpT. The d(G*pT*) and d(G*-T*) cross-linked products derived from the di- and tri-oligonucleotides, respectively, were used as standards for identifying the analogous lesions in calf thymus DNA by isotope dilution LC-MS/MS methods in the selected reaction-monitoring mode. The Nim and 8nitroG are the major products formed (~ 0.05% each), and lesser amounts of 8-oxoG (~ 0.02%), and d(G*pT*) and d(G*-T*) enzymatic digestion products (~ 0.002% each) were found. It is shown that the formation of d(G*pT*) enzyme digestion product can arise only from intrastrand cross-links, whereas d(G*-T*) can arise from both

  9. LET dependence of DNA-protein cross-links

    International Nuclear Information System (INIS)

    Blakely, E.A.; Chang, P.Y.; Bjornstad, K.A.

    1995-08-01

    We have preliminary data indicating a fluence-dependent yield of particle-induced protein cross-links (DPC's) with a dependency on LET and particle residual energy. Our data indicate that the DPC yield for hamster fibroblasts in vitro irradiated at 32 keV/μm is similar to that reported for hamster cells irradiated with cobalt-60 gamma rays. At 100-120 keV/μm there is some evidence for an enhanced DPC yield with increasing particle fluence, but there are differences in the yields that are dependent on particle track structure

  10. Role of special cross-links in structure formation of bacterial DNA polymer

    Science.gov (United States)

    Agarwal, Tejal; Manjunath, G. P.; Habib, Farhat; Lakshmi Vaddavalli, Pavana; Chatterji, Apratim

    2018-01-01

    Using data from contact maps of the DNA-polymer of Escherichia coli (E. Coli) (at kilobase pair resolution) as an input to our model, we introduce cross-links between monomers in a bead-spring model of a ring polymer at very specific points along the chain. Via suitable Monte Carlo simulations, we show that the presence of these cross-links leads to a particular organization of the chain at large (micron) length scales of the DNA. We also investigate the structure of a ring polymer with an equal number of cross-links at random positions along the chain. We find that though the polymer does get organized at the large length scales, the nature of the organization is quite different from the organization observed with cross-links at specific biologically determined positions. We used the contact map of E. Coli bacteria which has around 4.6 million base pairs in a single circular chromosome. In our coarse-grained flexible ring polymer model, we used 4642 monomer beads and observed that around 80 cross-links are enough to induce the large-scale organization of the molecule accounting for statistical fluctuations caused by thermal energy. The length of a DNA chain even of a simple bacterial cell such as E. Coli is much longer than typical proteins, hence we avoided methods used to tackle protein folding problems. We define new suitable quantities to identify the large scale structure of a polymer chain with a few cross-links.

  11. Activity of the DNA minor groove cross-linking agent SG2000 (SJG-136) against canine tumours.

    Science.gov (United States)

    Mellinas-Gomez, Maria; Spanswick, Victoria J; Paredes-Moscosso, Solange R; Robson, Matthew; Pedley, R Barbara; Thurston, David E; Baines, Stephen J; Stell, Anneliese; Hartley, John A

    2015-08-19

    Cancer is the leading cause of death in older dogs and its prevalence is increasing. There is clearly a need to develop more effective anti-cancer drugs in dogs. SG2000 (SJG-136) is a sequence selective DNA minor groove cross-linking agent. Based on its in vitro potency, the spectrum of in vivo and clinical activity against human tumours, and its tolerability in human patients, SG2000 has potential as a novel therapeutic against spontaneously occurring canine malignancies. In vitro cytotoxicity was assessed using SRB and MTT assays, and in vivo activity was assessed using canine tumour xenografts. DNA interstrand cross-linking (ICL) was determined using a modification of the single cell gel electrophoresis (comet) assay. Effects on cell cycle distribution were assessed by flow cytometry and measurement of γ-H2AX by immunofluorescence and immunohistochemistry. SG2000 had a multi-log differential cytotoxic profile against a panel of 12 canine tumour cell lines representing a range of common tumour types in dogs. In the CMeC-1 melanoma cell line, DNA ICLs increased linearly with dose following a 1 h treatment. Peak ICL was achieved within 1 h and no removal was observed over 48 h. A relationship between DNA ICL formation and cytotoxicity was observed across cell lines. The formation of γ-H2AX foci was slow, becoming evident after 4 h and reaching a peak at 24 h. SG2000 exhibited significant anti-tumour activity against two canine melanoma tumour models in vivo. Anti-tumour activity was observed at 0.15 and 0.3 mg/kg given i.v. either once, or weekly x 3. Dose-dependent DNA ICL was observed in tumours (and to a lower level in peripheral blood mononuclear cells) at 2 h and persisted at 24 h. ICL increased following the second and third doses in a repeated dose schedule. At 24 h, dose dependent γ-H2AX foci were more numerous than at 2 h, and greater in tumours than in peripheral blood mononuclear cells. SG2000-induced H2AX phosphorylation measured by

  12. Cross-linking between thymine and indolyl radical: possible mechanisms for cross-linking of DNA and tryptophan-containing peptides.

    Science.gov (United States)

    Mitrasinovic, Petar M

    2005-01-01

    As experimentally observed in gamma-irradiated aqueous solutions of tryptophan-containing peptides in the presence of DNA, a fast electron (or hydrogen atom) transfer from the DNA restores an intact tryptophan residue at the expense of the DNA integrity. Alternatively, addition of the deprotonated electron-deficient indolyl radical to the DNA, followed by subsequent rearrangement, may lead toward DNA/tryptophan-containing peptide cross-linking. Herein, possible reaction mechanisms for thymine-indolyl radical cross-linking are proposed. The consistent use of the contact spin density distribution is the key virtue of this work. The Becke 3, Lee, Yang, and Parr (B3LYP) density functional theory (DFT) method is employed to investigate the feasibility of the proposed cross-linking mechanisms. A possible complete reaction mechanism consists of a combination of the C(5)-hydroxylated thymine and indolyl radicals forming the initial cross-linked product, a hydrogen transfer within the initial cross-linked product by use of a bridging water molecule, and a dehydration step. The overall thermodynamics of the free energy profiles at 0 and 298 K are similar and display differences of magnitude for the hydrogen-transfer reaction. Temperature may be a key factor influencing the overall mechanism. The skeletal structures and contact spin densities on the heavy atoms of the tryptophan side chain and indolyl radicals are essentially equal. Hence, it is believed that a direct combination of the C(5)-hydroxylated thymine and tryptophan radicals should form the initial cross-linked product, as far as addition of the tryptophan radical to the DNA is concerned.

  13. Mass spectrometric analysis of a UV-cross-linked protein-DNA complex: tryptophans 54 and 88 of E. coli SSB cross-link to DNA

    DEFF Research Database (Denmark)

    Steen, Hanno; Petersen, Jørgen; Mann, Matthias

    2001-01-01

    acid and peptide entities present in such heteroconjugates. Sample preparation of the peptide-nucleic acid heteroconjugates is, therefore, a crucial step in any mass spectrometry-based analytical procedure. This study demonstrates the performance of four different MS-based strategies to characterize E....... coli single-stranded DNA binding protein (SSB) that was UV-cross-linked to a 5-iodouracil containing DNA oligomer. Two methods were optimized to circumvent the need for standard liquid chromatography and gel electrophoresis, thereby dramatically increasing the overall sensitivity of the analysis...

  14. UV induced DNA-protein cross links in vitro and in vivo

    International Nuclear Information System (INIS)

    Kornhauser, A.

    1976-01-01

    The review was not intended to cover all the past year's literature in this field; only selective material published in 1974 and 1975 has been surveyed. Covalent linkage of DNA and RNA to proteins induced by UV is considered, but DNA-membrade attachment, amino acids covalently bound to DNA as functions of growth conditions and protein non-covalently bound to DNA involved in cell regulation are excluded. Studies of DNA-protein cross-links upon UV irradiation in chemical model systems, bacteria and tissue culture systems, and an in vivo mammalian system are all surveyed. (U.K.)

  15. Molecular Dynamics Insights into Polyamine-DNA Binding Modes: Implications for Cross-Link Selectivity.

    Science.gov (United States)

    Bignon, Emmanuelle; Chan, Chen-Hui; Morell, Christophe; Monari, Antonio; Ravanat, Jean-Luc; Dumont, Elise

    2017-09-18

    Biogenic polyamines, which play a role in DNA condensation and stabilization, are ubiquitous and are found at millimolar concentration in the nucleus of eukaryotic cells. The interaction modes of three polyamines-putrescine (Put), spermine (Spm), and spermidine (Spd)-with a self-complementary 16 base pair (bp) duplex, are investigated by all-atom explicit-solvent molecular dynamics. The length of the amine aliphatic chain leads to a change of the interaction mode from minor groove binding to major groove binding. Through all-atom dynamics, noncovalent interactions that stabilize the polyamine-DNA complex and prefigure the reactivity, leading to the low-barrier formation of deleterious DNA-polyamine cross-links, after one-electron oxidation of a guanine nucleobase, are unraveled. The binding strength is quantified from the obtained trajectories by molecular mechanics generalized Born surface area post-processing (MM-GBSA). The values of binding free energies provide the same affinity order, PutDNA-polyamine cross-link formation through the extraction of average approaching distances between the C8 atom of guanines and the ammonium group. These results imply that the formation of DNA-polyamine cross-links involves deprotonation of the guanine radical cation to attack the polyamines, which must be positively charged to lie in the vicinity of the B-helix. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. A Dominant Mutation in Human RAD51 Reveals Its Function in DNA Interstrand Crosslink Repair Independent of Homologous Recombination.

    Science.gov (United States)

    Wang, Anderson T; Kim, Taeho; Wagner, John E; Conti, Brooke A; Lach, Francis P; Huang, Athena L; Molina, Henrik; Sanborn, Erica M; Zierhut, Heather; Cornes, Belinda K; Abhyankar, Avinash; Sougnez, Carrie; Gabriel, Stacey B; Auerbach, Arleen D; Kowalczykowski, Stephen C; Smogorzewska, Agata

    2015-08-06

    Repair of DNA interstrand crosslinks requires action of multiple DNA repair pathways, including homologous recombination. Here, we report a de novo heterozygous T131P mutation in RAD51/FANCR, the key recombinase essential for homologous recombination, in a patient with Fanconi anemia-like phenotype. In vitro, RAD51-T131P displays DNA-independent ATPase activity, no DNA pairing capacity, and a co-dominant-negative effect on RAD51 recombinase function. However, the patient cells are homologous recombination proficient due to the low ratio of mutant to wild-type RAD51 in cells. Instead, patient cells are sensitive to crosslinking agents and display hyperphosphorylation of Replication Protein A due to increased activity of DNA2 and WRN at the DNA interstrand crosslinks. Thus, proper RAD51 function is important during DNA interstrand crosslink repair outside of homologous recombination. Our study provides a molecular basis for how RAD51 and its associated factors may operate in a homologous recombination-independent manner to maintain genomic integrity. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Sharp kink of DNA at psoralen-cross-link site deduced from crystal structure of psoralen-thymine monoadduct

    International Nuclear Information System (INIS)

    Kim, S.H.; Peckler, S.; Graves, B.; Kanne, D.; Rapoport, H.; Hearst, J.E.

    1983-01-01

    Light-induced cross-linking of double-stranded nucleic acids by psoralens has been exploited to locate, in vivo or in vitro, those double-helical regions of DNA or RNA that can accommodate any structural changes caused by the psoralen cross-links. To determine three-dimensional structural parameters of the cross-link, we have solved the crystal structure of the psoralen-thymine monoadduct formed in photoreaction between calf thymus DNA and 8-methoxypsoralen (8MOP). There are eight possible configurations for psoralen-thymine monoadducts and 64 for diadducts. We describe here the structural details of a psoralen-thymine monoadduct obtained in a biological environment and the consequences of the photo-cross-link between 8MOP and double-helical DNA

  18. Induction of SCE by DNA cross-links in human fibroblasts exposed to 8-MOP and UVA irradiation

    International Nuclear Information System (INIS)

    Bredberg, A.; Lambert, B.

    1983-01-01

    To study the SCE-inducing effect of psoralen cross-links in the DNA of normal, human fibroblasts, cell cultures were exposed to PUVA (0.2-1 μg of 8-MOP per ml, followed by UVA irradiation at 0.04 J/cm 2 ) and carefully washed to remove non-covalently bound psoralen. Some cell cultures were then given a second dose of UVA (1.1 J/cm 2 ), either immediately after PUVA or 1-3 days later. By this type of treatment, cells with different proportions of DNA cross-links are obtained. The initial PUVA treatment will mainly give rise to psoralen monoadducts and only few cross-links in the DNA, and the second UVA irradiation will convert a number of the psoralen monoadducts into cross-links. (orig./AJ)

  19. ZATT (ZNF451)-mediated resolution of topoisomerase 2 DNA-protein cross-links.

    Science.gov (United States)

    Schellenberg, Matthew J; Lieberman, Jenna Ariel; Herrero-Ruiz, Andrés; Butler, Logan R; Williams, Jason G; Muñoz-Cabello, Ana M; Mueller, Geoffrey A; London, Robert E; Cortés-Ledesma, Felipe; Williams, R Scott

    2017-09-29

    Topoisomerase 2 (TOP2) DNA transactions proceed via formation of the TOP2 cleavage complex (TOP2cc), a covalent enzyme-DNA reaction intermediate that is vulnerable to trapping by potent anticancer TOP2 drugs. How genotoxic TOP2 DNA-protein cross-links are resolved is unclear. We found that the SUMO (small ubiquitin-related modifier) ligase ZATT (ZNF451) is a multifunctional DNA repair factor that controls cellular responses to TOP2 damage. ZATT binding to TOP2cc facilitates a proteasome-independent tyrosyl-DNA phosphodiesterase 2 (TDP2) hydrolase activity on stalled TOP2cc. The ZATT SUMO ligase activity further promotes TDP2 interactions with SUMOylated TOP2, regulating efficient TDP2 recruitment through a "split-SIM" SUMO2 engagement platform. These findings uncover a ZATT-TDP2-catalyzed and SUMO2-modulated pathway for direct resolution of TOP2cc. Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

  20. Mismatch repair and nucleotide excision repair proteins cooperate in the recognition of DNA interstrand crosslinks

    Science.gov (United States)

    Zhao, Junhua; Jain, Aklank; Iyer, Ravi R.; Modrich, Paul L.; Vasquez, Karen M.

    2009-01-01

    DNA interstrand crosslinks (ICLs) are among the most cytotoxic types of DNA damage, thus ICL-inducing agents such as psoralen, are clinically useful chemotherapeutics. Psoralen-modified triplex-forming oligonucleotides (TFOs) have been used to target ICLs to specific genomic sites to increase the selectivity of these agents. However, how TFO-directed psoralen ICLs (Tdp-ICLs) are recognized and processed in human cells is unclear. Previously, we reported that two essential nucleotide excision repair (NER) protein complexes, XPA–RPA and XPC–RAD23B, recognized ICLs in vitro, and that cells deficient in the DNA mismatch repair (MMR) complex MutSβ were sensitive to psoralen ICLs. To further investigate the role of MutSβ in ICL repair and the potential interaction between proteins from the MMR and NER pathways on these lesions, we performed electrophoretic mobility-shift assays and chromatin immunoprecipitation analysis of MutSβ and NER proteins with Tdp-ICLs. We found that MutSβ bound to Tdp-ICLs with high affinity and specificity in vitro and in vivo, and that MutSβ interacted with XPA–RPA or XPC–RAD23B in recognizing Tdp-ICLs. These data suggest that proteins from the MMR and NER pathways interact in the recognition of ICLs, and provide a mechanistic link by which proteins from multiple repair pathways contribute to ICL repair. PMID:19468048

  1. RPA activates the XPF‐ERCC1 endonuclease to initiate processing of DNA interstrand crosslinks

    KAUST Repository

    Abdullah, Ummi B

    2017-06-13

    During replication‐coupled DNA interstrand crosslink (ICL) repair, the XPF‐ERCC1 endonuclease is required for the incisions that release, or “unhook”, ICLs, but the mechanism of ICL unhooking remains largely unknown. Incisions are triggered when the nascent leading strand of a replication fork strikes the ICL. Here, we report that while purified XPF‐ERCC1 incises simple ICL‐containing model replication fork structures, the presence of a nascent leading strand, modelling the effects of replication arrest, inhibits this activity. Strikingly, the addition of the single‐stranded DNA (ssDNA)‐binding replication protein A (RPA) selectively restores XPF‐ERCC1 endonuclease activity on this structure. The 5′–3′ exonuclease SNM1A can load from the XPF‐ERCC1‐RPA‐induced incisions and digest past the crosslink to quantitatively complete the unhooking reaction. We postulate that these collaborative activities of XPF‐ERCC1, RPA and SNM1A might explain how ICL unhooking is achieved in vivo.

  2. Bifunctional alkylating agent-mediated MGMT-DNA cross-linking and its proteolytic cleavage in 16HBE cells

    International Nuclear Information System (INIS)

    Cheng, Jin; Ye, Feng; Dan, Guorong; Zhao, Yuanpeng; Wang, Bin; Zhao, Jiqing; Sai, Yan; Zou, Zhongmin

    2016-01-01

    Nitrogen mustard (NM), a bifunctional alkylating agent (BAA), contains two alkyl arms and can act as a cross-linking bridge between DNA and protein to form a DNA-protein cross-link (DPC). O 6 -methylguanine–DNA methyltransferase (MGMT), a DNA repair enzyme for alkyl adducts removal, is found to enhance cell sensitivity to BAAs and to promote damage, possibly due to its stable covalent cross-linking with DNA mediated by BAAs. To investigate MGMT-DNA cross-link (mDPC) formation and its possible dual roles in NM exposure, human bronchial epithelial cell line 16HBE was subjected to different concentrations of HN2, a kind of NM, and we found mDPC was induced by HN2 in a concentration-dependent manner, but the mRNA and total protein of MGMT were suppressed. As early as 1 h after HN2 treatment, high mDPC was achieved and the level maintained for up to 24 h. Quick total DPC (tDPC) and γ-H2AX accumulation were observed. To evaluate the effect of newly predicted protease DVC1 on DPC cleavage, we applied siRNA of MGMT and DVC1, MG132 (proteasome inhibitor), and NMS-873 (p97 inhibitor) and found that proteolysis plays a role. DVC1 was proven to be more important in the cleavage of mDPC than tDPC in a p97-dependent manner. HN2 exposure induced DVC1 upregulation, which was at least partially contributed to MGMT cleavage by proteolysis because HN2-induced mDPC level and DNA damage was closely related with DVC1 expression. Homologous recombination (HR) was also activated. Our findings demonstrated that MGMT might turn into a DNA damage promoter by forming DPC when exposed to HN2. Proteolysis, especially DVC1, plays a crucial role in mDPC repair. - Highlights: • Nitrogen mustard-induced MGMT-DNA cross-linking was detected in a living cell. • Concentration- and time-dependent manners of MGMT-DNA cross-linking were revealed. • Proteolysis played an important role in protein (MGMT)-DNA cross-linking repair. • DVC1 acts as a proteolytic enzyme in cross-linking repair in a p

  3. Dependence of DNA-protein cross-linking via guanine oxidation upon local DNA sequence as studied by restriction endonuclease inhibition.

    Science.gov (United States)

    Madison, Amanda L; Perez, Zitadel A; To, Phuong; Maisonet, Tiffany; Rios, Eunice V; Trejo, Yuri; Ochoa-Paniagua, Carmen; Reno, Anita; Stemp, Eric D A

    2012-01-10

    Oxidative damage plays a causative role in many diseases, and DNA-protein cross-linking is one important consequence of such damage. It is known that GG and GGG sites are particularly prone to one-electron oxidation, and here we examined how the local DNA sequence influences the formation of DNA-protein cross-links induced by guanine oxidation. Oxidative DNA-protein cross-linking was induced between DNA and histone protein via the flash quench technique, a photochemical method that selectively oxidizes the guanine base in double-stranded DNA. An assay based on restriction enzyme cleavage was developed to detect the cross-linking in plasmid DNA. Following oxidation of pBR322 DNA by flash quench, several restriction enzymes (PpuMI, BamHI, EcoRI) were then used to probe the plasmid surface for the expected damage at guanine sites. These three endonucleases were strongly inhibited by DNA-protein cross-linking, whereas the AT-recognizing enzyme AseI was unaffected in its cleavage. These experiments also reveal the susceptibility of different guanine sites toward oxidative cross-linking. The percent inhibition observed for the endonucleases, and their pBR322 cleavage sites, decreased in the order: PpuMI (5'-GGGTCCT-3' and 5'-AGGACCC-3') > BamHI (5'-GGATCC-3') > EcoRI (5'-GAATTC-3'), a trend consistent with the observed and predicted tendencies for guanine to undergo one-electron oxidation: 5'-GGG-3' > 5'-GG-3' > 5'-GA-3'. Thus, it appears that in mixed DNA sequences the guanine sites most vulnerable to oxidative cross-linking are those that are easiest to oxidize. These results further indicate that equilibration of the electron hole in the plasmid DNA occurs on a time scale faster than that of cross-linking.

  4. Formation of platinated GG cross-links on DNA by photoactivation of a Pt(IV) azide complex

    Czech Academy of Sciences Publication Activity Database

    Kašpárková, Jana; Mackay, F. S.; Brabec, Viktor; Sadler, P. J.

    2003-01-01

    Roč. 8, č. 7 (2003), s. 741-745 ISSN 0949-8257 R&D Projects: GA AV ČR IAA5004101; GA AV ČR KJB5004301 Institutional research plan: CEZ:AV0Z5004920 Keywords : cross-links * DNA * guanine Subject RIV: BO - Biophysics Impact factor: 3.905, year: 2003

  5. Wavelength dependence for the photoreactions of DNA-psoralen monoadducts. 2. Photo-cross-linking of monoadducts

    International Nuclear Information System (INIS)

    Shi, Y.; Hearst, J.E.

    1987-01-01

    The photoreactions of HMT [4'-(hydroxymethyl)-4,5',8-trimethylpsoralen] monoadducts in double-stranded DNA have been studied with complementary oligonucleotides. The HMT was first attached to the thymidine residue in the oligonucleotide 5'-GAAGCTACGAGC-3' as either a furan-side monoadduct or a pyrone-side monoadduct. The HMT-monoadducted oligonucleotide was then hybridized to the complementary oligonucleotide 5'-GCTCGTAGCTTC-3' and irradiated with monochromatic light. In the case of the pyrone-side monoadducted oligonucleotide, photoreversal was the predominant reaction, and very little cross-link was formed at all wavelengths. The course of the photoreaction of the double-stranded furan-side monoadducted oligonucleotide was dependent on the irradiation wavelength. At wavelengths below 313 nm, both photoreversal and photo-cross-linking occurred. At wavelengths above 313 nm, photoreversal of the monoadduct could not be detected, and photo-cross-linking occurred efficiently with a quantum yield of 2,4 x 10 -2

  6. Evaluation of mutagenic effects of formocresol: detection of DNA-protein cross-links and micronucleus in mouse bone marrow.

    Science.gov (United States)

    Ramos, Maria Emília Santos Pereira; Cavalcanti, Bruno Coêlho; Lotufo, Letícia Veras Costa; de Moraes, Manoel Odorico; Cerqueira, Eneida de Moraes Marcílio; Pessoa, Cláudia

    2008-03-01

    The genotoxic potential of formocresol was assessed by comet assay on human peripheral blood lymphocytes and in vivo micronucleus in mice. Peripheral blood lymphocytes, obtained from healthy donors, were exposed directly with different dilutions of formocresol for 45 minutes at 37 degrees C. To verify the possibility of formocresol to induce DNA-protein cross-links, treated lymphocytes were incubated with proteinase K. Micronucleus test was performed on male Swiss mice treated with several dilutions of formocresol by single intraperitoneal injection. After treatment, bone marrow was sampled 24 and 48 hours after formocresol administration. Formocresol did not produce detectable DNA damage as evaluated by comet assay. However, after proteinase K exposure, a dose-dependent increase of DNA migration was observed. Formocresol induced a significant increase in micronucleus frequencies at the highest dilution only at 24 hours after administration. Formocresol induced DNA-protein cross-links and an increased frequency of micronucleus.

  7. Ultraviolet irradiation produces cytotoxic synergy and increased DNA interstrand crosslinking with cis- and trans-diamminedichloroplatinum(II)

    International Nuclear Information System (INIS)

    Swinnen, L.J.; Erickson, L.C.

    1989-01-01

    The excision-repair mechanism responsible for the removal of UV-induced thymine dimers may also play a role in the repair of cis-diamminedichloroplatinum(II) (cis-DDP)-induced DNA adducts in both bacteria and mammalian cells. It was hypothesized that UV dimers and cis-DDP adducts, when present simultaneously, might compete for a common repair system. Colony survival assays were performed in HT-29 human colon carcinoma cells exposed either to cis-DDP alone or to cis-DDP immediately followed by UV exposure. Progressively greater cytotoxic synergy with both increasing UV dose and cis-DDP dose was observed, to a point of saturation beyond which further toxicity was purely additive. An approximate doubling in DNA crosslink frequency, relative to cis-DDP alone, was found in cells exposed to cis-DDP plus UV. Since cis-DDP produces both inter- and intrastrand DNA crosslinks similar studies were performed with trans-DDP, which is incapable of producing intrastrand crosslinks, but does produce interstrand crosslinks. Cytotoxic synergy and increased interstrand crosslinking again resulted from the addition of UV exposure, but not to the same extent as seen with cis-DDP. (author)

  8. DNA-induced inter-particle cross-linking during expanded bed adsorption chromatography - Impact on future support design

    DEFF Research Database (Denmark)

    Theodossiou, Irini; Thomas, Owen R. T.

    2002-01-01

    We have investigated the effects of adsorbent size, ionic capacity and surface immobilised polymers on dynamic capacity and changes occurring to beds of anion-exchangers during the binding of DNA. During application of low concentrations of "3-20 kilobase" calf thymus DNA feeds to expanded beds...... of anion-exchangers, the bed heights dropped progressively as DNA molecules physically cross-linked neighbouring adsorbent particles together, to form severely aggregated fluidised beds. In plots of dynamic binding capacities and absolute changes in bed porosity at maximum contraction, against the inverse...

  9. Induction and repair of DNA cross-links induced by sulfur mustard in the A-549 cell line followed by a comet assay.

    Science.gov (United States)

    Jost, Petr; Svobodova, Hana; Stetina, Rudolf

    2015-07-25

    Sulfur mustard is a highly toxic chemical warfare agent with devastating impact on intoxicated tissues. DNA cross-links are probably the most toxic DNA lesions induced in the cell by sulfur mustard. The comet assay is a very sensitive method for measuring DNA damage. In the present study using the A-549 lung cell line, the comet assay protocol was optimized for indirect detection of DNA cross-links induced by sulfur mustard. The method is based on the additional treatment of the assayed cells containing cross-links with the chemical mutagen, styrene oxide. Alkali-labile adducts of styrene oxide cause DNA breaks leading to the formation of comets. A significant dose-dependent reduction of DNA migration of the comet's tail was found after exposing cells to sulfur mustard, indicative of the amount of sulfur mustard induced cross-links. The remarkable decrease of % tail DNA could be observed as early as 5min following exposure to sulfur mustard and the maximal effect was found after 30min, when DNA migration was reduced to the minimum. Sulfur mustard preincubated in culture medium without cells lost its ability to induce cross-links and had a half-life of about 15min. Pre-incubation longer than 30min does not lead to a significant increase in cross-links when applied to cells. However, the amount of cross-links is decreased during further incubation due to repair. The current modification of the comet assay provides a useful tool for detecting DNA cross-links induced by sulfur mustard and could be used for detection of other DNA cross-linking agents such as chemotherapeutic drugs. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  10. Thermodynamic stability and energetics of DNA duplexes containing major intrastrand cross-links of second-generation antitumor dinuclear Pt(II) complexes.

    Science.gov (United States)

    Florian, Jakub; Kasparkova, Jana; Farrell, Nicholas P; Brabec, Viktor

    2012-02-01

    The effects of major DNA intrastrand cross-links of antitumor dinuclear Pt(II) complexes [{trans-PtCl(NH(3))(2)}(2)-μ-{trans-(H(2)N(CH(2))(6)NH(2)(CH(2))(2)NH(2)(CH(2))(6)NH(2))}](4+) (1) and [{PtCl(DACH)}(2)-μ-{H(2)N(CH(2))(6)NH(2)(CH(2))(2)NH(2)(CH(2))(6)NH(2))}](4+) (2) (DACH is 1,2-diaminocyclohexane) on DNA stability were studied with emphasis on thermodynamic origins of that stability. Oligodeoxyribonucleotide duplexes containing the single 1,2, 1,3, or 1,5 intrastrand cross-links at guanine residues in the central TGGT, TGTGT, or TGTTTGT sequences, respectively, were prepared and analyzed by differential scanning calorimetry. The unfolding of the platinated duplexes was accompanied by unfavorable free energy terms. The efficiency of the cross-links to thermodynamically destabilize the duplex depended on the number of base pairs separating the platinated bases. The trend was 1,5→1,2→1,3 cross-link of 1 and 1,5→1,3→1,2 cross-link of 2. Interestingly, the results showed that the capability of the cross-links to reduce the thermodynamic stability of DNA (ΔG(298)(0)) correlated with the extent of conformational distortions induced in DNA by various types of intrastrand cross-links of 1 or 2 determined by chemical probes of DNA conformation. We also examined the efficiency of the mammalian nucleotide excision repair systems to remove from DNA the intrastrand cross-links of 1 or 2. The efficiency of the excinucleases to remove the cross-links from DNA depended on the length of the cross-link; the trend was identical to that observed for the efficiency of the intrastrand cross-links to thermodynamically destabilize the duplex. Thus, the results are consistent with the thesis that an important factor that determines the susceptibility of the intrastrand cross-links of dinuclear platinum complexes 1 and 2 to be removed from DNA by nucleotide excision repair is the efficiency of these lesions to thermodynamically destabilize DNA.

  11. Proteomics reveals dynamic assembly of repair complexes during bypass of DNA cross-links

    DEFF Research Database (Denmark)

    Räschle, Markus; Smeenk, Godelieve; Hansen, Rebecca K

    2015-01-01

    a technique called chromatin mass spectrometry (CHROMASS) to study protein recruitment dynamics during perturbed DNA replication in Xenopus egg extracts. Using CHROMASS, we systematically monitored protein assembly and disassembly on ICL-containing chromatin. Among numerous prospective DNA repair factors, we...... to damaged DNA in vertebrate cells....

  12. Adenosine Triphosphate-Triggered Release of Macromolecular and Nanoparticle Loads from Aptamer/DNA-Cross-Linked Microcapsules.

    Science.gov (United States)

    Liao, Wei-Ching; Lu, Chun-Hua; Hartmann, Raimo; Wang, Fuan; Sohn, Yang Sung; Parak, Wolfgang J; Willner, Itamar

    2015-09-22

    The synthesis of stimuli-responsive DNA microcapsules acting as carriers for different payloads, and being dissociated through the formation of aptamer-ligand complexes is described. Specifically, stimuli-responsive anti-adenosine triphosphate (ATP) aptamer-cross-linked DNA-stabilized microcapsules loaded with tetramethylrhodamine-modified dextran (TMR-D), CdSe/ZnS quantum dots (QDs), or microperoxidase-11 (MP-11) are presented. In the presence of ATP as trigger, the microcapsules are dissociated through the formation of aptamer-ATP complexes, resulting in the release of the respective loads. Selective unlocking of the capsules is demonstrated, and CTP, GTP, or TTP do not unlock the pores. The ATP-triggered release of MP-11 from the microcapsules enables the MP-11-catalyzed oxidation of Amplex UltraRed by H2O2 to the fluorescent product resorufin.

  13. Radiation-induced cross-link DNA damages: synthesis, measurement and insertion into oligonucleotides for replication and enzymatic repair studies

    International Nuclear Information System (INIS)

    Bellon, Sophie

    2003-01-01

    This research thesis addresses the synthesis, measurement and study of the biological impact of radio-induced DNA double damages. In the first part, the author reports the study of the reactivity and fate of the 5-(2'-desoxy-uridilyl)methyl radical which is one of the intermediates formed by oxidizing photo-sensitisation of thymine. The next part reports results of the formation and measurement of double damages of isolated and cellular DNA, notably in the case of γ irradiation. The third part reports the study of in vitro replication of one of the double damages. The behaviour of different polymerases with respect to the damage is reported. Finally, the modified oligonucleotide has been used as a substrate to highlight possible activities of enzymatic repair for this type of cross-link damages by purified proteins or proteins present within cellular extracts [fr

  14. Vinylsulfonamide and Acrylamide Modification of DNA for Cross-linking with Proteins

    Czech Academy of Sciences Publication Activity Database

    Daďová, Jitka; Orság, Petr; Pohl, Radek; Brázdová, Marie; Fojta, Miroslav; Hocek, Michal

    2013-01-01

    Roč. 52, č. 40 (2013), s. 10515-10518 ISSN 1433-7851 R&D Projects: GA ČR GA203/09/0317; GA ČR(CZ) GAP301/11/2076 Institutional support: RVO:61388963 ; RVO:68081707 Keywords : bioorthogonal chemistry * DNA * DNA polymerase * Michael additions * proteins Subject RIV: CC - Organic Chemistry Impact factor: 11.336, year: 2013

  15. Sequence selectivity of azinomycin B in DNA alkylation and cross-linking: a QM/MM study.

    Science.gov (United States)

    Senthilnathan, Dhurairajan; Kalaiselvan, Anbarasan; Venuvanalingam, Ponnambalam

    2013-01-01

    Azinomycin B--a well-known antitumor drug--forms cross-links with DNA through alkylation of purine bases and blocks tumor cell growth. This reaction has been modeled using the ONIOM (B3LYP/6-31+g(d):UFF) method to understand the mechanism and sequence selectivity. ONIOM results have been checked for reliability by comparing them with full quantum mechanics calculations for selected paths. Calculations reveal that, among the purine bases, guanine is more reactive and is alkylated by aziridine ring through the C10 position, followed by alkylation of the epoxide ring through the C21 position of Azinomycin B. While the mono alkylation is controlled kinetically, bis-alkylation is controlled thermodynamically. Solvent effects were included using polarized-continuum-model calculations and no significant change from gas phase results was observed.

  16. Detection of DNA damage induced in vivo by a cross-linking agent with a circular channel crucible oscillating viscometer.

    Science.gov (United States)

    Balbi, C; Abelmoschi, M L; Roner, R; Giaretti, W; Parodi, S; Santi, L

    1985-11-01

    DNA damage induced in vivo by the cross-linking agent mitomycin C (MMC) was investigated with a new oscillating crucible viscometer. Viscosity was measured by lysing rat liver nuclei in an alkaline lysing solution (pH 12.5; 25 degrees C). In control samples the viscosity increased very slowly with time, reaching a plateau only after 10-12 h. The process was accelerated and the maximum viscosity was decreased by alkaline single-stranded breaks arising from methylation and subsequent depurination of DNA in vitro with dimethylsulphate (DMS). MMC, when given alone, had no evident effect on the time needed for reaching plateau viscosity but it induced a small increase in maximum viscosity. When MMC was given in association with DMS, the time of disentanglement remained unchanged (accelerated) but maximum viscosity was increased in a dose dependent way. We conclude that these data clearly confirm that the slow steady increase of the viscosity of control DNA with time reflects mainly the process of unwinding of the two strands. The speed of this process seems to depend only from the number of unwinding points in DNA (breaks).

  17. Impact of estrogenic compounds on DNA integrity in human spermatozoa: Evidence for cross-linking and redox cycling activities

    Energy Technology Data Exchange (ETDEWEB)

    Bennetts, L.E.; De Iuliis, G.N.; Nixon, B.; Kime, M.; Zelski, K. [ARC Centre of Excellence in Biotechnology and Development and Discipline of Biological Sciences, University of Newcastle, NSW (Australia); McVicar, C.M.; Lewis, S.E. [Obstetrics and Gynaecology, Queen' s University, Belfast (United Kingdom); Aitken, R.J. [ARC Centre of Excellence in Biotechnology and Development and Discipline of Biological Sciences, University of Newcastle, NSW (Australia)], E-mail: jaitken@mail.newcastle.edu.au

    2008-05-10

    A great deal of circumstantial evidence has linked DNA damage in human spermatozoa with adverse reproductive outcomes including reduced fertility and high rates of miscarriage. Although oxidative stress is thought to make a significant contribution to DNA damage in the male germ line, the factors responsible for creating this stress have not been elucidated. One group of compounds that are thought to be active in this context are the estrogens, either generated as a result of the endogenous metabolism of androgens within the male reproductive tract or gaining access to the latter as a consequence of environmental exposure. In this study, a wide variety of estrogenic compounds were assessed for their direct effects on human spermatozoa in vitro. DNA integrity was assessed using the Comet and TUNEL assays, lesion frequencies were quantified by QPCR using targets within the mitochondrial and nuclear ({beta}-globin) genomes, DNA adducts were characterized by mass spectrometry and redox activity was monitored using dihydroethidium (DHE) as the probe. Of the estrogenic and estrogen analogue compounds evaluated, catechol estrogens, quercetin, diethylstilbestrol and pyrocatechol stimulated intense redox activity while genistein was only active at the highest doses tested. Other estrogens and estrogen analogues, such as 17{beta}-estradiol, nonylphenol, bisphenol A and 2,3-dihydroxynaphthalene were inactive. Estrogen-induced redox activity was associated with a dramatic loss of motility and, in the case of 2-hydroxyestradiol, the induction of significant DNA fragmentation. Mass spectrometry also indicated that catechol estrogens were capable of forming dimers that can cross-link the densely packed DNA strands in sperm chromatin, impairing nuclear decondensation. These results highlight the potential importance of estrogenic compounds in creating oxidative stress and DNA damage in the male germ line and suggest that further exploration of these compounds in the aetiology of

  18. Recognition of double-stranded DNA using energetically activated duplexes with interstrand zippers of 1-, 2-or 4-pyrenyl-functionalized O2 '-alkylated RNA monomers

    DEFF Research Database (Denmark)

    Karmakar, Saswata; Madsen, Andreas Stahl; Guenther, Dale C.

    2014-01-01

    Despite advances with triplex-forming oligonucleotides, peptide nucleic acids, polyamides and more recently engineered proteins, there remains an urgent need for synthetic ligands that enable specific recognition of double-stranded (ds) DNA to accelerate studies aiming at detecting, regulating......'-alkylated uridine monomers X-Z by means of thermal denaturation experiments, optical spectroscopy, force-field simulations and recognition experiments using DNA hairpins as model targets. We demonstrate that Invaders with +1 interstrand zippers of X or Y monomers efficiently recognize mixed-sequence DNA...

  19. DNA double strand breaks but not interstrand crosslinks prevent progress through meiosis in fully grown mouse oocytes.

    Directory of Open Access Journals (Sweden)

    Wai Shan Yuen

    Full Text Available There is some interest in how mammalian oocytes respond to different types of DNA damage because of the increasing expectation of fertility preservation in women undergoing chemotherapy. Double strand breaks (DSBs induced by ionizing radiation and agents such as neocarzinostatin (NCS, and interstrand crosslinks (ICLs induced by alkylating agents such as mitomycin C (MMC, are toxic DNA lesions that need to be repaired for cell survival. Here we examined the effects of NCS and MMC treatment on oocytes collected from antral follicles in mice, because potentially such oocytes are readily collected from ovaries and do not need to be in vitro grown to achieve meiotic competency. We found that oocytes were sensitive to NCS, such that this ionizing radiation mimetic blocked meiosis I and caused fragmented DNA. In contrast, MMC had no impact on the completion of either meiosis I or II, even at extremely high doses. However, oocytes treated with MMC did show γ-H2AX foci and following their in vitro maturation and parthenogenetic activation the development of the subsequent embryos was severely compromised. Addition of MMC to 1-cell embryos caused a similarly poor level of development, demonstrating oocytes have eventual sensitivity to this ICL-inducing agent but this does not occur during their meiotic division. In oocytes, the association of Fanconi Anemia protein, FANCD2, with sites of ICL lesions was not apparent until entry into the embryonic cell cycle. In conclusion, meiotic maturation of oocytes is sensitive to DSBs but not ICLs. The ability of oocytes to tolerate severe ICL damage and yet complete meiosis, means that this type of DNA lesion goes unrepaired in oocytes but impacts on subsequent embryo quality.

  20. Conformation of DNA GG intrastrand cross-link of antitumor oxaliplatin and its enantiomeric analog

    Czech Academy of Sciences Publication Activity Database

    Malina, Jaroslav; Nováková, Olga; Vojtíšková, Marie; Natile, G.; Brabec, Viktor

    2007-01-01

    Roč. 93, č. 11 (2007), s. 3950-3962 ISSN 0006-3495 R&D Projects: GA ČR(CZ) GA305/05/2030; GA ČR(CZ) GA203/06/1239; GA AV ČR(CZ) 1QS500040581; GA AV ČR(CZ) KAN200200651; GA MŠk(CZ) LC06030; GA AV ČR(CZ) KJB400040601 Institutional research plan: CEZ:AV0Z50040507; CEZ:AV0Z50040702 Keywords : DNA * oxaliplatina * cancer Subject RIV: BO - Biophysics Impact factor: 4.627, year: 2007

  1. Mechanism of the formation of DNA-protein cross-links by antitumor cisplatin

    Czech Academy of Sciences Publication Activity Database

    Chválová, Kateřina; Brabec, Viktor; Kašpárková, Jana

    2007-01-01

    Roč. 35, č. 6 (2007), s. 1812-1821 ISSN 0305-1048 R&D Projects: GA ČR(CZ) GA305/05/2030; GA ČR(CZ) GD204/03/H016; GA MZd(CZ) NR8562; GA AV ČR(CZ) 1QS500040581; GA AV ČR(CZ) KAN200200651; GA MŠk(CZ) LC06030; GA MŠk(CZ) 1P05OC070; GA MŠk(CZ) 1P05OC072 Institutional research plan: CEZ:AV0Z50040702 Keywords : DNA * crosslinks * cancer Subject RIV: BO - Biophysics Impact factor: 6.954, year: 2007

  2. Raman spectroscopy of DNA modified by intrastrand cross-links of antitumor cisplatin

    Czech Academy of Sciences Publication Activity Database

    Vrána, Oldřich; Mašek, Vlastimil; Dražan, Viktor; Brabec, Viktor

    2007-01-01

    Roč. 159, č. 1 (2007), s. 1-8 ISSN 1047-8477 R&D Projects: GA ČR(CZ) GA203/05/2032; GA ČR(CZ) GD204/03/H016; GA ČR(CZ) GA305/05/2030; GA AV ČR(CZ) 1QS500040581; GA AV ČR(CZ) KAN200200651; GA MŠk(CZ) LC06030; GA MŠk(CZ) 1P05OC070; GA MŠk(CZ) 1P05OC072 Institutional research plan: CEZ:AV0Z50040507; CEZ:AV0Z50040702 Keywords : DNA * platinum * cancer Subject RIV: BO - Biophysics Impact factor: 3.677, year: 2007

  3. Transplatin is cytotoxic when photoactivated: Enhanced formation of DNA cross-links

    Czech Academy of Sciences Publication Activity Database

    Heringová, Pavla; Woods, J.; Mackay, F.S.; Kašpárková, Jana; Sadler, P.J.; Brabec, Viktor

    2006-01-01

    Roč. 49, č. 26 (2006), s. 7792-7798 ISSN 0022-2623 R&D Projects: GA ČR(CZ) GA305/05/2030; GA ČR(CZ) GD204/03/H016; GA MZd(CZ) NR8562; GA AV ČR(CZ) 1QS500040581; GA MŠk(CZ) LC06030; GA MŠk(CZ) 1P05OC070; GA MŠk(CZ) 1P05OC071; GA MŠk(CZ) 1P05OC072; GA AV ČR(CZ) KAN200200651 Institutional research plan: CEZ:AV0Z50040507 Keywords : cancer * platinum * DNA Subject RIV: BO - Biophysics Impact factor: 5.115, year: 2006

  4. Alkali reversal of psoralen cross-link for the targeted delivery of psoralen monoadduct lesion

    International Nuclear Information System (INIS)

    Yeung, A.T.; Dinehart, W.J.; Jones, B.K.

    1988-01-01

    Psoralen intercalates into double-stranded DNA and photoreacts mainly with thymines to form monoadducts and interstrand cross-links. The authors used an oligonucleotide model to demonstrate a novel mechanism: the reversal of psoralen cross-links by base-catalyzed rearrangement at 90 0 C (BCR). The BCR reaction is more efficient than the photoreversal reaction. They show that the BCR occurs predominantly on the furan side of a psoralen cross-link. The cleavage does not result in the breaking of the DNA backbone, and the thymine based freed from the cross-link by the cleavage reaction appears to be unmodified. Similarly, BCR of the furan-side monoadduct of psoralen removed the psoralen molecule and regenerated the unaltered native oligonucleotide. The pyrone-side psoralen monoadduct is relatively resistant to BCR. One can use BCR to perform efficient oligonucleotide-directed, site-specific delivery of a psoralen monoadduct. As a demonstration of this approach, they have hybridized a 19 base long oligonucleotide vehicle containing a furan-side psoralen monoadduct to a 56 base long complementary oligonucleotide target strand and formed a specific cross-link at the target site with 365-nm UV. Subsequent BCR released the oligonucleotide vehicle and deposited the psoralen at the target site

  5. The Bis-Electrophile Diepoxybutane Cross-links DNA to Human Histones but Does Not Result in Enhanced Mutagenesis in Recombinant Systems

    Science.gov (United States)

    Loecken, Elisabeth M.; Dasari, Surendra; Hill, Salisha; Tabb, David L.; Guengerich, F. Peter

    2009-01-01

    1,2-Dibromoethane and 1,3-butadiene are cancer suspects present in the environment and have been used widely in industry. The mutagenic properties of 1,2-dibromoethane and the 1,3-butadiene oxidation product diepoxybutane are thought to be related to the bis-electrophilic character of these chemicals. The discovery that overexpression of O6-alkylguanine alkyltransferase (AGT) enhances bis-electrophile-induced mutagenesis prompted a search for other proteins that may act by a similar mechanism. A human liver screen for nuclear proteins that cross-link with DNA in the presence of 1,2-dibromoethane identified histones H2b and H3 as candidate proteins. Treatment of isolated histones H2b and H3 with diepoxybutane resulted in DNA-protein cross-links and produced protein adducts, and DNA-histone H2b cross-links were identified (immunochemically) in Escherichia coli cells expressing histone H2b. However, heterologous expression of histone H2b in E. coli failed to enhance bis-electrophile-induced mutagenesis. These results are similar to those found with the cross-link candidate glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Loecken, E. M. and Guengerich, F. P. (2008) Chem. Res. Toxicol. 21, 453−458) but, in contrast to GAPDH, histone H2b bound DNA with even higher affinity than AGT. The extent of DNA cross-linking of isolated histone H2b was similar to that of AGT, suggesting that differences in post-cross-linking events explain the difference in mutagenesis. PMID:19364102

  6. Study of Photochemical Cytosine to Uracil Transition via Ultrafast Photo-Cross-Linking Using Vinylcarbazole Derivatives in Duplex DNA.

    Science.gov (United States)

    Sethi, Siddhant; Nakamura, Shigetaka; Fujimoto, Kenzo

    2018-04-04

    Gene therapies, including genome editing, RNAi, anti-sense technology and chemical DNA editing are becoming major methods for the treatment of genetic disorders. Techniques like CRISPR-Cas9, zinc finger nuclease (ZFN) and transcription activator-like effector-based nuclease (TALEN) are a few such enzymatic techniques. Most enzymatic genome editing techniques have their disadvantages. Thus, non-enzymatic and non-invasive technologies for nucleic acid editing has been reported in this study which might possess some advantages over the older methods of DNA manipulation. 3-cyanovinyl carbazole ( CNV K) based nucleic acid editing takes advantage of photo-cross-linking between a target pyrimidine and the CNV K to afford deamination of cytosine and convert it to uracil. This method previously required the use of high temperatures but, in this study, it has been optimized to take place at physiological conditions. Different counter bases (inosine, guanine and cytosine) complementary to the target cytosine were used, along with derivatives of CNV K ( NH2V K and OHV K) to afford the deamination at physiological conditions.

  7. Study of Photochemical Cytosine to Uracil Transition via Ultrafast Photo-Cross-Linking Using Vinylcarbazole Derivatives in Duplex DNA

    Directory of Open Access Journals (Sweden)

    Siddhant Sethi

    2018-04-01

    Full Text Available Gene therapies, including genome editing, RNAi, anti-sense technology and chemical DNA editing are becoming major methods for the treatment of genetic disorders. Techniques like CRISPR-Cas9, zinc finger nuclease (ZFN and transcription activator-like effector-based nuclease (TALEN are a few such enzymatic techniques. Most enzymatic genome editing techniques have their disadvantages. Thus, non-enzymatic and non-invasive technologies for nucleic acid editing has been reported in this study which might possess some advantages over the older methods of DNA manipulation. 3-cyanovinyl carbazole (CNVK based nucleic acid editing takes advantage of photo-cross-linking between a target pyrimidine and the CNVK to afford deamination of cytosine and convert it to uracil. This method previously required the use of high temperatures but, in this study, it has been optimized to take place at physiological conditions. Different counter bases (inosine, guanine and cytosine complementary to the target cytosine were used, along with derivatives of CNVK (NH2VK and OHVK to afford the deamination at physiological conditions.

  8. Hoechst 33258 dye generates DNA-protein cross-links during ultraviolet light-induced photolysis of bromodeoxyuridine in replicated and repaired DNA

    Energy Technology Data Exchange (ETDEWEB)

    Guo Xicang; Morgan, W.F.; Cleaver, J.E.

    1986-08-01

    Substitution of bromodeoxyuridine for thymidine in the DNA of mammalian cells sensitizes them to a range of wavelengths of ultraviolet light. Cells are also sensitized to photochemical reactions involving dyes such as Hoechst 33258, which is used to produce differential staining of chromatids according to their bromodeoxyuridine content. Irradiation with 313 nm light of human and hamster cells containing bromodeoxyuridine in their DNA produced single-strand breaks but no DNA-protein cross-links. Irradiation with 360 nm light in the presence of Hoechst 33258 produced extensive DNA-protein cross-linkage as well as single-strand breaks. These cross-links were observed in DNA containing bromodeoxyuridine incorporated by either semiconservative or repair replication. When the protein was removed with proteinase K, bromodeoxyuridine in repair patches after irradiation by doses of ultraviolet (254 nm) light as low as 0.26 J/m/sup 2/ could readily be detected. Hoechst 33258-mediated photolysis, therefore, provides a sensitive new technique for measuring repair replication after ultraviolet light irradiation.

  9. Recognition of double-stranded DNA using energetically activated duplexes with interstrand zippers of 1-, 2- or 4-pyrenyl-functionalized O2'-alkylated RNA monomers.

    Science.gov (United States)

    Karmakar, Saswata; Madsen, Andreas S; Guenther, Dale C; Gibbons, Bradley C; Hrdlicka, Patrick J

    2014-10-21

    Despite advances with triplex-forming oligonucleotides, peptide nucleic acids, polyamides and--more recently--engineered proteins, there remains an urgent need for synthetic ligands that enable specific recognition of double-stranded (ds) DNA to accelerate studies aiming at detecting, regulating and modifying genes. Invaders, i.e., energetically activated DNA duplexes with interstrand zipper arrangements of intercalator-functionalized nucleotides, are emerging as an attractive approach toward this goal. Here, we characterize and compare Invaders based on 1-, 2- and 4-pyrenyl-functionalized O2'-alkylated uridine monomers X-Z by means of thermal denaturation experiments, optical spectroscopy, force-field simulations and recognition experiments using DNA hairpins as model targets. We demonstrate that Invaders with +1 interstrand zippers of X or Y monomers efficiently recognize mixed-sequence DNA hairpins with single nucleotide fidelity. Intercalator-mediated unwinding and activation of the double-stranded probe, coupled with extraordinary stabilization of probe-target duplexes (ΔT(m)/modification up to +14.0 °C), provides the driving force for dsDNA recognition. In contrast, Z-modified Invaders show much lower dsDNA recognition efficiency. Thus, even very conservative changes in the chemical makeup of the intercalator-functionalized nucleotides used to activate Invader duplexes, affects dsDNA-recognition efficiency of the probes, which highlights the importance of systematic structure-property studies. The insight from this study will guide future design of Invaders for applications in molecular biology and nucleic acid diagnostics.

  10. The bis-electrophile diepoxybutane cross-links DNA to human histones but does not result in enhanced mutagenesis in recombinant systems.

    Science.gov (United States)

    Loecken, Elisabeth M; Dasari, Surendra; Hill, Salisha; Tabb, David L; Guengerich, F Peter

    2009-06-01

    1,2-Dibromoethane and 1,3-butadiene are cancer suspects present in the environment and have been used widely in industry. The mutagenic properties of 1,2-dibromoethane and the 1,3-butadiene oxidation product diepoxybutane are thought to be related to the bis-electrophilic character of these chemicals. The discovery that overexpression of O(6)-alkylguanine alkyltransferase (AGT) enhances bis-electrophile-induced mutagenesis prompted a search for other proteins that may act by a similar mechanism. A human liver screen for nuclear proteins that cross-link with DNA in the presence of 1,2-dibromoethane identified histones H2b and H3 as candidate proteins. Treatment of isolated histones H2b and H3 with diepoxybutane resulted in DNA-protein cross-links and produced protein adducts, and DNA-histone H2b cross-links were identified (immunochemically) in Escherichia coli cells expressing histone H2b. However, heterologous expression of histone H2b in E. coli failed to enhance bis-electrophile-induced mutagenesis. These results are similar to those found with the cross-link candidate glyceraldehyde 3-phosphate dehydrogenase (GAPDH) [ Loecken , E. M. and Guengerich , F. P. ( 2008 ) Chem. Res. Toxicol. 21 , 453 - 458 ], but in contrast to GAPDH, histone H2b bound DNA with even higher affinity than AGT. The extent of DNA cross-linking of isolated histone H2b was similar to that of AGT, suggesting that differences in postcross-linking events explain the difference in mutagenesis.

  11. Crystal structure of a nucleoside model for the interstrand cross-link formed by the reaction of 2′-deoxyguanosine and an abasic site in duplex DNA

    Directory of Open Access Journals (Sweden)

    Michael J. Catalano

    2016-05-01

    Full Text Available The title compound, 9-[(2R,4S,5R-4-hydroxy-5-(hydroxymethyltetrahydrofuran-2-yl]-2-{[(2R,4S,5R-4-methoxy-5-(methoxymethyltetrahydrofuran-2-yl]amino}-1H-purin-6(9H-one, C17H25N5O7, crystallizes with two independent molecules (A and B in the asymmetric unit. In the crystal, the guanosine moieties of molecules A and B are linked by N—H...N and O—H...N hydrogen-bonding interactions, forming ribbons which are stacked to form columns along [100]. These columns are then linked by O—H...O hydrogen bonds between the ribose moieties and numerous C—H...O interactions to complete the three-dimensional structure.

  12. Sequence Specificity, Corformation, and Recognition by HMG1 Protein of Major DNA Interstrand Cross-links of Antitumor Dinuclear Platinum Complexes

    Czech Academy of Sciences Publication Activity Database

    Kašpárková, Jana; Farrell, N.; Brabec, Viktor

    2000-01-01

    Roč. 275, - (2000), s. 15789-15798 ISSN 0021-9258 R&D Projects: GA ČR GA305/99/0695; GA ČR GA307/97/P029; GA AV ČR IAA5004702; GA MŠk ME 152 Institutional research plan: CEZ:A17/98:Z5-004-9-ii Subject RIV: BO - Biophysics Impact factor: 7.368, year: 2000

  13. Human XPC-hHR23B interacts with XPA-RPA in the recognition of triplex-directed psoralen DNA interstrand crosslinks

    DEFF Research Database (Denmark)

    Thoma, Brian S; Wakasugi, Mitsuo; Christensen, Jesper

    2005-01-01

    DNA interstrand crosslinks (ICLs) represent a severe form of damage that blocks DNA metabolic processes and can lead to cell death or carcinogenesis. The repair of DNA ICLs in mammals is not well characterized. We have reported previously that a key protein complex of nucleotide excision repair...... (NER), XPA-RPA, recognizes DNA ICLs. We now report the use of triplex technology to direct a site-specific psoralen ICL to a target DNA substrate to determine whether the human global genome NER damage recognition complex, XPC-hHR23B, recognizes this lesion. Our results demonstrate that XPC-hHR23B...... recognizes psoralen ICLs, which have a structure fundamentally different from other lesions that XPC-hHR23B is known to bind, with high affinity and specificity. XPC-hHR23B and XPA-RPA protein complexes were also observed to bind psoralen ICLs simultaneously, demonstrating not only that psoralen ICLs...

  14. Recognition of double-stranded DNA using energetically activated duplexes with interstrand zippers of 1-, 2- or 4-pyrenyl-functionalized O2′-alkylated RNA monomers†

    Science.gov (United States)

    Karmakar, Saswata; Madsen, Andreas S.; Guenther, Dale C.; Gibbons, Bradley C.; Hrdlicka, Patrick J.

    2014-01-01

    Despite advances with triplex-forming oligonucleotides, peptide nucleic acids, polyamides and - more recently - engineered proteins, there remains an urgent need for synthetic ligands that enable specific recognition of double-stranded (ds) DNA to accelerate studies aiming at detecting, regulating and modifying genes. Invaders, i.e., energetically activated DNA duplexes with interstrand zipper arrangements of intercalator-functionalized nucleotides, are emerging as an attractive approach toward this goal. Here, we characterize and compare Invaders based on 1-, 2- and 4-pyrenyl-functionalized O2′-alkylated uridine monomers X–Z by means of thermal denaturation experiments, optical spectroscopy, force-field simulations and recognition experiments using DNA hairpins as model targets. We demonstrate that Invaders with +1 interstrand zippers of X or Y monomers efficiently recognize mixed-sequence DNA hairpins with single nucleotide fidelity. Intercalator-mediated unwinding and activation of the double-stranded probe, coupled with extraordinary stabilization of probe-target duplexes (ΔTm/modification up to +14.0 °C), provides the driving force for dsDNA recognition. In contrast, Z-modified Invaders show much lower dsDNA recognition efficiency. Thus, even very conservative changes in the chemical makeup of the intercalator-functionalized nucleotides used to activate Invader duplexes, affects dsDNA-recognition efficiency of the probes, which highlights the importance of systematic structure-property studies. The insight from this study will guide future design of Invaders for applications in molecular biology and nucleic acid diagnostics. PMID:25144705

  15. DNA Condensing Effects and Sequence Selectivity of DNA Binding of Antitumor Noncovalent Polynuclear Platinum Complexes

    Czech Academy of Sciences Publication Activity Database

    Malina, Jaroslav; Farrell, N. P.; Brabec, Viktor

    2014-01-01

    Roč. 53, č. 3 (2014), s. 1662-1671 ISSN 0020-1669 R&D Projects: GA ČR(CZ) GA13-08273S; GA MŠk(CZ) LH13096 Institutional support: RVO:68081707 Keywords : INTERSTRAND CROSS-LINKS * LASER-LIGHT SCATTERING * CALF THYMUS DNA Subject RIV: BO - Biophysics Impact factor: 4.762, year: 2014

  16. Human MLH1 protein participates in genomic damage checkpoint signaling in response to DNA interstrand crosslinks, while MSH2 functions in DNA repair.

    Directory of Open Access Journals (Sweden)

    Qi Wu

    2008-09-01

    Full Text Available DNA interstrand crosslinks (ICLs are among the most toxic types of damage to a cell. For this reason, many ICL-inducing agents are effective therapeutic agents. For example, cisplatin and nitrogen mustards are used for treating cancer and psoralen plus UVA (PUVA is useful for treating psoriasis. However, repair mechanisms for ICLs in the human genome are not clearly defined. Previously, we have shown that MSH2, the common subunit of the human MutSalpha and MutSbeta mismatch recognition complexes, plays a role in the error-free repair of psoralen ICLs. We hypothesized that MLH1, the common subunit of human MutL complexes, is also involved in the cellular response to psoralen ICLs. Surprisingly, we instead found that MLH1-deficient human cells are more resistant to psoralen ICLs, in contrast to the sensitivity to these lesions displayed by MSH2-deficient cells. Apoptosis was not as efficiently induced by psoralen ICLs in MLH1-deficient cells as in MLH1-proficient cells as determined by caspase-3/7 activity and binding of annexin V. Strikingly, CHK2 phosphorylation was undetectable in MLH1-deficient cells, and phosphorylation of CHK1 was reduced after PUVA treatment, indicating that MLH1 is involved in signaling psoralen ICL-induced checkpoint activation. Psoralen ICLs can result in mutations near the crosslinked sites; however, MLH1 function was not required for the mutagenic repair of these lesions, and so its signaling function appears to have a role in maintaining genomic stability following exposure to ICL-induced DNA damage. Distinguishing the genetic status of MMR-deficient tumors as MSH2-deficient or MLH1-deficient is thus potentially important in predicting the efficacy of treatment with psoralen and perhaps with other ICL-inducing agents.

  17. Chemoprotective effect of taurine on potassium bromate-induced DNA damage, DNA-protein cross-linking and oxidative stress in rat intestine.

    Directory of Open Access Journals (Sweden)

    Mir Kaisar Ahmad

    Full Text Available Potassium bromate (KBrO3 is widely used as a food additive and is a major water disinfection by-product. It induces multiple organ toxicity in humans and experimental animals and is a probable human carcinogen. The present study reports the protective effect of dietary antioxidant taurine on KBrO3-induced damage to the rat intestine. Animals were randomly divided into four groups: control, KBrO3 alone, taurine alone and taurine+ KBrO3. Administration of KBrO3 alone led to decrease in the activities of intestinal brush border membrane enzymes while those of antioxidant defence and carbohydrate metabolism were also severely altered. There was increase in DNA damage and DNA-protein cross-linking. Treatment with taurine, prior to administration of KBrO3, resulted in significant attenuation in all these parameters but the administration of taurine alone had no effect. Histological studies supported these biochemical results showing extensive intestinal damage in KBrO3-treated animals and greatly reduced tissue injury in the taurine+ KBrO3 group. These results show that taurine ameliorates bromate induced tissue toxicity and oxidative damage by improving the antioxidant defence, tissue integrity and energy metabolism. Taurine can, therefore, be potentially used as a therapeutic/protective agent against toxicity of KBrO3 and related compounds.

  18. Chemoprotective effect of taurine on potassium bromate-induced DNA damage, DNA-protein cross-linking and oxidative stress in rat intestine.

    Science.gov (United States)

    Ahmad, Mir Kaisar; Khan, Aijaz Ahmed; Ali, Shaikh Nisar; Mahmood, Riaz

    2015-01-01

    Potassium bromate (KBrO3) is widely used as a food additive and is a major water disinfection by-product. It induces multiple organ toxicity in humans and experimental animals and is a probable human carcinogen. The present study reports the protective effect of dietary antioxidant taurine on KBrO3-induced damage to the rat intestine. Animals were randomly divided into four groups: control, KBrO3 alone, taurine alone and taurine+ KBrO3. Administration of KBrO3 alone led to decrease in the activities of intestinal brush border membrane enzymes while those of antioxidant defence and carbohydrate metabolism were also severely altered. There was increase in DNA damage and DNA-protein cross-linking. Treatment with taurine, prior to administration of KBrO3, resulted in significant attenuation in all these parameters but the administration of taurine alone had no effect. Histological studies supported these biochemical results showing extensive intestinal damage in KBrO3-treated animals and greatly reduced tissue injury in the taurine+ KBrO3 group. These results show that taurine ameliorates bromate induced tissue toxicity and oxidative damage by improving the antioxidant defence, tissue integrity and energy metabolism. Taurine can, therefore, be potentially used as a therapeutic/protective agent against toxicity of KBrO3 and related compounds.

  19. Chemoprotective Effect of Taurine on Potassium Bromate-Induced DNA Damage, DNA-Protein Cross-Linking and Oxidative Stress in Rat Intestine

    Science.gov (United States)

    Ahmad, Mir Kaisar; Khan, Aijaz Ahmed; Ali, Shaikh Nisar; Mahmood, Riaz

    2015-01-01

    Potassium bromate (KBrO3) is widely used as a food additive and is a major water disinfection by-product. It induces multiple organ toxicity in humans and experimental animals and is a probable human carcinogen. The present study reports the protective effect of dietary antioxidant taurine on KBrO3-induced damage to the rat intestine. Animals were randomly divided into four groups: control, KBrO3 alone, taurine alone and taurine+ KBrO3. Administration of KBrO3 alone led to decrease in the activities of intestinal brush border membrane enzymes while those of antioxidant defence and carbohydrate metabolism were also severely altered. There was increase in DNA damage and DNA-protein cross-linking. Treatment with taurine, prior to administration of KBrO3, resulted in significant attenuation in all these parameters but the administration of taurine alone had no effect. Histological studies supported these biochemical results showing extensive intestinal damage in KBrO3-treated animals and greatly reduced tissue injury in the taurine+ KBrO3 group. These results show that taurine ameliorates bromate induced tissue toxicity and oxidative damage by improving the antioxidant defence, tissue integrity and energy metabolism. Taurine can, therefore, be potentially used as a therapeutic/protective agent against toxicity of KBrO3 and related compounds. PMID:25748174

  20. Arginine-rich cross-linking peptides with different SV40 nuclear localization signal content as vectors for intranuclear DNA delivery.

    Science.gov (United States)

    Bogacheva, Mariia; Egorova, Anna; Slita, Anna; Maretina, Marianna; Baranov, Vladislav; Kiselev, Anton

    2017-11-01

    The major barriers for intracellular DNA transportation by cationic polymers are their toxicity, poor endosomal escape and inefficient nuclear uptake. Therefore, we designed novel modular peptide-based carriers modified with SV40 nuclear localization signal (NLS). Core peptide consists of arginine, histidine and cysteine residues for DNA condensation, endosomal escape promotion and interpeptide cross-linking, respectively. We investigated three polyplexes with different NLS content (10 mol%, 50 mol% and 90 mol% of SV40 NLS) as vectors for intranuclear DNA delivery. All carriers tested were able to condense DNA, to protect it from DNAase I and were not toxic to the cells. We observed that cell cycle arrest by hydroxyurea did not affect transfection efficacy of NLS-modified carriers which we confirmed using quantitative confocal microscopy analysis. Overall, peptide carrier modified with 90 mol% of SV40 NLS provided efficient transfection and nuclear uptake in non-dividing cells. Thus, incorporation of NLS into arginine-rich cross-linking peptides is an adequate approach to the development of efficient intranuclear gene delivery vehicles. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Characterization of hepatic DNA damage induced in rats by the pyrrolizidine alkaloid monocrotaline

    Energy Technology Data Exchange (ETDEWEB)

    Petry, T.W.; Bowden, G.T.; Huxtable, R.J.; Sipes, I.G.

    1984-04-01

    Hepatic DNA damage induced by the pyrrolizidine alkaloid monocrotaline was evaluated following i.p. administration to adult male Sprague-Dawley rats. Animals were treated with various doses ranging upward from 5 mg/kg, and hepatic nuclei were isolated 4 hr later. Hepatic nuclei were used as the DNA source in all experiments. DNA damage was characterized by the alkaline elution technique. A mixture of DNA-DNA interstrand cross-links and DNA-protein cross-links was induced. Following an injection of monocrotaline, 30 mg/kg i.p., DNA-DNA interstrand cross-linking reached a maximum within 12 hr or less and thereafter decreased over a protracted period of time. By 96 hr postadministration, the calculated cross-linking factor was no longer statistically different from zero. No evidence for the induction of DNA single-strand breaks was observed, although the presence of small numbers of DNA single-strand breaks could have been masked by the overwhelming predominance of DNA cross-links. These DNA cross-links may be related to the hepatocarcinogenic, hepatotoxic, and/or antimitotic effects of monocrotaline.

  2. Interplay between human high mobility group protein 1 and replication protein A on psoralen-cross-linked DNA

    DEFF Research Database (Denmark)

    Reddy, Madhava C; Christensen, Jesper; Vasquez, Karen M

    2005-01-01

    Human high mobility group box (HMGB) 1 and -2 proteins are highly conserved and abundant chromosomal proteins that regulate chromatin structure and DNA metabolism. HMGB proteins bind preferentially to DNA that is bent or underwound and to DNA damaged by agents such as cisplatin, UVC radiation...

  3. Drosophila DNA polymerase theta utilizes both helicase-like and polymerase domains during microhomology-mediated end joining and interstrand crosslink repair.

    Science.gov (United States)

    Beagan, Kelly; Armstrong, Robin L; Witsell, Alice; Roy, Upasana; Renedo, Nikolai; Baker, Amy E; Schärer, Orlando D; McVey, Mitch

    2017-05-01

    Double strand breaks (DSBs) and interstrand crosslinks (ICLs) are toxic DNA lesions that can be repaired through multiple pathways, some of which involve shared proteins. One of these proteins, DNA Polymerase θ (Pol θ), coordinates a mutagenic DSB repair pathway named microhomology-mediated end joining (MMEJ) and is also a critical component for bypass or repair of ICLs in several organisms. Pol θ contains both polymerase and helicase-like domains that are tethered by an unstructured central region. While the role of the polymerase domain in promoting MMEJ has been studied extensively both in vitro and in vivo, a function for the helicase-like domain, which possesses DNA-dependent ATPase activity, remains unclear. Here, we utilize genetic and biochemical analyses to examine the roles of the helicase-like and polymerase domains of Drosophila Pol θ. We demonstrate an absolute requirement for both polymerase and ATPase activities during ICL repair in vivo. However, similar to mammalian systems, polymerase activity, but not ATPase activity, is required for ionizing radiation-induced DSB repair. Using a site-specific break repair assay, we show that overall end-joining efficiency is not affected in ATPase-dead mutants, but there is a significant decrease in templated insertion events. In vitro, Pol θ can efficiently bypass a model unhooked nitrogen mustard crosslink and promote DNA synthesis following microhomology annealing, although ATPase activity is not required for these functions. Together, our data illustrate the functional importance of the helicase-like domain of Pol θ and suggest that its tethering to the polymerase domain is important for its multiple functions in DNA repair and damage tolerance.

  4. BO-1055, a novel DNA cross-linking agent with remarkable low myelotoxicity shows potent activity in sarcoma models

    OpenAIRE

    Ambati, Srikanth R.; Shieh, Jae-Hung; Pera, Benet; Lopes, Eloisi Caldas; Chaudhry, Anisha; Wong, Elissa W.P.; Saxena, Ashish; Su, Tsann-Long; Moore, Malcolm A.S.

    2016-01-01

    DNA damaging agents cause rapid shrinkage of tumors and form the basis of chemotherapy for sarcomas despite significant toxicities. Drugs having superior efficacy and wider therapeutic windows are needed to improve patient outcomes. We used cell proliferation and apoptosis assays in sarcoma cell lines and benign cells; ?-H2AX expression, comet assay, immunoblot analyses and drug combination studies in vitro and in patient derived xenograft (PDX) models. BO-1055 caused apoptosis and cell death...

  5. BO-1055, a novel DNA cross-linking agent with remarkable low myelotoxicity shows potent activity in sarcoma models.

    Science.gov (United States)

    Ambati, Srikanth R; Shieh, Jae-Hung; Pera, Benet; Lopes, Eloisi Caldas; Chaudhry, Anisha; Wong, Elissa W P; Saxena, Ashish; Su, Tsann-Long; Moore, Malcolm A S

    2016-07-12

    DNA damaging agents cause rapid shrinkage of tumors and form the basis of chemotherapy for sarcomas despite significant toxicities. Drugs having superior efficacy and wider therapeutic windows are needed to improve patient outcomes. We used cell proliferation and apoptosis assays in sarcoma cell lines and benign cells; γ-H2AX expression, comet assay, immunoblot analyses and drug combination studies in vitro and in patient derived xenograft (PDX) models. BO-1055 caused apoptosis and cell death in a concentration and time dependent manner in sarcoma cell lines. BO-1055 had potent activity (submicromolar IC50) against Ewing sarcoma and rhabdomyosarcoma, intermediate activity in DSRCT (IC50 = 2-3μM) and very weak activity in osteosarcoma (IC50 >10μM) cell lines. BO-1055 exhibited a wide therapeutic window compared to other DNA damaging drugs. BO-1055 induced more DNA double strand breaks and γH2AX expression in cancer cells compared to benign cells. BO-1055 showed inhibition of tumor growth in A673 xenografts and caused tumor regression in cyclophosphamide resistant patient-derived Ewing sarcoma xenografts and A204 xenografts. Combination of BO-1055 and irinotecan demonstrated synergism in Ewing sarcoma PDX models. Potent activity on sarcoma cells and its relative lack of toxicity presents a strong rationale for further development of BO-1055 as a therapeutic agent.

  6. ATR-p53 restricts homologous recombination in response to replicative stress but does not limit DNA interstrand crosslink repair in lung cancer cells.

    Directory of Open Access Journals (Sweden)

    Bianca M Sirbu

    Full Text Available Homologous recombination (HR is required for the restart of collapsed DNA replication forks and error-free repair of DNA double-strand breaks (DSB. However, unscheduled or hyperactive HR may lead to genomic instability and promote cancer development. The cellular factors that restrict HR processes in mammalian cells are only beginning to be elucidated. The tumor suppressor p53 has been implicated in the suppression of HR though it has remained unclear why p53, as the guardian of the genome, would impair an error-free repair process. Here, we show for the first time that p53 downregulates foci formation of the RAD51 recombinase in response to replicative stress in H1299 lung cancer cells in a manner that is independent of its role as a transcription factor. We find that this downregulation of HR is not only completely dependent on the binding site of p53 with replication protein A but also the ATR/ATM serine 15 phosphorylation site. Genetic analysis suggests that ATR but not ATM kinase modulates p53's function in HR. The suppression of HR by p53 can be bypassed under experimental conditions that cause DSB either directly or indirectly, in line with p53's role as a guardian of the genome. As a result, transactivation-inactive p53 does not compromise the resistance of H1299 cells to the interstrand crosslinking agent mitomycin C. Altogether, our data support a model in which p53 plays an anti-recombinogenic role in the ATR-dependent mammalian replication checkpoint but does not impair a cell's ability to use HR for the removal of DSB induced by cytotoxic agents.

  7. Preparation of a hyper-cross-linked polymer monolithic column and its application to the sensitive determination of genomic DNA methylation.

    Science.gov (United States)

    Chen, Ming-Luan; Liu, Yu-Li; Xing, Xi-Wen; Zhou, Xiang; Feng, Yu-Qi; Yuan, Bi-Feng

    2013-01-14

    A hyper-cross-linked polymer monolithic column, poly(methacrylatoethyl trimethyl ammonium-co-vinylbenzene chloride-co-divinylbenzene) (MATE-co-VBC-co-DVB) with phenyl and quaternary ammonium groups was successfully prepared in the current study. The prepared monolith possesses large specific surface area, narrow mesopore size distribution and high column efficiency. The poly(MATE-co-VBC-co-DVB) monolithic column was demonstrated to have strong anion exchange/reversed-phase (SAX/RP) mixed-mode retention for analytes on capillary liquid chromatography (cLC). By using this monolithic column, we developed a rapid and sensitive method for the detection of DNA methylation. Our results showed that six nucleobases (adenine, guanine, cytosine, thymine, uracil, and 5-methylcytosine (5-mC)) can be baseline separated within 15 min by electrostatic repulsion and hydrophobic interactions between nucleobases and the monolithic stationary phase. The limit of detection (LOD, signal/noise = 3) of 5-mC is 0.014 pmol and endogenous 5-mC can be distinctly detected by using only 10 ng genomic DNA, which is comparable to that obtained by mass spectrometry analysis. Furthermore, by using the method developed here, we found that DNA methylation inhibitor 5-azacytidine (5-aza-C) and 5-aza-2'-deoxycytidine (5-aza-CdR) could induce a significant decrease of genome-wide DNA methylation in human lung carcinoma cells (A549) and cervical carcinoma cells (HeLa). Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Genetic inactivation of the Fanconi anemia gene FANCC identified in the hepatocellular carcinoma cell line HuH-7 confers sensitivity towards DNA-interstrand crosslinking agents

    Directory of Open Access Journals (Sweden)

    Bassermann Florian

    2010-05-01

    Full Text Available Abstract Background Inactivation of the Fanconi anemia (FA pathway through defects in one of 13 FA genes occurs at low frequency in various solid cancer entities among the general population. As FA pathway inactivation confers a distinct hypersensitivity towards DNA interstrand-crosslinking (ICL-agents, FA defects represent rational targets for individualized therapeutic strategies. Except for pancreatic cancer, however, the prevalence of FA defects in gastrointestinal (GI tumors has not yet been systematically explored. Results A panel of GI cancer cell lines was screened for FA pathway inactivation applying FANCD2 monoubiquitination and FANCD2/RAD51 nuclear focus formation and a newly identified FA pathway-deficient cell line was functionally characterized. The hepatocellular carcinoma (HCC line HuH-7 was defective in FANCD2 monoubiquitination and FANCD2 nuclear focus formation but proficient in RAD51 focus formation. Gene complementation studies revealed that this proximal FA pathway inactivation was attributable to defective FANCC function in HuH-7 cells. Accordingly, a homozygous inactivating FANCC nonsense mutation (c.553C > T, p.R185X was identified in HuH-7, resulting in partial transcriptional skipping of exon 6 and leading to the classic cellular FA hypersensitivity phenotype; HuH-7 cells exhibited a strongly reduced proliferation rate and a pronounced G2 cell cycle arrest at distinctly lower concentrations of ICL-agents than a panel of non-isogenic, FA pathway-proficient HCC cell lines. Upon retroviral transduction of HuH-7 cells with FANCC cDNA, FA pathway functions were restored and ICL-hypersensitivity abrogated. Analyses of 18 surgical HCC specimens yielded no further examples for genetic or epigenetic inactivation of FANCC, FANCF, or FANCG in HCC, suggesting a low prevalence of proximal FA pathway inactivation in this tumor type. Conclusions As the majority of HCC are chemoresistant, assessment of FA pathway function in HCC could

  9. Câncer e agentes antineoplásicos ciclo-celular específicos e ciclo-celular não específicos que interagem com o DNA: uma introdução Cancer and cell cicle-specific and cell cicle nonspecific anticancer DNA-interactive agents: an introduction

    Directory of Open Access Journals (Sweden)

    Vera Lúcia de Almeida

    2005-02-01

    Full Text Available The chemotherapy agents against cancer may be classified as "cell cycle-specific" or "cell cycle-nonspecific". Nevertheless, several of them have their biological activity related to any kind of action on DNA such as: antimetabolic agents (DNA synthesis inhibition, inherently reactive agents (DNA alkylating electrophilic traps for macromolecular nucleophiles from DNA through inter-strand cross-linking - ISC - alkylation and intercalating agents (drug-DNA interactions inherent to the binding made due to the agent penetration in to the minor groove of the double helix. The earliest and perhaps most extensively studied and most heavily employed clinical anticancer agents in use today are the DNA inter-strand cross-linking agents.

  10. Electron transfer in DNA duplexes containing 2-methyl-1,4-naphthoquinone

    OpenAIRE

    Bergeron, François; Houde, Daniel; Hunting, Darel J.; Wagner, J. Richard

    2004-01-01

    2-Methyl-1,4-naphthoquinone (menadione, MQ) was linked to synthetic oligonucleotides and exposed to near-UV light to generate base radical cations in DNA. This model system of electron transfer induced alkali-labile breaks at GG doublets, similar to anthraquinone and metallointercalators systems. In sharp contrast to other systems, the photolysis of MQ–DNA duplexes gave interstrand cross-links and alkali-labile breaks at bases on the complementary strand opposite the MQ moiety. For sequences ...

  11. 1,2-GG intrastrand cross-link of antitumor dinuclear bifunctional platinum compound with spermidine linker inhibits DNA polymerization more effectively than the cross-link of conventional cisplatin

    Czech Academy of Sciences Publication Activity Database

    Moriarity, B.; Nováková, Olga; Farrell, N.; Brabec, Viktor; Kašpárková, Jana

    2007-01-01

    Roč. 459, č. 2 (2007), s. 264-272 ISSN 0003-9861 R&D Projects: GA ČR(CZ) GA305/05/2030; GA ČR(CZ) GA203/06/1239; GA MZd(CZ) NR8562; GA MŠk(CZ) LC06030; GA AV ČR(CZ) 1QS500040581; GA AV ČR(CZ) KAN200200651 Institutional research plan: CEZ:AV0Z50040702 Keywords : cancer * DNA * platinum Subject RIV: BO - Biophysics Impact factor: 2.578, year: 2007

  12. Interstrand resistance of SSC magnets

    International Nuclear Information System (INIS)

    Kovachev, V.T.; Neal, M.J.; Capone, D.W. II; Carr, W.J. Jr.; Swenson, C.

    1994-01-01

    In situ interstrand resistance measurements were conducted on selected section of the inner coil of a full size (15 m) and a short (1 m) model SSC magnets. A model for evaluating single contacts resistance between two strands was developed. Using this model analyses of adjacent and non-adjacent strand contacts were performed. The interstrand resistance distribution throughout the coil was found to correlate with the quench location data as well as with the multipoles decay characteristics of the magnet. An anisotropic continuum based model for evaluation of cable eddy current losses was developed and results were compared with the experimental data

  13. Interstrand resistance of SSC magnets

    Energy Technology Data Exchange (ETDEWEB)

    Kovachev, V.T.; Neal, M.J.; Capone, D.W. II [Superconducting Super Collider Lab., Dallas, TX (United States); Carr, W.J. Jr.; Swenson, C. [Westinghouse Electric Corp., Pittsburgh, PA (United States). Science and Technology Center

    1994-01-01

    In situ interstrand resistance measurements were conducted on selected section of the inner coil of a full size (15 m) and a short (1 m) model SSC magnets. A model for evaluating single contacts resistance between two strands was developed. Using this model analyses of adjacent and non-adjacent strand contacts were performed. The interstrand resistance distribution throughout the coil was found to correlate with the quench location data as well as with the multipoles decay characteristics of the magnet. An anisotropic continuum based model for evaluation of cable eddy current losses was developed and results were compared with the experimental data.

  14. NanoHPLC-nanoESI(+)-MS/MS quantitation of bis-N7-guanine DNA-DNA cross-links in tissues of B6C3F1 mice exposed to subppm levels of 1,3-butadiene.

    Science.gov (United States)

    Sangaraju, Dewakar; Goggin, Melissa; Walker, Vernon; Swenberg, James; Tretyakova, Natalia

    2012-02-07

    1,3-Butadiene (BD) is an important industrial chemical and a common environmental pollutant present in urban air. BD is classified as a human carcinogen based on epidemiological evidence for an increased incidence of leukemia in workers occupationally exposed to BD and its potent carcinogenicity in laboratory mice. A diepoxide metabolite of BD, 1,2,3,4-diepoxybutane (DEB), is considered the ultimate carcinogenic species of BD due to its ability to form genotoxic DNA-DNA cross-links. We have previously employed capillary HPLC-ESI(+)-MS/MS (liquid chromatography-electrospray ionization tandem mass spectrometry) methods to quantify DEB-induced DNA-DNA conjugates, e.g. 1,4-bis-(guan-7-yl)-2,3-butanediol (bis-N7G-BD), 1-(guan-7-yl)-4-(aden-1-yl)-2,3-butanediol (N7G-N1A-BD), and 1,N(6)-(1-hydroxymethyl-2-hydroxypropan-1,3-diyl)-2'-deoxyadenosine (1,N(6)-HMHP-dA), in tissues of laboratory mice exposed to 6.25-625 ppm BD (Goggin et al. Cancer Res. 2009, 69(6), 2479-2486). However, typical BD human exposure levels are 0.01 to 3.2 ppb in urban air and 1-2.0 ppm in an occupational setting, requiring greater detection sensitivity for these critical lesions. In the present study, a nanoHPLC-nanoESI(+)-MS/MS method was developed for ultrasensitive, accurate, and precise quantitation of bis-N7G-BD in tissues of laboratory mice treated with low ppm and subppm concentrations of BD. The LOD value of the new method is 0.5 fmol/100 μg DNA, and the LOQ is 1.0 fmol/100 μg DNA, making it possible to quantify bis-N7G-BD adducts present at concentrations of 3 per 10(9) nucleotides. Bis-N7G-BD adduct amounts in liver tissues of mice exposed to 0.5, 1.0, and 1.5 ppm BD for 2 weeks were 5.7 ± 3.3, 9.2 ± 1.5, and 18.6 ± 6.9 adducts per 10(9) nucleotides, respectively, suggesting that bis-N7G-BD adduct formation is more efficient under low exposure conditions. To our knowledge, this is the first quantitative analysis of DEB specific DNA adducts following low ppm and subppm exposure to BD

  15. Collagen cross linking: Current perspectives

    Directory of Open Access Journals (Sweden)

    Srinivas K Rao

    2013-01-01

    Full Text Available Keratoconus is a common ectatic disorder occurring in more than 1 in 1,000 individuals. The condition typically starts in adolescence and early adulthood. It is a disease with an uncertain cause and its progression is unpredictable, but in extreme cases, vision deteriorates and can require corneal transplant surgery. Corneal collagen cross-linking (CCL with riboflavin (C3R is a recent treatment option that can enhance the rigidity of the cornea and prevent disease progression. Since its inception, the procedure has evolved with newer instrumentation, surgical techniques, and is also now performed for expanded indications other than keratoconus. With increasing experience, newer guidelines regarding optimization of patient selection, the spectrum of complications and their management, and combination procedures are being described. This article in conjunction with the others in this issue, will try and explore the uses of collagen cross-linking (CXL in its current form.

  16. DNA-reactive protein monoepoxides induce cell death and mutagenesis in mammalian cells.

    Science.gov (United States)

    Tretyakova, Natalia Y; Michaelson-Richie, Erin D; Gherezghiher, Teshome B; Kurtz, Jamie; Ming, Xun; Wickramaratne, Susith; Campion, Melissa; Kanugula, Sreenivas; Pegg, Anthony E; Campbell, Colin

    2013-05-07

    Although cytotoxic alkylating agents possessing two electrophilic reactive groups are thought to act by cross-linking cellular biomolecules, their exact mechanisms of action have not been established. In cells, these compounds form a mixture of DNA lesions, including nucleobase monoadducts, interstrand and intrastrand cross-links, and DNA-protein cross-links (DPCs). Interstrand DNA-DNA cross-links block replication and transcription by preventing DNA strand separation, contributing to toxicity and mutagenesis. In contrast, potential contributions of drug-induced DPCs are poorly understood. To gain insight into the biological consequences of DPC formation, we generated DNA-reactive protein reagents and examined their toxicity and mutagenesis in mammalian cells. Recombinant human O(6)-alkylguanine DNA alkyltransferase (AGT) protein or its variants (C145A and K125L) were treated with 1,2,3,4-diepoxybutane to yield proteins containing 2-hydroxy-3,4-epoxybutyl groups on cysteine residues. Gel shift and mass spectrometry experiments confirmed that epoxide-functionalized AGT proteins formed covalent DPC but no other types of nucleobase damage when incubated with duplex DNA. Introduction of purified AGT monoepoxides into mammalian cells via electroporation generated AGT-DNA cross-links and induced cell death and mutations at the hypoxanthine-guanine phosphoribosyltransferase gene. Smaller numbers of DPC lesions and reduced levels of cell death were observed when using protein monoepoxides generated from an AGT variant that fails to accumulate in the cell nucleus (K125L), suggesting that nuclear DNA damage is required for toxicity. Taken together, these results indicate that AGT protein monoepoxides produce cytotoxic and mutagenic DPC lesions within chromosomal DNA. More generally, these data suggest that covalent DPC lesions contribute to the cytotoxic and mutagenic effects of bis-electrophiles.

  17. Photoreactivities and thermal properties of psoralen cross-links

    International Nuclear Information System (INIS)

    Yeung, A.T.; Jones, B.K.; Chu, C.T.

    1988-01-01

    The authors have studied the photoreaction of 8-methoxypsoralen (8-MOP), 4,5',8-trimethylpsoralen (TMP), and 4'-(hydroxymethyl)-4,5',8-trimethylpsoralen (HMT) with a pair of 18-base-long oligonucleotides in which a 14-base region is complementary. Only one 5'TpA site, favored for both monoadduct and cross-link formation with psoralen, is present in this oligonucleotide pair. They have used this model system to demonstrate, for the first time, strand specificity in the photoreaction of psoralen with DNA. They found that the two types of cross-links which form at this site have large differences in thermal stabilities. In addition, the denaturation of each cross-links isomer duplex occurred in at least three stages, which can be visualized as three bands in thermal equilibrium under the conditions of a denaturing polyacrylamide gel. This novel observation suggests that there are several domains differing in thermal stability in a psoralen cross-link

  18. Electrospinning formaldehyde cross-linked zein solutions

    Science.gov (United States)

    In order to develop zein fibers with improved physical properties and solvent resistance, formaldehyde was used as the cross-linking reagent before spinning. The cross-linking reaction was carried out in either acetic acid or ethanolic-HCl where the amount of cross-linking reagent was between 1 and...

  19. Chemical cross-linking of Chlamydia trachomatis

    DEFF Research Database (Denmark)

    Birkelund, Svend; Lundemose, AG; Christiansen, Gunna

    1988-01-01

    Purified elementary bodies (EBs) of Chlamydia trachomatis serovar L2 were analyzed by chemical cross-linking with disuccinimidyl selenodipropionate. The effect of the cross-linking was analyzed by immunoblotting sodium dodecyl sulfate-polyacrylamide gel electrophoresis-separated components which...

  20. Cross-linking of polymeric materials

    International Nuclear Information System (INIS)

    Bloom, L.I.; Du Plessis, T.A.; Meij, G.O.

    1991-01-01

    The invention provides a method of producing a cured polymeric artifact from a polymeric thermoplastic starting material, the material of the artifact having reduced thermoplasticity relative to the starting material and exhibiting an enhanced degree of cross-linking relative to the starting material. The method includes subjecting a polymeric thermoplastic starting material, which is capable of being cross-linked by irradiation, to sufficient irradiation partially to cross-linked the starting material to produce a thermoplastic partially cross-linked intermediate material. The thermoplasticity of the intermediate material is then reduced by heating it to raise its melting point. The invention also provides a method of making a partially cross-linked feedstocks and a master batch for use in making such artifacts

  1. Analysis of Translesion DNA Synthesis by the Mitochondrial DNA Polymerase γ.

    Science.gov (United States)

    Copeland, William C; Kasiviswanathan, Rajesh; Longley, Matthew J

    2016-01-01

    Mitochondrial DNA is replicated by the nuclear-encoded DNA polymerase γ (pol γ) which is composed of a single 140 kDa catalytic subunit and a dimeric 55 kDa accessory subunit. Mitochondrial DNA is vulnerable to various forms of damage, including several types of oxidative lesions, UV-induced photoproducts, chemical adducts from environmental sources, as well as alkylation and inter-strand cross-links from chemotherapy agents. Although many of these lesions block DNA replication, pol γ can bypass some lesions by nucleotide incorporation opposite a template lesion and further extension of the DNA primer past the lesion. This process of translesion synthesis (TLS) by pol γ can occur in either an error-free or an error-prone manner. Assessment of TLS requires extensive analysis of oligonucleotide substrates and replication products by denaturing polyacrylamide sequencing gels. This chapter presents protocols for the analysis of translesion DNA synthesis.

  2. A Photoactivatable Platinum( IV) Complex Targeting Genomic DNA and Histone Deacetylases

    Czech Academy of Sciences Publication Activity Database

    Kašpárková, Jana; Kostrhunová, Hana; Nováková, Olga; Křikavová, R.; Vančo, J.; Trávníček, Z.; Brabec, Viktor

    2015-01-01

    Roč. 54, č. 48 (2015), s. 14478-14482 ISSN 1433-7851 Institutional support: RVO:68081707 Keywords : INTERSTRAND CROSS-LINKS * CANCER-CELLS * ANTICANCER COMPLEXES Subject RIV: BO - Biophysics Impact factor: 11.709, year: 2015

  3. Cross-linking reconsidered : binding and cross-linking fields and the cellular response

    NARCIS (Netherlands)

    Sulzer, B.; Boer, R.J. de; Perelson, A.S.

    1996-01-01

    We analyze a model for the reversible cross-linking of cell surface receptors by a collection of bivalent ligands with different affinities for the receptor as would be found in a polyclonal anti-receptor serum. We assume that the amount of cross-linking determines, via a monotonic function, the

  4. Cross-linked structure of network evolution

    International Nuclear Information System (INIS)

    Bassett, Danielle S.; Wymbs, Nicholas F.; Grafton, Scott T.; Porter, Mason A.; Mucha, Peter J.

    2014-01-01

    We study the temporal co-variation of network co-evolution via the cross-link structure of networks, for which we take advantage of the formalism of hypergraphs to map cross-link structures back to network nodes. We investigate two sets of temporal network data in detail. In a network of coupled nonlinear oscillators, hyperedges that consist of network edges with temporally co-varying weights uncover the driving co-evolution patterns of edge weight dynamics both within and between oscillator communities. In the human brain, networks that represent temporal changes in brain activity during learning exhibit early co-evolution that then settles down with practice. Subsequent decreases in hyperedge size are consistent with emergence of an autonomous subgraph whose dynamics no longer depends on other parts of the network. Our results on real and synthetic networks give a poignant demonstration of the ability of cross-link structure to uncover unexpected co-evolution attributes in both real and synthetic dynamical systems. This, in turn, illustrates the utility of analyzing cross-links for investigating the structure of temporal networks

  5. Characterization of Cross-Linked Lipase Aggregates

    DEFF Research Database (Denmark)

    Prabhavathi Devi, Bethala Lakshmi Anu; Guo, Zheng; Xu, Xuebing

    2009-01-01

    Commercially available microbial lipases from different sources were immobilized as cross-linked enzyme aggregates (CLEAs) using different precipitants and glutaraldehyde as cross-linkers. These CLEAs were assayed based on esterification between lauric acid and n-propanol in solvent-free systems...

  6. Cross-linked structure of network evolution

    Energy Technology Data Exchange (ETDEWEB)

    Bassett, Danielle S., E-mail: dsb@seas.upenn.edu [Department of Bioengineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104 (United States); Department of Physics, University of California, Santa Barbara, California 93106 (United States); Sage Center for the Study of the Mind, University of California, Santa Barbara, California 93106 (United States); Wymbs, Nicholas F.; Grafton, Scott T. [Department of Psychology and UCSB Brain Imaging Center, University of California, Santa Barbara, California 93106 (United States); Porter, Mason A. [Oxford Centre for Industrial and Applied Mathematics, Mathematical Institute, University of Oxford, Oxford OX2 6GG (United Kingdom); CABDyN Complexity Centre, University of Oxford, Oxford, OX1 1HP (United Kingdom); Mucha, Peter J. [Carolina Center for Interdisciplinary Applied Mathematics, Department of Mathematics, University of North Carolina, Chapel Hill, North Carolina 27599 (United States); Department of Applied Physical Sciences, University of North Carolina, Chapel Hill, North Carolina 27599 (United States)

    2014-03-15

    We study the temporal co-variation of network co-evolution via the cross-link structure of networks, for which we take advantage of the formalism of hypergraphs to map cross-link structures back to network nodes. We investigate two sets of temporal network data in detail. In a network of coupled nonlinear oscillators, hyperedges that consist of network edges with temporally co-varying weights uncover the driving co-evolution patterns of edge weight dynamics both within and between oscillator communities. In the human brain, networks that represent temporal changes in brain activity during learning exhibit early co-evolution that then settles down with practice. Subsequent decreases in hyperedge size are consistent with emergence of an autonomous subgraph whose dynamics no longer depends on other parts of the network. Our results on real and synthetic networks give a poignant demonstration of the ability of cross-link structure to uncover unexpected co-evolution attributes in both real and synthetic dynamical systems. This, in turn, illustrates the utility of analyzing cross-links for investigating the structure of temporal networks.

  7. Electrospinning covalently cross-linking biocompatible hydrogelators

    Science.gov (United States)

    Schultz, Kelly M.; Campo-Deaño, Laura; Baldwin, Aaron D.; Kiick, Kristi L.; Clasen, Christian; Furst, Eric M.

    2012-01-01

    Many hydrogel materials of interest are homogeneous on the micrometer scale. Electrospinning, the formation of sub-micrometer to micrometer diameter fibers by a jet of fluid formed under an electric field, is one process being explored to create rich microstructures. However, electrospinning a hydrogel system as it reacts requires an understanding of the gelation kinetics and corresponding rheology near the liquid-solid transition. In this study, we correlate the structure of electrospun fibers of a covalently cross-linked hydrogelator with the corresponding gelation transition and kinetics. Polyethylene oxide (PEO) is used as a carrier polymer in a chemically cross-linking poly(ethylene glycol)-high molecular weight heparin (PEG-HMWH) hydrogel. Using measurements of gelation kinetics from multiple particle tracking microrheology (MPT), we correlate the material rheology with the the formation of stable fibers. An equilibrated, cross-linked hydrogel is then spun and the PEO is dissolved. In both cases, microstructural features of the electrospun fibers are retained, confirming the covalent nature of the network. The ability to spin fibers of a cross-linking hydrogel system ultimately enables the engineering of materials and microstructural length scales suitable for biological applications. PMID:23459473

  8. [Biophysical principles of collagen cross-linking].

    Science.gov (United States)

    Spörl, E; Raiskup-Wolf, F; Pillunat, L E

    2008-02-01

    The reduced mechanical stability of the cornea in keratoconus or in keratectasia after Lasik may be increased by photooxidative cross-linking of corneal collagen. The biophysical principles are compiled for the safe and effective application of this new treatment method. The setting of the therapy parameters should be elucidated from the absorption behaviour of the cornea. The safety of the method for the endothelium cells and the lens will be discussed. The induced cross-links are shown to be the result of changes in the physico-chemical properties of the cornea. To reach a high absorption of the irradiation energy in the cornea, riboflavin of a concentration of 0.1% and UV light of a wavelength of 370 nm, corresponding to the relative maximum of absorption of riboflavin, were used. An irradiance of 3 mW/cm(2) and an irradiation time of 30 min lead to an increase of the mechanical stiffness. The endothelium cells will be protected due to the high absorption within the cornea, that means the damaging threshold of the endothelium cells will not be reached in a 400 microm thick stroma. As evidence for cross-links we can consider the increase of the biomechanical stiffness, the increased resistance against enzymatic degradation, a higher shrinkage temperature, a lower swelling rate and an increased diameter of collagen fibres. The therapy parameters were tested experimentally and have been proven clinically in the corneal collagen cross-linking. These parameters should be respected to reach a safe cross-linking effect without damage of the adjacent tissues.

  9. Method of preparing cross-linked enzyme particles

    NARCIS (Netherlands)

    Mateo, C.; Van Langen, L.M.; Van Rantwijk, F.

    2004-01-01

    The invention relates to a method of preparing cross-linked enzyme particles using a cross-linking agent. According to the invention, the enzyme particles are formed and subsequently cross-linked using a cross-linking agent having at least n reactive groups where N>=3 and a molecular weight of

  10. The cross linking of EPDM and NBR rubber

    Directory of Open Access Journals (Sweden)

    Samardžija-Jovanović Suzana

    2005-01-01

    Full Text Available In the process of macromolecule cross linking, the choice of type and quantity of the components and the experimental conditions are important to obtain the new cross linked materials with better mechanical and chemical characteristics. The cross linking method depends on the rubber type and structure. Intermolecular cross linking results in the formation elastomer network. The basis of the cross linking process, between ethylene propylene diene rubber (EPDM and acrylonitrile butadiene rubber (NBR, is a chemical reaction. Fillers and other additives are present in different mass ratios in the material. The exploitation properties of the cross linked materials depend on the quantity of additive in the cross linked systems.

  11. Cross-Linking Studies of Lysozyme Nucleation

    Science.gov (United States)

    Forsythe, Elizabeth; Pusey, Marc

    2000-01-01

    Tetragonal chicken egg white crystals consist of 4(sub 3) helices running in alternating directions, the helix rows having a two fold symmetry with each other. The unit cell consists of one complete tetrameric turn from each of two adjacent helices (an octamer). PBC analysis indicates that the helix intermolecular bonds are the strongest in the crystal, therefore likely formed first. AFM analysis of the (110) surface shows only complete helices, no half steps or bisected helices being found, while AFM line scans to measure the growth step increments show that they are multiples of the 4(sub 3) helix tetramer dimensions. This supports our thesis that the growth units are in fact multiples of the four molecule 4(sub 3) helix unit, the "average" growth unit size for the (110) face being an octamer (two turns about the helix) and the (101) growth unit averaging about the size of a hexamer. In an effort to better understand the species involved in the crystal nucleation and growth process, we have initiated an experimental program to study the species formed in solution compared to what is found in the crystal through covalent cross-linking studies. These experiments use the heterobifunctional cross-linking agent aminoethyl-4-azidonitroanaline (AEANA). An aliphatic amine at one end is covalently attached to the protein by a carbodiimide-mediated reaction, and a photo reactive group at the other can be used to initiate crosslinking. Modifications to the parent structure can be used to alter the distance between the two reactive groups and thus the cross-linking agents "reach". In practice, the cross-linking agent is first coupled to the asp101 side chain through the amine group. Asp101 lies within the active site cleft, and previous work with fluorescent probes had shown that derivatives at this site still crystallize in the tetragonal space group. This was also found to be the case with the AEANA derivative, which gave red tetragonal crystals. The protein now has a

  12. Thermal Analyse sof Cross-Linked Polyethylene

    Directory of Open Access Journals (Sweden)

    Radek Polansky

    2007-01-01

    Full Text Available The paper summarizes results obtained during the structural analyses measurements (Differential Scanning Calorimetry DSC, Thermogravimetry TG, Thermomechanical analysis TMA and Fourier transform infrared spectroscopy FT-IR. The samples of cross-linked polyethylene cable insulation were tested via these analyses. The DSC and TG were carried out using simultaneous thermal analyzer TA Instruments SDT Q600 with connection of Fourier transform infrared spectrometer Nicolet 380. Thermomechanical analysis was carried out by TMA Q400EM TA Instruments apparatus.

  13. Collagen cross-linking in thin corneas

    Directory of Open Access Journals (Sweden)

    Prema Padmanabhan

    2013-01-01

    Full Text Available Collagen cross-linking (CXL has become the standard of care for progressive keratoconus, after numerous clinical studies have established its efficacy and safety in suitably selected eyes. The standard protocol is applicable in eyes which have a minimum corneal thickness of 400 μm after epithelial debridement. This prerequisite was stipulated to protect the corneal endothelium and intraocular tissues from the deleterious effect of ultraviolet-A (UVA radiation. However, patients with keratoconus often present with corneal thickness of less than 400 μm and could have otherwise benefited from this procedure. A few modifications of the standard procedure have been suggested to benefit these patients without a compromise in safety. Transepithelial cross-linking, pachymetry-guided epithelial debridement before cross-linking, and the use of hypoosmolar riboflavin are some of the techniques that have been attempted. Although clinical data is limited at the present time, these techniques are worth considering in patients with thin corneas. Further studies are needed to scientifically establish their efficacy and safety.

  14. Positive tone cross-linked resists based on photoacid inhibition of cross linking

    Science.gov (United States)

    Lawson, Richard A.; Chun, Jun Sung; Neisser, Mark; Tolbert, Laren M.; Henderson, Clifford L.

    2014-03-01

    A resist imaging design that utilizes photoacid inhibition of cationic polymerization and cross-linking during a postexposure bake step has been studied. The key to the design approach is the use of two different polymerization catalysts/initiators: (1) a photoacid produced from a photoacid generator (PAG) upon exposure of the resist that can result in polymerization and cross-linking of the resist matrix and (2) a thermal cross-linking catalyst (TCC) designed to thermally catalyze epoxide-phenol cross-linking. The TCC can be chosen from a variety of compounds such as triphenylphosphine (TPP) or imidazole. When only one of these catalysts (e.g TPP or photoacid) is present in an epoxide and phenol containing resist matrix, it will individually catalyze cross-linking. When they are present together, they effectively quench one another and little to no cross-linking occurs. This approach can be used to switch the tone of a resist from negative (photoacid catalyzed) to positive (TCC catalyzed and photoacid inhibited). The effect of the ratio of TCC:PAG was examined and the optimal ratio for positive tone behavior was determined. Resist contrast can be modified by optimization of epoxide:phenol ratio in the formulation. Dual tone behavior with positive tone at low dose and negative tone at higher doses can be observed in certain formulation conditions. Initial EUV patterning shows poor results, but the source of the poor imaging is not yet understood.

  15. Structure of the FANCI-FANCD2 Complex: Insights into the Fanconi Anemia DNA Repair Pathway

    Energy Technology Data Exchange (ETDEWEB)

    Joo, Woo; Xu, Guozhou; Persky, Nicole S.; Smogorzewska, Agata; Rudge, Derek G.; Buzovetsky, Olga; Elledge, Stephen J.; Pavletich, Nikola P. (Harvard-Med); (Cornell); (MSKCC)

    2011-08-29

    Fanconi anemia is a cancer predisposition syndrome caused by defects in the repair of DNA interstrand cross-links (ICLs). Central to this pathway is the Fanconi anemia I-Fanconi anemia D2 (FANCI-FANCD2) (ID) complex, which is activated by DNA damage-induced phosphorylation and monoubiquitination. The 3.4 angstrom crystal structure of the {approx}300 kilodalton ID complex reveals that monoubiquitination and regulatory phosphorylation sites map to the I-D interface, suggesting that they occur on monomeric proteins or an opened-up complex and that they may serve to stabilize I-D heterodimerization. The 7.8 angstrom electron-density map of FANCI-DNA crystals and in vitro data show that each protein has binding sites for both single- and double-stranded DNA, suggesting that the ID complex recognizes DNA structures that result from the encounter of replication forks with an ICL.

  16. Structure of the FANCI-FANCD2 Complex: Insights into the Fanconi Anemia DNA Repair Pathway

    Energy Technology Data Exchange (ETDEWEB)

    W Joo; G Xu; n Persky; A Smogorzewska; D Rudge; O Buzovetsky; S Elledge; N Pavletich

    2011-12-31

    Fanconi anemia is a cancer predisposition syndrome caused by defects in the repair of DNA interstrand cross-links (ICLs). Central to this pathway is the Fanconi anemia I-Fanconi anemia D2 (FANCI-FANCD2) (ID) complex, which is activated by DNA damage-induced phosphorylation and monoubiquitination. The 3.4 angstrom crystal structure of the {approx}300 kilodalton ID complex reveals that monoubiquitination and regulatory phosphorylation sites map to the I-D interface, suggesting that they occur on monomeric proteins or an opened-up complex and that they may serve to stabilize I-D heterodimerization. The 7.8 angstrom electron-density map of FANCI-DNA crystals and in vitro data show that each protein has binding sites for both single- and double-stranded DNA, suggesting that the ID complex recognizes DNA structures that result from the encounter of replication forks with an ICL.

  17. Contraction of cross-linked actomyosin bundles

    Science.gov (United States)

    Yoshinaga, Natsuhiko; Marcq, Philippe

    2012-08-01

    Cross-linked actomyosin bundles retract when severed in vivo by laser ablation, or when isolated from the cell and micromanipulated in vitro in the presence of ATP. We identify the timescale for contraction as a viscoelastic time τ, where the viscosity is due to (internal) protein friction. We obtain an estimate of the order of magnitude of the contraction time τ ≈ 10-100 s, consistent with available experimental data for circumferential microfilament bundles and stress fibers. Our results are supported by an exactly solvable, hydrodynamic model of a retracting bundle as a cylinder of isotropic, active matter, from which the order of magnitude of the active stress is estimated.

  18. Cross-linking for microbial keratitis

    Directory of Open Access Journals (Sweden)

    Jayesh Vazirani

    2013-01-01

    Full Text Available The success of collagen cross-linking as a clinical modality to modify the clinical course in keratoconus seems to have fueled the search for alternative applications for this treatment. Current clinical data on its efficacy is limited and laboratory data seems to indicate that it performs poorly against resistant strains of bacteria and against slow growing organisms. However, the biological plausibility of crosslinking and the lack of effective strategies in managing infections with these organisms continue to focus attention on this potential treatment. Well-conducted experimental and clinical studies with controls are required to answer the questions of its efficacy in future.

  19. Second-generation method for analysis of chromatin binding with formaldehyde-cross-linking kinetics.

    Science.gov (United States)

    Zaidi, Hussain; Hoffman, Elizabeth A; Shetty, Savera J; Bekiranov, Stefan; Auble, David T

    2017-11-24

    Formaldehyde-cross-linking underpins many of the most commonly used experimental approaches in the chromatin field, especially in capturing site-specific protein-DNA interactions. Extending such assays to assess the stability and binding kinetics of protein-DNA interactions is more challenging, requiring absolute measurements with a relatively high degree of physical precision. We previously described an experimental framework called the cross-linking kinetics (CLK) assay, which uses time-dependent formaldehyde-cross-linking data to extract kinetic parameters of chromatin binding. Many aspects of formaldehyde behavior in cells are unknown or undocumented, however, and could potentially affect CLK data analyses. Here, we report biochemical results that better define the properties of formaldehyde-cross-linking in budding yeast cells. These results have the potential to inform interpretations of "standard" chromatin assays, including chromatin immunoprecipitation. Moreover, the chemical complexity we uncovered resulted in the development of an improved method for measuring binding kinetics with the CLK approach. Optimum conditions included an increased formaldehyde concentration and more robust glycine-quench conditions. Notably, we observed that formaldehyde-cross-linking rates can vary dramatically for different protein-DNA interactions in vivo Some interactions were cross-linked much faster than the in vivo macromolecular interactions, making them suitable for kinetic analysis. For other interactions, we found the cross-linking reaction occurred on the same time scale or slower than binding dynamics; for these interactions, it was sometimes possible to compute the in vivo equilibrium-binding constant but not binding on- and off-rates. This improved method yields more accurate in vivo binding kinetics estimates on the minute time scale. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  20. Cross-linked polyethylenimine–tripolyphosphate nanoparticles for gene delivery

    Science.gov (United States)

    Huang, Xianzhang; Shen, Sujing; Zhang, Zhanfeng; Zhuang, Junhua

    2014-01-01

    The high transfection efficiency of polyethylenimine (PEI) makes it an attractive potential nonviral genetic vector for gene delivery and therapy. However, the highly positive charge of PEI leads to cytotoxicity and limits its application. To reduce the cytotoxicity of PEI, we prepared anion-enriched nanoparticles that combined PEI with tripolyphosphate (TPP). We then characterized the PEI-TPP nanoparticles in terms of size, zeta potential, and Fourier-transform infrared (FTIR) spectra, and assessed their transfection efficiency, cytotoxicity, and ability to resist deoxyribonuclease (DNase) I digestion. The cellular uptake of PEI-TPP with phosphorylated internal ribosome entry site–enhanced green fluorescent protein C1 or FAM (fluorouracil, Adriamycin [doxorubicin] and mitomycin)-labeled small interfering ribonucleic acids (siRNAs) was monitored by fluorescence microscopy and confocal laser microscopy. The efficiency of transfected delivery of plasmid deoxyribonucleic acid (DNA) and siRNA in vitro was 1.11- to 4.20-fold higher with the PEI-TPP particles (7.6% cross-linked) than with the PEI, at all N:P ratios (nitrogen in PEI to phosphorus in DNA) tested. The cell viability of different cell lines was more than 90% at the chosen N:P ratios of PEI-TPP/DNA complexes. Moreover, PEI-TPP nanoparticles resisted digestion by DNase I for more than 2 hours. The time-dependent absorption experiment showed that 7.6% of cross-linked PEI-TPP particles were internalized by 293T cells within 1 hour. In summary, PEI-TPP nanoparticles effectively transfected cells while conferring little or no toxicity, and thus have potential application in gene delivery. PMID:25342902

  1. Corneal cross-linking treatment of keratoconus

    Directory of Open Access Journals (Sweden)

    Mahgol Farjadnia

    2015-01-01

    Full Text Available Keratoconus as the most common cause of ectasia is one of the leading cause of corneal transplants worldwide. The current available therapies do not modify the underlying pathogenesis of the disease, and none of the available approaches but corneal transplant hinder the ongoing ectasia. Several studies document Crosslink defect between collagen fibrils in the pathogenesis of keratoconus. Collagen cross link is a relatively new approach that with the application of the riboflavin and ultraviolet A, new covalent bands reform. Subjective and objective results following this method seem to be promising. Endothelial damage besides other deep structural injury, which is the major concern of this technique have not yet been reported, when applying the standard method.

  2. Transplatin enhances effect of cisplatin on both single DNA molecules and live tumor cells.

    Science.gov (United States)

    Liu, Yu-Ru; Ji, Chao; Zhang, Hong-Yan; Dou, Shuo-Xing; Xie, Ping; Wang, Wei-Chi; Wang, Peng-Ye

    2013-08-01

    Cisplatin is the main platinum antitumor drug applied in clinical settings. However, its trans isomer, transplatin, is known to have an ineffective antitumor activity. Despite intensive studies in this field, the structural and biophysical properties of DNA molecules reacting with these two platinum complexes have not been fully elucidated. In the present study, we observed that transplatin made efficient cross-linking of DNA in the vicinity of cisplatin adducts. High-resolution atomic force microscopy studies revealed that the transplatin-induced cross-linkings of nucleotides flanking cisplatin adducts were characterized by kinked-loop structures with rod-like shapes of nanometer scales (∼10-60nm). The results were further confirmed by denaturing gel electrophoresis and single-molecule experiment using magnetic tweezers. In vivo studies revealed that transplatin and cisplatin co-treatment could induce a considerable amount of kinked loops with smaller sizes (∼15nm) in cellular DNA. Furthermore, compared with cisplatin treatment alone, the co-treatment resulted in enhanced cytotoxicity, increased amount of interstrand cross-links, and cell lesions more reluctant to cellular repair system. The results of the present study provide a new clue for understanding the stepwise reactions of DNA with platinum drugs and might serve as a basis for the development of a new antitumor strategy. Copyright © 2013 Elsevier Inc. All rights reserved.

  3. Cross-link guided molecular modeling with ROSETTA.

    Directory of Open Access Journals (Sweden)

    Abdullah Kahraman

    Full Text Available Chemical cross-links identified by mass spectrometry generate distance restraints that reveal low-resolution structural information on proteins and protein complexes. The technology to reliably generate such data has become mature and robust enough to shift the focus to the question of how these distance restraints can be best integrated into molecular modeling calculations. Here, we introduce three workflows for incorporating distance restraints generated by chemical cross-linking and mass spectrometry into ROSETTA protocols for comparative and de novo modeling and protein-protein docking. We demonstrate that the cross-link validation and visualization software Xwalk facilitates successful cross-link data integration. Besides the protocols we introduce XLdb, a database of chemical cross-links from 14 different publications with 506 intra-protein and 62 inter-protein cross-links, where each cross-link can be mapped on an experimental structure from the Protein Data Bank. Finally, we demonstrate on a protein-protein docking reference data set the impact of virtual cross-links on protein docking calculations and show that an inter-protein cross-link can reduce on average the RMSD of a docking prediction by 5.0 Å. The methods and results presented here provide guidelines for the effective integration of chemical cross-link data in molecular modeling calculations and should advance the structural analysis of particularly large and transient protein complexes via hybrid structural biology methods.

  4. FancJ regulates interstrand crosslinker induced centrosome amplification through the activation of polo-like kinase 1

    Directory of Open Access Journals (Sweden)

    Jianqiu Zou

    2013-08-01

    DNA damage response (DDR and the centrosome cycle are two of the most critical processes for maintaining a stable genome in animals. Sporadic evidence suggests a connection between these two processes. Here, we report our findings that six Fanconi Anemia (FA proteins, including FancI and FancJ, localize to the centrosome. Intriguingly, we found that the localization of FancJ to the mother centrosome is stimulated by a DNA interstrand crosslinker, Mitomycin C (MMC. We further show that, in addition to its role in interstrand crosslinking (ICL repair, FancJ also regulates the normal centrosome cycle as well as ICL induced centrosome amplification by activating the polo-like kinase 1 (PLK1. We have uncovered a novel function of FancJ in centrosome biogenesis and established centrosome amplification as an integral part of the ICL response.

  5. Grafted Cross-Linked Polyolefin Substrates for Peptide Synthesis and Assays

    DEFF Research Database (Denmark)

    1999-01-01

    suited for use in solid-phase biosystems, notably bioassays, such as immunoassays, DNA hybridization assays or PCR amplification. The grafted chains may bear substituents which are such that the polymer-grafted cross-linked polyolefin substrate is swellable by water or aqueous media, in other words...

  6. Insights into the Structure of Intrastrand Cross-Link DNA Lesion-Containing Oligonucleotides: G[8-5m]T and G[8-5]C from Molecular Dynamics Simulations

    Czech Academy of Sciences Publication Activity Database

    Dumont, E.; Dršata, Tomáš; Guerra, C. F.; Lankaš, Filip

    2015-01-01

    Roč. 54, č. 5 (2015), s. 1259-1267 ISSN 0006-2960 R&D Projects: GA ČR(CZ) GA14-21893S Institutional support: RVO:61388963 Keywords : hydrogen bonds * abasic sites * duplex DNA Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.876, year: 2015

  7. Riboflavin for corneal cross-linking.

    Science.gov (United States)

    O'Brart, D P S

    2016-06-01

    Corneal collagen cross-linking (CXL) with riboflavin and ultraviolet A (UVA) radiation is the first therapeutic modality that appears to arrest the progression of keratoconus and other corneal ectasias. Riboflavin is central to the process, acting as a photosensitizer for the production of oxygen singlets and riboflavin triplets. These free radicals drive the CXL process within the proteins of the corneal stroma, altering its biomechanical properties. Riboflavin also absorbs the majority of the UVA radiation, which is potentially cytotoxic and mutagenic, within the anterior stroma, preventing damage to internal ocular structures, such as the corneal endothelium, lens and retina. Clinical studies report cessation of ectatic progression in over 90% of cases and the majority document significant improvements in visual, keratometric and topographic parameters. Clinical follow-up is limited to 5-10 years, but suggests sustained stability and enhancement in corneal shape. Sight-threatening complications are rare. The optimal stromal riboflavin dosage for CXL is as yet undetermined. Copyright 2016 Prous Science, S.A.U. or its licensors. All rights reserved.

  8. Cross-linking for microbial keratitis.

    Science.gov (United States)

    Chan, Tommy C Y; Agarwal, Tushar; Vajpayee, Rasik B; Jhanji, Vishal

    2016-07-01

    Microbial keratitis is one of the leading causes of ocular morbidity. The standard treatment consists of antibiotics, which is intensive and is fraught with risks of antibiotic resistance. Corneal collagen cross-linking (CXL) has recently been advocated as an adjunctive therapy for management of microbial keratitis. The addition of CXL to ongoing antimicrobial treatment can have a potential effect on overall duration of the disease, need for corneal transplantation, final visual outcome, and long-term impact on drug resistance pattern. CXL has been used in cases with bacterial, fungal as well as amoebic keratitis. However, so far the reported results have been variable and the evidence is largely anecdotal. The debate over the safety and efficacy of this modality continues especially with regards to its utilization in early phases of the disease when the corneal involvement is limited to the anterior stroma. CXL appears to be a promising adjunctive treatment in selective cases of mild to moderate bacterial keratitis. Its efficacy in fungal and amoebic keratitis is questionable. Treatment protocols in microbial keratitis need to be individualized. Long-term, prospective, randomized trials are needed to determine its usefulness in microbial keratitis.

  9. Cross-Linking Aromatic Polymers With Ionizing Radiation

    Science.gov (United States)

    Bell, Vernon L.; Havens, Stephen J.

    1987-01-01

    Resistance to heat and solvents increased. Certain aromatic polymers containing radiation-sensitive methylene groups cross-linked through methylene groups upon exposure to ionizing radiation. Cross-linked polymers resistant to most organic solvents and generally more resistant to high temperatures, with less tendency to creep under load. No significant embrittlement of parts fabricated from these polymers when degree of cross-linking, as controlled by irradiation dose, kept at moderate level.

  10. P-glycoprotein attenuates DNA repair activity in multidrug-resistant cells by acting through the Cbp-Csk-Src cascade.

    Science.gov (United States)

    Lin, Li-Fang; Wu, Ming-Hsi; Pidugu, Vijaya Kumar; Ho, I-Ching; Su, Tsann-Long; Lee, Te-Chang

    2017-07-11

    Recent studies have demonstrated that P-glycoprotein (P-gp) expression impairs DNA interstrand cross-linking agent-induced DNA repair efficiency in multidrug-resistant (MDR) cells. To date, the detailed molecular mechanisms underlying how P-gp interferes with Src activation and subsequent DNA repair activity remain unclear. In this study, we determined that the C-terminal Src kinase-binding protein (Cbp) signaling pathway involved in the negative control of Src activation is enhanced in MDR cells. We also demonstrated that cells that ectopically express P-gp exhibit reduced activation of DNA damage response regulators, such as ATM, Chk2, Braca1 and Nbs1 and hence attenuated DNA double-strand break repair capacity and become more susceptible than vector control cells to DNA interstrand cross-linking (ICL) agents. Moreover, we demonstrated that P-gp can not only interact with Cbp and Src but also enhance the formation of inhibitory C-terminal Src kinase (Csk)-Cbp complexes that reduce phosphorylation of the Src activation residue Y416 and increase phosphorylation of the Src negative regulatory residue Y527. Notably, suppression of Cbp expression in MDR cells restores cisplatin-induced Src activation, improves DNA repair capacity, and increases resistance to ICL agents. Ectopic expression of Cbp attenuates cisplatin-induced Src activation and increases the susceptibility of cells to ICL agents. Together, the current results indicate that P-gp inhibits DNA repair activity by modulating Src activation via Cbp-Csk-Src cascade. These results suggest that DNA ICL agents are likely to have therapeutic potential against MDR cells with P-gp-overexpression.

  11. Cross-linking of streptomycin to the 16S ribosomal RNA of Escherichia coli

    International Nuclear Information System (INIS)

    Gravel, M.; Melancon, P.; Barkier-Gingras, L.

    1987-01-01

    [ 3 H]Dihydrostreptomycin was cross-linked to the 30S ribosomal subunit from Escherichia coli with the bifunctional reagent nitrogen mustard. The cross-linking primarily involved the 16S RNA. To localize the site of cross-linking of streptomycin to the 16S RNA, the authors hybridized RNA labeled with streptomycin to restriction fragments of the 16S RNA gene. Labeled RNA hybridized to DNA fragments corresponding to bases 892-917 and bases 1394-1415. These two segments of the ribosomal RNA must by juxtaposed in the ribosome, since there is a single binding site for streptomycin. This region has been implicated both in the decoding site and in the binding of initiation factor IF-3, indicating its functional importance

  12. Corneal collagen cross-linking: ectasia and beyond.

    Science.gov (United States)

    Suri, Kunal; Hammersmith, Kristin M; Nagra, Parveen K

    2012-07-01

    Corneal collagen cross-linking has recently emerged as a novel approach for management of ectasia. This article reviews the literature published in the past 3 years about the expanding spectrum of cross-linking as a therapeutic modality and its complications. Recent studies have confirmed the beneficial effects of cross-linking in stabilization and to a lesser extent, regression of keratoconus and postrefractive surgery ectasia. Other applications include cross-linking as a combined procedure with intracorneal ring segments, and photorefractive keratectomy for ectasia, corneal edema, and infectious keratitis. Animal studies of chemical cross-linking of sclera as a potential treatment for progressive myopia have also been performed. Various modifications of the technique to increase the safety profile of cross-linking have been reported, including the use of hypoosmolar riboflavin, transepithelial cross-linking, customized epithelial debridement, and higher fluence shorter duration ultraviolet A light exposure. Reported complications include keratitis, corneal haze, endothelial cell loss and failure of treatment. Cross-linking has been shown to be an effective modality for corneal ectasia, the regression being less in patients with postrefractive ectasia than keratoconus. In a few studies, it has been found to be effective in symptomatic improvement of bullous keratopathy, and infectious keratitis but further studies are required. Cross-linking with epithelial debridement is found to be most effective but various modifications are being investigated for an improved, and better safety outcome.

  13. Biodegradable cyclen-based linear and cross-linked polymers as non-viral gene vectors.

    Science.gov (United States)

    Li, Shuo; Wang, Yu; Wang, Shan; Zhang, Ji; Wu, Shi-Fei; Wang, Bo-Lin; Zhu, Wen; Yu, Xiao-Qi

    2012-02-15

    Several 1,4,7,10-tetraazacyclododecane (cyclen)-based linear (3a-c) and cross-linked (8a-d) polymers containing biodegradable ester or disulfide bonds were described. These polymeric compounds were prepared by ring-opening polymerization from various diol glycidyl ethers. The molecular weights of the title polymers were measured by GPC. Agarose gel retardation assays showed that these compounds have good DNA-binding ability and can completely retard plasmid DNA (pDNA) at weight ratio of 20 for linear polymers and 1.2 for cross-linked polymers. The degradation of these polymers was confirmed by GPC. The formed polyplexes have appropriate sizes around 400 nm and zeta-potential values about 15-40 mV. The cytotoxicities of 8 assayed by MTT are much lower than that of 25 KDa PEI. In vitro transfection toward A549 and 293 cells showed that the transfection efficiency (TE) of 8c-DNA polyplex is close to that of 25 kDa PEI at 8c/DNA weight ratio of 4. Structure-activity relationships (SAR) of these linear and cross-linked polymers were discussed in their DNA-binding, cytotoxicity, and transfection studies. In addition, in the presence of serum, the TE of 8/DNA polyplexes could be improved by introducing chloroquine or Ca(2+) to pretreated cells. Copyright © 2012 Elsevier Ltd. All rights reserved.

  14. Collagen Cross-linking for Microbial Keratitis

    Science.gov (United States)

    Garg, Prashant; Das, Sujata; Roy, Aravind

    2017-01-01

    Collagen cross-linking is gaining popularity not only for arresting the progression of keratoconus but also other indications including management of corneal infections. In this review article, we analyzed the published literature to understand the level of evidence for its use in corneal ulcer. Photoactivated riboflavin and ultraviolet A light are known to possess antimicrobial properties. The treatment also induces formation of inter- and intra-fibrillar bonds, thereby making the corneal collagen resistant to the action of proteases arresting stromal melt. Both properties are well documented in in vitro experiments. The antimicrobial action is seen against bacteria, fungi, and parasites. The animal experiments have documented its efficacy against bacterial and fungal keratitis models. The literature on its application in human corneal infection is highly variable and comprises case reports, case series, and comparative nonrandomized and randomized trials. The treatment has been used as primary treatment, adjunctive treatment along with antibiotics, as the first line of treatment as well as for failed medical treatment cases. Even the cases included are of variable severity caused by a variety of microorganisms including culture-negative cases. Furthermore, the treatment protocols are also variable. While most reports show beneficial effects for bacterial corneal ulcer cases, especially those with superficial infiltrate, the effect has been mixed for fungal and parasitic keratitis. In view of these characteristics, we infer that the level of evidence for its use in corneal ulcer is at most weak. We need well-characterized, high-quality, clinical trials of sufficient power to assess its true value. PMID:28546688

  15. Cross-linked polyethylenimine–tripolyphosphate nanoparticles for gene delivery

    Directory of Open Access Journals (Sweden)

    Huang XZ

    2014-10-01

    Full Text Available Xianzhang Huang,1 Sujing Shen,2 Zhanfeng Zhang,1 Junhua Zhuang1 1Department of Laboratory Science, Second Affiliated Hospital of Guangzhou University of Chinese Medicine, 2Department of Laboratory Science, Guangdong Second Provincial Traditional Chinese Medicine Hospital, Guangzhou, People’s Republic of China Abstract: The high transfection efficiency of polyethylenimine (PEI makes it an attractive potential nonviral genetic vector for gene delivery and therapy. However, the highly positive charge of PEI leads to cytotoxicity and limits its application. To reduce the cytotoxicity of PEI, we prepared anion-enriched nanoparticles that combined PEI with tripolyphosphate (TPP. We then characterized the PEI-TPP nanoparticles in terms of size, zeta potential, and Fourier-transform infrared (FTIR spectra, and assessed their transfection efficiency, cytotoxicity, and ability to resist deoxyribonuclease (DNase I digestion. The cellular uptake of PEI-TPP with phosphorylated internal ribosome entry site–enhanced green fluorescent protein C1 or FAM (fluorouracil, Adriamycin [doxorubicin] and mitomycin-labeled small interfering ribonucleic acids (siRNAs was monitored by fluorescence microscopy and confocal laser microscopy. The efficiency of transfected delivery of plasmid deoxyribonucleic acid (DNA and siRNA in vitro was 1.11- to 4.20-fold higher with the PEI-TPP particles (7.6% cross-linked than with the PEI, at all N:P ratios (nitrogen in PEI to phosphorus in DNA tested. The cell viability of different cell lines was more than 90% at the chosen N:P ratios of PEI-TPP/DNA complexes. Moreover, PEI-TPP nanoparticles resisted digestion by DNase I for more than 2 hours. The time-dependent absorption experiment showed that 7.6% of cross-linked PEI-TPP particles were internalized by 293T cells within 1 hour. In summary, PEI-TPP nanoparticles effectively transfected cells while conferring little or no toxicity, and thus have potential application in gene

  16. Cytotoxic platinum coordination compounds. DNA binding agents

    Czech Academy of Sciences Publication Activity Database

    Brabec, Viktor; Hrabina, O.; Kašpárková, Jana

    2017-01-01

    Roč. 351, č. 2017 (2017), s. 2-31 ISSN 0010-8545 R&D Projects: GA ČR GA17-09436S Institutional support: RVO:68081707 Keywords : interstrand cross-links * nucleotide excision-repair * pt-ii complex Subject RIV: CE - Biochemistry OBOR OECD: Biochemistry and molecular biology Impact factor: 13.324, year: 2016

  17. Covalently cross-linked polyetheretherketone proton exchange membrane for DMFC

    CSIR Research Space (South Africa)

    Luo, H

    2009-05-01

    Full Text Available The proton exchange membrane was prepared by covalent cross-linking sulfonated-sulfinated polyetheretherketone. The cross-linked membrane showed high proton conductivity (0.04 S/cm) with suitable water uptake, low methanol permeability (2.21 × 10...

  18. Hydrogels Prepared from Cross-Linked Nanofibrillated Cellulose

    Science.gov (United States)

    Sandeep S. Nair; J.Y. Zhu; Yulin Deng; Arthur J. Ragauskas

    2014-01-01

    Nanocomposite hydrogels were developed by cross-linking nanofibrillated cellulose with poly(methyl vinyl ether-co-maleic acid) and polyethylene glycol. The cross-linked hydrogels showed enhanced water absorption and gel content with the addition of nanocellulose. In addition, the thermal stability, mechanical strength, and modulus increased with an increase in the...

  19. Recent advances in corneal collagen cross-linking

    Directory of Open Access Journals (Sweden)

    Gitansha Shreyas Sachdev

    2017-01-01

    Full Text Available Corneal collagen cross-linking has become the preferred modality of treatment for corneal ectasia since its inception in late 1990s. Numerous studies have demonstrated the safety and efficacy of the conventional protocol. Our understanding of the cross-linking process is ever evolving, with its wide implications in the form of accelerated and pulsed protocols. Newer advancements in technology include various riboflavin formulations and the ability to deliver higher fluence protocols with customised irradiation patterns. A greater degree of customisation is likely the path forward, which will aim at achieving refractive improvements along with disease stability. The use of cross-linking for myopic correction is another avenue under exploration. Combination of half fluence cross-linking with refractive correction for high errors to prevent post LASIK regression is gaining interest. This review aims to highlight the various advancements in the cross-linking technology and its clinical applications.

  20. Differential repair of platinum-DNA adducts in human bladder and testicular tumor continuous cell lines

    International Nuclear Information System (INIS)

    Bedford, P.; Fichtinger-Schepman, A.M.; Shellard, S.A.; Walker, M.C.; Masters, J.R.; Hill, B.T.

    1988-01-01

    The formation and removal of four platinum-DNA adducts were immunochemically quantitated in cultured cells derived from a human bladder carcinoma cell line (RT112) and from two lines derived from germ cell tumors of the testis (833K and SUSA), following exposure in vitro to 16.7 microM (5 micrograms/ml) cisplatin. RT112 cells were least sensitive to the drug and were proficient in the repair of all four adducts, whereas SUSA cells, which were 5-fold more sensitive, were deficient in the repair of DNA-DNA intrastrand cross-links in the sequences pApG and pGpG. Despite expressing a similar sensitivity to SUSA cells, 833K cells were proficient in the repair of all four adducts, although less so than the RT112 bladder tumor cells. In addition, SUSA cells were unable to repair DNA-DNA interstrand cross-links whereas 50-85% of these lesions were removed in RT112 and 833K cells 24 h following drug exposure. It is possible that the inability of SuSa cells to repair platinated DNA may account for their hypersensitivity to cisplatin

  1. Manufacture of polyethylene foam by electron beam cross-linking

    International Nuclear Information System (INIS)

    Tamai, Isamu

    1976-01-01

    The manufacturing process of polyethylene foam, comparison between electron beam cross-linking process and chemical cross-linking process, the electron beam irradiation technique for continuous sheets, the characteristics and uses of polyethylene foam are reviewed. The pore diameter can be controlled by selecting the dose rate, because there is strong relationship between the pore diameter and the dose rate. As the dose if higher, the foam becomes finer. The electron accelerators having large capacity show the lowest cost as the radiation source, and are applicable industrially. If the production capacity exceeds about 200 tons per month, the costs of electron beam irradiation process may be more advantageous than that of chemical process according to the circumstances. It is difficult to obtain the uniform distribution of absorption dose in the direction of thickness. General characteristics of cross-linked polyethylene foam are listed. The special feature of electron beam process is that the degree of cross-linking can be controlled arbitrarily before foaming. The products obtained by the electron beam cross-linking process have finer foams and smoother surfaces than those obtained by the chemical process, because the separation of the decomposition of foaming agents from that of cross-linking agents in the chemical cross-linking is difficult. (Iwakiri, K.)

  2. Immunogenic Display of Purified Chemically Cross-Linked HIV-1 Spikes

    Science.gov (United States)

    Leaman, Daniel P.; Lee, Jeong Hyun; Ward, Andrew B.

    2015-01-01

    ABSTRACT HIV-1 envelope glycoprotein (Env) spikes are prime vaccine candidates, at least in principle, but suffer from instability, molecular heterogeneity and a low copy number on virions. We anticipated that chemical cross-linking of HIV-1 would allow purification and molecular characterization of trimeric Env spikes, as well as high copy number immunization. Broadly neutralizing antibodies bound tightly to all major quaternary epitopes on cross-linked spikes. Covalent cross-linking of the trimer also stabilized broadly neutralizing epitopes, although surprisingly some individual epitopes were still somewhat sensitive to heat or reducing agent. Immunodepletion using non-neutralizing antibodies to gp120 and gp41 was an effective method for removing non-native-like Env. Cross-linked spikes, purified via an engineered C-terminal tag, were shown by negative stain EM to have well-ordered, trilobed structure. An immunization was performed comparing a boost with Env spikes on virions to spikes cross-linked and captured onto nanoparticles, each following a gp160 DNA prime. Although differences in neutralization did not reach statistical significance, cross-linked Env spikes elicited a more diverse and sporadically neutralizing antibody response against Tier 1b and 2 isolates when displayed on nanoparticles, despite attenuated binding titers to gp120 and V3 crown peptides. Our study demonstrates display of cross-linked trimeric Env spikes on nanoparticles, while showing a level of control over antigenicity, purity and density of virion-associated Env, which may have relevance for Env based vaccine strategies for HIV-1. IMPORTANCE The envelope spike (Env) is the target of HIV-1 neutralizing antibodies, which a successful vaccine will need to elicit. However, native Env on virions is innately labile, as well as heterogeneously and sparsely displayed. We therefore stabilized Env spikes using a chemical cross-linker and removed non-native Env by immunodepletion with non

  3. Current status of accelerated corneal cross-linking

    Directory of Open Access Journals (Sweden)

    Michael Mrochen

    2013-01-01

    Full Text Available Corneal cross-linking with riboflavin is a technique to stabilize or reduce corneal ectasia, in diseases such as keratoconus and post-laser-assisted in situ keratomileusis (LASIK ectasia. There is an interest by patient as well as clinicians to reduce the overall treatment time. Especially, the introduction of corneal cross-linking in combination with corneal laser surgery demands a shorter treatment time to assure a sufficient patient flow. The principles and techniques of accelerated corneal cross-linking is discussed.

  4. Interstrand resistance of selected sections of DCA312

    Energy Technology Data Exchange (ETDEWEB)

    Kovachev, V.; Neal, M.; Seuntjens, J.; Qin, J.; Cline, P.; Capone, D. II [Superconducting Super Collider Lab., Dallas, TX (United States); Swenson, C. [Westinghouse Electric Corp., Round Rock, TX (United States)

    1993-11-01

    The quench current ramp rate sensitivity of the full length SSC dipole DCA312 was strongly A-type and the eddy current losses for this magnet were the largest of all ASST magnets. The quench current remained constant with ramp rate up to approximately 20 A/s. At higher ramp rates the quench current decreased dramatically. At a current ramp rate of 200 A/s the quench current was only 2000 A. The eddy current losses of DCA312 at 60 A/s for a monopolar cycle of 500A-5000A-500A were approximately 3600 J/Cycle, a factor of 5--7 larger than for magnets DCA311, DCA319, and DCA318. In order to find the cause of the anomalous behavior of DCA312, it was decided that the magnet would be cut into sections and examined at WMSD and SSCL. The general goals of the DCA312 autopsy included verification of theoretical models for A-type behavior in SSC magnets as well as correlation of fabrication and assembly procedures to the quench behavior of A-type magnets. One of the important aspects of the magnet autopsy plan was the in-situ measurement of interstrand resistance in collared magnet sections. Four sections of DCA312 were designated for interstrand resistance measurements. Two of those sections (C and M) contained quench sites (100 A/s and 200 A/s respectively) and two of them (E and J) were sections representative of the magnet where quenches did not occur. The aim of the present work was to measure and analyze the interstrand resistance in each quadrant of these four sections of DCA312 and to find a correlation between the measured interstrand resistance, the actual resistance of individual interstrand contacts, and the quench performance of the magnet.

  5. Mutagenicity of acrolein and acrolein-induced DNA adducts.

    Science.gov (United States)

    Liu, Xing-yu; Zhu, Mao-xiang; Xie, Jian-ping

    2010-01-01

    Acrolein mutagenicity relies on DNA adduct formation. Reaction of acrolein with deoxyguanosine generates alpha-hydroxy-1, N(2)-propano-2'-deoxyguanosine (alpha-HOPdG) and gamma-hydroxy-1, N(2)-propano-2'-deoxyguanosine (gamma-HOPdG) adducts. These two DNA adducts behave differently in mutagenicity. gamma-HOPdG is the major DNA adduct and it can lead to interstrand DNA-DNA and DNA-peptide/protein cross-links, which may induce strong mutagenicity; however, gamma-HOPdG can be repaired by some DNA polymerases complex and lessen its mutagenic effects. alpha-HOPdG is formed much less than gamma-HOPdG, but difficult to be repaired, which contributes to accumulation in vivo. Results of acrolein mutagenicity studies haven't been confirmed, which is mainly due to the conflicting mutagenicity data of the major acrolein adduct (gamma-HOPdG). The minor alpha-HOPdG is mutagenic in both in vitro and in vivo test systems. The role of alpha-HOPdG in acrolein mutagenicity needs further investigation. The inconsistent result of acrolein mutagenicity can be attributed, at least partially, to a variety of acrolein-DNA adducts formation and their repair in diverse detection systems. Recent results of detection of acrolein-DNA adduct in human lung tissues and analysis of P53 mutation spectra in acrolein-treated cells may shed some light on mechanisms of acrolein mutagenicity. These aspects are covered in this mini review.

  6. AUTOCLAVABLE HIGHLY CROSS-LINKED POLYURETHANE NETWORKS IN OPHTHALMOLOGY

    NARCIS (Netherlands)

    BRUIN, P; MEEUWSEN, EAJ; VANANDEL, MV; WORST, JGF; PENNINGS, AJ

    1993-01-01

    Highly cross-linked aliphatic polyurethane networks have been prepared by the bulk step reaction of low molecular weight polyols and hexamethylenediisocyanate (HDI). These polyurethane networks are optically transparent, colourless and autoclavable amorphous glassy thermosets, which are suited for

  7. 21 CFR 177.1211 - Cross-linked polyacrylate copolymers.

    Science.gov (United States)

    2010-04-01

    ... Basic Components of Single and Repeated Use Food Contact Surfaces § 177.1211 Cross-linked polyacrylate... shall be determined using size exclusion chromatography or an equivalent method. When conducting the...

  8. Enzymatic digestibility of peptides cross-linked by ionizing radiation

    International Nuclear Information System (INIS)

    Dizdaroglu, M.; Gajewski, E.; Simic, M.G.

    1984-01-01

    Digestibility by proteolytic enzymes of peptides cross-linked by ionizing radiation was investigated. Small peptides of alanine and phenylalanine were chosen as model compounds and aminopeptidases and carboxypeptidases were used as proteolytic enzymes. Peptides exposed to γ-radiation in aqueous solution were analysed by high-performance liquid chromatography before and after hydrolysis by aminopeptidase M, leucine aminopeptidase carboxypeptidase A and carboxypeptidase Y. The results obtained clearly demonstrate the different actions of these enzymes on cross-linked aliphatic and aromatic peptides. Peptide bonds of cross-linked dipeptides of alanine were completely resistant to enzymatic hydrolysis whereas the enzymes, except for carboxypeptidase Y, cleaved all peptide bonds of cross-linked peptides of phenylalanine. The actions of the enzymes on these particular compounds are discussed in detail. (author)

  9. Complications Following Corneal Cross-Linking Treatment in Keratectasia

    Directory of Open Access Journals (Sweden)

    Faik Oruçoğlu

    2013-01-01

    Full Text Available Pur po se: To report complications of corneal cross-linking treatment in keratoconus and secondary ectatic eyes. Ma te ri al and Met hod: Cases that were treated between February 2007 and February 2011, had a minimum follow-up of one month, and developed complications following corneal cross-linking were retrospectively evaluated. Re sults: Complication data were present in eight cases. Scar formation developed in three cases. Scar density has decreased in one patient during long-term follow-up. Keratometric flattening was observed associated with scar development. Cross-linking was performed on the second eye of the affected patient, and no scar development was observed. Corneal clouding was observed in one patient three days after treatment. Corneal clouding has decreased in the following days. Local corneal edema has developed in one patient, and improvement was achieved in the following days. Uveitis has developed in one case with total improvement after adding topical steroids. Post-LASIK ectasia eye showed deep lamellar keratitis after cross-linking. In one patient, epithelization was completed in 14 days. Dis cus si on: Corneal scars, corneal clouding and edema, uveitis, and late epithelization are possible complications after corneal cross-linking. Scar density was decreased, while other complications showed total improvement. (Turk J Ophthalmol 2013; 43: 1-6

  10. Formaldehyde cross-linking and structural proteomics: Bridging the gap.

    Science.gov (United States)

    Srinivasa, Savita; Ding, Xuan; Kast, Juergen

    2015-11-01

    Proteins are dynamic entities constantly moving and altering their structures based on their functions and interactions inside and outside the cell. Formaldehyde cross-linking combined with mass spectrometry can accurately capture interactions of these rapidly changing biomolecules while maintaining their physiological surroundings. Even with its numerous established uses in biology and compatibility with mass spectrometry, formaldehyde has not yet been applied in structural proteomics. However, formaldehyde cross-linking is moving toward analyzing tertiary structure, which conventional cross-linkers have already accomplished. The purpose of this review is to describe the potential of formaldehyde cross-linking in structural proteomics by highlighting its applications, characteristics and current status in the field. Copyright © 2015 Elsevier Inc. All rights reserved.

  11. DNA excision repair in cell extracts from human cell lines exhibiting hypersensitivity to DNA-damaging agents

    International Nuclear Information System (INIS)

    Hansson, J.; Keyse, S.M.; Lindahl, T.; Wood, R.D.

    1991-01-01

    Whole cell extracts from human lymphoid cell lines can perform in vitro DNA repair synthesis in plasmids damaged by agents including UV or cis-diamminedichloroplatinum(II) (cis-DDP). Extracts from xeroderma pigmentosum (XP) cells are defective in repair synthesis. We have now studied in vitro DNA repair synthesis using extracts from lymphoblastoid cell lines representing four human hereditary syndromes with increased sensitivity to DNA-damaging agents. Extracts of cell lines from individuals with the sunlight-sensitive disorders dysplastic nevus syndrome or Cockayne's syndrome (complementation groups A and B) showed normal DNA repair synthesis in plasmids with UV photoproducts. This is consistent with in vivo measurements of the overall DNA repair capacity in such cell lines. A number of extracts were prepared from two cell lines representing the variant form of XP (XP-V). Half of the extracts prepared showed normal levels of in vitro DNA repair synthesis in plasmids containing UV lesions, but the remainder of the extracts from the same cell lines showed deficient repair synthesis, suggesting the possibility of an unusually labile excision repair protein in XP-V. Fanconi's anemia (FA) cells show cellular hypersensitivity to cross-linking agents including cis-DDP. Extracts from cell lines belonging to two different complementation groups of FA showed normal DNA repair synthesis in plasmids containing cis-DDP or UV adducts. Thus, there does not appear to be an overall excision repair defect in FA, but the data do not exclude a defect in the repair of interstrand DNA cross-links

  12. Scleral lens tolerance after corneal cross-linking for keratoconus

    NARCIS (Netherlands)

    Visser, Esther Simone; Soeters, Nienke; Tahzib, Nayyirih G.

    2015-01-01

    Purpose. Subjective and objective evaluation of scleral lens tolerance and fitting before and after corneal cross-linking (CXL) for progressive keratoconus. Methods. In this prospective cohort, evaluations were made of 18 unilateral eyes in patients who underwent CXL and had been wearing scleral

  13. Cross-linked enzyme aggregates (CLEAs) : Stable and recyclable biocatalysts

    NARCIS (Netherlands)

    Sheldon, R.A.

    2007-01-01

    The key to obtaining an optimum performance of an enzyme is often a question of devising an effective method for its immobilization. In the present review, we describe a novel, versatile and effective methodology for enzyme immobilization as CLEAs (cross-linked enzyme aggregates). The method is

  14. Chitosan Cross-linked Reconstituted Amniotic Collagen Membrane ...

    Indian Academy of Sciences (India)

    First page Back Continue Last page Overview Graphics. Chitosan Cross-linked Reconstituted Amniotic Collagen Membrane – An Excellent Cell Substratum. The KERATINOCYTE proliferation and Differentiation into multiple layers is due to the presence of type - IV collagen in the amnion. Cultured FIBROBLASTS had good ...

  15. Preparation and adsorption property of aminated cross linking ...

    Indian Academy of Sciences (India)

    Administrator

    Preparation and adsorption property of aminated cross linking microbeads of GMA/EGDMA for bilirubin. ZHIPING CHEN, BAOJIAO GAO* and XIAOFENG YANG. Department of Chemical Engineering, North University of China, Taiyuan, 030051, China e-mail: gaobaojiao@126.com. MS received 17 December 2008; revised ...

  16. Functionalisation of cross-linked polyethylenimine for the removal of ...

    African Journals Online (AJOL)

    Cross-linked polyethylenimine (CPEI) was phosphonated by reaction with phosphorous acid and formaldehyde. The functionalised polymer was used as an adsorbent for the removal of arsenic as an oxo-anion. The binding affinity of the synthesised polymer to abstract As from synthetic solutions and wastewater samples ...

  17. Hyper-cross-linked, hybrid membranes via interfacial polymerization

    NARCIS (Netherlands)

    Raaijmakers, Michiel

    2015-01-01

    Hyper-cross-linked, hybrid membranes consist of covalent networks of alternating organic and inorganic, or biological groups. This thesis reports on the preparation of such hybrid networks via interfacial polymerization. The structure-property relationships of the hybrid networks depend strongly on

  18. Thermoreversibly Cross-Linked EPM Rubber Nanocomposites with Carbon Nanotubes

    NARCIS (Netherlands)

    Polgar, Lorenzo Massimo; Criscitiello, Francesco; van Essen, Machiel; Araya-Hermosilla, Rodrigo; Migliore, Nicola; Lenti, Mattia; Raffa, Patrizio; Picchioni, Francesco; Pucci, Andrea

    2018-01-01

    Conductive rubber nanocomposites were prepared by dispersing conductive nanotubes (CNT) in thermoreversibly cross-linked ethylene propylene rubbers grafted with furan groups (EPM-g-furan) rubbers. Their features were studied with a strong focus on conductive and mechanical properties relevant for

  19. Subunit topography of RNA polymerase from Escherichia coli. A cross-linking study with bifunctional reagents.

    Science.gov (United States)

    Hillel, Z; Wu, C W

    1977-07-26

    The quaternary structures of Escherichia coli DNA-dependent RNA polymerase holenzyme (alpha 2 beta beta' sigma) and core enzyme (alpha 2 beta beta') have been investigated by chemical cross-linking with a cleavable bifunctional reagent, methyl 4-mercaptobutyrimidate, and noncleavable reagents, dimethyl suberimidate and N,N'-(1,4-phenylene)bismaleimide. A model of the subunit organization deduced from cross-linked subunit neighbors identified by dodecyl sulfate-polyacrylamide gel electrophoresis indicates that the large beta and beta' subunits constitute the backbone of both core and holoenzyme, while sigma and two alpha subunits interact with this structure along the contact domain of beta and beta' subunits. In holoenzyme, sigma subunit is in the vicinity of at least one alpha subunit. The two alpha subunits are close to each other in holoenzyme, core enzyme, and the isolated alpha 2 beta complex. Cross-linking of the "premature" core and holoenzyme intermediates in the in vitro reconstitution of active enzyme from isolated subunits suggests that these species are composed of subunit complexes of molecular weight lower than that of native core and holoenzyme, respectively. The structural information obtained for RNA polymerase and its subcomplexes has important implications for the enzyme-promoter recognition as well as the mechanism of subunit assembly of the enzyme.

  20. A comparison of UV cross-linking and vacuum baking for nucleic acid immobilization and retention

    International Nuclear Information System (INIS)

    Nierzwicki-Bauer, S.A.; Gebhardt, J.S.; Linkkila, L.; Walsh, K.

    1990-01-01

    The effectiveness of UV cross-linking and in vacuo baking for the immobilization and retention of DNA to various solid supports was investigated. Optimal immobilization treatments for supported and unsupported nitrocellulose and nylon membranes were: UV cross-linking at 254 nm with an exposure of 120 milliJoules/cm 2 , or baking in vacuo for two hours at 80 degrees C. UV-immobilized nitrocellulose-based membranes showed no increase in sensitivity when compared to baked membranes. An increase in sensitivity was observed for UV-immobilized nylon membranes as compared with baked nylon membranes in some instances, although this varied within lots of the membranes tested. Repeated strippings and heterologous reprobings resulted in loss of target DNA from UV-immobilized nylon membranes as compared to baked nylon membranes. Loss of target DNA from UV-immobilized nitrocellulose-based membranes due to repeated strippings and reprobings was even more pronounced. In vacuo baking of supported and unsupported nitrocellulose and nylon membranes was more effective for immobilization, and more importantly, for retention of target DNA through many reprobings of the same blot

  1. Quantitative PCR analysis of diepoxybutane and epihalohydrin damage to nuclear versus mitochondrial DNA

    International Nuclear Information System (INIS)

    LaRiviere, Frederick J.; Newman, Adam G.; Watts, Megan L.; Bradley, Sharonda Q.; Juskewitch, Justin E.; Greenwood, Paul G.; Millard, Julie T.

    2009-01-01

    The bifunctional alkylating agents diepoxybutane (DEB) and epichlorohydrin (ECH) are linked to the elevated incidence of certain cancers among workers in the synthetic polymer industry. Both compounds form interstrand cross-links within duplex DNA, an activity suggested to contribute to their cytotoxicity. To assess the DNA targeting of these compounds in vivo, we assayed for damage within chicken erythro-progenitor cells at three different sites: one within mitochondrial DNA, one within expressed nuclear DNA, and one within unexpressed nuclear DNA. We determined the degree of damage at each site via a quantitative polymerase chain reaction, which compares amplification of control, untreated DNA to that from cells exposed to the agent in question. We found that ECH and the related compound epibromohydrin preferentially target nuclear DNA relative to mitochondrial DNA, whereas DEB reacts similarly with the two genomes. Decreased reactivity of the mitochondrial genome could contribute to the reduced apoptotic potential of ECH relative to DEB. Additionally, formation of lesions by all agents occurred at comparable levels for unexpressed and expressed nuclear loci, suggesting that alkylation is unaffected by the degree of chromatin condensation.

  2. The spectra character of photodegraded the pyridinoline cross-links by Hypocrellin B

    International Nuclear Information System (INIS)

    Zhang Jucheng; Chen Rui; Liu Wei; Chen Zhuo; Shu Lidan; Liu Yingji

    2011-01-01

    Pyridinoline cross-links is one of the cross-link formation in collagen which in cell matrix, many research shown that this cross-link cause the fibrosis. Hypocrellin B (HB) is one of the nature photosensitizers, this work investigated the pyridinoline cross-link in collagen was photodegraded by HB. The result shown HB can degrade the pyridinoline cross-link with photo. This is to say, HB may be use as the photodynamic reagent to study the fibrosis.

  3. Transient Anisocoria after Corneal Collagen Cross-Linking

    Directory of Open Access Journals (Sweden)

    George D. Kymionis

    2014-01-01

    Full Text Available Purpose. To report a case with transient anisocoria after corneal collagen cross-linking (CXL. Methods. Case report. Results. A 24-year-old male underwent corneal collagen cross-linking (CXL in his right eye for keratoconus. At the end of the procedure, the pupil of the treated eye was irregular and dilated, while the pupil of the fellow eye was round, regular, and reactive (anisocoria. The following day, pupils were round, regular, and reactive in both eyes. Conclusion. Anisocoria may be a transient and innocuous complication after CXL. A possible cause for this complication might be the anesthetic drops used before and during the surgical procedure or/and the ultraviolet A irradiation during the treatment.

  4. Cross-linking reveals laminin coiled-coil architecture

    Science.gov (United States)

    Armony, Gad; Jacob, Etai; Moran, Toot; Levin, Yishai; Mehlman, Tevie; Levy, Yaakov; Fass, Deborah

    2016-01-01

    Laminin, an ∼800-kDa heterotrimeric protein, is a major functional component of the extracellular matrix, contributing to tissue development and maintenance. The unique architecture of laminin is not currently amenable to determination at high resolution, as its flexible and narrow segments complicate both crystallization and single-particle reconstruction by electron microscopy. Therefore, we used cross-linking and MS, evaluated using computational methods, to address key questions regarding laminin quaternary structure. This approach was particularly well suited to the ∼750-Å coiled coil that mediates trimer assembly, and our results support revision of the subunit order typically presented in laminin schematics. Furthermore, information on the subunit register in the coiled coil and cross-links to downstream domains provide insights into the self-assembly required for interaction with other extracellular matrix and cell surface proteins. PMID:27815530

  5. Computer simulation of randomly cross-linked polymer networks

    International Nuclear Information System (INIS)

    Williams, Timothy Philip

    2002-01-01

    In this work, Monte Carlo and Stochastic Dynamics computer simulations of mesoscale model randomly cross-linked networks were undertaken. Task parallel implementations of the lattice Monte Carlo Bond Fluctuation model and Kremer-Grest Stochastic Dynamics bead-spring continuum model were designed and used for this purpose. Lattice and continuum precursor melt systems were prepared and then cross-linked to varying degrees. The resultant networks were used to study structural changes during deformation and relaxation dynamics. The effects of a random network topology featuring a polydisperse distribution of strand lengths and an abundance of pendant chain ends, were qualitatively compared to recent published work. A preliminary investigation into the effects of temperature on the structural and dynamical properties was also undertaken. Structural changes during isotropic swelling and uniaxial deformation, revealed a pronounced non-affine deformation dependant on the degree of cross-linking. Fractal heterogeneities were observed in the swollen model networks and were analysed by considering constituent substructures of varying size. The network connectivity determined the length scales at which the majority of the substructure unfolding process occurred. Simulated stress-strain curves and diffraction patterns for uniaxially deformed swollen networks, were found to be consistent with experimental findings. Analysis of the relaxation dynamics of various network components revealed a dramatic slowdown due to the network connectivity. The cross-link junction spatial fluctuations for networks close to the sol-gel threshold, were observed to be at least comparable with the phantom network prediction. The dangling chain ends were found to display the largest characteristic relaxation time. (author)

  6. [Complications and postoperative therapeutic strategies in cross-linking].

    Science.gov (United States)

    Kohlhaas, M

    2017-08-01

    The reduced corneal mechanical stability in keratoconus and similar collagen diseases can lead to a progressive and irregular corneal shape and decrease of visual acuity. A progression of keratectatic diseases can be shown with corneal topography. Keratoconus can be treated by photo-oxidative cross-linking of the corneal collagen. In order to achieve a high absorption of irradiation energy in the cornea, riboflavin at a concentration of 0.1% and UVA light at a wavelength of 370 nm corresponding to the relative maximum absorption of riboflavin (vitamin B2) are used. Evidence for corneal cross-linking are the increase of biomechanical stiffness, the increased resistance against enzymatic degradation, a higher shrinkage temperature, a lower swelling rate and an increased diameter of collagen fibers. The currently available data demonstrate that the therapeutic cross-linking procedure is safe when respecting the important theoretical and clinical parameters and that a progression of the keratoconus can be avoided. In 80% of cases an average levelling of the curvature of approximately 2 dpt can be achieved, which leads not only to stabilization but also to an increase in visual acuity of approximately 1.2 lines. In a Cochrane review from 2015 publications about complications and results were reviewed. Complication rates ranged from 1-10% depending on the initial situation, comorbidities and stage of the keratoconus. The most important complications are early epithelial wound healing problems as well as extremely rare perforations. Corneal cross-linking is a well-established and safe procedure but is not free of complications.

  7. Cytokines and growth factors cross-link heparan sulfate

    Science.gov (United States)

    Migliorini, Elisa; Thakar, Dhruv; Kühnle, Jens; Sadir, Rabia; Dyer, Douglas P.; Li, Yong; Sun, Changye; Volkman, Brian F.; Handel, Tracy M.; Coche-Guerente, Liliane; Fernig, David G.; Lortat-Jacob, Hugues; Richter, Ralf P.

    2015-01-01

    The glycosaminoglycan heparan sulfate (HS), present at the surface of most cells and ubiquitous in extracellular matrix, binds many soluble extracellular signalling molecules such as chemokines and growth factors, and regulates their transport and effector functions. It is, however, unknown whether upon binding HS these proteins can affect the long-range structure of HS. To test this idea, we interrogated a supramolecular model system, in which HS chains grafted to streptavidin-functionalized oligoethylene glycol monolayers or supported lipid bilayers mimic the HS-rich pericellular or extracellular matrix, with the biophysical techniques quartz crystal microbalance (QCM-D) and fluorescence recovery after photobleaching (FRAP). We were able to control and characterize the supramolecular presentation of HS chains—their local density, orientation, conformation and lateral mobility—and their interaction with proteins. The chemokine CXCL12α (or SDF-1α) rigidified the HS film, and this effect was due to protein-mediated cross-linking of HS chains. Complementary measurements with CXCL12α mutants and the CXCL12γ isoform provided insight into the molecular mechanism underlying cross-linking. Fibroblast growth factor 2 (FGF-2), which has three HS binding sites, was also found to cross-link HS, but FGF-9, which has just one binding site, did not. Based on these data, we propose that the ability to cross-link HS is a generic feature of many cytokines and growth factors, which depends on the architecture of their HS binding sites. The ability to change matrix organization and physico-chemical properties (e.g. permeability and rigidification) implies that the functions of cytokines and growth factors may not simply be confined to the activation of cognate cellular receptors. PMID:26269427

  8. Hyper-cross-linked, hybrid membranes via interfacial polymerization

    OpenAIRE

    Raaijmakers, Michiel

    2015-01-01

    Hyper-cross-linked, hybrid membranes consist of covalent networks of alternating organic and inorganic, or biological groups. This thesis reports on the preparation of such hybrid networks via interfacial polymerization. The structure-property relationships of the hybrid networks depend strongly on the type, size and flexibility of the constituents. The collection of polymers that can be synthesized via interfacial polymerization includes polyamides, polyurethanes, polyureas, polyanilines, po...

  9. The cellular response to transglutaminase-cross-linked collagen.

    Science.gov (United States)

    Chau, David Y S; Collighan, Russell J; Verderio, Elisabetta A M; Addy, Victoria L; Griffin, Martin

    2005-11-01

    Collagen, type I, is a highly abundant natural protein material which has been cross-linked by a variety of methods including chemical agents, physical heating and UV irradiation with the aim of enhancing its physical characteristics such as mechanical strength, thermal stability, resistance to proteolytic breakdown, thus increasing its overall biocompatibility. However, in view of the toxicity of residual cross-linking agents, or impracticability at large scales, it would be more useful if the collagen could be cross-linked by a milder, efficient and more practical means by using enzymes as biological catalysts. We demonstrate that on treating native collagen type I (from bovine skin) with both tissue transglutaminase (TG2; tTG) and microbial transglutaminase (mTG; Streptoverticillium mobaraense) leads to an enhancement in cell attachment, spreading and proliferation of human osteoblasts (HOB) and human foreskin dermal fibroblasts (HFDF) when compared to culture on native collagen. The transglutaminase-treated collagen substrates also showed a greater resistance to cell-mediated endogenous protease degradation than the native collagen. In addition, the HOB cells were shown to differentiate at a faster rate than on native collagen when assessed by measurement of alkaline phosphatase activity and osteopontin expression.

  10. Cross-linked polymeric membranes for carbon dioxide separation

    Energy Technology Data Exchange (ETDEWEB)

    Hong, Tao; Chatterjee, Sabornie; Mahurin, Shannon Mark; Long, Brian Keith; Jiang, De-en; Mays, Jimmy Wayne; Sokolov, Alexei P.; Saito, Tomonori

    2018-01-23

    A membrane useful in gas separation, the membrane comprising a cross-linked polysiloxane structure having a cross-link density of about 0.1.times.10.sup.-5 mol/cm.sup.3 to about 6.times.10.sup.-5 mol/cm.sup.3, where, in particular embodiments, the cross-linked polysiloxane structure has the following general structure: ##STR00001## wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, and R.sup.6 are independently selected from hydrocarbon groups having at least 1 and up to 6 carbon atoms; A.sup.1 and A.sup.2 are independently selected from cyclic hydrocarbon groups; L.sup.1 and L.sup.2 are linking groups or covalent bonds; n is an integer of at least 1; r and s are independently selected from integers of at least 1; and p is an integer of at least 10. The invention also includes methods for making and using the above-described membranes for gas separation.

  11. Homogeneous UVA system for corneal cross-linking treatment

    Science.gov (United States)

    Ayres Pereira, Fernando R.; Stefani, Mario A.; Otoboni, José A.; Richter, Eduardo H.; Ventura, Liliane

    2010-02-01

    The treatment of keratoconus and corneal ulcers by collagen cross-linking using ultraviolet type A irradiation, combined with photo-sensitizer Riboflavin (vitamin B2), is a promising technique. The standard protocol suggests instilling Riboflavin in the pre-scratched cornea every 5min for 30min, during the UVA irradiation of the cornea at 3mW/cm2 for 30 min. This process leads to an increase of the biomechanical strength of the cornea, stopping the progression, or sometimes, even reversing Keratoconus. The collagen cross-linking can be achieved by many methods, but the utilization of UVA light, for this purpose, is ideal because of its possibility of a homogeneous treatment leading to an equal result along the treated area. We have developed a system, to be clinically used for treatment of unhealthy corneas using the cross-linking technique, which consists of an UVA emitting delivery device controlled by a closed loop system with high homogeneity. The system is tunable and delivers 3-5 mW/cm2, at 365nm, for three spots (6mm, 8mm and 10mm in diameter). The electronics close loop presents 1% of precision, leading to an overall error, after the calibration, of less than 10% and approximately 96% of homogeneity.

  12. Kinetics of carboplatin-DNA binding in genomic DNA and bladder cancer cells as determined by accelerator mass spectrometry

    International Nuclear Information System (INIS)

    Hah, S S; Stivers, K M; Vere White, R; Henderson, P T

    2005-01-01

    Cisplatin and carboplatin are platinum-based drugs that are widely used in cancer chemotherapy. The cytotoxicity of these drugs is mediated by platinum-DNA monoadducts and intra- and interstrand diadducts, which are formed following uptake of the drug into the nucleus of cells. The pharmacodynamics of carboplatin display fewer side effects than for cisplatin, albeit with less potency, which may be due to differences in rates of DNA adduct formation. We report the use of accelerator mass spectrometry (AMS), a sensitive detection method often used for radiocarbon quantitation, to measure both the kinetics of [ 14 C]carboplatin-DNA adduct formation with genomic DNA and drug uptake and DNA binding in T24 human bladder cancer cells. Only carboplatin-DNA monoadducts contain radiocarbon in the platinated DNA, which allowed for calculation of kinetic rates and concentrations within the system. The percent of radiocarbon bound to salmon sperm DNA in the form of monoadducts was measured by AMS over 24 h. Knowledge of both the starting concentration of the parent carboplatin and the concentration of radiocarbon in the DNA at a variety of time points allowed calculation of the rates of Pt-DNA monoadduct formation and conversion to toxic cross-links. Importantly, the rate of carboplatin-DNA monoadduct formation was approximately 100-fold slower than that reported for the more potent cisplatin analogue, which may explain the lower toxicity of carboplatin. T24 human bladder cancer cells were incubated with a subpharmacological dose of [ 14 C]carboplatin, and the rate of accumulation of radiocarbon in the cells and nuclear DNA was measured by AMS. The lowest concentration of radiocarbon measured was approximately 1 amol/10 (micro)g of DNA. This sensitivity may allow the method to be used for clinical applications

  13. Inhibition of nuclear factor kappaB proteins-platinated DNA interactions correlates with cytotoxic effectiveness of the platinum complexes

    Czech Academy of Sciences Publication Activity Database

    Brabec, Viktor; Kašpárková, Jana; Kostrhunová, Hana; Farrell, N.P.

    2016-01-01

    Roč. 6, AUG2016 (2016), č. článku 28474. ISSN 2045-2322 R&D Projects: GA MŠk(CZ) LH13096 Institutional support: RVO:68081707 Keywords : interstrand cross-links * group domain proteins Subject RIV: BO - Biophysics Impact factor: 4.259, year: 2016

  14. Preparation and properties of silk sericin/cellulose cross-linking films

    Directory of Open Access Journals (Sweden)

    Wang Kunyan

    2017-01-01

    Full Text Available Silk sericin/cellulose cross-linked films were successfully prepared using glutaraldehyde as cross-linkinger. FTIR was applied to characterize the chemical structure of films. Cross-linked silk sericin film was found the peak intensity of FTIR for cross-linked film decreased markedly compared to pure silk sericin, which indicating cross-linking reaction has been occurred. The increasing value of swelling ratio also indicated the cross-linking has been happened. The cross-linking reaction increased the thermal decomposition temperature.

  15. Cisplatin adducts on a GGG sequence within a DNA duplex studied by NMR spectroscopy and molecular dynamics simulations.

    Science.gov (United States)

    Téletchéa, Stéphane; Skauge, Tormod; Sletten, Einar; Kozelka, Jirí

    2009-11-16

    The antitumor drug cisplatin(cis-[PtCl2(NH3)2]) reacts with cellular DNA to form GG intrastrand adducts between adjacent guanines as predominant lesions. GGG sites have been shown to be hotspots of platination. To study the structural perturbation induced by binding of cisplatin to two adjacent guanines of a GGG trinucleotide,we examined here the decanucleotide duplex d[(G1C2C3G*4 G*5 G6T7-C8G9C10).d(G11C12G13A14C15C16C17G18-G19C20)] (dsCG*G*G) intrastrand cross-linked at the G* guanines by cis-{Pt(NH3)2}2+ using NMR spectroscopy and molecular dynamics (MD) simulations.The NMR spectra of dsCG*G*G were found to be similar to those of previously characterized DNA duplexes cross-linked by cisplatin at apyG*G*X site (py=pyrimidine; X=C,T, A). This similarity of NMR spectra indicates that the base at the 3'-side of the G*G*-Pt cross-link does not affect the structure to a large extent. An unprecedented reversible isomerization between the duplex dsCG*G*G (bearing a G*4 G*5 -Pt chelate) and duplex dsGG*G*T (bearing a G*5 G*6 -Pt chelate)was observed, which yielded a 40:60 equilibrium between the two intrastrand GG-Pt cross-links. No formation of interstrand cross-links was observed.NMR spectroscopic data of dsCG*G*G indicated that the deoxyribose of the 5'-G* adopts an N-type conformation, and the cytidines C3, C15,and C16 have average phase angles intermediate between S and N. The NMR spectroscopic chemical shifts of dsGG*G*T showed some fundamental differences to those of pyG*G*-platinum adducts but were in agreement with the NMR spectra reported previously for the DNA duplexes crosslinked at an AG*G*C sequence by cisplatin or oxaliplatin. The presence of apurine instead of a pyrimidine at the 5'-side of the G*G* cross-link seems therefore to affect the structure of the XG* step significantly.

  16. Probing actin polymerization by intermolecular cross-linking

    OpenAIRE

    1988-01-01

    We have used N,N'-1,4-phenylenebismaleimide, a bifunctional sulfhydryl cross-linking reagent, to probe the oligomeric state of actin during the early stages of its polymerization into filaments. We document that one of the first steps in the polymerization of globular monomeric actin (G-actin) under a wide variety of ionic conditions is the dimerization of a significant fraction of the G-actin monomer pool. As polymerization proceeds, the yield of this initial dimer ("lower" dimer with an app...

  17. Newer protocols and future in collagen cross-linking

    Directory of Open Access Journals (Sweden)

    Arthur B Cummings

    2013-01-01

    Full Text Available Corneal Cross-Linking (CXL is an established surgical procedure for the treatment of corneal disorders such as corneal ectasia and keratoconus. This method of treatment stabilises the corneal structure and increases rigidity, reducing the requirement for corneal transplantation. Since its development, many scientific studies have been conducted to investigate ways of improving the procedure. Biomechanical stability of the cornea after exposure to UV-A light, and the effect of shortening procedure time has been some of the many topics explored

  18. Mapping protein structural changes by quantitative cross-linking

    Czech Academy of Sciences Publication Activity Database

    Kukačka, Zdeněk; Strohalm, Martin; Kavan, Daniel; Novák, Petr

    2015-01-01

    Roč. 89, NOV 2015 (2015), s. 112-120 ISSN 1046-2023 R&D Projects: GA MŠk(CZ) EE2.3.20.0055; GA MŠk(CZ) EE2.3.30.0003; GA MŠk(CZ) ED1.1.00/02.0109 Grant - others:OPPC(XE) CZ.2.16/3.1.00/24023 Institutional support: RVO:61388971 Keywords : Chemical cross-linking * Proteolysis * Mass spectrometry Subject RIV: CE - Biochemistry Impact factor: 3.503, year: 2015

  19. Peripheral hepatic arterial embolization with cross-linked collagen fibers

    International Nuclear Information System (INIS)

    Daniels, J.R.; Kerlan, R.K. Jr.; Dodds, L.; McLaughlin, P.; La Berge, J.M.; Harrington, D.; Daniels, A.M.; Ring, E.J.

    1986-01-01

    Hepatic artery embolization with a nonimmunogenic, cross-linked collagen preparation (Angiostat, collagen for embolization, Target Therapeutics) was studied in mongrel dogs. Flow-directed technique was used to achieve complete distal arterial occlusion. Serial liver function evaluation demonstrated marked alterations at 48 to 72 hours, partial correction at 1 week, and resolution of abnormalities by 1 month. Restoration of large-vessel blood flow was angiographically demonstrable at 1 week. Recanalization, achieved by migration of endothelial cells around the collagen, resulted in complete restoration of normal hepatic vascular and tissue anatomy at 1 month. Repeated embolization at biweekly intervals was well tolerated

  20. Double cross-linked polyetheretherketone proton exchange membrane for fuel cell

    CSIR Research Space (South Africa)

    Luo, H

    2012-04-01

    Full Text Available The proton exchange membrane based on polyetheretherketone was prepared via two steps of cross-linking. The properties of the double cross-linked membrane (water uptake, proton conductivity, methanol permeability and thermal stability) have been...

  1. Collagen Cross-Linking: Current Status and Future Directions

    Directory of Open Access Journals (Sweden)

    Marine Hovakimyan

    2012-01-01

    Full Text Available Collagen cross-linking (CXL using UVA light and riboflavin (vitamin B2 was introduced as a clinical application to stabilize the cornea by inducing cross-links within and between collagen fibers. CXL has been investigated extensively and has been shown clinically to arrest the progression of keratoconic or post-LASIK ectasia. With its minimal cost, simplicity, and proven positive clinical outcome, CXL can be regarded as a useful approach to reduce the number of penetrating keratoplasties performed. Small case series have also indicated that CXL is beneficial in corneal edema by reducing stromal swelling behavior and in keratitis by inhibiting pathogen growth. Despite these encouraging results, CXL remains a relatively new method that is potentially associated with complications. Aspects such as side effects and recurrence rates have still to be elucidated. In light of the growing interest in CXL, our paper summarizes present knowledge about this promising approach. We have intentionally endeavored to include the more relevant studies from the recent literature to provide an overview of the current status of CXL.

  2. Damage and fatigue in cross-linked rubbers

    Science.gov (United States)

    Melnikov, Alexei

    Damage and fatigue of elastomers have not been fundamentally understood because of the complex nature of these materials. All currently existing models are completely phenomenological. Therefore two problems have been investigated in this research to address those fundamental issues. The first problem was creating an innovative concept with a mathematical modeling, which would be able to describe the damage using molecular characteristics of elastomers. The second problem is developing new approaches to study fatigue, and especially impact fatigue of elastomers. The following results have been obtained in this research. A theoretical model of damage has been developed which involves the basic molecular characteristics of cross-linked elastomers and takes into account the effects of viscoelasticity and stress-induced crystallization. This model was found very reliable and successful in description of numerous quasi-static simple extension experiments for monotonous and repeating loadings. It also roughly predicts in molecular terms the failure of elastomers with various degrees of cross-linking. Quasi-impact fatigue tests with different geometry of an indenter have also been performed. Some microscopic features of rubber damage have been investigated using optical microscopy and SEM. In particular, the accumulation of a completely de-vulcanized, liquid-like substance was observed under intense, multi-cycle impacts. All the findings discovered in quasi-impact experiments are consistent with the damage model predictions.

  3. Liquid droplets of cross-linked actin filaments

    Science.gov (United States)

    Weirich, Kimberly; Banerjee, Shiladitya; Dasbiswas, Kinjal; Vaikuntanathan, Suriyanarayan; Gardel, Margaret

    Soft materials constructed from biomolecules self-assemble into a myriad of structures that work in concert to support cell physiology. One critical soft material is the actin cytoskeleton, a viscoelastic gel composed of cross-linked actin filaments. Although actin networks are primarily known for their elastic properties, which are crucial to regulating cell mechanics, the viscous behavior has been theorized to enable shape changes and flows. We experimentally demonstrate a fluid phase of cross-linked actin, where cross-linker condenses dilute short actin filaments into spindle-shaped droplets, or tactoids. Tactoids have shape dynamics consistent with a continuum model of liquid crystal droplets. The cross-linker, which acts as a long range attractive interaction, analogous to molecular cohesion, controls the tactoid shape and dynamics, which reports on the liquid's interfacial tension and viscosity. We investigate how the cross-linker properties and filament length influence the liquid properties. These results demonstrate a novel mechanism to control organization of the actin cytoskeleton and provide insight into design principles for complex, macromolecular liquid phases.

  4. Collagen cross-linking: Strengthening the unstable cornea

    Directory of Open Access Journals (Sweden)

    Oren Tomkins

    2008-05-01

    Full Text Available Oren Tomkins, Hanna J GarzoziDepartment of Ophthalmology, Bnai Zion Medical Center, Haifa, IsraelAbstract: Corneal ectasia, a weakening of corneal integrity, occurs both due to acquired and congenital conditions such as keratoconus. It is a progressing condition that affects both visual acuity, and corneal stability. Various methods exist for correcting this impairment, however none address the inherit pathology, an increase laxity of the corneal stroma. Collagen crosslinking, a new, minimally invasive method, aims to strengthen the stroma by inducing cross links between neighboring collagen fibers. This method results in an increase in corneal tensile strength, with no medium term adverse effects on its normal architecture. Clinically, treated patients display improvement in both visual acuity and keratometric readings. This method may provide clinicians with easily accessible tools to stop the progression, and even correct visual deterioration due to corneal ectasia. Here we review the current information regarding this new method, as well as discuss its potential benefits and downfalls.Keywords: corneal cross-linking, corneal ectasia, keratoconus, stroma, cornea

  5. Tea derived galloylated polyphenols cross-link purified gastrointestinal mucins.

    Directory of Open Access Journals (Sweden)

    Pantelis Georgiades

    Full Text Available Polyphenols derived from tea are thought to be important for human health. We show using a combination of particle tracking microrheology and small-angle neutron scattering that polyphenols acts as cross-linkers for purified gastrointestinal mucin, derived from the stomach and the duodenum. Both naturally derived purified polyphenols, and green and black tea extracts are shown to act as cross-linkers. The main active cross-linking component is found to be the galloylated forms of catechins. The viscosity, elasticity and relaxation time of the mucin solutions experience an order of magnitude change in value upon addition of the polyphenol cross-linkers. Similarly small-angle neutron scattering experiments demonstrate a sol-gel transition with the addition of polyphenols, with a large increase in the scattering at low angles, which is attributed to the formation of large scale (>10 nm heterogeneities during gelation. Cross-linking of mucins by polyphenols is thus expected to have an impact on the physicochemical environment of both the stomach and duodenum; polyphenols are expected to modulate the barrier properties of mucus, nutrient absorption through mucus and the viscoelastic microenvironments of intestinal bacteria.

  6. Ultra-sonication assisted cross-linking of cellulose polymers.

    Science.gov (United States)

    Udoetok, Inimfon A; Wilson, Lee D; Headley, John V

    2018-04-01

    Cross-linked cellulose-epichlorohydrin polymers were synthesized by a conventional heating with stirring (C-EP heating) and a parallel process using ultra-sonication (C-EP sonication) in the presence of aqueous ammonia. Structural characterization of modified cellulose was carried out using FTIR/ 13 C solid state NMR spectroscopy and thermal methods (DSC and TGA). Evidence of products with variable textural properties and morphology was supported by nitrogen gas adsorption, solvent swelling, and microscopy (SEM, TEM) results. C-EP sonication possess greater cross-linker content judging by the loss of the cellulose fibril structure which was facilitated by acoustic cavitation effects due to ultra-sonication. Equilibrium sorption studies in aqueous solution with 2-naphthoxy acetic acid (NAA) revealed that C-EP heating had slightly greater sorption capacity than C-EP sonication at alkaline pH. By contrast, C-EP sonication had greater uptake of NAA at acidic pH. Kinetic uptake studies at pH 3 is described by the pseudo-second order model, where the surface sites of C-EP heating became saturated within ca. 75 min; whereas, ca. 350 min occurred for C-EP sonication. This study demonstrates that the yield of sonication assisted cross-linking of cellulose is greater with improved adsorption properties. The study also reveals the utility of sonication assisted synthesis for the valorization and utilization of cellulose modified materials. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Cross-linked comb-shaped anion exchange membranes with high base stability

    Energy Technology Data Exchange (ETDEWEB)

    Li, NW; Wang, LZ; Hickner, M

    2014-01-01

    A unique one-step cross-linking strategy that connects quaternary ammonium centers using Grubbs II-catalyzed olefin metathesis was developed. The cross-linked anion exchange membranes showed swelling ratios of less than 10% and hydroxide conductivities of 18 to 40 mS cm(- 1). Cross-linking improved the membranes' stability to hydroxide degradation compared to their non-cross-linked analogues.

  8. Small Strain Topological Effects of Biopolymer Networks with Rigid Cross-Links

    NARCIS (Netherlands)

    Zagar, G.; Onck, P. R.; Van der Giessen, E.; Garikipati, K; Arruda, EM

    2010-01-01

    Networks of cross-linked filamentous biopolymers form topological structures characterized by L, T and X cross-link types of connectivity 2, 3 and 4, respectively. The distribution of cross-links over these three types proofs to be very important for the initial elastic shear stiffness of isotropic

  9. Cross-linking of wheat gluten using a water-soluble carbodiimide

    NARCIS (Netherlands)

    Tropini, V.; Lens, J.P.; Mulder, W.J.; Silvestre, F.

    2000-01-01

    Wheat gluten was cross-linked using water-soluble 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide HCl (EDC). To enhance cross-linking, N-hydroxysuccinimide (NHS) was added to the reaction mixture. The cross-linking efficiency was evaluated by the decrease in the amount of amino groups, the solubility

  10. Oxidative cross-linking of casein by horseradish peroxidase and its ...

    African Journals Online (AJOL)

    The central composite design using response surface methodology was used to optimize cross-linking conditions of casein. The optimal cross-linking conditions of casein ... Cross-linking of food proteins induced by horseradish peroxidase might serve as an alternative approach to modify functional property of the proteins.

  11. Permanent Set of Cross-Linking Networks: Comparison of Theory with Molecular Dynamics Simulations

    DEFF Research Database (Denmark)

    Rottach, Dana R.; Curro, John G.; Budzien, Joanne

    2006-01-01

    The permanent set of cross-linking networks is studied by molecular dynamics. The uniaxial stress for a bead-spring polymer network is investigated as a function of strain and cross-link density history, where cross-links are introduced in unstrained and strained networks. The permanent set is fo...

  12. Toxicity in tumor cells, DNA binding mode, and resistance to decomposition by sulfur nucleophiles of new dinuclear bifunctional trans-Pt-II complexes containing long alkane linkers

    Czech Academy of Sciences Publication Activity Database

    Prachařová, J.; Nováková, Olga; Kašpárková, Jana; Gibson, J.; Brabec, Viktor

    2013-01-01

    Roč. 85, č. 2 (2013), s. 343-354 ISSN 0033-4545 R&D Projects: GA ČR(CZ) GD301/09/H004 Institutional support: RVO:68081707 Keywords : PLATINUM ANTITUMOR COMPLEXES * INTERSTRAND CROSS-LINKS * GEOMETRIC ISOMERISM Subject RIV: BO - Biophysics Impact factor: 3.112, year: 2013

  13. Mapping the miRNA interactome by cross-linking ligation and sequencing of hybrids (CLASH).

    Science.gov (United States)

    Helwak, Aleksandra; Tollervey, David

    2014-03-01

    RNA-RNA interactions have critical roles in many cellular processes, but studying them is difficult and laborious. Here we describe an experimental procedure, termed cross-linking ligation and sequencing of hybrids (CLASH), which allows high-throughput identification of sites of RNA-RNA interaction. During CLASH, a tagged bait protein is UV-cross-linked in cell cultures to stabilize RNA interactions, and it is purified under denaturing conditions. RNAs associated with the bait protein are partially truncated, and the ends of RNA duplexes are ligated together. After linker addition, cDNA library preparation and high-throughput sequencing, the ligated duplexes give rise to chimeric cDNAs, which unambiguously identify RNA-RNA interaction sites independent of bioinformatic predictions. This protocol is optimized for studying miRNA targets bound by Argonaute (AGO) proteins, but it should be easily adapted for other RNA-binding proteins and classes of RNA. The protocol requires ∼5 d to complete, excluding the time required for high-throughput sequencing and bioinformatic analyses.

  14. Sequence context effects on 8-methoxypsoralen photobinding to defined DNA fragments

    International Nuclear Information System (INIS)

    Sage, E.; Moustacchi, E.

    1987-01-01

    The photoreaction of 8-methoxypsoralen (8-MOP) with DNA fragments of defined sequence was studied. The authors took advantage of the blockage by bulky adducts of the 3'-5'-exonuclease activity associated with the T4 DNA polymerase. The action of the exonuclease is stopped by biadducts as well as by monoadducts. The termination products were analyzed on sequencing gels. A strong sequence specificity was observed in the DNA photobinding of 8-MOP. The exonuclease terminates its digestion near thymine residues, mainly at potentially cross-linkable sites. There is an increasing reactivity of thymine residues in the order T < TT << TTT in a GC environment. For thymine residues in cross-linkable sites, the reactivity follows the order AT << TA ∼ TAT << ATA < ATAT < ATATAA. Repeated A-T sequences are hot spots for the photochemical reaction of 8-MOP with DNA. Both monoadducts and interstrand cross-links are formed preferentially in 5'-TpA sites. The results highlight the role of the sequence and consequently of the conformation around a potential site in the photobinding of 8-MOP to DNA

  15. Reduced cross-linking demarcation line depth at the peripheral cornea after corneal collagen cross-linking.

    Science.gov (United States)

    Yam, Jason C S; Cheng, Arthur C K

    2013-01-01

    To compare the corneal collagen cross-linking (CXL) demarcation line depth between the central and peripheral cornea after cross-linking using anterior segment optical coherence tomography. Retrospective interventional case series of 38 eyes with keratoconus or postoperative LASIK ectasia treated with riboflavin ultraviolet A CXL (UV-X, IROC). CXL demarcation line depth, corneal thickness, and the ratio of the CXL demarcation line depth to the corneal thickness were measured using anterior segment optical coherence tomography at the central cornea and at 2 and 4 mm from the corneal center in four regions: temporal, nasal, superior, and inferior. The CXL demarcation line depths at the center and periphery were compared using the Friedman test. The CXL demarcation line was deepest in the central cornea (302 μm; range: 180 to 397 μm) and was reduced progressively toward the peripheral cornea, at nasal 2 mm (289.5 μm; range: 125 to 370 μm), at nasal 4 mm (206.5 μm; range: 100 to 307 μm), at temporal 2 mm (278.5 μm; range: 128 to 375 μm), and at temporal 4 mm (194 μm; range: 80 to 325 μm) (Pcorneal thickness were greater at the central cornea than the peripheral cornea. Copyright 2013, SLACK Incorporated.

  16. Nek1 silencing slows down DNA repair and blocks DNA damage-induced cell cycle arrest.

    Science.gov (United States)

    Pelegrini, Alessandra Luíza; Moura, Dinara Jaqueline; Brenner, Bethânia Luise; Ledur, Pitia Flores; Maques, Gabriela Porto; Henriques, João Antônio Pegas; Saffi, Jenifer; Lenz, Guido

    2010-09-01

    Never in mitosis A (NIMA)-related kinases (Nek) are evolutionarily conserved proteins structurally related to the Aspergillus nidulans mitotic regulator NIMA. Nek1 is one of the 11 isoforms of the Neks identified in mammals. Different lines of evidence suggest the participation of Nek1 in response to DNA damage, which is also supported by the interaction of this kinase with proteins involved in DNA repair pathways and cell cycle regulation. In this report, we show that cells with Nek1 knockdown (KD) through stable RNA interference present a delay in DNA repair when treated with methyl-methanesulfonate (MMS), hydrogen peroxide (H(2)O(2)) and cisplatin (CPT). In particular, interstrand cross links induced by CPT take much longer to be resolved in Nek1 KD cells when compared to wild-type (WT) cells. In KD cells, phosphorylation of Chk1 in response to CPT was strongly reduced. While WT cells accumulate in G(2)/M after DNA damage with MMS and H(2)O(2), Nek1 KD cells do not arrest, suggesting that G(2)/M arrest induced by the DNA damage requires Nek1. Surprisingly, CPT-treated Nek1 KD cells arrest with a 4N DNA content similar to WT cells. This deregulation in cell cycle control in Nek1 KD cells leads to an increased sensitivity to genotoxic agents when compared to WT cells. These results suggest that Nek1 is involved in the beginning of the cellular response to genotoxic stress and plays an important role in preventing cell death induced by DNA damage.

  17. Transepithelial corneal collagen cross-linking in ultrathin keratoconic corneas

    Directory of Open Access Journals (Sweden)

    Spadea L

    2012-11-01

    Full Text Available Leopoldo Spadea,1 Rita Mencucci21University of L'Aquila, Department of Biotechnological and Applied Clinical Sciences, Eye Clinic, L'Aquila, 2University of Florence, Department of Oto-Neuro-Ophthalmological Surgical Sciences, Eye Clinic, Florence, ItalyBackground: The purpose of this paper was to report the results of transepithelial corneal collagen cross-linking (CXL with modified riboflavin and ultraviolet A irradiation in patients affected by keratoconus, each with thinnest pachymetry values of less than 400 µm (with epithelium and not treatable using standard de-epithelialization techniques.Methods: Sixteen patients affected by progressive keratoconus with thinnest pachymetry values ranging from 331 µm to 389 µm underwent transepithelial CXL in one eye using a riboflavin 0.1% solution in 15% Dextran T500 containing ethylenediamine tetra-acetic acid 0.01% and trometamol to enhance epithelial penetration. The patients underwent complete ophthalmological examination, including endothelial cell density measurements and computerized videokeratography, before CXL and at one day, one week, and one, 6, and 12 months thereafter.Results: Epithelial healing was complete in all patients after one day of use of a soft bandage contact lens. No side effects or damage to the limbal region was observed during the follow-up period. All patients showed slightly improved uncorrected and spectacle-corrected visual acuity; keratometric astigmatism showed reductions (up to 5.3 D and apical ectasia power decreased (Kmax values reduced up to 4.3 D. Endothelial cell density was unchanged.Conclusion: Application of transepithelial CXL using riboflavin with substances added to enhance epithelial permeability was safe, seemed to be moderately effective in keratoconic eyes with ultrathin corneas, and applications of the procedure could be extended to patients with advanced keratoconus.Keywords: keratoconus, pachymetry, topography, transepithelial corneal collagen

  18. Yield and Failure Behavior Investigated for Cross-Linked Phenolic Resins Using Molecular Dynamics

    Science.gov (United States)

    Monk, Joshua D.; Lawson, John W.

    2016-01-01

    Molecular dynamics simulations were conducted to fundamentally evaluate the yield and failure behavior of cross-linked phenolic resins at temperatures below the glass transition. Yield stress was investigated at various temperatures, strain rates, and degrees of cross-linking. The onset of non-linear behavior in the cross-linked phenolic structures was caused by localized irreversible molecular rearrangements through the rotation of methylene linkers followed by the formation or annihilation of neighboring hydrogen bonds. The yield stress results, with respect to temperature and strain rate, could be fit by existing models used to describe yield behavior of amorphous glasses. The degree of cross-linking only indirectly influences the maximum yield stress through its influence on glass transition temperature (Tg), however there is a strong relationship between the degree of cross-linking and the failure mechanism. Low cross-linked samples were able to separate through void formation, whereas the highly cross-linked structures exhibited bond scission.

  19. Photo-cross-linking of amniotic membranes for limbal epithelial cell cultivation

    Energy Technology Data Exchange (ETDEWEB)

    Lai, Jui-Yang, E-mail: jylai@mail.cgu.edu.tw

    2014-12-01

    In the present study, we developed photo-cross-linked amniotic membrane (AM) as a limbal stem cell niche. After ultraviolet (UV) irradiation for varying time periods, the biological tissues were studied by determinations of cross-linking structure, degradability, and nutrient permeation ability. Our results showed that the number of cross-links per unit mass of AM significantly increased with increasing illumination time from 5 to 50 min. However, the cross-link formation was inhibited by longer irradiation time (i.e., 150 min), probably due to the scission of tissue collagen chains through irradiation. The biological stability and matrix permeability of photo-cross-linked AM materials strongly depended on their cross-linking densities affected by the UV irradiation. In vitro biocompatibility studies including cell viability and pro-inflammatory gene expression analyses demonstrated that, irrespective of the irradiation time employed, the physically cross-linked biological tissues exhibited negligible cytotoxicity and similar interleukin-6 (IL-6) mRNA levels. The data clearly indicate that these AM matrices do not cause potential harm to the corneal epithelial cells. After the growth of limbal epithelial cells (LECs) on AM substrates, Western blot analyses were conducted to examine the expression of ABCG2. It was found that the ability of UV-irradiated AM to maintain the undifferentiated precursor cell phenotype was significantly enhanced with increasing extent of photo-cross-linking. In summary, the UV irradiation time may have a profound influence on the fabrication of photo-cross-linked AM matrices for LEC cultivation. - Highlights: • We report the development of photo-cross-linked AM as a limbal stem cell niche. • Cross-linked structure of tissue materials was controlled by UV irradiation time. • Biostability and matrix permeability of AM depended on cross-linking density. • All the studied photo-cross-linked AM showed good in vitro biocompatibility.

  20. Cross-linking da cornea: protocolo padrão

    Directory of Open Access Journals (Sweden)

    Marcony R. Santhiago

    Full Text Available RESUMO O objetivo desta revisão é de determinar as indicações e eficácia da cirurgia que promove novas ligações covalentes entre as fibras de colágeno da córnea, conhecida como Cross-Linking (CXL, assim como esclarecer seus objetivos. O ceratocone é uma doença ectasica da córnea, bilateral, assimétrica, que, principalmente, cursa com encurvamento e afinamentos progressivo, e se inicia em geral na segunda década de vida. O uso primário do CXL tem sido na interrupção da progressão do Ceratocone. Apesar do conhecido encurvamento no estroma da córnea ocorrer nesses pacientes, a fisiopatologia por trás do ceratocone ainda é desconhecida e parece ser multifatorial. Pela evidencia literária disponível até o momento, o CXL da córnea esta, portanto indicado nos pacientes com doença em progressão. Concluímos que existe evidencia suficiente para afirmar que o CXL da córnea é eficaz na estabilização da doença ectásica da cornea.

  1. Memory Dynamics in Cross-linked Actin Networks

    Science.gov (United States)

    Scheff, Danielle; Majumdar, Sayantan; Gardel, Margaret

    Cells demonstrate the remarkable ability to adapt to mechanical stimuli through rearrangement of the actin cytoskeleton, a cross-linked network of actin filaments. In addition to its importance in cell biology, understanding this mechanical response provides strategies for creation of novel materials. A recent study has demonstrated that applied stress can encode mechanical memory in these networks through changes in network geometry, which gives rise to anisotropic shear response. Under later shear, the network is stiffer in the direction of the previously applied stress. However, the dynamics behind the encoding of this memory are unknown. To address this question, we explore the effect of varying either the rigidity of the cross-linkers or the length of actin filament on the time scales required for both memory encoding and over which it later decays. While previous experiments saw only a long-lived memory, initial results suggest another mechanism where memories relax relatively quickly. Overall, our study is crucial for understanding the process by which an external stress can impact network arrangement and thus the dynamics of memory formation.

  2. Pyridinium cross-links in heritable disorders of collagen

    Energy Technology Data Exchange (ETDEWEB)

    Pasquali, M.; Still, M.J.; Dembure, P.P. [Emory Univ., Atlanta, GA (United States)] [and others

    1995-12-01

    Ehlers-Danlos syndrome (EDS) is a heterogeneous group of inherited disorders of collagen that is characterized by skin fragility, skin hyperextensibility, and joint hypermobility. EDS type VI is caused by impaired collagen lysyl hydroxylase (procollagen-lysine, 2-oxoglutarate 5-dioxygenase; E.C.1.14.11.4), the ascorbate-dependent enzyme that hydroxylates lysyl residues on collagen neopeptides. Different alterations in the gene for collagen lysyl hydroxylase have been reported in families with EDS type VI. In EDS type VI, impairment of collagen lysyl hydroxylase results in a low hydroxylysine content in mature collagen. Hydroxylysine is a precursor of the stable, covalent, intermolecular cross-links of collagen, pyridinoline (Pyr), and deoxypyridinoline (Dpyr). Elsewhere we reported in preliminary form that patients with EDS type VI had a distinctive alteration in the urinary excretion of Pyr and Dpyr. In the present study, we confirm that the increased Dpyr/Pyr ratio is specific for EDS type VI and is not observed in other inherited or acquired collagen disorders. In addition, we find that skin from patients with EDS type VI has reduced Pyr and increased Dpyr, which could account for the organ pathology. 19 refs., 1 tab.

  3. Energy harvesting from vibration with cross-linked polypropylene piezoelectrets

    Directory of Open Access Journals (Sweden)

    Xiaoqing Zhang

    2015-07-01

    Full Text Available Piezoelectret films are prepared by modification of the microstructure of polypropylene foam sheets cross-linked by electronic irradiation (IXPP, followed by proper corona charging. Young’s modulus, relative permittivity, and electromechanical coupling coefficient of the fabricated films, determined by dielectric resonance spectra, are about 0.7 MPa, 1.6, and 0.08, respectively. Dynamic piezoelectric d33 coefficients up to 650 pC/N at 200 Hz are achieved. The figure of merit (FOM, d33 ⋅ g33 for a more typical d33 value of 400 pC/N is about 11.2 GPa−1. Vibration-based energy harvesting with one-layer and two-layer stacks of these films is investigated at various frequencies and load resistances. At an optimum load resistance of 9 MΩ and a resonance frequency of 800 Hz, a maximum output power of 120 μW, referred to the acceleration g due to gravity, is obtained for an energy harvester consisting of a one-layer IXPP film with an area of 3.14 cm2 and a seismic mass of 33.7 g. The output power can be further improved by using two-layer stacks of IXPP films in electric series. IXPP energy harvesters could be used to energize low-power electronic devices, such as wireless sensors and LED lights.

  4. Energy harvesting from vibration with cross-linked polypropylene piezoelectrets

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Xiaoqing [Shanghai Key Laboratory of Special Artificial Microstructure Materials and Technology & School of Physics Science and Engineering, Tongji University, Shanghai 200092 (China); Institute for Telecommunications Technology, Merckstrasse 25, 64283 Darmstadt (Germany); Wu, Liming [Shanghai Key Laboratory of Special Artificial Microstructure Materials and Technology & School of Physics Science and Engineering, Tongji University, Shanghai 200092 (China); Sessler, Gerhard M., E-mail: g.sessler@nt.tu-darmstadt.de [Institute for Telecommunications Technology, Merckstrasse 25, 64283 Darmstadt (Germany)

    2015-07-15

    Piezoelectret films are prepared by modification of the microstructure of polypropylene foam sheets cross-linked by electronic irradiation (IXPP), followed by proper corona charging. Young’s modulus, relative permittivity, and electromechanical coupling coefficient of the fabricated films, determined by dielectric resonance spectra, are about 0.7 MPa, 1.6, and 0.08, respectively. Dynamic piezoelectric d{sub 33} coefficients up to 650 pC/N at 200 Hz are achieved. The figure of merit (FOM, d{sub 33} ⋅ g{sub 33}) for a more typical d{sub 33} value of 400 pC/N is about 11.2 GPa{sup −1}. Vibration-based energy harvesting with one-layer and two-layer stacks of these films is investigated at various frequencies and load resistances. At an optimum load resistance of 9 MΩ and a resonance frequency of 800 Hz, a maximum output power of 120 μW, referred to the acceleration g due to gravity, is obtained for an energy harvester consisting of a one-layer IXPP film with an area of 3.14 cm{sup 2} and a seismic mass of 33.7 g. The output power can be further improved by using two-layer stacks of IXPP films in electric series. IXPP energy harvesters could be used to energize low-power electronic devices, such as wireless sensors and LED lights.

  5. Controlled Light Cross-Linking Technique to Prepare Healable Materials

    Directory of Open Access Journals (Sweden)

    Soliman Abdalla

    2017-06-01

    Full Text Available Detection of defects, damages and cracks in structural polymers is very difficult, and even if they are detected, they will be very hard to be repaired. This is because different kinds of stress can reduce the mechanical efficiency of structural and functional thermosetting composite materials and they can damage the polymer matrix, thus reducing the purposed properties. General healing processes use thermal energy “alone” to heal these materials, thus impairing the intended properties of the materials. Therefore, we present a thermal healing ability that can be switched-on and/or -off at desire using illumination by photon energy (visible and ultra violet. By this technique, one can control local heal while keeping the efficiency of the material nearly unchanged. Furan-based cross-linker chemically reacts (forward- and reverse-reaction with short-chains of maleimide-substituted poly(lauryl methacrylate to form robust chemical bonds. This permits us to perform local control over thermally induced de- and/or re-cross-linking techniques. One can extend and apply this technique to cover micro-devices, coating-techniques, fine lithography, micro- and nano-fabrication processes, etc. Therefore, the present work developed a suitable technology with structural polymeric material, which has the ability to self-heal cracks (and damages and recover structural function.

  6. Cross-linking of polytetrafluoroethylene during room-temperature irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Pugmire, David L [Los Alamos National Laboratory; Wetteland, Chris J [Los Alamos National Laboratory; Duncan, Wanda S [Los Alamos National Laboratory; Lakis, Rollin E [Los Alamos National Laboratory; Schwartz, Daniel S [Los Alamos National Laboratory

    2008-01-01

    Exposure of polytetrafluoroethylene (PTFE) to {alpha}-radiation was investigated to detennine the physical and chemical effects, as well as to compare and contrast the damage mechanisms with other radiation types ({beta}, {gamma}, or thermal neutron). A number of techniques were used to investigate the chemical and physical changes in PTFE after exposure to {alpha}-radiation. These techniques include: Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and fluorescence spectroscopy. Similar to other radiation types at low doses, the primary damage mechanism for the exposure of PTFE to {alpha}-radiation appears to be chain scission. Increased doses result in a change-over of the damage mechanism to cross-linking. This result is not observed for any radiation type other than {alpha} when irradiation is performed at room temperature. Finally, at high doses, PTFE undergoes mass-loss (via smallfluorocarbon species evolution) and defluorination. The amount and type of damage versus sample depth was also investigated. Other types of radiation yield damage at depths on the order of mm to cm into PTFE due to low linear energy transfer (LET) and the correspondingly large penetration depths. By contrast, the {alpha}-radiation employed in this study was shown to only induce damage to a depth of approximately 26 {mu}m, except at very high doses.

  7. Corneal cross-linking in the treatment of corneal ulcers.

    Science.gov (United States)

    Price, Marianne O; Price, Francis W

    2016-05-01

    New treatments for corneal ulcers are needed to address challenges with antibiotic resistance, cost, and specificity requiring timely pathogen identification. This review assesses the evidence regarding safety and efficacy of corneal cross-linking (CXL) as an adjunct or stand-alone treatment. To date approximately 200 clinical cases of CXL used with various types of infectious keratitis have been reported in about 30 publications. Most employed the CXL protocol developed for keratoconus as an adjunct to antibiotics for resistant ulcers, and a number of cases resolved after this intervention. However, a few studies raised concerns about resurgence and perforation when CXL was utilized with deep fungal infections. The infiltrate depth is an important consideration, because the standard CXL treatment is cytotoxic (to keratocytes) to a depth of approximately 200-300 μm and 50% of the energy is absorbed within the first 100 μm. CXL was used successfully as a monotherapy in approximately 16 eyes with early bacterial or shallow fungal infections. Further work is needed to develop optimized CXL protocols for treatment of corneal ulcers, define the appropriate conditions for use, and determine the safety and efficacy relative to standard antibiotic treatments.

  8. Homologous and non-homologous recombination differentially affect DNA damage repair in mice.

    NARCIS (Netherlands)

    J. Essers (Jeroen); H. van Steeg (Harry); J. de Wit (Jan); M. Vermeij (Marcel); J.H.J. Hoeijmakers (Jan); R. Kanaar (Roland); S.M.A. Swagemakers (Sigrid)

    2000-01-01

    textabstractIonizing radiation and interstrand DNA crosslinking compounds provide important treatments against cancer due to their extreme genotoxicity for proliferating cells. Both the efficacies of such treatments and the mutagenic potential of these agents are modulated by

  9. Tailoring the properties of cholecyst-derived extracellular matrix using carbodiimide cross-linking.

    LENUS (Irish Health Repository)

    Burugapalli, Krishna

    2009-01-01

    Modulation of properties of extracellular matrix (ECM) based scaffolds is key for their application in the clinical setting. In the present study, cross-linking was used as a tool for tailoring the properties of cholecyst-derived extracellular matrix (CEM). CEM was cross-linked with varying cross-linking concentrations of N,N-(3-dimethyl aminopropyl)-N\\'-ethyl carbodiimide (EDC) in the presence of N-hydroxysuccinimide (NHS). Shrink temperature measurements and ATR-FT-IR spectra were used to determine the degree of cross-linking. The effect of cross-linking on degradation was tested using the collagenase assay. Uniaxial tensile properties and the ability to support fibroblasts were also evaluated as a function of cross-linking. Shrink temperature increased from 59 degrees C for non-cross-linked CEM to 78 degrees C for the highest EDC cross-linking concentration, while IR peak area ratios for the free -NH(2) group at 3290 cm(-1) to that of the amide I band at 1635 cm(-1) decreased with increasing EDC cross-linking concentration. Collagenase assay demonstrated that degradation rates for CEM can be tailored. EDC concentrations 0 to 0.0033 mmol\\/mg CEM were the cross-linking concentration range in which CEM showed varied susceptibility to collagenase degradation. Furthermore, cross-linking concentrations up to 0.1 mmol EDC\\/mg CEM did not have statistically significant effect on the uniaxial tensile strength, as well as morphology, viability and proliferation of fibroblasts on CEM. In conclusion, the degradation rates of CEM can be tailored using EDC-cross-linking, while maintaining the mechanical properties and the ability of CEM to support cells.

  10. Chemical cross-linking with thiol-cleavable reagents combined with differential mass spectrometric peptide mapping--a novel approach to assess intermolecular protein contacts

    DEFF Research Database (Denmark)

    Bennett, K L; Kussmann, M; Björk, P

    2000-01-01

    The intermolecular contact regions between monomers of the homodimeric DNA binding protein ParR and the interaction between the glycoproteins CD28 and CD80 were investigated using a strategy that combined chemical cross-linking with differential MALDI-MS analyses. ParR dimers were modified in vitro...... a "head-to-tail" arrangement of the monomers in the dimeric complex. Glycoprotein fusion constructs CD28-IgG and CD80-Fab were cross-linked in vitro by DTSSP, characterized by nonreducing SDS-PAGE, digested in situ with trypsin and analyzed by MALDI-MS peptide mapping (+/- thiol reagent). The data...... revealed the presence of an intermolecular cross-link between the receptor regions of the glycoprotein constructs, as well as a number of unexpected but nonetheless specific interactions between the fusion domains of CD28-IgG and the receptor domain of CD80-Fab. The strategy of chemical cross...

  11. Analysis of protein-nucleic acid interactions by photochemical cross-linking and mass spectrometry

    DEFF Research Database (Denmark)

    Steen, Hanno; Jensen, Ole Nørregaard

    2002-01-01

    Photochemical cross-linking is a commonly used method for studying the molecular details of protein-nucleic acid interactions. Photochemical cross-linking aids in defining nucleic acid binding sites of proteins via subsequent identification of cross-linked protein domains and amino acid residues...... and for sequencing of peptide-nucleic acid heteroconjugates. The combination of photochemical cross-linking and MS provides a fast screening method to gain insights into the overall structure and formation of protein-oligonucleotide complexes. Because the analytical methods are continuously refined and protein...

  12. G-Quadruplexes Involving Both Strands of Genomic DNA Are Highly Abundant and Colocalize with Functional Sites in the Human Genome.

    Directory of Open Access Journals (Sweden)

    Andrzej S Kudlicki

    Full Text Available The G-quadruplex is a non-canonical DNA structure biologically significant in DNA replication, transcription and telomere stability. To date, only G4s with all guanines originating from the same strand of DNA have been considered in the context of the human nuclear genome. Here, I discuss interstrand topological configurations of G-quadruplex DNA, consisting of guanines from both strands of genomic DNA; an algorithm is presented for predicting such structures. I have identified over 550,000 non-overlapping interstrand G-quadruplex forming sequences in the human genome--significantly more than intrastrand configurations. Functional analysis of interstrand G-quadruplex sites shows strong association with transcription initiation, the results are consistent with the XPB and XPD transcriptional helicases binding only to G-quadruplex DNA with interstrand topology. Interstrand quadruplexes are also enriched in origin of replication sites. Several topology classes of interstrand quadruplex-forming sequences are possible, and different topologies are enriched in different types of structural elements. The list of interstrand quadruplex forming sequences, and the computer program used for their prediction are available at the web address http://moment.utmb.edu/allquads.

  13. Inter-strand resistance measurements in the termination of the ITER SULTAN samples

    International Nuclear Information System (INIS)

    Cau, F; Bruzzone, P

    2009-01-01

    In cabled conductors a perfect uniformity of the current among the strands is hardly reached, due to the non-homogeneity of the contact resistance distribution between the strands and the copper of the electrical terminations. In the case of large current unbalance, the overloaded strands hit the critical surface at high field early, developing a current sharing voltage, which drives the redistribution of the current, mainly in the electrical terminations where the inter-strand resistance is lower than in the high field conductor. If the inter-strand resistance in the termination is low, the voltage levels are sufficiently low to allow an effective redistribution of the current to the less loaded strands. The inter-strand resistance of three different termination layouts of ITER short length samples is measured to make a database available which can be used to qualify the layout of the joints and their capability of redistributing the current among the strands.

  14. The theory and art of corneal cross-linking

    Directory of Open Access Journals (Sweden)

    Rebecca McQuaid

    2013-01-01

    Full Text Available Before the discovery of corneal cross-linking (CXL, patients with keratoconus would have had to undergo corneal transplantation, or wear rigid gas permeable lenses (RGPs that would temporarily flatten the cone, thereby improving the vision. The RGP contact lens (CL would not however alter the corneal stability and if the keratoconus was progressive, the continued steepening of the cone would occur under the RGP CL. To date, the Siena Eye has been the largest study to investigate long term effects of standard CXL. Three hundred and sixty-three eyes were treated and monitored over 4 years, producing reliable long-term results proving long-term stability of the cornea by halting the progression of keratoconus, and proving the safety of the procedure. Traditionally, CXL requires epithelial removal prior to corneal soakage of a dextran-based 0.1% riboflavin solution, followed by exposure of ultraviolet-A (UV-A light for 30 min with an intensity of 3 mW/cm2. A series of in vitro investigations on human and porcine corneas examined the best treatment parameters for standard CXL, such as riboflavin concentration, intensity, wavelength of UV-A light, and duration of treatment. Photochemically, CXL is achieved by the generation of chemical bonds within the corneal stroma through localized photopolymerization, strengthening the cornea whilst minimizing exposure to the surrounding structures of the eye. In vitro studies have shown that CXL has an effect on the biomechanical properties of the cornea, with an increased corneal rigidity of approximately 70%. This is a result of the creation of new chemical bonds within the stroma.

  15. Unconventional metal organic frameworks: porous cross-linked phosphonates.

    Science.gov (United States)

    Clearfield, Abraham

    2008-11-28

    The past decade has witnessed an exponential growth of metal organic framework compounds (MOFs). The defining character of these compounds is their porosity. However, in many cases no effort was made to show evidence that a stable porous structure has been achieved and that the pores may be accessed. In the present paper we describe recent work on porous pillared zirconium diphosphonates, and the newer and in many respects different characteristics of tin(iv) phosphonates. The Sn(IV) monophosphonates form spherical globules that exhibit very high surface areas. The surface area arises from their nano-sized particles that pack in a "house of cards" arrangement. Also, it is shown that the 1,4-monophenyldiphosphonic acid forms highly porous (250-400 m2 g(-1)) materials with Sn(IV) when prepared in alcohol-water media. This is not the case with analogous Zr(IV) compounds. The many variations in the syntheses of both the zirconium and tin aryl- and alkyldiphosphonate pillars and their combinations with spacers such as methyl- and monophenylphosphonic acid have created a variety of highly porous materials that are stable to 400 degrees C in air, highly stable in acid media, do not collapse when de-solvated, and can be post and presynthesis altered to include functional groups. Several new directions taken by other researchers are also described. However, it is emphasized in this presentation that the cross-linked compounds form particles that precipitate rapidly into nanoparticles that exhibit only short range order. Therefore, they differ from the more conventional MOFs in that they are not amenable to structure solution by X-ray or neutron diffraction techniques. Rather, they must be understood on the basis of modeling and indirect data from EM, NMR, and additional spectroscopic and textural studies.

  16. Order-Disorder Transitions in Cross-Linked Block Copolymer Solids

    Energy Technology Data Exchange (ETDEWEB)

    Das, J.

    2005-01-12

    With a view toward creating solid block copolymers wherein the order-disorder transition can be accessed many times they investigated the nature of order-disorder transitions in cross-linked diblock copolymer melts using synergistic theory and experiment. A mean-field theory based on a coarse grained free-energy and the Random Phase Approximation (RPA) is developed for the system of interest. The quenched distribution of cross-links is averaged using the replica method. The phase behavior of a particular A-B block copolymer melt with a randomly cross-linked B-Block is determined as a function of the Florry-Huggins interaction parameter ({chi}) and the average number of cross-links per chain N{sub c}. They find for a cross-link density greater than N*{sub c} the B monomers are localized within a region of size {zeta} {approx} (N{sub c} - N*{sub c}){sup -1/2}. The cross-links strongly oppose ordering in the system as {zeta} becomes comparable to the radius of gyration of the block copolymer chain. As such the order-disorder transition temperature T{sub ODT} decreases precipitously when N{sub c} > N*{sub c}. When N{sub c} < N*{sub c}, T{sub ODT} increases weakly with N{sub c}. Experiments were conducted on cross-linked polystyrene-block-polyisoprene copolymer samples wherein the polyisoprene block was selectively cross-linked at a temperature well above the order-disorder transition temperature of the pure block copolymer. Small angle X-ray scattering (SAXS) and birefringence measurements on the cross-linked samples are consistent with the theoretical prediction. T{sub ODT} decreases rapidly when the cross-linking density exceeds the critical cross-linking density.

  17. Analytical characterisation of glutardialdehyde cross-linking products in gelatine-gum arabic complex coacervates

    International Nuclear Information System (INIS)

    Fuguet, Elisabet; Platerink, Chris van; Janssen, Hans-Gerd

    2007-01-01

    Encapsulates having shells of cross-linked mixtures of proteins and polysaccharides are widely used in the food and pharmaceutical industry for controlled release of actives and flavour compounds. In order to be able to predict the behaviour and the release characteristics of the microcapsules, a better understanding of the nature and extent of the cross-linking reaction is needed. Several analytical techniques were applied for the characterisation of glutardialdehyde (GDA) cross-linked encapsulates made of gelatine and gum arabic. To allow the use of sensitive, high-resolution methods such as chromatography and mass spectrometry, the sample first had to be hydrolysed. In this way, a mixture of amino acids, small peptides and the cross-link moieties was obtained. High-resolution liquid chromatography coupled to high-resolution mass spectrometry (HPLC-MS) was applied to detect possible cross-link markers through a comparison of HPLC-MS mass-chromatograms obtained for cross-linked and non-cross-linked coacervates. HPLC-MS/MS was used to identify the species responsible for the differences. Cross-linking occurred between GDA molecules and lysine and hydroxylysine ε-amino groups, and up to eight cross-link products of different nature could be identified. They included pyridinium ions and Schiff bases, and also unreacted GDA condensation products. Next, based on the insight gained in the possible chemical structures present in the cross-link markers, methods for selective labelling of these functionalities were employed to allow easier detection of related reaction products. Both liquid chromatography (LC) and gas chromatography (GC) were used in these experiments. Unfortunately, these approaches failed to detect new cross-link markers, most likely as a result of the low levels at which these are present

  18. Investigation of anisotropic thermal transport in cross-linked polymers

    Science.gov (United States)

    Simavilla, David Nieto

    Thermal transport in lightly cross-linked polyisoprene and polybutadine subjected to uniaxial elongation is investigated experimentally. We employ two experimental techniques to assess the effect that deformation has on this class of materials. The first technique, which is based on Forced Rayleigh Scattering (FRS), allows us to measure the two independent components of the thermal diffusivity tensor as a function of deformation. These measurements along with independent measurements of the tensile stress and birefringence are used to evaluate the stress-thermal and stress-optic rules. The stress-thermal rule is found to be valid for the entire range of elongations applied. In contrast, the stress-optic rule fails for moderate to large stretch ratios. This suggests that the degree of anisotropy in thermal conductivity depends on both orientation and tension in polymer chain segments. The second technique, which is based on infrared thermography (IRT), allows us to measure anisotropy in thermal conductivity and strain induced changes in heat capacity. We validate this method measurements of anisotropic thermal conductivity by comparing them with those obtained using FRS. We find excellent agreement between the two techniques. Uncertainty in the infrared thermography method measurements is estimated to be about 2-5 %. The accuracy of the method and its potential application to non-transparent materials makes it a good alternative to extend current research on anisotropic thermal transport in polymeric materials. A second IRT application allows us to investigate the dependence of heat capacity on deformation. We find that heat capacity increases with stretch ratio in polyisoprene specimens under uniaxial extension. The deviation from the equilibrium value of heat capacity is consistent with an independent set of experiments comparing anisotropy in thermal diffusivity and conductivity employing FRS and IRT techniques. We identify finite extensibility and strain

  19. Oxidative cross-linking of casein by horseradish peroxidase and its ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-10-19

    Oct 19, 2009 ... lyzed by HRP was studied. The cross-linking of casein treated with HRP was demonstrated by the analysis of the casein samples with capillary zone electrophoresis (CZE). The degree of cross-linking of casein in the casein sam- ples was also measured by CZE with peak area normali- zation methodology.

  20. SECONDARY CYTOTOXICITY OF CROSS-LINKED DERMAL SHEEP COLLAGENS DURING REPEATED EXPOSURE TO HUMAN FIBROBLASTS

    NARCIS (Netherlands)

    VANLUYN, MJA; VANWACHEM, PB; DAMINK, LHHO; DIJKSTRA, PJ; FEIJEN, J; NIEUWENHUIS, P

    1992-01-01

    We investigated commercially available dermal sheep collagen either cross-linked with hexamethylenedlisocyanate, or cross-linked with glutaraldehyde. In previous in vitro studies we could discriminate primary, i.e. extractable, and secondary cytotoxicity, due to cell-biomaterial interactions, i.e.

  1. Peroxidase-mediated cross-linking of bovine a-lactalbumin

    NARCIS (Netherlands)

    Heijnis, W.H.

    2010-01-01

    The research presented in this thesis aimed at controlling the horseradish peroxidase-catalyzed cross-linking of bovine α lactalbumin and the implications of this cross-linking for the foam stabilizing properties. Attention is also given to microreactors and their potential to control the enzymatic

  2. Optimized fragmentation schemes and data analysis strategies for proteome-wide cross-link identification

    NARCIS (Netherlands)

    Liu, Fan; Lössl, Philip; Scheltema, Richard; Viner, Rosa; Heck, Albert J R

    2017-01-01

    We describe optimized fragmentation schemes and data analysis strategies substantially enhancing the depth and accuracy in identifying protein cross-links using non-restricted whole proteome databases. These include a novel hybrid data acquisition strategy to sequence cross-links at both MS2 and MS3

  3. Models for stiffening in cross-linked biopolymer networks : A comparative study

    NARCIS (Netherlands)

    van Dillen, T.; Onck, P. R.; Van der Giessen, E.

    In a recent publication, we studied the mechanical stiffening behavior in two-dimensional (2D) cross-linked networks of semiflexible biopolymer filaments under simple shear [Onck, P.R., Koeman, T., Van Dillen, T., Van der Giessen, E., 2005. Alternative explanation of stiffening in cross-linked

  4. Effects of Supercritical CO 2 Conditioning on Cross-Linked Polyimide Membranes

    KAUST Repository

    Kratochvil, Adam M.

    2010-05-25

    The effects of supercritical CO2 (scCO2) conditioning on high-performance cross-linked polyimide membranes is examined through gas permeation and sorption experiments. Under supercritical conditions, the cross-linked polymers do not exhibit a structural reorganization of the polymer matrix that was observed in the non-cross-linkable, free acid polymer. Pure gas permeation isotherms and mixed gas permeabilities and selectivities show the cross-linked polymers to be much more stable to scCO2 conditioning than the free acid polymer. In fact, following scCO2 conditioning, the mixed gas CO2 permeabilities of the cross-linked polymers increased while the CO2/CH4 separation factors remained relatively unchanged. This response highlights the stability and high performance of these cross-linked membranes in aggressive environments. In addition, this response reveals the potential for the preconditioning of cross-linked polymer membranes to enhance productivity without sacrificing efficiency in practical applications which, in effect, provides another tool to \\'tune\\' membrane properties for a given separation. Finally, the dual mode model accurately describes the sorption and dilation characteristics of the cross-linked polymers. The changes in the dual mode sorption model parameters before and after the scCO2 exposure also provide insights into the alterations in the different glassy samples due to the cross-linking and scCO2 exposure. © 2010 American Chemical Society.

  5. Peroxidase-mediated cross-linking of a tyrosine-containing peptide with ferulic acid

    NARCIS (Netherlands)

    Oudgenoeg, G.; Hilhorst, H.; Piersma, S.R.; Boeriu, C.G.; Gruppen, H.; Voragen, A.G.J.; Laane, C.

    2001-01-01

    The tyrosine-containing peptide Gly-Tyr-Gly (GYG) was oxidatively cross-linked by horseradish peroxidase in the presence of hydrogen peroxide. As products, covalently coupled di- to pentamers of the peptide were identified by LC-MS. Oxidative cross-linking of ferulic acid with horseradish peroxidase

  6. Carboxymethyl starch cross-linked by electron beam radiation in presence of acrylic acid sensitizer

    International Nuclear Information System (INIS)

    Doan Binh; Nguyen Thanh Duoc; Pham Thi Thu Hong

    2013-01-01

    Carboxymethyl starch (CMS) can be cross-linked by electron beam radiation to form a biocompatible and environment-friendly hydrogel at a high absorbed dose and a condensed CMS concentration. Acrylic acid (AAc) can be used as a sensitizer in order to reduce the absorbed doses to an acceptable certain level. At an absorbed dose of 3-4 kGy, the gel content of crosslinked CMS can be obtained about 50% with 5% (w/w) AAc concentration used. The compressive strength of CMS samples increased with increasing their cross-linked densities due to raising absorbed doses. The swelling ratio of cross-linked CMS was also attainable at a maximum of 50 times in the distilled water. The enzymatic degradation of cross-linked CMS was carried out in acetate buffer pH 4.6 with 0.1% α-amylase enzymatic solution incubated at 40℃ for 6 h. The crosslinked CMS samples were degraded slower than uncrosslinked CMS ones. The results indicated that the highly cross-linked CMS was almost fully degradable when the enzymatic hydrolysis was performed during 6 h. The FT IR spectra of cross-linked CMS in the presence of AAc were examined to observe the carboxyl group of AAc in the structure of cross-linked CMS. The hydrophilic of cross-linked CMS surface was determined by a contact-angle analysis. (authors)

  7. Cross-linked PEEK-WC proton exchange membrane for fuel cell

    CSIR Research Space (South Africa)

    Lou, H

    2009-10-01

    Full Text Available The low cost proton exchange membrane was prepared by cross-linking water soluble sulfonated-sulfinated poly(oxa-p-phenylene-3,3-phthalido-p-phenylene-oxa-p-phenylene-oxyphenylene) (SsPEEK-WC). The prepared cross-linked membrane became insoluble...

  8. In vitro cross-linking of elastin peptides and molecular characterization of the resultant biomaterials

    DEFF Research Database (Denmark)

    Heinz, Andrea; Ruttkies, Christoph K H; Jahreis, Günther

    2013-01-01

    BACKGROUND: Elastin is a vital protein and the major component of elastic fibers which provides resilience to many vertebrate tissues. Elastin's structure and function are influenced by extensive cross-linking, however, the cross-linking pattern is still unknown. METHODS: Small peptides containin...

  9. Durability and mechanical properties of silane cross-linked wood thermoplastic composites

    Science.gov (United States)

    Magnus Bengtsson; Nicole M. Stark; Kristiina Oksman

    2007-01-01

    In this study, silane cross-linked wood–polyethylene composite profiles were manufactured by reactive extrusion. These composites were evaluated regarding their durability and mechanical properties in comparison with two non-cross-linked wood– polyethylene composites. An addition of only 2% w/w of silane solution during manufacturing was enough to achieve almost 60%...

  10. Covalently Cross-Linked Sulfone Polybenzimidazole Membranes with Poly(Vinylbenzyl Chloride) for Fuel Cell Applications

    DEFF Research Database (Denmark)

    Yang, Jingshuai; Aili, David; Li, Qingfeng

    2013-01-01

    Covalently cross-linked polymer membranes were fabricated from poly(aryl sulfone benzimidazole) (SO(2) PBI) and poly(vinylbenzyl chloride) (PVBCl) as electrolytes for high-temperature proton-exchange-membrane fuel cells. The cross-linking imparted organo insolubility and chemical stability against...... radical attack to the otherwise flexible SO(2) PBI membranes. Steady phosphoric acid doping of the cross-linked membranes was achieved at elevated temperatures with little swelling. The acid-doped membranes exhibited increased mechanical strength compared to both pristine SO(2) PBI and poly[2,2'-(m......-phenylene)-5,5'-bibenzimidazole] (mPBI). The superior characteristics of the cross-linked SO(2) PBI membranes allowed higher acid doping levels and, therefore, higher proton conductivity. Fuel-cell tests with the cross-linked membranes demonstrated a high open circuit voltage and improved power performance...

  11. Interrelation of electret properties of polyethylene foam from the method of cross-linking

    Science.gov (United States)

    Gilmanov, I. R.; Galikhanov, M. F.; Gilmanova, A. R.

    2017-09-01

    The electret properties of chemically cross-linked polyethylene foam and physically cross-linked polyethylene foam have been studied. It has been shown that chemically cross-linked polyethylene foam has higher surface potential, effective surface charge density and electric field strength compared to physically bonded polyethylene foam. This is due to the presence of molecules and fragments of dicumyl peroxide, which can play the role of traps for injection charge carriers, a greater degree of cross-linking and with the oxidation of polyethylene, which occurs during irradiation during physical cross-linking. When the foam is deformed, its electret properties are reduced, and when the volume is relaxed, they are restored. This is due to the partial mutual compensation of homo- and heterocharge during compression and the return of the structure of the gas-filled polymer to its former position when the load is removed.

  12. Corneal changes following collagen cross linking and simultaneous topography guided photoablation with collagen cross linking for keratoconus

    Directory of Open Access Journals (Sweden)

    Prema Padmanabhan

    2014-01-01

    Full Text Available Purpose: To compare the outcome of Collagen cross-linking (CXL with that following topography-guided customized ablation treatment (T-CAT with simultaneous CXL in eyes with progressive keratoconus. Materials and Methods: This was a prospective, non-randomized single centre study of 66 eyes with progressive keratoconus. Of these, 40 eyes underwent CXL and 26 eyes underwent T-CAT + CXL. The refractive, topographic, tomographic and aberrometric changes measured at baseline, 1, 3 and 6 months post-operatively were compared between both groups. Results: After a mean follow-up of 7.7 ± 1.3 months, the mean retinoscopic cylinder decreased by 1.02 ± 3.16 D in the CXL group ( P = 0.1 and 2.87 ± 3.22 D in the T-CAT + CXL group ( P = 0.04. The Best corrected visual acuity increased by 2 lines or more in 10% of eyes in the CXL group and in 23.3% of eyes in the T-CAT + CXL group. The mean steepest-K reduced by 0.40 ± 3.71 D ( P = 0.77 in the CXL group and by 2.91 ± 2.01D ( P = 0.03 in the T-CAT + CXL group. The sag factor and surface asymmetry index showed no significant change in the CXL group but reduced by 3.59 ± 5.94 D ( P = 0.01 and 0.72 ± 1.18 ( P = 0.02 respectively in the T-CAT + CXL group. There was a significant increase in the highest posterior corneal elevation in both groups (9.57 ± 14.93 μ in the CXL group and 7.85 ± 9.25 μ in the T-CAT + CXL group, P ≤ 0.001 for both. There was significantly greater reduction of mean coma ( P < 0.001 and mean higher-order aberrations ( P = 0.01 following T-CAT + CXL compared to CXL. Conclusions: CAT + CXL is an effective approach to confer biomechanical stability and to improve the corneal contour in eyes with keratoconus and results in better refractive, topographic and aberrometric outcomes than CXL alone.

  13. Study on interstrand coupling losses in Rutherford-type superconducting cables

    International Nuclear Information System (INIS)

    Lei, Y.Z.; Shintomi, T.; Terashima, A.; Hirabayashi, H.

    1993-02-01

    Two sets of experimental apparatus for measuring the AC losses in superconducting strands and Rutherford-type cable conductors have been constructed. A few strand samples and a number of compacted cable samples with and without a CuMn matrix have been measured. The hysteresis loss, loss from coupling within strands and loss from coupling between strands in cables have been distinguished from each other. The results show that, even for Rutherford cables without any soldering and coating, their AC losses may be quite different from each other due to the variation of the interstrand coupling loss. For cables without a CuMn matrix, interstrand coupling loss increases nearly according to a geometrical series with an increase of curing temperature simulating coil fabrication. However, cables with the CuMn matrix show a relatively small curing temperature dependence. For most of the samples, losses do not show any evident dependence on the mechanical pressure. Interstrand resistances in one of these cables have also been measured; the results indicate that the tendency for a decrease in the interstrand resistances is consistent with the results of AC loss measurements. (author)

  14. On the Reproducibility of Label-Free Quantitative Cross-Linking/Mass Spectrometry

    Science.gov (United States)

    Müller, Fränze; Fischer, Lutz; Chen, Zhuo Angel; Auchynnikava, Tania; Rappsilber, Juri

    2018-02-01

    Quantitative cross-linking/mass spectrometry (QCLMS) is an emerging approach to study conformational changes of proteins and multi-subunit complexes. Distinguishing protein conformations requires reproducibly identifying and quantifying cross-linked peptides. Here we analyzed the variation between multiple cross-linking reactions using bis[sulfosuccinimidyl] suberate (BS3)-cross-linked human serum albumin (HSA) and evaluated how reproducible cross-linked peptides can be identified and quantified by LC-MS analysis. To make QCLMS accessible to a broader research community, we developed a workflow that integrates the established software tools MaxQuant for spectra preprocessing, Xi for cross-linked peptide identification, and finally Skyline for quantification (MS1 filtering). Out of the 221 unique residue pairs identified in our sample, 124 were subsequently quantified across 10 analyses with coefficient of variation (CV) values of 14% (injection replica) and 32% (reaction replica). Thus our results demonstrate that the reproducibility of QCLMS is in line with the reproducibility of general quantitative proteomics and we establish a robust workflow for MS1-based quantitation of cross-linked peptides.

  15. Interrelation of Hydration, Collagen Cross-Linking Treatment, and Biomechanical Properties of the Cornea.

    Science.gov (United States)

    Hatami-Marbini, Hamed; Rahimi, Abdolrasol

    2016-05-01

    The present study was designed to investigate the effects of hydration and collagen cross-linking treatment on biomechanical properties of the cornea. The original corneal collagen cross-linking protocol was used to induce cross-links in bovine corneas. The thickness of samples was used as a measure of their hydration and five different thickness groups (n = 5 each) were considered. The cross-linked corneal strips were allowed to hydrate/dehydrate until their thickness reached 500, 700, 900, 1100, and 1500 μm. The tensile behavior of specimens in each thickness group was characterized by conducting uniaxial tensile experiments. The experiments were done in mineral oil in order to keep the thickness of samples constant and minimize hydration changes. It was observed that collagen cross-linking treatment significantly increased both the maximum tensile stress and the equilibrium (relaxed) stress of the bovine cornea (p  0.99), respectively. Hydration and collagen cross-linking treatment concomitantly affect biomechanical properties of the cornea. Therefore, an accurate estimate of stiffening effects of collagen cross-linking treatment option using uniaxial tensile experiments is only possible if the hydration of specimens is fully controlled.

  16. Modified gum arabic cross-linked gelatin scaffold for biomedical applications

    Energy Technology Data Exchange (ETDEWEB)

    Sarika, P.R. [Department of Chemistry, Indian Institute of Space Science and Technology, Valiamala, Thiruvananthapuram, Kerala 695 547 (India); Cinthya, Kuriakose [Tissue Culture Laboratory, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Poojappura, Thiruvananthapuram, Kerala 695 012 (India); Jayakrishnan, A. [Department of Biotechnology, Indian Institute of Technology Madras, Chennai 600 036 (India); Anilkumar, P.R., E-mail: anilkumarpr@sctimst.ac.in [Tissue Culture Laboratory, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Poojappura, Thiruvananthapuram, Kerala 695 012 (India); James, Nirmala Rachel, E-mail: nirmala@iist.ac.in [Department of Chemistry, Indian Institute of Space Science and Technology, Valiamala, Thiruvananthapuram, Kerala 695 547 (India)

    2014-10-01

    The present work deals with development of modified gum arabic cross-linked gelatin scaffold for cell culture. A new biocompatible scaffold was developed by cross-linking gelatin (Gel) with gum arabic, a polysaccharide. Gum arabic was subjected to periodate oxidation to obtain gum arabic aldehyde (GAA). GAA was reacted with gelatin under appropriate pH to prepare the cross-linked hydrogel. Cross-linking occurred due to Schiff's base reaction between aldehyde groups of oxidized gum arabic and amino groups of gelatin. The scaffold prepared from the hydrogel was characterized by swelling properties, degree of cross-linking, in vitro degradation and scanning electron microscopy (SEM). Cytocompatibility evaluation using L-929 and HepG2 cells confirmed non-cytotoxic and non-adherent nature of the scaffold. These properties are essential for generating multicellular spheroids and hence the scaffold is proposed to be a suitable candidate for spheroid cell culture. - Highlights: • Gum arabic cross-linked gelatin scaffold was developed for tissue engineering. • Cross-linking was achieved by Schiff's base reaction. • The scaffold is non-cytotoxic and non adherent to fibroblast and hepatocytes. • The scaffolds are potential candidates for spheroid cell culture.

  17. On the Reproducibility of Label-Free Quantitative Cross-Linking/Mass Spectrometry

    Science.gov (United States)

    Müller, Fränze; Fischer, Lutz; Chen, Zhuo Angel; Auchynnikava, Tania; Rappsilber, Juri

    2017-12-01

    Quantitative cross-linking/mass spectrometry (QCLMS) is an emerging approach to study conformational changes of proteins and multi-subunit complexes. Distinguishing protein conformations requires reproducibly identifying and quantifying cross-linked peptides. Here we analyzed the variation between multiple cross-linking reactions using bis[sulfosuccinimidyl] suberate (BS3)-cross-linked human serum albumin (HSA) and evaluated how reproducible cross-linked peptides can be identified and quantified by LC-MS analysis. To make QCLMS accessible to a broader research community, we developed a workflow that integrates the established software tools MaxQuant for spectra preprocessing, Xi for cross-linked peptide identification, and finally Skyline for quantification (MS1 filtering). Out of the 221 unique residue pairs identified in our sample, 124 were subsequently quantified across 10 analyses with coefficient of variation (CV) values of 14% (injection replica) and 32% (reaction replica). Thus our results demonstrate that the reproducibility of QCLMS is in line with the reproducibility of general quantitative proteomics and we establish a robust workflow for MS1-based quantitation of cross-linked peptides. [Figure not available: see fulltext.

  18. Epoxides cross-linked hexafluoropropylidene polybenzimidazole membranes for application as high temperature proton exchange membranes

    International Nuclear Information System (INIS)

    Yang, Jingshuai; Xu, Yixin; Liu, Peipei; Gao, Liping; Che, Quantong; He, Ronghuan

    2015-01-01

    Covalently cross-linked hexafluoropropylidene polybenzimidazole (F 6 PBI) was prepared and used to fabricate high temperature proton exchange membranes with enhanced mechanical strength against thermoplastic distortion. Three different epoxides, i.e. bisphenol A diglycidyl ether (R 1 ), bisphenol A propoxylate diglycidyl ether (R 2 ) and poly(ethylene glycol) diglycidyl ether (R 3 ), were chosen as the cross-linkers to investigate the influence of their structures on the properties of the cross-linked F 6 PBI membranes. All the cross-linked F 6 PBI membranes displayed excellent stability towards the radical oxidation. Comparing with the pure F 6 PBI membrane, the cross-linked F 6 PBI membranes showed high acid doping level but less swelling after doping phosphoric acid at elevated temperatures. The mechanical strength at 130 °C was improved from 0.4 MPa for F 6 PBI membrane to a range of 0.8–2.0 MPa for the cross-linked F 6 PBI membranes with an acid doping level as high as around 14, especially for that crosslinking with the epoxide (R 3 ), which has a long linear structure of alkyl ether. The proton conductivity of the cross-linked membranes was increased accordingly due to the high acid doping levels. Fuel cell tests demonstrated the technical feasibility of the acid doped cross-linked F 6 PBI membranes for high temperature proton exchange membrane fuel cells

  19. The Synergy of Double Cross-linking Agents on the Properties of Styrene Butadiene Rubber Foams

    Science.gov (United States)

    Shao, Liang; Ji, Zhan-You; Ma, Jian-Zhong; Xue, Chao-Hua; Ma, Zhong-Lei; Zhang, Jing

    2016-11-01

    Sulfur (S) cross-linking styrene butadiene rubber (SBR) foams show high shrinkage due to the cure reversion, leading to reduced yield and increased processing cost. In this paper, double cross-linking system by S and dicumyl peroxide (DCP) was used to decrease the shrinkage of SBR foams. Most importantly, the synergy of double cross-linking agents was reported for the first time to our knowledge. The cell size and its distribution of SBR foams were investigated by FESEM images, which show the effect of DCP content on the cell structure of the SBR foams. The relationships between shrinkage and crystalline of SBR foams were analyzed by the synergy of double cross-linking agents, which were demonstrated by FTIR, Raman spectra, XRD, DSC and TGA. When the DCP content was 0.6 phr, the SBR foams exhibit excellent physical and mechanical properties such as low density (0.223 g/cm3), reduced shrinkage (2.25%) and compression set (10.96%), as well as elevated elongation at break (1.78 × 103%) and tear strength (54.63 N/mm). The results show that these properties are related to the double cross-linking system of SBR foams. Moreover, the double cross-linking SBR foams present high electromagnetic interference (EMI) shielding properties compared with the S cross-linking SBR foams.

  20. Modified gum arabic cross-linked gelatin scaffold for biomedical applications

    International Nuclear Information System (INIS)

    Sarika, P.R.; Cinthya, Kuriakose; Jayakrishnan, A.; Anilkumar, P.R.; James, Nirmala Rachel

    2014-01-01

    The present work deals with development of modified gum arabic cross-linked gelatin scaffold for cell culture. A new biocompatible scaffold was developed by cross-linking gelatin (Gel) with gum arabic, a polysaccharide. Gum arabic was subjected to periodate oxidation to obtain gum arabic aldehyde (GAA). GAA was reacted with gelatin under appropriate pH to prepare the cross-linked hydrogel. Cross-linking occurred due to Schiff's base reaction between aldehyde groups of oxidized gum arabic and amino groups of gelatin. The scaffold prepared from the hydrogel was characterized by swelling properties, degree of cross-linking, in vitro degradation and scanning electron microscopy (SEM). Cytocompatibility evaluation using L-929 and HepG2 cells confirmed non-cytotoxic and non-adherent nature of the scaffold. These properties are essential for generating multicellular spheroids and hence the scaffold is proposed to be a suitable candidate for spheroid cell culture. - Highlights: • Gum arabic cross-linked gelatin scaffold was developed for tissue engineering. • Cross-linking was achieved by Schiff's base reaction. • The scaffold is non-cytotoxic and non adherent to fibroblast and hepatocytes. • The scaffolds are potential candidates for spheroid cell culture

  1. Interactions of antimicrobial compounds with cross-linking agents of alginate dressings.

    Science.gov (United States)

    Goh, Cheong Hian; Heng, Paul Wan Sia; Huang, Esther Pei En; Li, Benny Kai Hui; Chan, Lai Wah

    2008-07-01

    The aim of this study was to investigate the antimicrobial activities of calcium ions and other cross-linking agents of alginate dressings, as well as their compatibility with commonly used topical antimicrobials. The antimicrobial activities of cross-linking agents and antimicrobials (five antibiotics and four antiseptics) were evaluated by the broth dilution method. The interactions between individual cross-linking agents and antimicrobials were evaluated using the chequerboard test against common skin pathogens, Staphylococcus aureus and Pseudomonas aeruginosa. From the MIC determined, antibiotics were the most active, followed by the antiseptics and cross-linking agents. Calcium ions, which are commonly used to cross-link alginate, exhibited very weak antimicrobial activity and higher fractional inhibitory concentration than the other cross-linking agents. The use of calcium and gentamicin resulted in antagonism against S. aureus. In contrast, aluminium, zinc and copper ions exhibited higher antimicrobial activities but insignificant interactions with the antimicrobials. Commonly used topical antimicrobials that are active against the skin pathogens S. aureus and P. aeruginosa could be potentially incompatible with calcium alginate dressings. Copper, zinc and aluminium ions are more suitable cross-linking agents for alginate as they do not show antagonism with the antimicrobials and could impart antimicrobial property to the resultant dressing.

  2. Riboflavin/UVA collagen cross-linking-induced changes in normal and keratoconus corneal stroma.

    Directory of Open Access Journals (Sweden)

    Sally Hayes

    Full Text Available To determine the effect of Ultraviolet-A collagen cross-linking with hypo-osmolar and iso-osmolar riboflavin solutions on stromal collagen ultrastructure in normal and keratoconus ex vivo human corneas.Using small-angle X-ray scattering, measurements of collagen D-periodicity, fibril diameter and interfibrillar spacing were made at 1 mm intervals across six normal post-mortem corneas (two above physiological hydration (swollen and four below (unswollen and two post-transplant keratoconus corneal buttons (one swollen; one unswollen, before and after hypo-osmolar cross-linking. The same parameters were measured in three other unswollen normal corneas before and after iso-osmolar cross-linking and in three pairs of swollen normal corneas, in which only the left was cross-linked (with iso-osmolar riboflavin.Hypo-osmolar cross-linking resulted in an increase in corneal hydration in all corneas. In the keratoconus corneas and unswollen normal corneas, this was accompanied by an increase in collagen interfibrillar spacing (p<0.001; an increase in fibril diameter was also seen in two out of four unswollen normal corneas and one unswollen keratoconus cornea (p<0.001. Iso-osmolar cross-linking resulted in a decrease in tissue hydration in the swollen normal corneas only. Although there was no consistent treatment-induced change in hydration in the unswollen normal samples, iso-osmolar cross-linking of these corneas did result in a compaction of collagen fibrils and a reduced fibril diameter (p<0.001; these changes were not seen in the swollen normal corneas. Collagen D-periodicity was not affected by either treatment.The observed structural changes following Ultraviolet-A cross-linking with hypo-osmolar or iso-osmolar riboflavin solutions are more likely a consequence of treatment-induced changes in tissue hydration rather than cross-linking.

  3. Characterization of solid UV cross-linked PEGDA for biological applications

    KAUST Repository

    Castro, David

    2013-10-20

    This paper reports on solid UV cross-linked Poly(ethylene)-glycol-diacrylate (PEGDA) as a material for microfluidic devices for biological applications. We have evaluated biocompatibility of PEGDA through two separate means: 1) by examining cell viability and attachment on cross-linked PEGDA surfaces for cell culture applications, and 2) by determining if cross-linked PEGDA inhibits the polymerase chain reaction (PCR) processes for on-chip PCR. Through these studies a correlation has been found between degree of curing and cell viability, attachment, as well as on PCR outcome.

  4. Novel cross-linked polyacrylamide matrices: an investigation using gradient gel electrophoresis.

    Science.gov (United States)

    Caulfield, M J; Purss, H K; Solomon, D H

    2001-12-01

    Gradient gel electrophoresis was used to examine the separation properties of novel cross-linking compounds for polyacrylamide (PAAm). At low %T and at the same %C protein migration difference is accentuated for bismethacrylamide cross-linked networks relative to bisacrylamide cross-linked networks. Similar properties were observed for cyclic monomers at low %T. This trend is maintained throughout the gradient. However, at higher %T migration differential relative to N,N'-methylenebisacrylamide (Bis) was less pronounced. Evidence from gradient gels suggests that reactivity and functionality of vinyl groups impose an overriding control over network formation.

  5. Trapping of a Cross-link Formed by a Major Purine Adduct of a Metabolite of the Carcinogen N-Nitrosomorpholine by Inorganic and Biological Reductants

    Science.gov (United States)

    Koissi, Niangoran; Fishbein, James C.

    2013-01-01

    3-Hydroperoxy-N-nitrosomorpholine in buffered aqueous media in the presence of calf thymus DNA was treated with a phosphine reductant to generate the transient α-hydroxynitrosamine and subsequent diazonium ion that alkylated the DNA, as previously reported. Subsequent addition of hydride donors, for 30 min, followed by acid hydrolysis of the mixture allowed detection and quantification of 6-(2-(2-((9H-purin-6-yl)amino)ethoxy)ethoxy)-9H-purin-2-amine, the reduced cross-link formed from deposition, via the diazonium ion, of a 3-oxa-pentanal fragment on O6-Gua, and condensation with N6-Ade, presumably in the vicinity. Decreasing temperature of the reactions and decreasing pH modestly increased the yields of trapped crosslink. Among three borohydride reductants, NaNCBH3 is superior, being ∼4 times more effective on a molar basis, as opposed to a hydride equivalent basis, than NaBH4 or Na(AcO)3BH. For trapping with NaNCBH3, it is deduced that the reaction likely occurs with the iminium ion that is in protonic equilibrium with its conjugate base imine. In an experiment in which the hydroperoxide was decomposed and NaNCBH3 was introduced after various times, the amount of cross-link was observed to increase, nearly linearly, by about four-fold over one week. These data indicate that there are a minimum of 2 populations of cross-links, one that forms rapidly, in minutes, and another that grows in with time, over days. Reduced nicotinamide co-factors and ascorbate are observed to effect reduction (over 3 days) of the cross-links confirming the possibility that otherwise reversible cross-links might be immortalized under biological conditions. PMID:23587048

  6. Vitamin E-diffused highly cross-linked UHMWPE particles induce less osteolysis compared to highly cross-linked virgin UHMWPE particles in vivo

    DEFF Research Database (Denmark)

    Bichara, David A; Malchau, Erik; Sillesen, Nanna H

    2014-01-01

    when compared to virgin gamma irradiated cross-linked UHMWPE. Groups received equal amount of particulate debris overlaying the calvarium for 10 days. Calvarial bone was examined using high resolution micro-CT and histomorphometric analyses. There was a statistically significant difference between...

  7. Cross-linking proteins with bimetallic tetracarboxylate compounds of transition metals

    Science.gov (United States)

    Kostic, N.M.; Chen, J.

    1991-03-05

    Stable cross-linked complexes of transition-metal tetracarboxylates and proteins are formed. The preferred transition-metal is rhodium. The protein may be collagen or an enzyme such as a proteolytic enzyme. No Drawings

  8. CrossWork: Software-assisted identification of cross-linked peptides

    DEFF Research Database (Denmark)

    Rasmussen, Morten; Refsgaard, Jan; Peng, Li

    2011-01-01

    The increased interest in chemical cross-linking for probing protein structure and interaction has led to a large increase in literature describing new cross-linkers and search programs. However, this has not led to a corresponding increase in the analysis of large and complex proteins. A major...... obstacle is that the new cross-linkers are either not readily available and/or have a low reactivity. In combination with aging search programs that are slow and have low sensitivity, or new search programs that are described but not released, these efforts do little to advance the field of cross......-linking. Here we present a method pipeline for chemical cross-linking, using two standard cross-linkers, BS3 and BS2G, combined with our freely available CrossWork search program. By this approach we generate cross-link data sufficient to derive structural information for large and complex proteins. Cross...

  9. A Review of Collagen Cross-Linking in Cornea and Sclera

    Directory of Open Access Journals (Sweden)

    Xiao Zhang

    2015-01-01

    Full Text Available Riboflavin/UVA cross-linking is a technique introduced in the past decades for the treatment of keratoconus, keratectasia, and infectious keratitis. Its efficacy and safety have been investigated with clinical and laboratory studies since its first clinical application by Wollensak for the treatment of keratoconus. Although its complications are encountered during clinical practice, such as infection inducing risk, minimal invasion merits a further investigation on its future application in clinical practice. Recently, collagen cross-linking in sclera shows a promising prospect. In present study, we summarized the representative studies describing the clinical and laboratory application of collagen cross-linking published in past decades and provided our opinion on the positive and negative results of cross-linking in the treatment of ophthalmic disorders.

  10. Cross-linked enzyme aggregates of recombinant Pseudomonas putida nitrilase for enantioselective nitrile hydrolysis.

    Science.gov (United States)

    Kumar, Sandeep; Mohan, Utpal; Kamble, Ashwini L; Pawar, Sandip; Banerjee, Uttam C

    2010-09-01

    The cross-linked enzyme aggregate (CLEA) method is used for the dual purpose of combining both the purification and immobilization of enzyme in one step. The present work involved the preparation of a carrier-free, highly active reusable biocatalyst (nitrilase) which encounters least mass-transfer limitations with higher thermal and storage stability. The effect of type of aggregating agent, its concentration as well as that of cross-linking agent was studied. Nitrilase aggregates were prepared using ammonium sulphate (35%) precipitation followed by cross-linking with glutaraldehyde (125 mM) which rendered 70% activity retention. The various cross-linking parameters were optimized in order to increase the activity retention. Stability in terms of temperature, reusability and leaching were also examined. The CLEA preparation showed residual nitrilase activity on repeated use. A highly stable CLEA of nitrilase was finally prepared with maximum activity recovery. (c) 2010 Elsevier Ltd. All rights reserved.

  11. Determination of the cross-linking effect of adipic acid dihydrazide on glycoconjugate preparation.

    Science.gov (United States)

    Bystrický, S; Machová, E; Malovíková, A; Kogan, G

    1999-11-01

    The cross-linking effect of adipic acid dihydrazide (ADH) on polysaccharide derivatization can be evaluated by applying combination of elemental analysis and colorimetric assay. Elemental analysis is used for estimation of total ADH bound to polysaccharide and a colorimetric trinitrobenzene sulfonic acid assay is used to determine the part of ADH not involved in cross-linking. The difference of values expressed as molar ratios (per repeating unit) provides information on the amount of ADH involved in cross-linking the polysaccharides. Carboxymethylated polysaccharides were derivatized with different amounts of ADH to test the procedure. Analytical results showed that excess of ADH in the reaction only slightly decreased the cross-linking. The number of carboxyl groups remained unmodified even at high excess of ADH and high concentration of carbodiimide (EDC) coupling reagent.

  12. Comparative study of PBI Cross Linked Utilizing Agents of Varying Steric Configurations

    DEFF Research Database (Denmark)

    Kirkebcek, Andreas; Aili, David; Li, Qingfeng

    2016-01-01

    is dependent on the amount of PA present within the membrane. However mechanical properties are reduced are significantly reduced due to the plasticizing effect shown by PA [1]. This effect is due to PBI chain displacement. This effect can be lessened by use of cross linking [2-4]. This can be obtained using......The high thermal and chemical stability of poly[2,2'-(m-phenylene)-5,5' bibenzimidazole] (PBI) accounts for its wise spread use in high temperature polymer electrolyte membrane fuel cells (HT- PEMFC). By doping the membrane with phosphoric acid (PA) ionic conductivity is obtained. Thus conductivity...... ionic or covalent cross linking. When considering such, little attention is devoted to explore the effect of the sterical configuration of the cross linking agent. In this contribution three different cross linking agents are utilized to evaluate how these affects final membrane properties....

  13. An Investigation on Rheology of Peroxide Cross-linking of Low Density Polyethylene

    DEFF Research Database (Denmark)

    Ghasemi, Ismaeil; Rasmussen, Henrik K.; Szabo, Peter

    2005-01-01

    One of the most important post-reactor modifications of polyethylene is cross-linking. It improves some properties of polyethylene such as environmental stress cracking resistance, chemical and abrasion resistance, and service temperature. In this study, the effect of peroxide cross-linking on th......One of the most important post-reactor modifications of polyethylene is cross-linking. It improves some properties of polyethylene such as environmental stress cracking resistance, chemical and abrasion resistance, and service temperature. In this study, the effect of peroxide cross......-linking on the rheological behaviour of low density polyethylene was investigated by using a combination of creep test and differential scanning calorimeter (DSC) in isotherm condition. The used peroxide was di-cumyl peroxide and its concentration was 2 wt%. The experiments were carried out at 150,160, and 170 degrees C...

  14. Novel cross-linked sulfonated poly (arylene ether ketone) membranes for direct methanol fuel cell

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Chengji; Lin, Haidan; Na, Hui [Alan G. MacDiarmid Institute, College of Chemistry, Jilin University, Changchun, Jilin 130012 (China)

    2010-03-15

    To prepare a cross-linked proton exchange membrane with low methanol permeability and high proton conductivity, poly (vinyl alcohol) is first blended with sulfonated poly (arylene ether ketone) bearing carboxylic acid groups (SPAEK-C) and then heated to induce a cross-linking reaction between the carboxyl groups in SPAEK-C and the hydroxyl groups in PVA. Fourier transform infrared spectroscopy is used to characterize and confirm the structure of SPAEK-C and the cross-linked membranes. The proton conductivity of the cross-linked membrane with 15% PVA in weight reaches up to 0.18 S cm{sup -1} at 80 C (100% relative humidity), which is higher than that of Nafion membrane, while the methanol permeability is nearly five times lower than Nafion. The ion-exchange capacity, water uptake and thermal stability are investigated to confirm their applicability in fuel cells. (author)

  15. Stabilized sulfonated aromatic polymers by in situ solvothermal cross-linking

    Directory of Open Access Journals (Sweden)

    Maria Luisa eDi Vona

    2014-10-01

    Full Text Available The cross-link reaction via sulfone bridges of sulfonated polyetheretherketone (SPEEK by thermal treatment at 180 °C in presence of dimethylsulfoxide (DMSO is discussed. The modifications of properties subsequent to the cross-linking are presented. The mechanical strength as well as the hydrolytic stability increased with the thermal treatment time, i.e., with the degree of cross-linking. The proton conductivity was determined as function of temperature, IEC, degree of cross-linking and hydration number. The memory effect, which is the membrane ability to remember the water uptake reached at high temperature also at lower temperature, is exploited in order to achieve high values of conductivity. Membranes swelled at 110 °C can reach a conductivity of 0.14 S/cm at 80°C with a hydration number ( of 73.

  16. Assessment of UVA-Riboflavin Corneal Cross-Linking Using Small Amplitude Oscillatory Shear Measurements.

    Science.gov (United States)

    Aslanides, Ioannis M; Dessi, Claudia; Georgoudis, Panagiotis; Charalambidis, Georgios; Vlassopoulos, Dimitris; Coutsolelos, Athanassios G; Kymionis, George; Mukherjee, Achyut; Kitsopoulos, Theofanis N

    2016-04-01

    The effect of ultraviolet (UV)-riboflavin cross-linking (CXL) has been measured primarily using the strip extensometry technique. We propose a simple and reliable methodology for the assessment of CXL treatment by using an established rheologic protocol based on small amplitude oscillatory shear (SAOS) measurements. It provides information on the average cross-link density and the elastic modulus of treated cornea samples. Three fresh postmortem porcine corneas were used to study the feasibility of the technique, one serving as control and two receiving corneal collagen cross-linking treatment. Subsequently, five pairs of fresh postmortem porcine corneas received corneal collagen cross-linking treatment with riboflavin and UVA-irradiation (370 nm; irradiance of 3 mW/cm2) for 30 minutes (Dresden protocol); the contralateral porcine corneas were used as control samples. After the treatment, the linear viscoelastic moduli of the corneal samples were measured using SAOS measurements and the average cross-linking densities extracted. For all cases investigated, the dynamic moduli of the cross-linked corneas were higher compared to those of the corresponding control samples. The increase of the elastic modulus of the treated samples was between 122% and 1750%. The difference was statistically significant for all tested samples (P = 0.018, 2-tailed t-test). We report a simple and accurate methodology for quantifying the effects of cross-linking on porcine corneas treated with the Dresden protocol by means of SAOS measurements in the linear regime. The measured dynamic moduli, elastic and viscous modulus, represent the energy storage and energy dissipation, respectively. Hence, they provide a means to assess the changing physical properties of the cross-linked collagen networks after CXL treatment.

  17. Computational exploration of polymer nanocomposite mechanical property modification via cross-linking topology

    Science.gov (United States)

    Lacevic, Naida; Gee, Richard H.; Saab, Andrew; Maxwell, Robert

    2008-09-01

    Molecular dynamics simulations have been performed in order to study the effects of nanoscale filler cross-linking topologies and loading levels on the mechanical properties of a model elastomeric nanocomposite. The model system considered here is constructed from octafunctional polyhedral oligomeric silsesquioxane (POSS) dispersed in a poly(dimethylsiloxane) (PDMS) matrix. Shear moduli, G, have been computed for pure and for filled and unfilled PDMS as a function of cross-linking density, POSS fill loading level, and polymer network topology. The results reported here show that G increases as the cross-linking (covalent bonds formed between the POSS and the PDMS network) density increases. Further, G is found to have a strong dependence on cross-linking topology. The increase in shear modulus, G, for POSS filled PDMS is significantly higher than that for unfilled PDMS cross-linked with standard molecular species, suggesting an enhanced reinforcement mechanism for POSS. In contrast, in blended systems (POSS/PDMS mixture with no cross-linking) G was not observed to significantly increase with POSS loading. Finally, we find intriguing differences in the structural arrangement of bond strains between the cross-linked and the blended systems. In the unfilled PDMS the distribution of highly strained bonds appears to be random, while in the POSS filled system, the strained bonds form a netlike distribution that spans the network. Such a distribution may form a structural network "holding" the composite together and resulting in increases in G compared to an unfilled, cross-linked system. These results are of importance for engineering of new POSS-based multifunctional materials with tailor-made mechanical properties.

  18. Novel thermoplastic vulcanizates (TPVs based on silicone rubber and polyamide exploring peroxide cross-linking

    Directory of Open Access Journals (Sweden)

    K. Naskar

    2014-04-01

    Full Text Available Novel thermoplastic vulcanizates (TPVs based on silicone rubber (PDMS and polyamide (PA12 have been prepared by dynamic vulcanization process. The effect of dynamic vulcanization and influence of various types of peroxides as cross-linking agents were studied in detail. All the TPVs were prepared at a ratio of 50/50 wt% of silicone rubber and polyamide. Three structurally different peroxides, namely dicumyl peroxide (DCP, 3,3,5,7,7-pentamethyl 1,2,4-trioxepane (PMTO and cumyl hydroperoxide (CHP were taken for investigation. Though DCP was the best option for curing the silicone rubber, at high temperature it suffers from scorch safety. An inhibitor 2,2,6,6-tetramethylpiperidinyloxyl (TEMPO was added with DCP to stabilize the radicals in order to increase the scorch time. Though CHP (hydroperoxide had higher half life time than DCP at higher temperature, it has no significant effect on cross-linking of silicone rubber. PMTO showed prolonged scorch safety and better cross-linking efficiency rather than the other two. TPVs of DCP and PMTO were made up to 11 minutes of mixing. Increased values of tensile strength and elongation at break of PMTO cross-linked TPV indicate the superiority of PMTO. Scanning electron micrographs correlate with mechanical properties of the TPVs. High storage modulus (E' and lower loss tangent value of the PMTO cross-linked TPV indicate the higher degree of cross-linking which is also well supported by the overall cross-link density value. Thus PMTO was found to be the superior peroxide for cross-linking of silicone rubber at high temperature.

  19. Recyclable cross-linked anion exchange membrane for alkaline fuel cell application

    Science.gov (United States)

    Hou, Jianqiu; Liu, Yazhi; Ge, Qianqian; Yang, Zhengjin; Wu, Liang; Xu, Tongwen

    2018-01-01

    Cross-linking can effectively solve the conductivity-swelling dilemma in anion exchange membranes (AEMs) but will generate solid wastes. To address this, we developed an AEM cross-linked via disulfide bonds, bearing quaternary ammonium groups, which can be easily recycled. The membrane (RC-QPPO) with IEC of 1.78 mmol g-1, when cross-linked, showed enhanced mechanical properties and good hydroxide conductivity (24.6 mS cm-1 at 30 °C). Even at higher IEC value (2.13 mmol g-1), it still has low water uptake, low swelling ratio and delivers a peak power density of 150 mW cm-2 at 65 °C. Exploiting the formation of disulfide bonds from -SH groups, the membrane can be readily cross-linked in alkaline condition and recycled by reversibly breaking disulfide bonds with dithiothreitol (DTT). The recycled membrane solution can be directly utilized to cast a brand-new AEM. By washing away the residual DTT with water and exposure to air, it can be cross-linked again and this process is repeatable. During the recycling and cross-linking processes, the membrane showed a slight IEC decrease of 5% due to functional group degradation. The strategy presented here is promising in enhancing AEM properties and reducing the impact of artificial polymers on the environment.

  20. Characterization of the receptor for endothelial cell growth factor (ECGF) by affinity cross-linking

    Energy Technology Data Exchange (ETDEWEB)

    Friesel, R.; Burgess, W.H.; Mehlman, T.; Maciag, T.

    1986-05-01

    The authors have demonstrated high affinity receptors for ECGF on endothelial cells by covalent cross-linking of (/sup 125/I)-ECGF with disuccinimidyl suberate and observe a single chain cross-linked polypeptide species with an apparent M/sub r/ of 170K. The M/sub r/ 170K species represents (/sup 125/I)-ECGF bound to its receptor since (i) excess unlabeled ECGF inhibits the cross-linking of (/sup 125/I)-ECGF, (ii) labeling of the M/sub r/170K species does not take place in the absence of cross-linker, (iii) cells previously shown to be refractory to ECGF and lack ECGF receptors do not yield a cross-linked species, (iv) the cross-linked species can be immunoprecipitated with anti-ECGF antibodies, and (v) preincubation of cells with ECGF at 37/sup 0/C significantly reduces cross-linking while incubation at 4/sup 0/C does not. These data demonstrate that ECGF induced cell proliferation occurs through the occupancy of a specific cell surface polypeptide receptor with an apparent M/sub r/ of 150K, and suggests that internalization of the receptor-ligand complex may be relevant to ECGF-induced signal transduction.

  1. Riboflavin/UVA Collagen Cross-Linking-Induced Changes in Normal and Keratoconus Corneal Stroma

    Science.gov (United States)

    Hayes, Sally; Boote, Craig; Kamma-Lorger, Christina S.; Rajan, Madhavan S.; Harris, Jonathan; Dooley, Erin; Hawksworth, Nicholas; Hiller, Jennifer; Terill, Nick J.; Hafezi, Farhad; Brahma, Arun K.; Quantock, Andrew J.; Meek, Keith M.

    2011-01-01

    Purpose To determine the effect of Ultraviolet-A collagen cross-linking with hypo-osmolar and iso-osmolar riboflavin solutions on stromal collagen ultrastructure in normal and keratoconus ex vivo human corneas. Methods Using small-angle X-ray scattering, measurements of collagen D-periodicity, fibril diameter and interfibrillar spacing were made at 1 mm intervals across six normal post-mortem corneas (two above physiological hydration (swollen) and four below (unswollen)) and two post-transplant keratoconus corneal buttons (one swollen; one unswollen), before and after hypo-osmolar cross-linking. The same parameters were measured in three other unswollen normal corneas before and after iso-osmolar cross-linking and in three pairs of swollen normal corneas, in which only the left was cross-linked (with iso-osmolar riboflavin). Results Hypo-osmolar cross-linking resulted in an increase in corneal hydration in all corneas. In the keratoconus corneas and unswollen normal corneas, this was accompanied by an increase in collagen interfibrillar spacing (priboflavin solutions are more likely a consequence of treatment-induced changes in tissue hydration rather than cross-linking. PMID:21850225

  2. Solution processed organic light-emitting diodes using the plasma cross-linking technology

    Energy Technology Data Exchange (ETDEWEB)

    He, Kongduo [Department of Light Sources and Illuminating Engineering, Fudan University, Shanghai 200433 (China); Liu, Yang [Department of Light Sources and Illuminating Engineering, Fudan University, Shanghai 200433 (China); Engineering Research Center of Advanced Lighting Technology, Ministry of Education, Shanghai 200433 (China); Gong, Junyi; Zeng, Pan; Kong, Xun; Yang, Xilu; Yang, Cheng; Yu, Yan [Department of Light Sources and Illuminating Engineering, Fudan University, Shanghai 200433 (China); Liang, Rongqing [Department of Light Sources and Illuminating Engineering, Fudan University, Shanghai 200433 (China); Engineering Research Center of Advanced Lighting Technology, Ministry of Education, Shanghai 200433 (China); Ou, Qiongrong, E-mail: qrou@fudan.edu.cn [Department of Light Sources and Illuminating Engineering, Fudan University, Shanghai 200433 (China); Engineering Research Center of Advanced Lighting Technology, Ministry of Education, Shanghai 200433 (China)

    2016-09-30

    Highlights: • Mixed acetylene and Ar plasma treatment makes the organic film surface cross-linked. • The plasma treatment for 30 s does not affect the performance of OLEDs. • Cross-linking surface can resist rinsing and corrosion of organic solvent. • The surface morphology is nearly unchanged after plasma treatment. • The plasma cross-linking method can realize solution processed multilayer OLEDs. - Abstract: Solution processed multilayer organic light-emitting diodes (OLEDs) present challenges, especially regarding dissolution of the first layer during deposition of a second layer. In this work, we first demonstrated a plasma cross-linking technology to produce a solution processed OLED. The surfaces of organic films can be cross-linked after mixed acetylene and Ar plasma treatment for several tens of seconds and resist corrosion of organic solvent. The film thickness and surface morphology of emissive layers (EMLs) with plasma treatment and subsequently spin-rinsed with chlorobenzene are nearly unchanged. The solution processed triple-layer OLED is successfully fabricated and the current efficiency increases 50% than that of the double-layer OLED. Fluorescent characteristics of EMLs are also observed to investigate factors influencing the efficiency of the triple-layer OLED. Plasma cross-linking technology may open up a new pathway towards fabrication of all-solution processed multilayer OLEDs and other soft electronic devices.

  3. Corneal collagen cross-linking. A review of its clinical applications.

    Science.gov (United States)

    Balparda, K; Maldonado, M J

    2017-04-01

    To perform a literature review of the current clinical applications of corneal collagen cross-linking. An exhaustive literature search was made, including the main biomedical databases, and encompassing all years since the introduction of cross-linking in ophthalmology practice. Corneal collagen cross-linking using UVA irradiation and riboflavin is a surgical technique that is currently being optimised, and is supported by a good amount of pre-clinical and clinical studies. These papers found show the beneficial effect of the surgery on preventing the progression of corneal ectasia, especially keratoconus, but also on pellucid marginal degeneration and keratectasia after refractive surgery. The effect of cross-linking on avoiding the occurrence of iatrogenic keratectasia when combined with a photo-ablative procedure is less clear to date. Additionally, it appears that cross-linking may have a considerable beneficial effect on controlling corneal infection caused by fungi, bacteria and amoebae. However, its effect on viral keratitis can be detrimental. The benefit on bullous keratopathy seems to be rather transient. Corneal collagen cross-linking may be used with relative safety and efficacy in patients with progressive keratoconus. Its use could also be considered in patients with other corneal ectasias or with corneal infections of non-viral origin. Currently, there is still a need for more studies as regards its effect on preventing iatrogenic keratectasia. Copyright © 2016 Sociedad Española de Oftalmología. Publicado por Elsevier España, S.L.U. All rights reserved.

  4. Fibromodulin Interacts with Collagen Cross-linking Sites and Activates Lysyl Oxidase*

    Science.gov (United States)

    Bihan, Dominique; Bonna, Arkadiusz; Rubin, Kristofer; Farndale, Richard W.

    2016-01-01

    The hallmark of fibrotic disorders is a highly cross-linked and dense collagen matrix, a property driven by the oxidative action of lysyl oxidase. Other fibrosis-associated proteins also contribute to the final collagen matrix properties, one of which is fibromodulin. Its interactions with collagen affect collagen cross-linking, packing, and fibril diameter. We investigated the possibility that a specific relationship exists between fibromodulin and lysyl oxidase, potentially imparting a specific collagen matrix phenotype. We mapped the fibromodulin-collagen interaction sites using the collagen II and III Toolkit peptide libraries. Fibromodulin interacted with the peptides containing the known collagen cross-linking sites and the MMP-1 cleavage site in collagens I and II. Interestingly, the interaction sites are closely aligned within the quarter-staggered collagen fibril, suggesting a multivalent interaction between fibromodulin and several collagen helices. Furthermore, we detected an interaction between fibromodulin and lysyl oxidase (a major collagen cross-linking enzyme) and mapped the interaction site to 12 N-terminal amino acids on fibromodulin. This interaction also increases the activity of lysyl oxidase. Together, the data suggest a fibromodulin-modulated collagen cross-linking mechanism where fibromodulin binds to a specific part of the collagen domain and also forms a complex with lysyl oxidase, targeting the enzyme toward specific cross-linking sites. PMID:26893379

  5. Fibromodulin Interacts with Collagen Cross-linking Sites and Activates Lysyl Oxidase.

    Science.gov (United States)

    Kalamajski, Sebastian; Bihan, Dominique; Bonna, Arkadiusz; Rubin, Kristofer; Farndale, Richard W

    2016-04-08

    The hallmark of fibrotic disorders is a highly cross-linked and dense collagen matrix, a property driven by the oxidative action of lysyl oxidase. Other fibrosis-associated proteins also contribute to the final collagen matrix properties, one of which is fibromodulin. Its interactions with collagen affect collagen cross-linking, packing, and fibril diameter. We investigated the possibility that a specific relationship exists between fibromodulin and lysyl oxidase, potentially imparting a specific collagen matrix phenotype. We mapped the fibromodulin-collagen interaction sites using the collagen II and III Toolkit peptide libraries. Fibromodulin interacted with the peptides containing the known collagen cross-linking sites and the MMP-1 cleavage site in collagens I and II. Interestingly, the interaction sites are closely aligned within the quarter-staggered collagen fibril, suggesting a multivalent interaction between fibromodulin and several collagen helices. Furthermore, we detected an interaction between fibromodulin and lysyl oxidase (a major collagen cross-linking enzyme) and mapped the interaction site to 12 N-terminal amino acids on fibromodulin. This interaction also increases the activity of lysyl oxidase. Together, the data suggest a fibromodulin-modulated collagen cross-linking mechanism where fibromodulin binds to a specific part of the collagen domain and also forms a complex with lysyl oxidase, targeting the enzyme toward specific cross-linking sites. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  6. Solution processed organic light-emitting diodes using the plasma cross-linking technology

    International Nuclear Information System (INIS)

    He, Kongduo; Liu, Yang; Gong, Junyi; Zeng, Pan; Kong, Xun; Yang, Xilu; Yang, Cheng; Yu, Yan; Liang, Rongqing; Ou, Qiongrong

    2016-01-01

    Highlights: • Mixed acetylene and Ar plasma treatment makes the organic film surface cross-linked. • The plasma treatment for 30 s does not affect the performance of OLEDs. • Cross-linking surface can resist rinsing and corrosion of organic solvent. • The surface morphology is nearly unchanged after plasma treatment. • The plasma cross-linking method can realize solution processed multilayer OLEDs. - Abstract: Solution processed multilayer organic light-emitting diodes (OLEDs) present challenges, especially regarding dissolution of the first layer during deposition of a second layer. In this work, we first demonstrated a plasma cross-linking technology to produce a solution processed OLED. The surfaces of organic films can be cross-linked after mixed acetylene and Ar plasma treatment for several tens of seconds and resist corrosion of organic solvent. The film thickness and surface morphology of emissive layers (EMLs) with plasma treatment and subsequently spin-rinsed with chlorobenzene are nearly unchanged. The solution processed triple-layer OLED is successfully fabricated and the current efficiency increases 50% than that of the double-layer OLED. Fluorescent characteristics of EMLs are also observed to investigate factors influencing the efficiency of the triple-layer OLED. Plasma cross-linking technology may open up a new pathway towards fabrication of all-solution processed multilayer OLEDs and other soft electronic devices.

  7. Comment on "Delivering a photosensitive transplatin prodrug to overcome cisplatin drug resistance'' by H. Song, W. Li, R. Qi, L. Yan, X. Jing, M. Zheng and H. Xiao, Chem. Commun., 2015, 51, 11493

    Czech Academy of Sciences Publication Activity Database

    Brabec, Viktor; Vrána, Oldřich; Nováková, Olga; Kašpárková, Jana

    2016-01-01

    Roč. 52, č. 21 (2016), s. 4096-4098 ISSN 1359-7345 Institutional support: RVO:68081707 Keywords : interstrand cross-links * dna * platinum Subject RIV: BO - Biophysics Impact factor: 6.319, year: 2016

  8. Decreased drug accumulation and increased tolerance to DNA damage in tumor cells with a low level of cisplatin resistance.

    Science.gov (United States)

    Lanzi, C; Perego, P; Supino, R; Romanelli, S; Pensa, T; Carenini, N; Viano, I; Colangelo, D; Leone, R; Apostoli, P; Cassinelli, G; Gambetta, R A; Zunino, F

    1998-04-15

    In an attempt to examine the cellular changes associated with cisplatin resistance, we selected a cisplatin-resistant (A43 1/Pt) human cervix squamous cell carcinoma cell line following continuous in vitro drug exposure. The resistant subline was characterized by a 2.5-fold degree of resistance. In particular, we investigated the expression of cellular defence systems and other cellular factors probably involved in dealing with cisplatin-induced DNA damage. Resistant cells exhibited decreased platinum accumulation and reduced levels of DNA-bound platinum and interstrand cross-link frequency after short-term drug exposure. Analysis of the effect of cisplatin on cell cycle progression revealed a cisplatin-induced G2M arrest in sensitive and resistant cells. Interestingly, a slowdown in S-phase transit was found in A431/Pt cells. A comparison of the ability of sensitive and resistant cells to repair drug-induced DNA damage suggested that resistant cells were able to tolerate higher levels of cisplatin-induced DNA damage than their parental counterparts. Analysis of the expression of proteins involved in DNA mismatch repair showed a decreased level of MSH2 in resistant cells. Since MSH2 seems to be involved in recognition of drug-induced DNA damage, this change may account for the increased tolerance to DNA damage observed in the resistant subline. In conclusion, the involvement of accumulation defects and the increased tolerance to cisplatin-induced DNA damage in these cisplatin-resistant cells support the notion that multiple changes contribute to confer a low level of cisplatin resistance.

  9. The involvement of DNA repair genes in the hypoxia-dependent NLCQ-1 (NSC 709257) toxicity and its synergistic interaction with cisplatin or melphalan

    International Nuclear Information System (INIS)

    Papadopoulou, M.V.; Xue, C.-J.; Bloomer, W.D.

    2003-01-01

    4-[3-(2-Nitro-1-imidazolyl)-propylamino]-7-chloro-quinoline hydrochloride (NLCQ-1) is a weakly DNA-intercalating hypoxia selective cytotoxin, which synergistically enhances the antitumor effect of several chemotherapeutic agents or radiation against mouse tumors or human xenografts. Synergy with melphalan (L-PAM) or cisplatin (cisPt) requires hypoxic pre-exposure of cells to NLCQ-1 or, in mice, administration of NLCQ-1 about 1 h before L-PAM or cisPt. This suggests that NLCQ-1 may cause DNA lesions upon reductive metabolism. To indirectly identify such lesions, rodent cell lines defective in specific DNA repair genes (EM9 and UV41) and their repair-proficient parental AA8, were exposed to NLCQ-1 alone and in combination with L-PAM or cisPt under hypoxic/aerobic conditions and appropriate routes, and assessed for clonogenicity. Selected comparisons with tirapazamine (TPZ) were also performed. DNA ssbs were identified by using the alkaline comet assay. Synergism was assessed by isobologramic analysis. EM9, which lack the functional XRCC1 gene and are unable to efficiently repair DNA ssbs, were 3.7x and 4.5x more sensitive to NLCQ-1 and TPZ, respectively, than the parental AA8 cells. Similarly, UV41, which are defective in the ERCC4/XPF gene and thus, hypersensitive to DNA cross-linking agents, were 4.1x more sensitive than AA8 cells to NLCQ-1. Equitoxic concentrations of NLCQ-1 and TPZ gave similar numbers of ssbs in AA8 and EM9 cells exposed to each compound for 1 h under hypoxic conditions. In combination with L-PAM or cisPt, synergy was observed in AA8 but not in EM9 or UV41 cells, with either NLCQ-1 or TPZ. These results suggest that NLCQ-1 is involved in the formation of DNA ssbs and interstrand crosslinks, with the latter being most likely responsible for NLCQ-1 hypoxic toxicity. The synergistic interaction of NLCQ-1 with L-PAM or cisPt is probably due to an enhancement in the L-PAM/cisPt-induced DNA interstrand crosslinks, possibly as a result of an inhibited

  10. Using response surface methodology to optimize process parameters and cross-linking agents for production of combined-cross-linked bovine serum albumin gels.

    Science.gov (United States)

    Gan, Chee-Yuen; Alkarkhi, Abbas F M; Easa, Azhar Mat

    2009-04-01

    D-optimal design was employed to optimize the mixture of cross-linking agents formulation: microbial transglutaminase (MTGase) and ribose, and the processing parameters (i.e. incubation and heating time) in the mixture in order to obtain combined-cross-linked bovine serum albumin gels that have high gel strength, pH close to neutral and yet medium in browning. Analysis of variance (ANOVA) showed that the contribution of quadratic term to the model over the linear was significant for pH and L* value, whereas linear model was significant for gel strength. Optimization study using response surface methodology (RSM) was performed to the mixture components and process variables and the optimum conditions obtained were: MTGase of 1.34-1.43 g/100 mL, ribose of 1.07-1.16 g/100 mL, incubation time of 5 h at 40 degrees C and heating time of 3 h at 90 degrees C.

  11. Cross-Linked Amylose Bio-Plastic: A Transgenic-Based Compostable Plastic Alternative.

    Science.gov (United States)

    Sagnelli, Domenico; Hooshmand, Kourosh; Kemmer, Gerdi Christine; Kirkensgaard, Jacob J K; Mortensen, Kell; Giosafatto, Concetta Valeria L; Holse, Mette; Hebelstrup, Kim H; Bao, Jinsong; Stelte, Wolfgang; Bjerre, Anne-Belinda; Blennow, Andreas

    2017-09-30

    Bio-plastics and bio-materials are composed of natural or biomass derived polymers, offering solutions to solve immediate environmental issues. Polysaccharide-based bio-plastics represent important alternatives to conventional plastic because of their intrinsic biodegradable nature. Amylose-only (AO), an engineered barley starch with 99% amylose, was tested to produce cross-linked all-natural bioplastic using normal barley starch as a control. Glycerol was used as plasticizer and citrate cross-linking was used to improve the mechanical properties of cross-linked AO starch extrudates. Extrusion converted the control starch from A-type to Vh- and B-type crystals, showing a complete melting of the starch crystals in the raw starch granules. The cross-linked AO and control starch specimens displayed an additional wide-angle diffraction reflection. Phospholipids complexed with Vh-type single helices constituted an integrated part of the AO starch specimens. Gas permeability tests of selected starch-based prototypes demonstrated properties comparable to that of commercial Mater-Bi © plastic. The cross-linked AO prototypes had composting characteristics not different from the control, indicating that the modified starch behaves the same as normal starch. The data shows the feasibility of producing all-natural bioplastic using designer starch as raw material.

  12. Synthesis, Characterization, and Antibacterial Activity of Cross-Linked Chitosan-Glutaraldehyde

    Directory of Open Access Journals (Sweden)

    Long-Biao Guo

    2013-05-01

    Full Text Available This present study deals with synthesis, characterization and antibacterial activity of cross-linked chitosan-glutaraldehyde. Results from this study indicated that cross-linked chitosan-glutaraldehyde markedly inhibited the growth of antibiotic-resistant Burkholderia cepacia complex regardless of bacterial species and incubation time while bacterial growth was unaffected by solid chitosan. Furthermore, high temperature treated cross-linked chitosan-glutaraldehyde showed strong antibacterial activity against the selected strain 0901 although the inhibitory effects varied with different temperatures. In addition, physical-chemical and structural characterization revealed that the cross-linking of chitosan with glutaraldehyde resulted in a rougher surface morphology, a characteristic Fourier transform infrared (FTIR band at 1559 cm−1, a specific X-ray diffraction peak centered at 2θ = 15°, a lower contents of carbon, hydrogen and nitrogen, and a higher stability of glucose units compared to chitosan based on scanning electron microscopic observation, FTIR spectra, X-ray diffraction pattern, as well as elemental and thermo gravimetric analysis. Overall, this study indicated that cross-linked chitosan-glutaraldehyde is promising to be developed as a new antibacterial drug.

  13. Cross-Linked Amylose Bio-Plastic: A Transgenic-Based Compostable Plastic Alternative

    Directory of Open Access Journals (Sweden)

    Domenico Sagnelli

    2017-09-01

    Full Text Available Bio-plastics and bio-materials are composed of natural or biomass derived polymers, offering solutions to solve immediate environmental issues. Polysaccharide-based bio-plastics represent important alternatives to conventional plastic because of their intrinsic biodegradable nature. Amylose-only (AO, an engineered barley starch with 99% amylose, was tested to produce cross-linked all-natural bioplastic using normal barley starch as a control. Glycerol was used as plasticizer and citrate cross-linking was used to improve the mechanical properties of cross-linked AO starch extrudates. Extrusion converted the control starch from A-type to Vh- and B-type crystals, showing a complete melting of the starch crystals in the raw starch granules. The cross-linked AO and control starch specimens displayed an additional wide-angle diffraction reflection. Phospholipids complexed with Vh-type single helices constituted an integrated part of the AO starch specimens. Gas permeability tests of selected starch-based prototypes demonstrated properties comparable to that of commercial Mater-Bi© plastic. The cross-linked AO prototypes had composting characteristics not different from the control, indicating that the modified starch behaves the same as normal starch. The data shows the feasibility of producing all-natural bioplastic using designer starch as raw material.

  14. Evaluation of objective visual quality after corneal cross-linking using double-pass analysis system

    Directory of Open Access Journals (Sweden)

    Meng-Meng Wang

    2016-06-01

    Full Text Available AIM: To evaluate the objective visual quality of patients who underwent corneal cross-linking for the keratoconus using double-pass analysis system. METHODS: Advanced keratoconus patients who underwent UV-riboflavin corneal cross-linking from January to July 2015 were included.The outcomes of their objective scattering index(OSI, predicted visual acuity(VA, the cut-off frequency of modulation transfer function(MTF cut-off, the Strehl ratio(SRwere compared before and 6mo after corneal cross-linking. RESULTS: A total of 13 patients(16 eyeswere included. There was no statistically significant difference between pre- and 6mo postoperative data in uncorrected visual acuity,best corrected visual acuity,refractions and mean value of Sim-k(P>0.05. Non-invasive average tear film break up time(NIAvg-BUTdetected by the Sirius system decreased after corneal cross-linking(PP>0.05. Tear Film Analysis Mean OSI increased at 6mo postoperatively(PCONCLUSION: The subjective visual quality isn't effected by corneal cross-linking. The tear stabilities of patients are influenced by these operations at 6mo postoperatively. More observations on long-term effect are needed to be taken in the future.

  15. Effects of collagen cross-linking on the interlamellar cohesive strength of porcine cornea.

    Science.gov (United States)

    Tao, Chen; Sun, Yong; Zhou, Chuanqing; Han, Zhaolong; Ren, Qiushi

    2013-02-01

    To determine the interlamellar cohesive strength (ICS) of porcine cornea before and after collagen cross-linking using riboflavin and ultraviolet A (UVA) irradiation. Corneal flaps, 130 μm thick, were created with a microkeratome. Cross-linking was accomplished with 0.1% riboflavin and UVA irradiation for 30 minutes (3 mW/cm, 5.4 J/cm) at a distance of 10 mm. Two experiments were performed. (1) The maximal ICS between corneal flap and the stroma was determined and histological examination was performed. (2) The mean stromal ICS was measured. Untreated eyes served as controls in all experiments. The mean maximum flap-stroma ICS was 0.088 ± 0.046 N/mm in the experimental group and 0.012 ± 0.004 N/mm in the control group (P = 0.009). In experiment 2, incomplete (2 mm long) corneal flaps were used and the mean stromal ICS was 0.750 ± 0.077 N/mm in the experimental group and 0.338 ± 0.046 N/mm in the control group (P cornea with cross-linking, but a smooth separation surface in the cornea without cross-linking. Collagen cross-linking activated by riboflavin/UVA increases ICS in porcine corneas.

  16. Effects of partial hydrolysis and subsequent cross-linking on wheat gluten physicochemical properties and structure.

    Science.gov (United States)

    Wang, Kaiqiang; Luo, Shuizhong; Cai, Jing; Sun, Qiaoqiao; Zhao, Yanyan; Zhong, Xiyang; Jiang, Shaotong; Zheng, Zhi

    2016-04-15

    The rheological behavior and thermal properties of wheat gluten following partial hydrolysis using Alcalase and subsequent microbial transglutaminase (MTGase) cross-linking were investigated. The wheat gluten storage modulus (G') and thermal denaturation temperature (Tg) were significantly increased from 2.26 kPa and 54.43°C to 7.76 kPa and 57.69°C, respectively, by the combined action of partial hydrolysis (DH 0.187%) and cross-linking. The free SH content, surface hydrophobicity, and secondary structure analysis suggested that an appropriate degree of Alcalase-based hydrolysis allowed the compact wheat gluten structure to unfold, increasing the β-sheet content and surface hydrophobicity. This improved its molecular flexibility and exposed additional glutamine sites for MTGase cross-linking. SEM images showed that a compact 3D network formed, while SDS-PAGE profiles revealed that excessive hydrolysis resulted in high-molecular-weight subunits degrading to smaller peptides, unsuitable for cross-linking. It was also demonstrated that the combination of Alcalase-based partial hydrolysis with MTGase cross-linking might be an effective method for modifying wheat gluten rheological behavior and thermal properties. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Effects of alginate hydrogel cross-linking density on mechanical and biological behaviors for tissue engineering.

    Science.gov (United States)

    Jang, Jinah; Seol, Young-Joon; Kim, Hyeon Ji; Kundu, Joydip; Kim, Sung Won; Cho, Dong-Woo

    2014-09-01

    An effective cross-linking of alginate gel was made through reaction with calcium carbonate (CaCO3). We used human chondrocytes as a model cell to study the effects of cross-linking density. Three different pore size ranges of cross-linked alginate hydrogels were fabricated. The morphological, mechanical, and rheological properties of various alginate hydrogels were characterized and responses of biosynthesis of cells encapsulated in each gel to the variation in cross-linking density were investigated. Desired outer shape of structure was maintained when the alginate solution was cross-linked with the applied method. The properties of alginate hydrogel could be tailored through applying various concentrations of CaCO3. The rate of synthesized GAGs and collagens was significantly higher in human chondrocytes encapsulated in the smaller pore structure than that in the larger pore structure. The expression of chondrogenic markers, including collagen type II and aggrecan, was enhanced in the smaller pore structure. It was found that proper structural morphology is a critical factor to enhance the performance and tissue regeneration. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. A General Method for Targeted Quantitative Cross-Linking Mass Spectrometry.

    Directory of Open Access Journals (Sweden)

    Juan D Chavez

    Full Text Available Chemical cross-linking mass spectrometry (XL-MS provides protein structural information by identifying covalently linked proximal amino acid residues on protein surfaces. The information gained by this technique is complementary to other structural biology methods such as x-ray crystallography, NMR and cryo-electron microscopy[1]. The extension of traditional quantitative proteomics methods with chemical cross-linking can provide information on the structural dynamics of protein structures and protein complexes. The identification and quantitation of cross-linked peptides remains challenging for the general community, requiring specialized expertise ultimately limiting more widespread adoption of the technique. We describe a general method for targeted quantitative mass spectrometric analysis of cross-linked peptide pairs. We report the adaptation of the widely used, open source software package Skyline, for the analysis of quantitative XL-MS data as a means for data analysis and sharing of methods. We demonstrate the utility and robustness of the method with a cross-laboratory study and present data that is supported by and validates previously published data on quantified cross-linked peptide pairs. This advance provides an easy to use resource so that any lab with access to a LC-MS system capable of performing targeted quantitative analysis can quickly and accurately measure dynamic changes in protein structure and protein interactions.

  19. Supercritical CO2 Foaming of Radiation Cross-Linked Isotactic Polypropylene in the Presence of TAIC

    Directory of Open Access Journals (Sweden)

    Chen-Guang Yang

    2016-12-01

    Full Text Available Since the maximum foaming temperature window is only about 4 °C for supercritical CO2 (scCO2 foaming of pristine polypropylene, it is important to raise the melt strength of polypropylene in order to more easily achieve scCO2 foaming. In this work, radiation cross-linked isotactic polypropylene, assisted by the addition of a polyfunctional monomer (triallylisocyanurate, TAIC, was employed in the scCO2 foaming process in order to understand the benefits of radiation cross-linking. Due to significantly enhanced melt strength and the decreased degree of crystallinity caused by cross-linking, the scCO2 foaming behavior of polypropylene was dramatically changed. The cell size distribution, cell diameter, cell density, volume expansion ratio, and foaming rate of radiation-cross-linked polypropylene under different foaming conditions were analyzed and compared. It was found that radiation cross-linking favors the foamability and formation of well-defined cell structures. The optimal absorbed dose with the addition of 2 wt % TAIC was 30 kGy. Additionally, the foaming temperature window was expanded to about 8 °C, making the handling of scCO2 foaming of isotactic polypropylene much easier.

  20. Surface characteristics determining the cell compatibility of ionically cross-linked alginate gels

    International Nuclear Information System (INIS)

    Machida-Sano, Ikuko; Hirakawa, Makoto; Matsumoto, Hiroki; Kamada, Mitsuki; Ogawa, Sakito; Satoh, Nao; Namiki, Hideo

    2014-01-01

    In this study we investigated differences in the characteristics determining the suitability of five types of ion (Fe 3+ , Al 3+ , Ca 2+ , Ba 2+ and Sr 2+ )-cross-linked alginate films as culture substrates for cells. Human dermal fibroblasts were cultured on each alginate film to examine the cell affinity of the alginates. Since cell behavior on the surface of a material is dependent on the proteins adsorbed to it, we investigated the protein adsorption ability and surface features (wettability, morphology and charge) related to the protein adsorption abilities of alginate films. We observed that ferric, aluminum and barium ion-cross-linked alginate films supported better cell growth and adsorbed higher amounts of serum proteins than other types. Surface wettability analysis demonstrated that ferric and aluminum ion-cross-linked alginates had moderate hydrophilic surfaces, while other types showed highly hydrophilic surfaces. The roughness was exhibited only on barium ion-cross-linked alginate surface. Surface charge measurements revealed that alginate films had negatively charged surfaces, and showed little difference among the five types of gel. These results indicate that the critical factors of ionically cross-linked alginate films determining the protein adsorption ability required for their cell compatibility may be surface wettability and morphology. (paper)

  1. Modification mechanism of sesbania gum, and preparation, property, adsorption of dialdehyde cross-linked sesbania gum.

    Science.gov (United States)

    Tang, Hongbo; Gao, Shiqi; Li, Yanping; Dong, Siqing

    2016-09-20

    This paper studied the modification mechanism of Sesbania gum (SG) by means of the variations in the numbers of surface hydroxyl groups on the granules, Schiff's agent coloration of aldehyde groups, Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD), energy dispersive spectrum (EDS), etc., and also examined the preparation, property and adsorption of dialdehyde cross-linked sesbania gum (DCLSG). The results showed that the surface hydroxyl numbers of cross-linked sesbania gum (CLSG) decreased with increasing the cross-linking degree. The distribution of the aldehyde groups on the DCLSG particles was nonuniform because most of aldehyde groups mainly located on the edge of particles. The cross-linking occurred only on the surface of SG particles. The oxidization occurred not only on the surface of SG particles, but also in the interior of particles. The cross-linking or oxidization changed the thermal properties, and reduced the swelling power, viscosity, alkali and acid resistance of SG. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Preparation of cross-linked hen-egg white lysozyme crystals free of cracks

    Science.gov (United States)

    Yan, Er-Kai; Lu, Qin-Qin; Zhang, Chen-Yan; Liu, Ya-Li; He, Jin; Chen, Da; Wang, Bo; Zhou, Ren-Bin; Wu, Ping; Yin, Da-Chuan

    2016-01-01

    Cross-linked protein crystals (CLPCs) are very useful materials in applications such as biosensors, catalysis, and X-ray crystallography. Hence, preparation of CLPCs is an important research direction. During the preparation of CLPCs, an often encountered problem is that cracks may appear in the crystals, which may finally lead to shattering of the crystals into small pieces and cause problem in practical applications. To avoid cross-link induced cracking, it is necessary to study the cracking phenomenon in the preparation process. In this paper, we present an investigation on how to avoid cracking during preparation of CLPCs. An orthogonal experiment was designed to study the phenomenon of cross-link induced cracking of hen-egg white lysozyme (HEWL) crystals against five parameters (temperature, solution pH, crystal growth time, glutaraldehyde concentration, and cross-linking time). The experimental results showed that, the solution pH and crystal growth time can significantly affect cross-link induced cracking. The possible mechanism was studied, and optimized conditions for obtaining crack-free CLPCs were obtained and experimentally verified. PMID:27703210

  3. Physico-chemical/biological properties of tripolyphosphate cross-linked chitosan based nanofibers

    International Nuclear Information System (INIS)

    Sarkar, Soumi Dey; Farrugia, Brooke L.; Dargaville, Tim R.; Dhara, Santanu

    2013-01-01

    In this study, chitosan-PEO blend, prepared in a 15 M acetic acid, was electrospun into nanofibers (∼ 78 nm diameter) with bead free morphology. While investigating physico-chemical parameters of blend solutions, effect of yield stress on chitosan based nanofiber fabrication was clearly evidenced. Architectural stability of nanofiber mat in aqueous medium was achieved by ionotropic cross-linking of chitosan by tripolyphosphate (TPP) ions. The TPP cross-linked nanofiber mat showed swelling up to ∼ 300% in 1 h and ∼ 40% degradation during 30 day study period. 3T3 fibroblast cells showed good attachment, proliferation and viability on TPP treated chitosan based nanofiber mats. The results indicate non-toxic nature of TPP cross-linked chitosan based nanofibers and their potential to be explored as a tissue engineering matrix. - Highlights: ► Chitosan based nanofiber fabrication through electrospinning. ► Roles of solution viscosity and yield stress on spinnability of chitosan evidenced. ► Tripolyphosphate (TPP) cross-linking rendered structural stability to nanofibers. ► TPP cross-linking also improved cellular response on chitosan based nanofibers. ► Thus, chitosan based nanofibers are suitable for tissue engineering application

  4. Synthesis of acrylic and allylic bifunctional cross-linking monomers derived from PET waste

    International Nuclear Information System (INIS)

    Cruz-Aguilar, A; Herrera-González, A M; Vázquez-García, R A; Coreño, J; Navarro-Rodríguez, D

    2013-01-01

    An acrylic and two novel allylic monomers synthesized from bis (hydroxyethyl) terephthalate, BHET, are reported. This was obtained by glycolysis of post-consumer PET with boiling ethylene glycol. The bifunctional monomer bis(2-(acryloyloxy)ethyl) terephthalate was obtained from acryloyl chloride, while the allylic monomers 2-(((allyloxi)carbonyl)oxy) ethyl (2-hydroxyethyl) terephthalate and bis(2-(((allyloxi)carbonyl)oxy)ethyl) terephthalate, from allyl chloroformate. Cross-linking was studied in bulk polymerization using two different thermal initiators. Monomers were analyzed by means of 1 H NMR and the cross-linked polymers by infrared spectroscopy. Gel content higher than 90% was obtained for the acrylic monomer. In the case of the mixture of the allylic monomers, the cross-linked polymer was 80 % using BPO initiator, being this mixture 24 times less reactive than the acrylic monomer.

  5. Abrasive wear and metallosis associated with cross-linked polyethylene in total hip arthroplasty.

    Science.gov (United States)

    O'Brien, Sean T; Burnell, Colin D; Hedden, David R; Brandt, Jan-M

    2013-01-01

    A 34-year-old female patient received a cobalt-chromium (CoCr) alloy femoral head on cross-linked polyethylene total hip replacement for the revision of her fractured ceramic-on-ceramic total hip replacement. The CoCr alloy femoral head became severely worn due to third-body abrasive wear by ceramic particles that could not be removed by synovectomy or irrigation at revision surgery. Ceramic particles were found embedded in the cross-linked polyethylene liner. The CoCr alloy femoral head exhibited a total mass loss of 14.2 g and the generated wear particles triggered metallosis in the patient. The present case study suggests not revising a fractured ceramic-on-ceramic total hip replacement with a CoCr alloy femoral head and a cross-linked polyethylene liner to avoid metallosis due to third-body abrasive wear. Crown Copyright © 2013. Published by Elsevier Inc. All rights reserved.

  6. Photochromic cross-link polymer for color changing and sensing surface

    Science.gov (United States)

    Fu, Richard; Shi, Jianmin; Forsythe, Eric; Srour, Merric

    2016-12-01

    Photochromic cross-link polymers were developed using patented ultraviolet (UV) photoinitiator and commercial photochromic dyes. The photochromic dyes have been characterized by measuring absorbance before and after UV activation using UV-visible (Vis) spectrometry with varying activation intensities and wavelengths. Photochromic cross-link polymers were characterized by a dynamic xenon and UV light activation and fading system. The curing processes on cloth were established and tested to obtain effective photochromic responses. Both PulseForge photonic curing and PulseForge plus heat surface curing processes had much better photochromic responses (18% to 19%, 16% to 25%, respectively) than the xenon lamp treatment (8%). The newly developed photochromic cross-link polymer showed remarkable coloration contrasts and fast and comparable coloration and fading rates. Those intelligent, controlled color changing and sensing capabilities will be used on flexible and "drapeable" surfaces, which will incorporate ultra-low power sensors, sensor indicators, and identifiers.

  7. A genetic anomaly of oriented collagen biosynthesis and cross-linking: Keratoconus.

    Science.gov (United States)

    Bourges, J L; Robert, A M; Robert, L

    2015-02-01

    Oriented collagen biosynthesis is one of the major mechanisms involved in tissue and organ formation during development. Corneal biogenesis is one example. Defects in this process lead to anomalies in tissue structure and function. The transparency of cornea and its achievement are a good example as well as its pathological modifications. Keratoconus is one example of this type of pathologies, involving also inappropriate cross-linking of collagen fibers. Among the tentatives to correct this anomaly, the riboflavin-potentiated UV-cross-linking (CXL) of keratoconus corneas appears clinically satisfactory, although none of the experiments and clinical results published prove effective cross-linking. The published results are reviewed in this article. Copyright © 2014. Published by Elsevier SAS.

  8. Vitamin E diffused highly cross-linked polyethylene in total hip arthroplasty at five years

    DEFF Research Database (Denmark)

    Nebergall, A. K.; Greene, M. E.; Laursen, M. B.

    2017-01-01

    Aims: The objective of this five-year prospective, blinded, randomised controlled trial (RCT) was to compare femoral head penetration into a Vitamin E diffused highly cross-linked polyethylene (HXLPE) liner with penetration into a medium cross-linked polyethylene control liner using......, ArComXL. This is the longest-term RCT comparing the wear performance and clinical outcome of Vitamin E diffused HXLPE with a previous generation of medium cross-linked polyethylene....... radiostereometric analysis. Patients and Methods: Patients scheduled for total hip arthroplasty (THA) were randomised to receive either the study E1 (32 patients) or the control ArComXL polyethylene (35 patients). The median age (range) of the overall cohort was 66 years (40 to 76). Results: The five-year median...

  9. Cross-linked PAN-based thin-film composite membranes for non-aqueous nanofiltration

    KAUST Repository

    Pérez-Manríquez, Liliana

    2015-01-01

    A new approach on the development of cross-linked PAN based thin film composite (TFC) membranes for non-aqueous application is presented in this work. Polypropylene backed neat PAN membranes fabricated by phase inversion process were cross-linked with hydrazine to get excellent solvent stability toward dimethylformamide (DMF). By interfacial polymerization a selective polyamide active layer was coated over the cross-linked PAN using N,N′-diamino piperazine (DAP) and trimesoyl chloride (TMC) as monomers. Permeation and molecular weight cut off (MWCO) experiments using various dyes were done to evaluate the performance of the membranes. Membranes developed by such method show excellent solvent stability toward DMF with a permeance of 1.7 L/m2 h bar and a molecular weight cut-off of less than 600 Da.

  10. Genipin Cross-Linked Polymeric Alginate-Chitosan Microcapsules for Oral Delivery: In-Vitro Analysis

    Directory of Open Access Journals (Sweden)

    Hongmei Chen

    2009-01-01

    Full Text Available We have previously reported the preparation of the genipin cross-linked alginate-chitosan (GCAC microcapsules composed of an alginate core with a genipin cross-linked chitosan membrane. This paper is the further investigation on their structural and physical characteristics. Results showed that the GCAC microcapsules had a smooth and dense surface and a networked interior. Cross-linking by genipin substantially reduced swelling and physical disintegration of microcapsules induced by nongelling ions and calcium sequestrants. Strong resistance to mechanical shear forces and enzymatic degradation was observed. Furthermore, the GCAC membranes were permeable to bovine serum albumin and maintained a molecular weight cutoff at 70 KD, analogous to the widely studied alginate-chitosan, and alginate-poly-L-lysine-alginate microcapsules. The release features and the tolerance of the GCAC microcapsules in the stimulated gastrointestinal environment were also investigated. This GCAC microcapsule formulation offers significant potential as a delivery vehicle for many biomedical applications.

  11. Production of Materials with Spatially-Controlled Cross-Link Density via Vat Photopolymerization.

    Science.gov (United States)

    Peterson, Gregory I; Schwartz, Johanna J; Zhang, Di; Weiss, Benjamin M; Ganter, Mark A; Storti, Duane W; Boydston, Andrew J

    2016-10-26

    We describe an efficient method to produce objects comprising spatially controlled and graded cross-link densities using vat photopolymerization additive manufacturing (AM). Using a commercially available diacrylate-based photoresin, 3D printer, and digital light processing (DLP) projector, we projected grayscale images to print objects in which the varied light intensity was correlated to controlled cross-link densities and associated mechanical properties. Cylinder and bar test specimens were used to establish correlations between light intensities used for printing and cross-link density in the resulting specimens. Mechanical testing of octet truss unit cells in which the properties of the crossbars and vertices were independently modified revealed unique mechanical responses from the different compositions. From the various test geometries, we measured changes in mechanical properties such as increased strain-to-break in inhomogeneous structures in comparison with homogeneous variants.

  12. Optimizing end-group cross-linking polymer electrolytes for fuel cell applications

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Yu Seung [Los Alamos National Laboratory; Lee, Kwan Soo [Los Alamos National Laboratory; Jeong, Myung - Hwan [GIST, KOREA; Lee, Jae - Suk [GIST, KOREA

    2009-01-01

    This paper demonstrates the optimization of proton conductivity and water uptake for cross-linkable polymer electrolytes through synthesis and characterization of end-group cross-linkable sulfonated poly(arylene ether) copolymers (ESF-BPs). The extent of reaction of cross-linking was controlled by reaction time resulting in a series of polymers with two, independent tunable parameters, degree of sulfonation (DS) and degree of cross-linking (DC). For the polymers presented, cross-linking improved proton conductivity while reducing water uptake, an uncommon trend in polymer electrolytes where water is critical for proton conduction. Other trends relating to changes are reported and the results yield insight into the role of DS and DC and how to optimize electrochemical properties and performance of polymer electrolytes through these tunable parameters. Select polymer electrolytes were tested in fuel cells where performance and durability with accelerated relative humidity cycling were compared with Nafion{reg_sign}.

  13. Physical Characterization Of High Amylose/Pectin Mixtures Cross-Linked With Sodium Trimetaphosphate

    International Nuclear Information System (INIS)

    Carbinatto, F.M.; Cury, B.S.F.; Evangelista, R.C.

    2010-01-01

    Some researches have reported that pectin and high amylose mixtures presented superior mechanical properties in relation to those of the isolated polymers. In this work, mixtures at different ratios (1:4; 1:1) of pectin and high amylose were crosslinked with sodium trimetaphosphate at different degrees by varying reaction conditions. All samples were characterized by rheological and X-ray diffraction analyses. Samples without cross-linker were prepared as control. The oscillatory dynamic tests showed that all samples exhibited predominant elastic behavior, although cross-linked samples presented higher G' values, suggesting that crosslinking by phosphorylation resulted in more strength structures. The diffractograms showed that cross-linked samples underwent structural modifications that resulted in increase of crystallinity due to cross-linking process. (author)

  14. Cross-linking and rheological changes of whey proteins treated with microbial transglutaminase.

    Science.gov (United States)

    Truong, Van-Den; Clare, Debra A; Catignani, George L; Swaisgood, Harold E

    2004-03-10

    Modification of the functionality of whey proteins using microbial transglutaminase (TGase) has been the subject of recent studies. However, changes in rheological properties of whey proteins as affected by extensive cross-linking with TGase are not well studied. The factors affecting cross-linking of whey protein isolate (WPI) using both soluble and immobilized TGase were examined, and the rheological properties of the modified proteins were characterized. The enzyme was immobilized on aminopropyl glass beads (CPG-3000) by selective adsorption of the biotinylated enzyme on avidin that had been previously immobilized. WPI (4 and 8% w/w) in deionized water, pH 7.5, containing 10 mM dithiothreitol was cross-linked using enzyme/substrate ratios of 0.12-10 units of activity/g WPI. The reaction was carried out in a jacketed bioreactor for 8 h at 40 degrees C with continuous circulation. The gel point temperature of WPI solutions treated with 0.12 unit of immobilized TGase/g was slightly decreased, but the gel strength was unaffected. However, increasing the enzyme/substrate ratio resulted in extensive cross-linking of WPI that was manifested by increases in apparent viscosity and changes in the gelation properties. For example, using 10 units of soluble TGase/g resulted in extensive cross-linking of alpha-lactalbumin and beta-lactoglobulin in WPI, as evidenced by SDS-PAGE and Western blotting results. Interestingly, the gelling point of WPI solutions increased from 68 to 94 degrees C after a 4-h reaction, and the gel strength was drastically decreased (lower storage modulus, G'). Thus, extensive intra- and interchain cross-linking probably caused formation of polymers that were too large for effective network development. These results suggest that a process could be developed to produce heat-stable whey proteins for various food applications.

  15. Biomimetic acellular detoxified glutaraldehyde cross-linked bovine pericardium for tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Mathapati, Santosh; Bishi, Dillip Kumar [Stem Cell and Molecular Biology Laboratory, Department of Biotechnology, Indian Institute of Technology Madras, Chennai (India); Frontier Lifeline Pvt Ltd. and Dr. K. M. Cherian Heart Foundation, Mogappair, Chennai (India); Healthcare and Energy Materials Laboratory, NUSNNI, Faculty of Engineering, National University of Singapore (Singapore); Guhathakurta, Soma [Departmet of Engineering Design, Indian Institute of Technology Madras, Chennai (India); Cherian, Kotturathu Mammen [Frontier Lifeline Pvt Ltd. and Dr. K. M. Cherian Heart Foundation, Mogappair, Chennai (India); Venugopal, Jayarama Reddy; Ramakrishna, Seeram [Healthcare and Energy Materials Laboratory, NUSNNI, Faculty of Engineering, National University of Singapore (Singapore); Verma, Rama Shanker, E-mail: vermars@iitm.ac.in [Stem Cell and Molecular Biology Laboratory, Department of Biotechnology, Indian Institute of Technology Madras, Chennai (India)

    2013-04-01

    Glutaraldehyde (GLUT) processing, cellular antigens, calcium ions in circulation, and phospholipids present in the native tissue are predominantly responsible for calcification, degeneration, and lack of natural microenvironment for host progenitor cell migration in tissue implants. The study presents an improved methodology for adhesion and proliferation of endothelial progenitor cells (EPCs) without significant changes in biomechanical and biodegradation properties of the processed acellular bovine pericardium. The anti-calcification potential of the processed tissue was enhanced by detoxification of GLUT-cross-linked bovine pericardium by decellularization, pretreating it with ethanol or removing the free aldehydes by citric acid treatment and lyophilization. The treated tissues were assessed for biomechanical properties, GLUT ligand quantification, adhesion, proliferation of EPCs, and biodegradability. The results indicate that there was no significant change in biomechanical properties and biodegradability when enzymatic hydrolysis (p > 0.05) is employed in detoxified acellular GLUT cross-linked tissue (DBP–G–CA–ET), compared with the native detoxified GLUT cross-linked bovine pericardium (NBP–G–CA–ET). DBP–G–CA–ET exhibited a significant (p > 0.05) increase in the viability of EPCs and cell adhesion as compared to acellular GLUT cross-linked bovine pericardium (p < 0.05). Lyophilized acellular detoxified GLUT cross-linked bovine pericardium, employed in our study as an alternative to conventional GLUT cross-linked bovine pericardium, might provide longer durability and better biocompatibility, and reduce calcification. The developed bovine pericardium patches could be used in cardiac reconstruction and repair, arteriotomy, soft tissue repair, and general surgical procedures with tissue regeneration dimensions. - Highlights: ► We improved the quality of patch biomaterial for cardiovascular surgical procedures. ► Bovine pericardium was

  16. CLMSVault: A Software Suite for Protein Cross-Linking Mass-Spectrometry Data Analysis and Visualization.

    Science.gov (United States)

    Courcelles, Mathieu; Coulombe-Huntington, Jasmin; Cossette, Émilie; Gingras, Anne-Claude; Thibault, Pierre; Tyers, Mike

    2017-07-07

    Protein cross-linking mass spectrometry (CL-MS) enables the sensitive detection of protein interactions and the inference of protein complex topology. The detection of chemical cross-links between protein residues can identify intra- and interprotein contact sites or provide physical constraints for molecular modeling of protein structure. Recent innovations in cross-linker design, sample preparation, mass spectrometry, and software tools have significantly improved CL-MS approaches. Although a number of algorithms now exist for the identification of cross-linked peptides from mass spectral data, a dearth of user-friendly analysis tools represent a practical bottleneck to the broad adoption of the approach. To facilitate the analysis of CL-MS data, we developed CLMSVault, a software suite designed to leverage existing CL-MS algorithms and provide intuitive and flexible tools for cross-platform data interpretation. CLMSVault stores and combines complementary information obtained from different cross-linkers and search algorithms. CLMSVault provides filtering, comparison, and visualization tools to support CL-MS analyses and includes a workflow for label-free quantification of cross-linked peptides. An embedded 3D viewer enables the visualization of quantitative data and the mapping of cross-linked sites onto PDB structural models. We demonstrate the application of CLMSVault for the analysis of a noncovalent Cdc34-ubiquitin protein complex cross-linked under different conditions. CLMSVault is open-source software (available at https://gitlab.com/courcelm/clmsvault.git ), and a live demo is available at http://democlmsvault.tyerslab.com/ .

  17. Covalent cross-linking of single fibers from rabbit psoas increases oscillatory power

    OpenAIRE

    Tawada, K.; Kawai, M.

    1990-01-01

    Single fibers from chemically skinned rabbit psoas muscle were treated with 1-ethyl-3-[3-dimethyl-amino)proyl]-carbodiimide (EDC) at 20 degrees C after rigor was induced. A 22-min treatment resulted in 18% covalent cross-linking between myosin heads and the thin filament as determined by stiffness measurements. This treatment also results in covalent cross-linking among rod portions of myosin molecules in the backbone of the thick filament. The fibers thus prepared are stable and do not disso...

  18. Kinetics of enzyme-catalyzed cross-linking of feruloylated arabinan from sugar beet

    DEFF Research Database (Denmark)

    Abang Zaidel, Dayang Norulfairuz; Arnous, Anis; Holck, Jesper

    2011-01-01

    the kinetics of HRP catalyzed cross-linking of FA esterified to α-(1,5)-linked arabinans are affected by the length of the arabinan chains carrying the feruloyl substitutions. The kinetics of the HRP-catalyzed cross-linking of four sets of arabinan samples from sugar beet pulp, having different molecular...... weights and hence different degrees of polymerization, were monitored by the disappearance of FA absorbance at 316 nm. MALDI-TOF/TOF-MS analysis confirmed that the sugar beet arabinans were feruloyl-substituted, and HPLC analysis verified that the amounts of diFAs increased when FA levels decreased...

  19. Cross-linked aromatic cationic polymer electrolytes with enhanced stability for high temperature fuel cell applications

    DEFF Research Database (Denmark)

    Ma, Wenjia; Zhao, Chengji; Yang, Jingshuai

    2012-01-01

    Diamine-cross-linked membranes were prepared from cross-linkable poly(arylene ether ketone) containing pendant cationic quaternary ammonium group (QPAEK) solution by a facile and general thermal curing method using 4,4′-diaminodiphenylmethane with rigid framework and 1,6-diaminohexane with flexible...... framework as cross-linker, respectively. Self-cross-linked cationic polymer electrolytes membranes were also prepared for comparison. The diamines were advantageously distributed within the polymeric matrix and its amine function groups interacted with the benzyl bromide of QPAEK, resulting in a double...

  20. Physical and mechanical properties of gamma radiation cross-linked polyethylene

    International Nuclear Information System (INIS)

    Gonzalez, Maria E.; Romero, G.; Smolko, Eduardo E.

    1999-01-01

    Granulated LDPE 2003 polyethylene was extruded and irradiated under nitrogen with 150, 200 and 300 kGy gamma rays doses to produce cross-linking. The study of the physical and mechanical properties shows that the product has a high degree of molecular cross-linking, can be heated up to 200 C for 2 hours without deformation and that the mechanical properties improve. Preliminary aging tests indicate that after heating at 60 C for 4 weeks no physical or mechanical deterioration can be observed. (author)

  1. Induction of DNA–protein cross-links by ionizing radiation and their elimination from the genome

    Energy Technology Data Exchange (ETDEWEB)

    Nakano, Toshiaki; Mitsusada, Yusuke [Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima 739-8526 (Japan); Salem, Amir M.H. [Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima 739-8526 (Japan); Department of Pathology, Medical Research Division, National Research Centre, El-Bohouth St., Dokki, Giza 12311 (Egypt); Shoulkamy, Mahmoud I. [Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima 739-8526 (Japan); Department of Zoology, Biological Science Building, Faculty of Science, Minia University, Minia 61519 (Egypt); Sugimoto, Tatsuya [Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima 739-8526 (Japan); Hirayama, Ryoichi; Uzawa, Akiko [Research Center for Charged Particle Therapy, National Institute of Radiological Sciences (NIRS), Chiba 263-8555 (Japan); Furusawa, Yoshiya [Development and Support Center, National Institute of Radiological Sciences (NIRS), Chiba 263-8555 (Japan); Ide, Hiroshi, E-mail: ideh@hiroshima-u.ac.jp [Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima 739-8526 (Japan)

    2015-01-15

    Highlights: • Normoxic and hypoxic mouse tumors were irradiated with X-rays and C-ion beams. • DNA–protein cross-links (DPCs) and DNA double-strand breaks (DSBs) were analyzed. • C-ion beams produced more DPCs than did X-rays in normoxic and hypoxic tumor cells. • DPCs were eliminated from the genome much more slowly than DSBs. • Persisting DPCs may have deleterious effects on cells in conjunction with DSBs. - Abstract: Ionizing radiation produces various types of DNA lesions, such as base damage, single-strand breaks, double-strand breaks (DSBs), and DNA–protein cross-links (DPCs). Of these, DSBs are the most critical lesions underlying the lethal effects of ionizing radiation. With DPCs, proteins covalently trapped in DNA constitute strong roadblocks to replication and transcription machineries, and hence can be lethal to cells. The formation of DPCs by ionizing radiation is promoted in the absence of oxygen, whereas that of DSBs is retarded. Accordingly, the contribution of DPCs to the lethal events in irradiated cells may not be negligible for hypoxic cells, such as those present in tumors. However, the role of DPCs in the lethal effects of ionizing radiation remains largely equivocal. In the present study, normoxic and hypoxic mouse tumors were irradiated with X-rays [low linear energy transfer (LET) radiation] and carbon (C)-ion beams (high LET radiation), and the resulting induction of DPCs and DSBs and their removal from the genome were analyzed. X-rays and C-ion beams produced more DPCs in hypoxic tumors than in normoxic tumors. Interestingly, the yield of DPCs was slightly but statistically significantly greater (1.3- to 1.5-fold) for C-ion beams than for X-rays. Both X-rays and C-ion beams generated two types of DPC that differed according to their rate of removal from the genome. This was also the case for DSBs. The half-lives of the rapidly removed components of DPCs and DSBs were similar (<1 h), but those of the slowly removed components

  2. DNA minor groove targeted alkylating agents based on bisbenzimidazole carriers: synthesis, cytotoxicity and sequence-specificity of DNA alkylation.

    Science.gov (United States)

    Smaill, J B; Fan, J Y; Denny, W A

    1998-12-01

    A series of bisbenzimidazoles bearing a variety of alkylating agents [ortho- and meta-mustards, imidazolebis(hydroxymethyl), imidazolebis(methylcarbamate) and pyrrolebis(hydroxymethyl)], appended by a propyl linker chain, were prepared and investigated for sequence-specificity of DNA alkylation and their cytotoxicity. Previous work has shown that, for para-aniline mustards, a propyl linker is optimal for cytotoxicity. Alkaline cleavage assays using a variety of different labelled oligonucleotides showed that the preferred sequences for adenine alkylation were 5'-TTTANANAANN and 5'-ATTANANAANN (underlined bases show the drug alkylation sites), with AT-rich sequences required on both the 5' and 3' sides of the alkylated adenine. The different aniline mustards showed little variation in alkylation pattern and similar efficiencies of DNA cross-link formation despite the changes in orientation and positioning of the mustard, suggesting that the propyl linker has some flexibility. The imidazole- and pyrrolebis(hydroxymethyl) alkylators showed no DNA strand cleavage following base treatment, indicating that no guanine or adenine N3 or N7 adducts were formed. Using the PCR-based polymerase stop assay, these alkylators showed PCR blocks at 5'-C*G sites (the * nucleotide indicates the blocked site), particularly at 5'-TAC*GA 5'-AGC*GGA, and 5'-AGCC*GGT sequences, caused by guanine 2-NH2 lesions on the opposite strand. Only the (more reactive) imidazolebis(methylcarbamoyl) and pyrrolebis(hydroxymethyl) alkylators demonstrated interstrand cross-linking ability. All of the bifunctional mustards showed large (approximately 100-fold) increases in cytotoxicity over chlorambucil, with the corresponding monofunctional mustards being 20- to 60-fold less cytotoxic. These results suggest that in the mustards the propyl linker provides sufficient flexibility to achieve delivery of the alkylator to favoured (adenine N3) sites in the minor groove, regardless of its exact geometry with

  3. Chloroacetamide-Linked Nucleotides and DNA for Cross-Linking with Peptides and Proteins

    Czech Academy of Sciences Publication Activity Database

    Olszewska, Agata; Pohl, Radek; Brázdová, Marie; Fojta, Miroslav; Hocek, Michal

    2016-01-01

    Roč. 27, č. 9 (2016), s. 2089-2094 ISSN 1043-1802 R&D Projects: GA ČR GBP206/12/G151 Institutional support: RVO:61388963 ; RVO:68081707 Keywords : tumor suppressor p53 * Diels-Alder reaction * reductive amination Subject RIV: CC - Organic Chemistry Impact factor: 4.818, year: 2016 http://pubs.acs.org/doi/full/10.1021/acs.bioconjchem.6b00342

  4. Constrained swelling of polymer networks: characterization of vapor-deposited cross-linked polymer thin films

    Czech Academy of Sciences Publication Activity Database

    Dušek, Karel; Choukourov, A.; Dušková-Smrčková, Miroslava; Biederman, H.

    2014-01-01

    Roč. 47, č. 13 (2014), s. 4417-4427 ISSN 0024-9297 R&D Projects: GA ČR GAP101/12/1306 Institutional support: RVO:61389013 Keywords : swelling * cross-linked polymer * elasticity Subject RIV: CD - Macromolecular Chemistry Impact factor: 5.800, year: 2014

  5. Sensitizing potential of enzymatically cross-linked peanut proteins in a mouse model of peanut allergy.

    NARCIS (Netherlands)

    Smit, Joost

    2014-01-01

    SCOPE: The cross-linking of proteins by enzymes to form high-molecular-weight protein, aggregates can be used to tailor the technological or physiological functionality of food products. Aggregation of dietary proteins by food processing may promote allergic sensitization, but the effects of

  6. Characterization and in vitro and in vivo evaluation of cross-linked ...

    Indian Academy of Sciences (India)

    2016-08-26

    Aug 26, 2016 ... Home; Journals; Bulletin of Materials Science; Volume 36; Issue 1. Characterization and in vitro and in vivo evaluation of cross-linked chitosan films as implant for controlled release of citalopram. Patit P Kundu Santosh Kumar Jindal Manish Goswami. Volume 36 Issue 1 February 2013 pp 175-182 ...

  7. Phosphoric acid doped polysulfone membranes with aminopyridine pendant groups and imidazole cross-links

    DEFF Research Database (Denmark)

    Hink, Steffen; Elsøe, Katrine; Cleemann, Lars Nilausen

    2015-01-01

    Udel polysulfone based membranes with 4-aminopyridine pendant groups and cross-linking imidazole units are synthesized in a simple two step reaction. The ratio of 4-aminopyridine and imidazole is varied and the materials are extensively characterized. The average phosphoric acid uptake (in 85 wt%...

  8. Polymers and Cross-Linking: A CORE Experiment to Help Students Think on the Submicroscopic Level

    Science.gov (United States)

    Bruce, Mitchell R. M.; Bruce, Alice E.; Avargil, Shirly; Amar, Francois G.; Wemyss, Thomas M.; Flood, Virginia J.

    2016-01-01

    The Polymers and Cross-Linking experiment is presented via a new three phase learning cycle: CORE (Chemical Observations, Representations, Experimentation), which is designed to model productive chemical inquiry and to promote a deeper understanding about the chemistry operating at the submicroscopic level. The experiment is built on two familiar…

  9. Facilitating identification of minimal protein binding domains by cross-linking mass spectrometry

    NARCIS (Netherlands)

    Liu, Qingyang; Remmelzwaal, Sanne; Heck, Albert J R; Akhmanova, Anna; Liu, Fan

    2017-01-01

    Characterization of protein interaction domains is crucial for understanding protein functions. Here we combine cross-linking mass spectrometry (XL-MS) with deletion analysis to accurately locate minimal protein interaction domains. As a proof of concept, we investigated in detail the binding

  10. Ionically cross-linked hyaluronic acid: wetting, lubrication, and viscoelasticity of a modified adhesion barrier gel

    Directory of Open Access Journals (Sweden)

    Katherine Vorvolakos

    2010-12-01

    Full Text Available Katherine Vorvolakos1, Irada S Isayeva1, Hoan-My Do Luu1, Dinesh V Patwardhan1, Steven K Pollack21Division of Chemistry and Material Science, 2Office of Science and Engineering Laboratories, Center for Devices and Radiological Health, Food and Drug Administration, Silver Spring, MD, USAAbstract: Hyaluronic acid (HA, in linear or cross-linked form, is a common component of cosmetics, personal care products, combination medical products, and medical devices. In all cases, the ability of the HA solution or gel to wet surfaces and/or disrupt and lubricate interfaces is a limiting feature of its mechanism of action. We synthesized ferric ion–cross-linked networks of HA based on an adhesion barrier, varied the degree of cross-linking, and performed wetting goniometry, viscometry, and dynamic mechanical analysis. As cross-linking increases, so do contact angle, viscosity, storage modulus, and loss modulus; thus, wetting and lubrication are compromised. These findings have implications in medical device materials, such as adhesion barriers and mucosal drug delivery vehicles.Keywords: hyaluron, adhesion barrier, wetting, contact angle, viscosity, lubrication, elasticity, viscoelastic, hydrogel, ferric

  11. Non-volatile transistor memory devices using charge storage cross-linked core-shell nanoparticles.

    Science.gov (United States)

    Lo, Chen-Tsyr; Watanabe, Yu; Oya, Hiroshi; Nakabayashi, Kazuhiro; Mori, Hideharu; Chen, Wen-Chang

    2016-06-07

    Solution processable cross-linked core-shell poly[poly(ethylene glycol)methylether methacrylate]-block-poly(2,5-dibromo-3-vinylthiophene) (poly(PEGMA)m-b-poly(DB3VT)n) nanoparticles are firstly explored as charge storage materials for transistor-type memory devices owing to their efficient and controllable ability in electric charge transfer and trapping.

  12. Thermogelling and chemoselectively cross-linked hydrogels with controlled mechanical properties and degradation behavior

    NARCIS (Netherlands)

    Boere, Kristel W M; Van Den Dikkenberg, Joep; Gao, Yuan; Visser, Jetze; Hennink, Wim E.; Vermonden, Tina

    2015-01-01

    Chemoselectively cross-linked hydrogels have recently gained increasing attention for the development of novel, injectable biomaterials given their limited side reactions. In this study, we compared the properties of hydrogels obtained by native chemical ligation (NCL) and its recently described

  13. Cross-Linking Treatment and Corneal Transplant in Refractory Acremonium Keratitis: Case Report.

    Science.gov (United States)

    Yagci, Ayse; Palamar, Melis; Polat Hilmioglu, Suleyha; Irkec, Murat

    2016-10-01

    To report a case of Acremonium keratitis treated with voriconazole, corneal collagen cross-linking, and corneal transplant. Case report. A 42-year-old woman who wore contact lenses daily was referred for refractory keratitis. Her main complaints were gritty sensation and pain. At slit lamp biomicroscopy, an infiltrate on the inferior paracentral cornea and an arcuate conjunctival ulceration were evident. The rest of the cornea was clear with no anterior chamber reaction. Scrapings from the corneal ulcer showed Candida parapsilosis and Acremonium species, which were sensitive to voriconazole. Despite the administration of topical, systemic, and intrastromal voriconazole for 1 month, repeat corneal scraping was positive for Acremonium, and clinical appearance and pain did not resolve. Therefore, corneal collagen cross-linking was applied. Although the pain resolved immediately after the procedure, the lesion showed no improvement. After a month of cross-linking, corneal transplant was performed for visual rehabilitation and treatment of the refractory lesion. The excised corneal button was negative for any microorganisms. Although corneal collagen cross-linking may be an effective treatment for Acremonium keratitis refractory to medical therapy, corneal transplant was required for visual gain and recovery.

  14. Identification of Pyridinoline Trivalent Collagen Cross-Links by Raman Microspectroscopy.

    Science.gov (United States)

    Gamsjaeger, Sonja; Robins, Simon P; Tatakis, Dimitris N; Klaushofer, Klaus; Paschalis, Eleftherios P

    2017-06-01

    Intermolecular cross-linking of bone collagen is intimately related to the way collagen molecules are arranged in a fibril, imparts certain mechanical properties to the fibril, and may be involved in the initiation of mineralization. Raman microspectroscopy allows the analysis of minimally processed bone blocks and provides simultaneous information on both the mineral and organic matrix (mainly type I collagen) components, with a spatial resolution of ~1 μm. The aim of the present study was to validate Raman spectroscopic parameters describing one of the major mineralizing type I trivalent cross-links, namely pyridinoline (PYD). To achieve this, a series of collagen cross-linked peptides with known PYD content (as determined by HPLC analysis), human bone, porcine skin, predentin and dentin animal model tissues were analyzed by Raman microspectroscopy. The results of the present study confirm that it is feasible to monitor PYD trivalent collagen cross-links by Raman spectroscopic analysis in mineralized tissues, exclusively through a Raman band ~1660 wavenumbers. This allows determination of the relative PYD content in undecalcified bone tissues with a spatial resolution of ~1 μm, thus enabling correlations with histologic and histomorphometric parameters.

  15. Cross-linked graphene oxide sheets via modified extracted cellulose with high metal adsorption.

    Science.gov (United States)

    Yakout, Amr A; El-Sokkary, Ramadan H; Shreadah, Mohamed A; Abdel Hamid, Omnia G

    2017-09-15

    We have studied the extraction of Cu(II) and Pb(II) ions from different types of aqueous solution by novel cross-linked graphene oxide sheets by modified extracted cellulose. The novel sorbent cellulose was extracted from the mangrove trees (Avicennia marina) and it was then grafted with acrylamide and immobilized by ethylenediamine for cross-linking process. The cross-linked graphene oxide sheets were identified by means of FT-IR, SEM and XRD. The adsorption studies of synthesized sorbent was optimized. Langmuir and Freundlich models were used for establish sorption equilibria. The cross-linked graphene oxide sheets showed maximum adsorption capacity 46.39 and 186.48mgg -1 for Cu(II) and Pb(II), respectively. The potential applications of this sorbent was applied to remove Cu(II) and Pb(II) metal ions from hard water samples by using a multi-stage micro-column technique. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Gelatin-glutaraldehyde cross-linking on silicone rubber to increase endothelial cell adhesion and growth.

    Science.gov (United States)

    Ai, Hua; Mills, David K; Jonathan, Alexander S; Jones, Steven A

    2002-10-01

    Silicone is a biomaterial that is widely used in many areas because of its high optical clarity, its durability, and the ease with which it can be cast. However, these advantages are counterbalanced by strong hydrophobicity. Gelatin cross-linking has been used as a hydrophilic coating on many biomaterials but not on silicone rubber. In this study, two gelatin glutaraldehyde (GA) cross-linking methods were used to coat a hydrophilic membrane on silicone rubber. In method I, gelatin and GA were mixed in three different proportions (64:1, 128:1, and 256:1) before coating. In method II, a newly formed 5% gelatin membrane was cross-linked with a 2.5% GA solution. All coatings were hydrophilic, as determined from the measurement of contact angle for a drop of water on the surface. Bovine coronary arterial endothelial cells were shown to grow well on the surface modified by method II at 72 h. In method I, the cells grew well for gelatin-GA proportions of 64:1 and 128:1 at 72 h. No cell attachment on untreated silicone rubber was observed by the third d of seeding. The results indicated that both methods of gelatin-GA cross-linking provided a hydrophilic surface on silicone for endothelial cell adhesion and growth in vitro.

  17. Substituent distribution within cross-linked and hydroxypropylated sweet potato starch and potato starch

    NARCIS (Netherlands)

    Zhao, J.; Schols, H.A.; Chen Zenghong,; Jin Zhengyu,; Buwalda, P.L.; Gruppen, H.

    2012-01-01

    Revealing the substituents distribution within starch can help to understand the changes of starch properties after modification. The distribution of substituents over cross-linked and hydroxypropylated sweet potato starch was investigated and compared with modified potato starch. The starches were

  18. Biocatalytic cross-linking of pectic polysaccharides for designed food functionality

    DEFF Research Database (Denmark)

    Zaidel, Dayang Norulfairuz Abang; Meyer, Anne S.

    2012-01-01

    Recent research has demonstrated how cross-linking of pectic polysaccharides to obtain gel formation can be promoted by enzymatic catalysis reactions, and provide opportunities for functional upgrading of pectic polysaccharides present in agro-industrial sidestreams. This review highlights the me...

  19. Cross-linking methods of electrospun fibrinogen scaffolds for tissue engineering applications

    Energy Technology Data Exchange (ETDEWEB)

    Sell, Scott A; Garg, Koyal; McClure, Michael J; Bowlin, Gary L [Department of Biomedical Engineering, Virginia Commonwealth University, Richmond, VA 23284-3067 (United States); Francis, Michael P [Department of Pathology, Virginia Commonwealth University, Richmond, VA 23298-0709 (United States); Simpson, David G [Department of Anatomy and Neurobiology, Virginia Commonwealth University, Richmond, VA 23298-0709 (United States)], E-mail: glbowlin@vcu.edu

    2008-12-15

    The purpose of this study was to enhance the mechanical properties and slow the degradation of an electrospun fibrinogen scaffold, while maintaining the scaffold's high level of bioactivity. Three different cross-linkers were used to achieve this goal: glutaraldehyde vapour, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) in ethanol and genipin in ethanol. Scaffolds with a fibrinogen concentration of 120 mg ml{sup -1} were electrospun and cross-linked with one of the aforementioned cross-linkers. Mechanical properties were determined through uniaxial tensile testing performed on scaffolds incubated under standard culture conditions for 1 day, 7 days and 14 days. Cross-linked scaffolds were seeded with human foreskin fibroblasts (BJ-GFP-hTERT) and cultured for 7, 14 and 21 days, with histology and scanning electron microscopy performed upon completion of the time course. Mechanical testing revealed significantly increased peak stress and modulus values for the EDC and genipin cross-linked scaffolds, with significantly slowed degradation. However, cross-linking with EDC and genipin was shown to have some negative effect on the bioactivity of the scaffolds as cell migration throughout the thickness of the scaffold was slowed.

  20. Baking Performance of Phosphorylated Cross-Linked Resistant Starch in Low-Moisture Bakery Goods

    Science.gov (United States)

    Phosphorylated cross-linked resistant starch (RS) is a type 4 RS, which can be used for enhancing the benefits of dietary fiber. The baking performance of the RS was explored using wire-cut cookie baking and benchtop chemically-leavened cracker baking methods to produce low-moisture baked goods (coo...

  1. Design and Preparation of Cross-Linked Polystyrene Nanoparticles for Elastomer Reinforcement

    Directory of Open Access Journals (Sweden)

    Ming Lu

    2010-01-01

    Full Text Available Cross-linked polystyrene (PS particles in a latex form were synthesized by free radical emulsion polymerization. The nano-PS-filled elastomer composites were prepared by the energy-saving latex compounding method. Results showed that the PS particles took a spherical shape in the size of 40–60 nm with a narrow size distribution, and the glass-transition temperature of the PS nanoparticles increased with the cross-linking density. The outcomes from the mechanical properties demonstrated that when filled into styrene-butadiene rubber (SBR, nitrile-butadiene rubber (NBR, and natural rubber (NR, the cross-linked PS nano-particles exhibited excellent reinforcing capabilities in all the three matrices, and the best in the SBR matrix. In comparison with that of the carbon black filled composites, another distinguished advantage of the cross-linked PS particles filled elastomer composites was found to be light weight in density, which could help to save tremendous amount of energy when put into end products.

  2. Preparation of Nanocellulose Reinforced Chitosan Films, Cross-Linked by Adipic Acid.

    Science.gov (United States)

    Falamarzpour, Pouria; Behzad, Tayebeh; Zamani, Akram

    2017-02-13

    Adipic acid, an abundant and nontoxic compound, was used to dissolve and cross-link chitosan. After the preparation of chitosan films through casting technique, the in situ amidation reaction was performed at 80-100 °C as verified by Fourier transform infrared (FT-IR). The reaction was accompanied by the release of water which was employed to investigate the reaction kinetics. Accordingly, the reaction rate followed the first-order model and Arrhenius equation, and the activation energy was calculated to be 18 kJ/mol. Furthermore, the mechanical properties of the chitosan films were comprehensively studied. First, optimal curing conditions (84 °C, 93 min) were introduced through a central composite design. In order to evaluate the effects of adipic acid, the mechanical properties of physically cross-linked (uncured), chemically cross-linked (cured), and uncross-linked (prepared by acetic acid) films were compared. The use of adipic acid improved the tensile strength of uncured and chemically cross-linked films more than 60% and 113%, respectively. Finally, the effect of cellulose nanofibrils (CNFs) on the mechanical performance of cured films, in the presence of glycerol as a plasticizer, was investigated. The plasticized chitosan films reinforced by 5 wt % CNFs showed superior properties as a promising material for the development of chitosan-based biomaterials.

  3. Mocaf cross-linking with gluten to improve the quality of mocaf dough

    Science.gov (United States)

    Raharja, Sapta; Udin, Faqih; Suparno, Ono; Febrianti, Faricha Helfi; Nuraisyah, Ani

    2017-03-01

    Crosslink between mocaf and gluten is conducted to increase the using of mocaf which has very big potential in Indonesia. The effort of cross-linking between mocaf and gluten is to get mocaf flour with better dough quality. This study aims to produce a cross-linked mocaf-gluten flour and to evaluate the influence of heating temperature (X1) and the addition of gluten concentration (X2) using completely randomized design factorial (RAFL). The cross-linking is carried out in alkaline solution with 10%, 20%, and 30% gluten addition and heating temperature at 50, 55, and 60 °C. The result showed that mocaf - gluten flour with the treatment of 30% gluten addition at 55 °C had the largest amount of protein and baking expansion (i.e 19.77% and 2.78 mL/g). Swelling power of the flour was increasing along with the increasing of water absorbing capacity of the mocaf - gluten flour. Birefringence properties of mocaf - gluten flour tended to be reduced as the increasing heating temperature. FTIR analysis of mocaf - gluten flour showed that there was peak strengthening of the infrared spectrum of the C - N bond at 1167-1159 cm-1 which was presumably resulted from the gluten addition and the cross-linking properties.

  4. Hygral Behavior of Superabsorbent Polymers with Various Particle Sizes and Cross-Linking Densities

    Directory of Open Access Journals (Sweden)

    Kyong-Ku Yun

    2017-11-01

    Full Text Available This study focuses on investigating the effects of particle size and cross-linking density on the hygral behavior of superabsorbent polymers (SAPs, which are increasingly used as an internal curing material for high-performance concrete. Four SAPs with different mean particle diameters and cross-linking densities were tested under controlled wetting and drying conditions to measure free absorption and desorption kinetics. Absorption capacities of SAPs under actual mixing conditions were additionally measured and verified by means of mortar flow and semi-adiabatic hydration heat measurements. In addition, the effects of SAP type and dosage (i.e., 0.2, 0.4, and 0.6% by mass of cement on the mechanical properties of hardened mortar were assessed. The results indicated that: (1 the absorption capacity increased with decreased cross-linking density and increased particle size under both load-free and mixing conditions; and (2 the greater the cross-linking density and the lower the particle size, the shorter the desorption time. It was also confirmed that while the early-age mechanical properties were more related with the gel strength of swollen SAP, the later-age mechanical properties were more affected by the water retention capacity and spatial distribution of SAP in the matrix.

  5. TGFβ affects collagen cross-linking independent of chondrocyte phenotype but strongly depending on physical environment

    NARCIS (Netherlands)

    Bastiaansen-Jenniskens, Y.M.; Koevoet, W.; Bart, A.C.W. de; Zuurmond, A.-M.; Bank, R.A.; Verhaar, J.A.N.; Groot, J. de; Osch, G.J.V.M. van

    2008-01-01

    Transforming growth factor beta (TGFβ) is often used in cartilage tissue engineering to increase matrix formation by cells with various phenotypes. However, adverse effects of TGFβ, such as extensive cross-linking in cultured fibroblasts, have also been reported. Our goal was to study effects of

  6. A minimal model for stabilization of biomolecules by hydrocarbon cross-linking

    Science.gov (United States)

    Hamacher, K.; Hübsch, A.; McCammon, J. A.

    2006-04-01

    Programmed cell death regulating protein motifs play an essential role in the development of an organism, its immune response, and disease-related cellular mechanisms. Among those motifs the BH3 domain of the BCL-2 family is found to be of crucial importance. Recent experiments showed how the isolated, otherwise unstructured BH3 peptide can be modified by a hydrocarbon linkage to regain function. We parametrized a reduced, dynamic model for the stability effects of such covalent cross-linking and confirmed that the model reproduces the reinforcement of the structural stability of the BH3 motif by cross-linking. We show that an analytically solvable model for thermostability around the native state is not capable of reproducing the stabilization effect. This points to the crucial importance of the peptide dynamics and the fluctuations neglected in the analytic model for the cross-linking system to function properly. This conclusion is supported by a thorough analysis of a simulated Gō model. The resulting model is suitable for rational design of generic cross-linking systems in silicio.

  7. Interactions of cross-linked and uncross-linked chitosan hydrogels ...

    African Journals Online (AJOL)

    The swelling equilibrium of Chitosan and sodium tripolyphosphate (NaTPP) cross-linked chitosan hydrogels in aqueous solutions of surfactants differing in structure and hydrophobicity at 250C is reported. Anionic surfactant sodium dodecylsulfate (SDS), the cationic surfactant hexadecyltrimethylammonium bromide (HTAB) ...

  8. Characterization and in vitro and in vivo evaluation of cross-linked ...

    Indian Academy of Sciences (India)

    suggested that cross-linked CH films of above said composition can be used as implant for long term application in depression and related disorders. Keywords. Chitosan film; citalopram; physical properties; mechanical properties; in vitro drug release. 1. Introduction. Depression and related disorders like schizophrenia, ...

  9. Mitochondrial protein interactome elucidated by chemical cross-linking mass spectrometry.

    Science.gov (United States)

    Schweppe, Devin K; Chavez, Juan D; Lee, Chi Fung; Caudal, Arianne; Kruse, Shane E; Stuppard, Rudy; Marcinek, David J; Shadel, Gerald S; Tian, Rong; Bruce, James E

    2017-02-14

    Mitochondrial protein interactions and complexes facilitate mitochondrial function. These complexes range from simple dimers to the respirasome supercomplex consisting of oxidative phosphorylation complexes I, III, and IV. To improve understanding of mitochondrial function, we used chemical cross-linking mass spectrometry to identify 2,427 cross-linked peptide pairs from 327 mitochondrial proteins in whole, respiring murine mitochondria. In situ interactions were observed in proteins throughout the electron transport chain membrane complexes, ATP synthase, and the mitochondrial contact site and cristae organizing system (MICOS) complex. Cross-linked sites showed excellent agreement with empirical protein structures and delivered complementary constraints for in silico protein docking. These data established direct physical evidence of the assembly of the complex I-III respirasome and enabled prediction of in situ interfacial regions of the complexes. Finally, we established a database and tools to harness the cross-linked interactions we observed as molecular probes, allowing quantification of conformation-dependent protein interfaces and dynamic protein complex assembly.

  10. Effect of dual modification with hydroxypropylation and cross-linking on physicochemical properties of taro starch.

    Science.gov (United States)

    Hazarika, Bidyut Jyoti; Sit, Nandan

    2016-04-20

    Dual modification of taro starch by hydroxypropylation and cross-linking was carried out and the properties of the modified starches were investigated. Two different levels of hydroxypropylation (5 and 10%) and cross-linking (0.05 and 0.10%) were used in different sequences. The amylose contents of the starch decreased due to single and dual modification. For the dual-modified starches, the swelling, solubility and clarity was found to increase with level of hydroxypropylation and decrease with level of cross-linking. The freeze-thaw stability of the dual-modified starches was also affected by the sequence of modification. The viscosities of the cross-linked and dual-modified starches were more than native and hydroxypropylated starches. The firmness of the dual-modified starches was also higher than native and single modified starches. The dual-modified starches have benefits of both type of modifications and could be used for specific purposes e.g. food products requiring high viscosity as well as freeze-thaw stability. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. ROMP-based thermosetting polymers from modified castor oil with various cross-linking agents

    Science.gov (United States)

    Ding, Rui

    Polymers derived from bio-renewable resources are finding an increase in global demand. In addition, polymers with distinctive functionalities are required in certain advanced fields, such as aerospace and civil engineering. In an attempt to meet both these needs, the goal of this work aims to develop a range of bio-based thermosetting matrix polymers for potential applications in multifunctional composites. Ring-opening metathesis polymerization (ROMP), which recently has been explored as a powerful method in polymer chemistry, was employed as a unique pathway to polymerize agricultural oil-based reactants. Specifically, a novel norbornyl-functionalized castor oil alcohol (NCA) was investigated to polymerize different cross-linking agents using ROMP. The effects of incorporating dicyclopentadiene (DCPD) and a norbornene-based crosslinker (CL) were systematically evaluated with respect to curing behavior and thermal mechanical properties of the polymers. Isothermal differential scanning calorimetry (DSC) was used to investigate the conversion during cure. Dynamic DSC scans at multiple heating rates revealed conversion-dependent activation energy by Ozawa-Flynn-Wall analysis. The glass transition temperature, storage modulus, and loss modulus for NCA/DCPD and NCA/CL copolymers with different cross-linking agent loading were compared using dynamic mechanical analysis. Cross-link density was examined to explain the very different dynamic mechanical behavior. Mechanical stress-strain curves were developed through tensile test, and thermal stability of the cross-linked polymers was evaluated by thermogravimetric analysis to further investigate the structure-property relationships in these systems.

  12. EGDMA–cross-linked polystyrene resin: An efficient support for gel ...

    Indian Academy of Sciences (India)

    cross-linked polystyrene resin: An efficient support for gel phase peptide synthesis. P K Ajikumar K S ... EGDMA-PS resin undergoes facile swelling in a variety of solvents, both polar and nonpolar, used in peptide synthesis. The polymer was ...

  13. Highly conductive carbon nanotube buckypapers with improved doping stability via conjugational cross-linking.

    Science.gov (United States)

    Chen, I-Wen Peter; Liang, Richard; Zhao, Haibo; Wang, Ben; Zhang, Chuck

    2011-12-02

    Carbon nanotube (CNT) sheets or buckypapers have demonstrated promising electrical conductivity and mechanical performance. However, their electrical conductivity is still far below the requirements for engineering applications, such as using as a substitute for copper mesh, which is currently used in composite aircraft structures for lightning strike protection. In this study, different CNT buckypapers were stretched to increase their alignment, and then subjected to conjugational cross-linking via chemical functionalization. The conjugationally cross-linked buckypapers (CCL-BPs) demonstrated higher electrical conductivity of up to 6200 S cm( - 1), which is more than one order increase compared to the pristine buckypapers. The CCL-BPs also showed excellent doping stability in over 300 h in atmosphere and were resistant to degradation at elevated temperatures. The tensile strength of the stretched CCL-BPs reached 220 MPa, which is about three times that of pristine buckypapers. We attribute these property improvements to the effective and stable conjugational cross-links of CNTs, which can simultaneously improve the electrical conductivity, doping stability and mechanical properties. Specifically, the electrical conductivity increase resulted from improving the CNT alignment and inter-tube electron transport capability. The conjugational cross-links provide effective 3D conductive paths to increase the mobility of electrons among individual nanotubes. The stable covalent bonding also enhances the thermal stability and load transfer. The significant electrical and mechanical property improvement renders buckypaper a multifunctional material for various applications, such as conducting composites, battery electrodes, capacitors, etc.

  14. Characterization of the network structure of carbodiimide cross-linked gelatin gels

    NARCIS (Netherlands)

    Kuijpers, A. J.; Engbers, G. H. M.; Feijen, J.; de Smedt, S. C.; Meyvis, T. K. L.; Demeester, J.; Krijgsveld, J.; Zaat, S. A. J.; Dankert, J.

    1999-01-01

    The network structure of native and carbodiimide cross-linked gelatin A and B gels was studied based on their rheological behavior. Gelatin A and B contain different numbers of carboxylic acid groups caused by different preparation conditions and had previously shown different characteristics in

  15. Cross-linking methods of electrospun fibrinogen scaffolds for tissue engineering applications

    International Nuclear Information System (INIS)

    Sell, Scott A; Garg, Koyal; McClure, Michael J; Bowlin, Gary L; Francis, Michael P; Simpson, David G

    2008-01-01

    The purpose of this study was to enhance the mechanical properties and slow the degradation of an electrospun fibrinogen scaffold, while maintaining the scaffold's high level of bioactivity. Three different cross-linkers were used to achieve this goal: glutaraldehyde vapour, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) in ethanol and genipin in ethanol. Scaffolds with a fibrinogen concentration of 120 mg ml -1 were electrospun and cross-linked with one of the aforementioned cross-linkers. Mechanical properties were determined through uniaxial tensile testing performed on scaffolds incubated under standard culture conditions for 1 day, 7 days and 14 days. Cross-linked scaffolds were seeded with human foreskin fibroblasts (BJ-GFP-hTERT) and cultured for 7, 14 and 21 days, with histology and scanning electron microscopy performed upon completion of the time course. Mechanical testing revealed significantly increased peak stress and modulus values for the EDC and genipin cross-linked scaffolds, with significantly slowed degradation. However, cross-linking with EDC and genipin was shown to have some negative effect on the bioactivity of the scaffolds as cell migration throughout the thickness of the scaffold was slowed.

  16. A general method for targeted quantitative cross-linking mass spectrometry

    Science.gov (United States)

    Chemical cross-linking mass spectrometry (XL-MS) provides protein structural information by identifying covalently linked proximal amino acid residues on protein surfaces. The information gained by this technique is complementary to other structural biology methods such as x-ray crystallography, NM...

  17. Pickering emulsions stabilized by whey protein nanoparticles prepared by thermal cross-linking

    NARCIS (Netherlands)

    Wu, Jiande; Shi, Mengxuan; Li, Wei; Zhao, Luhai; Wang, Ze; Yan, Xinzhong; Norde, Willem; Li, Yuan

    2015-01-01

    A Pickering (o/w) emulsion was formed and stabilized by whey protein isolate nanoparticles (WPI NPs). Those WPI NPs were prepared by thermal cross-linking of denatured WPI proteins within w/o emulsion droplets at 80. °C for 15. min. During heating of w/o emulsions containing 10% (w/v) WPI

  18. Core-cross-linked polymeric micelles: a versatile nanomedicine platform with broad applicability

    NARCIS (Netherlands)

    Hu, Q.

    2015-01-01

    This dissertation addresses the broad applicability of the nanomedicine platform core-cross-linked polymeric micelles (CCL-PMs) composed of thermosensitive mPEG-b-pHPMAmLacn block copolymers. In Chapter 1, a general introduction to nanomedicines is provided, with a particular focus on polymeric

  19. Protective effect of Withania somnifera (Solanaceae) on collagen glycation and cross-linking.

    Science.gov (United States)

    Babu, Pon Velayutham Anandh; Gokulakrishnan, Adikesavan; Dhandayuthabani, Rajendra; Ameethkhan, Dowlath; Kumar, Chandrasekara Vimal Pradeep; Ahamed, Md Iqbal Niyas

    2007-06-01

    Modification of collagen such as non-enzymatic glycation and cross-linking plays an important role in diabetic complications and age-related diseases. We evaluate the effect of Withania somnifera on glucose-mediated collagen glycation and cross-linking in vitro. Extent of glycation, viscosity, collagen-linked fluorescence and pepsin solubility were assessed in different experimental procedures to investigate the effect of W. somnifera. Tail tendons obtained from rats (Rattus norvegicus) weighing 250-275 g were incubated with 50 mM glucose and 100 mg of metformin or Withania root powder or ethanolic extract of Withania under physiological conditions of temperature and pH for 30 days. Formation of advanced glycation end products (AGE) was measured by fluorescent method whereas the cross-linking of collagen was assessed by pepsin digestion and viscosity measurements. Tendon collagen incubated with glucose showed an increase in glycation, AGE and cross-linking of collagen. The collagen incubated with W. somnifera and metformin ameliorates these modifications. The ethanolic extract of Withania showed more prominent effect than Withania root powder. The activity of ethanolic extract of Withania is comparable to metformin, a known antiglycating agent. In conclusion, Withania could have therapeutic role in the prevention of glycation induced pathogenesis in diabetes mellitus and aging.

  20. Preparation of Nanocellulose Reinforced Chitosan Films, Cross-Linked by Adipic Acid

    Directory of Open Access Journals (Sweden)

    Pouria Falamarzpour

    2017-02-01

    Full Text Available Adipic acid, an abundant and nontoxic compound, was used to dissolve and cross-link chitosan. After the preparation of chitosan films through casting technique, the in situ amidation reaction was performed at 80–100 °C as verified by Fourier transform infrared (FT-IR. The reaction was accompanied by the release of water which was employed to investigate the reaction kinetics. Accordingly, the reaction rate followed the first-order model and Arrhenius equation, and the activation energy was calculated to be 18 kJ/mol. Furthermore, the mechanical properties of the chitosan films were comprehensively studied. First, optimal curing conditions (84 °C, 93 min were introduced through a central composite design. In order to evaluate the effects of adipic acid, the mechanical properties of physically cross-linked (uncured, chemically cross-linked (cured, and uncross-linked (prepared by acetic acid films were compared. The use of adipic acid improved the tensile strength of uncured and chemically cross-linked films more than 60% and 113%, respectively. Finally, the effect of cellulose nanofibrils (CNFs on the mechanical performance of cured films, in the presence of glycerol as a plasticizer, was investigated. The plasticized chitosan films reinforced by 5 wt % CNFs showed superior properties as a promising material for the development of chitosan-based biomaterials.

  1. Isolation and removal of proteolytic enzymes with magnetic cross-linked erythrocytes

    International Nuclear Information System (INIS)

    Safarik, I. Ivo; Safarikova, Mirka

    2001-01-01

    New magnetic adsorbents for batch isolation and removal of various proteolytic enzymes were prepared by glutaraldehyde cross-linking of bovine, porcine and human erythrocytes in the presence of fine magnetic particles. Trypsin, chymotrypsin, alkaline bacterial protease and proteases present in various commercial enzyme preparations were efficiently adsorbed on these adsorbents; on the contrary, proteins without proteolytic activity were not adsorbed

  2. Free-radical-induced chain scission and cross-linking of polymers in aqueous solution. An overview

    International Nuclear Information System (INIS)

    Von Sonntag, C.

    2002-01-01

    Complete text of publication follows. In the radiolysis of N 2 O-saturated aqueous solutions OH are generated. In their reactions with polymers, they give rise to polymer-derived radicals. The kinetics of the formation and decay of these radicals are reviewed. The rate of reaction of a polymer with a reactive free radical is noticeably lower than that of an equivalent concentration of monomer due to the non-random distribution of the reaction sites. Once a larger number of radicals are formed on one polymer molecule, e.g. upon pulse radiolysis, close-by radicals recombine more rapidly while the more distant ones survive for much longer times than an equivalent concentration of freely diffusing radicals. Intermolecular cross-linking (between two polymer chains, increase in molecular weight) and intramolecular cross-linking (formation of small loops, no increase in polymer weight) are competing processes, and their relative yields thus depend on the dose rate and polymer concentration. Hydrogen-transfer reactions within the polymer, e.g. transformation of a secondary radical into a tertiary one, are common and facilitated by the high local density of reactive sites. Due to repulsive forces, the lifetime of radicals of charged polymers is substantially increased. This enables even relatively slow b-fragmentation reactions to become of importance. In the case of poly(methacrylic acid), where β-fragmentation is comparatively fast, this even leads to an unzipping, and as a consequence of the efficient release of methacrylic acid the situation of equilibrium polymerization is approached. Heterolytic β-fragmentation is possible when adequate leaving groups are available, e.g. in polynucleotides and DNA. In the presence of O 2 , chain scission occurs via oxyl radicals as intermediates. Some implications for technical applications are discussed

  3. Thermogelling and Chemoselectively Cross-Linked Hydrogels with Controlled Mechanical Properties and Degradation Behavior.

    Science.gov (United States)

    Boere, Kristel W M; van den Dikkenberg, Joep; Gao, Yuan; Visser, Jetze; Hennink, Wim E; Vermonden, Tina

    2015-09-14

    Chemoselectively cross-linked hydrogels have recently gained increasing attention for the development of novel, injectable biomaterials given their limited side reactions. In this study, we compared the properties of hydrogels obtained by native chemical ligation (NCL) and its recently described variation termed oxo-ester-mediated native chemical ligation (OMNCL) in combination with temperature-induced physical gelation. Triblock copolymers consisting of cysteine functionalities, thermoresponsive N-isopropylacrylamide (NIPAAm) units and degradable moieties were mixed with functionalized poly(ethylene glycol) (PEG) cross-linkers. Thioester or N-hydroxysuccinimide (NHS) functionalities attached to PEG reacted with cysteine residues of the triblock copolymers via either an NCL or OMNCL pathway. The combined physical and chemical cross-linking resulted in rapid network formation and mechanically strong hydrogels. Stiffness of the hydrogels was highest for thermogels that were covalently linked via OMNCL. Specifically, the storage modulus after 4 h reached a value of 26 kPa, which was over a 100 times higher than hydrogels formed by solely thermal physical interactions. Endothelial cells showed high cell viability of 98 ± 2% in the presence of OMNCL cross-linked hydrogels after 16 h of incubation, in contrast to a low cell viability (13 ± 7%) for hydrogels obtained by NCL cross-linking. Lysozyme was loaded in the gels and after 2 days more than 90% was released, indicating that the cross-linking reaction was indeed chemoselective as the protein was not covalently grafted to the hydrogel network. Moreover, the degradation rates of these hydrogels under physiological conditions could be tailored from 12 days up to 6 months by incorporation of a monomer containing a hydrolyzable lactone ring in the thermosensitive triblock copolymer. These results demonstrate a high tunability of mechanical properties and degradation rates of these in situ forming hydrogels that could be

  4. Evaluation of the efficacy of excimer laser ablation of cross-linked porcine cornea.

    Directory of Open Access Journals (Sweden)

    Shihao Chen

    Full Text Available BACKGROUND: Combination of riboflavin/UVA cross-linking (CXL and excimer laser ablation is a promising therapy for treating corneal ectasia. The cornea is strengthened by cross-linking, while the irregular astigmatism is reduced by laser ablation. This study aims to compare the efficacy of excimer laser ablation on porcine corneas with and without cross-linking. METHODS AND FINDINGS: The porcine cornea was de-epithelialized and treated with 0.1% riboflavin solution for 30 minutes. A half of the cornea was exposed to UVA-radiation for another 30 minutes while the controlled half of the cornea was protected from the UVA using a metal shield. Photo therapeutic keratectomy (PTK was then performed on the central cornea. Corneal thickness of 5 paired locations on the horizontal line, ± 0.5, ± 1.0, ± 1.5, ± 2.0, and ± 2.5 mm from the central spot, were measured using optical coherence tomography prior to and after PTK. The ablation depth was then determined by the corneal thickness. There was a 9% difference (P<0.001 in the overall ablation depth between the CXL-half corneas (158 ± 22 µm and the control-half corneas (174 ± 26 µm. The ablation depths of all 5 correspondent locations on the CXL-half were significantly smaller (P<0.001. CONCLUSION: The efficacy of the laser ablation seems to be lower in cross-linked cornea. Current ablation algorithms may need to be modified for cross-linked corneas.

  5. Acoustic radiation force for noninvasive evaluation of corneal biomechanical changes induced by cross-linking therapy.

    Science.gov (United States)

    Urs, Raksha; Lloyd, Harriet O; Silverman, Ronald H

    2014-08-01

    To noninvasively measure changes in corneal biomechanical properties induced by ultraviolet-activated riboflavin cross-linking therapy using acoustic radiation force (ARF). Cross-linking was performed on the right eyes of 6 rabbits, with the left eyes serving as controls. Acoustic radiation force was used to assess corneal stiffness before treatment and weekly for 4 weeks after treatment. Acoustic power levels were within US Food and Drug Administration guidelines for ophthalmic safety. Strain, determined from ARF-induced displacement of the front and back surfaces of the cornea, was fit to the Kelvin-Voigt model to determine the elastic modulus (E) and coefficient of viscosity (η). The stiffness factor, the ratio of E after treatment to E before treatment, was calculated for treated and control eyes. At the end of 4 weeks, ex vivo thermal shrinkage temperature analysis was performed for comparison with in vivo stiffness measurements. One-way analysis of variance and Student t tests were performed to test for differences in E, η, the stiffness factor, and corneal thickness. Biomechanical stiffening was immediately evident in cross-linking-treated corneas. At 4 weeks after treatment, treated corneas were 1.3 times stiffer and showed significant changes in E (P= .006) and η (P= .007), with no significant effect in controls. Corneal thickness increased immediately after treatment but did not differ significantly from the pretreatment value at 4 weeks. Our findings demonstrate a statistically significant increase in stiffness in cross-linking-treated rabbit corneas based on in vivo axial stress/strain measurements obtained using ARF. The capacity to noninvasively monitor corneal stiffness offers the potential for clinical monitoring of cross-linking therapy. © 2014 by the American Institute of Ultrasound in Medicine.

  6. Sodium tripolyphosphate cross-linked chitosan based sensor for enhacing sensing properties towards acetone

    Science.gov (United States)

    Nasution, T. I.; Asrosa, R.; Nainggolan, I.; Balyan, M.; Indah, R.; Wahyudi, A.

    2018-02-01

    In this report, sensing properties of sodium tripolyphosphate (TPP) cross-linked chitosan based sensor has been successfully enhanced towards acetone. Chitosan solutions were cross-linked with sodium TPP in variation of 0.1%, 0.5%, 1% and 1.5% w/v, respectively. The sensors were fabricated in film form using an electrochemical deposition method. The sensing properties of the sensors were observed by exposing the pure chitosan and sodium TPP cross-linked chitosan sensors towards acetone concentrations of 5, 10, 50, 100 and 200 ppm. The measurement results revealed that the maximum response in output voltage value of pure chitosan sensor was 0.35 V while sodium TPP crosslinked chitosan sensors were above 0.35 V towards 5 ppm acetone concentration. When the sensors were exposed towards acetone concentration of 200 ppm, the maximum response of pure chitosan was 0.45 V while sodium TPP crosslinked chitosan sensors were above 0.45 V. Amongst the variation of sodium TPP, the maximum response of 1% sodium TPP was the highest since the maximum response was 0.4 V and 0.6 V towards 5 ppm and 200 ppm acetone concentration, respectively. While the maximum responses of other sodium TPP concentrations were under 0.4 V and 0.6 V towards 5 ppm and 200 ppm acetone concentration. Moreover, 1% sodium TPP cross-linked chitosan based sensor showed good reproducibility and outstanding lifetime. Therefore, 1% sodium TPP cross-linked chitosan based sensor has exhibited remarkable sensing properties as a novel acetone sensor.

  7. CD24 cross-linking induces apoptosis in, and inhibits migration of, MCF-7 breast cancer cells

    International Nuclear Information System (INIS)

    Kim, Jong Bin; Bae, Ji-Yeon; Jee, Hyeon-Gun; Noh, Dong-Young; Ko, Eunyoung; Han, Wonshik; Lee, Jeong Eon; Lee, Kyung-Min; Shin, Incheol; Kim, Sangmin; Lee, Jong Won; Cho, Jihyoung

    2008-01-01

    The biological effects of CD24 (FL-80) cross-linking on breast cancer cells have not yet been established. We examined the impact of CD24 cross-linking on human breast cancer cell line MCF-7. MCF-7 and MDA-MB-231 cells were treated with anti-rabbit polyclonal IgG or anti-human CD24 rabbit polyclonal antibodies to induce cross-linking, and then growth was studied. Changes in cell characteristics such as cell cycle modulation, cell death, survival in three-dimensional cultures, adhesion, and migration ability were assayed after CD24 cross-linking in MCF-7. Expression of CD24 was analyzed by flow cytometry in MDA-MB-231 and MCF-7 cells where 2% and 66% expression frequencies were observed, respectively. CD24 cross-linking resulted in time-dependent proliferation reduction in MCF-7 cells, but no reduction in MDA-MB-231 cells. MCF-7 cell survival was reduced by 15% in three-dimensional culture after CD24 cross-linking. Increased MCF-7 cell apoptosis was observed after CD24 cross-linking, but no cell cycle arrest was observed in that condition. The migration capacity of MCF-7 cells was diminished by 30% after CD24 cross-linking. Our results showed that CD24 cross-linking induced apoptosis and inhibited migration in MCF-7 breast cancer cells. We conclude that CD24 may be considered as a novel therapeutic target for breast cancer

  8. Efeito terapêutico do "cross-linking" corneano na ceratopatia bolhosa sintomática Therapeutic effect of corneal cross-linking on symptomatic bullous keratophaty

    Directory of Open Access Journals (Sweden)

    Diego Nery Benevides Gadelha

    2009-08-01

    Full Text Available OBJETIVO: Avaliar o efeito do "cross-linking" corneano na ceratopatia bolhosa sintomática e seu impacto na acuidade visual, espessura corneana e sintomatologia dolorosa. MÉTODOS: Doze pacientes com ceratopatia bolhosa sintomática foram incluídos. Exame clínico com questionário específico para a pesquisa com escala de dor (escala visual analógica numérica, acuidade visual e mensuração da espessura corneana foi realizada pré "cross-linking" corneano, 7, 30 e 60 dias após. Em todos os pacientes o tratamento com UVA-crosslinking foi realizado após abrasão do epitélio corneano na lâmpada de fenda e instilação de solução de riboflavina 0,1% a cada 5 minutos por 30 minutos. Após esse período o paciente foi submetido à exposição à luz ultravioleta A (UVA, utilizando riboflavina e anestesia tópica a cada 5 minutos por 30 minutos. O paciente utilizou colírios de ofloxacina 0,3% e lágrima artificial até completa reepitelização. O teste de Friedman foi usado para comparar as médias das frequências da acuidade visual, sintomatologia dolorosa e espessura central da córnea. Os valores de "p" menor que 0,05 foram considerados como estatisticamente significantes. RESULTADOS: Doze olhos de 12 pacientes com erosões epiteliais recorrentes foram tratados. O tempo de seguimento foi de dois meses. Foi observada redução significante da dor (pPURPOSE: To evaluate the corneal cross-linking (CXL effect on symptomatic bullous keratopathy, and its impact on the visual acuity, corneal thickness and pain symptomatology. METHODS: Twelve patients with symptomatic bullous keratophaty were included. A clinical examination with a specific questionnaire (pain scale - visual numeric analog scale, visual acuity, and corneal thickness measures was performed before corneal cross-linking and 7, 30 and 60 days after the procedure. In all patients, the treatment with the UVA-crosslinking was done after corneal abrasion on the slit-lamp and

  9. Oegylated and cross-linking carbazole dendrons and dendrimers: Synthesis, characterization, assembly and thin film fabrication

    Science.gov (United States)

    Felipe, Mary Jane Legaspi

    2011-12-01

    Dendrimers and dendrons (fractional dendrimers) are macromolecular structures that have well-defined molecular weights and precise number of functional groups. Tailoring these structures has provided designer molecules that can be used for various applications including drug delivery, sensors, and anti-biofouling surfaces. Overall, this dissertation provides novel protocols for the understanding of molecular design, synthesis, and structure-property relationship of OEGylated and conjugated carbazole dendrons and dendrimers. In this design, the use of oligo(ethylene glycol) (OEG) allows for the fabrication of biocompatible materials and imparts hydrophilicity on the structure while the carbazole functionality allows the cross-linking of these designer molecules. Such fine-tuning of macromolecular structures leading to the fabrication of anti-biofouling thin films, nanostructuring at the air-water interface, and assembly into supramolecular superstructures are considered in this dissertation. Chapter 2 details the synthesis, characterization, and electrochemical cross-linking of OEGylated linear dendrons and "Janus-type" dendrimers. Cross-linking the carbazole moieties enables the deposition of these films on Au, indium tin oxide-coated glass, and doped silicon through cyclic voltammetry and provides films with secondary level of organization imparted by the inter- and intra-molecular interaction among the carbazole units. Chapter 3 describes the fabrication of nonspecific protein adsorption resistant surfaces through electrochemical grafting of three different dendrons on SAM carbazole-coated gold substrates. The predictable shape of each dendron and the ability to cross-link the carbazole units have enabled parametrization of OEG conformation and density on these interfaces. Chapter 4 demonstrates the fundamental architectural requirements for obtaining stable films with OEGylated linear dendron molecules providing a new architectural design of nanostructuring

  10. Dynamic constitutional frameworks for DNA biomimetic recognition.

    Science.gov (United States)

    Catana, Romina; Barboiu, Mihail; Moleavin, Ioana; Clima, Lilia; Rotaru, Alexandru; Ursu, Elena-Laura; Pinteala, Mariana

    2015-02-07

    Linear and cross-linked dynamic constitutional frameworks generated from reversibly interacting linear PEG/core constituents and cationic sites shed light on the dominant coiling versus linear DNA binding behaviours, closer to the histone DNA binding wrapping mechanism.

  11. Interstrand and AC-loss measurements on Rutherford-type cables for accelerator magnet applications

    CERN Document Server

    Otmani, R; Tixador, P

    2001-01-01

    One of the main issues for particle accelerator magnets is the control of interstrand resistances. Too low resistances result in large coupling currents during ramping, which distort field quality, while too large resistances may prevent current redistribution among cable strands, resulting in degraded quench performance. In this paper, we review a series of interstrand resistance and AC-loss measurements performed on four Rutherford-type cables. The four cables have the same number of strands and similar outer dimensions, corresponding to LHC quadrupole cable specifications. The first cable is made from NbTi strands, coated with silver-tin alloy, the second one is made from bare Nb/sub 3/Sn strands, the third one is made also from bare Nb/sub 3/Sn strands but includes a 25- mu m-thick stainless steel core between the strand layers, and the last one is made from Nb/sub 3/Sn strands plated with chromium. To cross-check the two measurement types and assess their consistency, we compare the coupling-current time...

  12. Comparative studies of photochemical cross-linking methods for stabilizing the bulk hetero-junction morphology in polymer solar cells

    DEFF Research Database (Denmark)

    Carlé, Jon Eggert; Andreasen, Birgitta; Tromholt, Thomas

    2012-01-01

    We are here presenting a comparative study between four different types of functionalities for cross-linking. With relatively simple means bromine, azide, vinyl and oxetane could be incorporated into the side chains of the low band-gap polymer TQ1. Cross-linking of the polymers was achieved by UV...

  13. Effects of Granule Size of Cross-Linked and Hydroxypropylated Sweet Potato Starches on Their Physicochemical Properties

    NARCIS (Netherlands)

    Zhao, J.; Chen Zenghong,; Jing, Z.; Buwalda, Piet; Gruppen, H.; Schols, H.A.

    2015-01-01

    Sweet potato starch was modified by cross-linking, hydroxypropylation, and combined cross-linking and hydroxypropylation, and the starches were subsequently sieved to obtain differently sized granule fractions. The effects of granule size of native and modified sweet potato starch fractions and all

  14. Identification and characterization of a pituitary corticotropin-releasing factor binding protein by chemical cross-linking

    DEFF Research Database (Denmark)

    Nishimura, E; Billestrup, Nils; Perrin, M

    1987-01-01

    A corticotropin-releasing factor (CRF) binding protein has been identified based on the chemical cross-linking of ovine [Nle21,m-125I-Tyr32]CRF (125I-oCRF) to bovine anterior pituitary membranes using disuccinimidyl suberate (DSS). The apparent molecular weight of the cross-linked complex...

  15. Cross-linking of β-lactoglobulin enhances allergic sensitization through changes in cellular uptake and processing

    NARCIS (Netherlands)

    Stojadinovic, Marija; Pieters, Raymond; Smit, Joost; Velickovic, Tanja Cirkovic

    Cross-linking of proteins has been exploited by the food industry to change food texture and functionality but the effects of these manipulations on food allergenicity still remain unclear. To model the safety assessment of these food biopolymers, we created cross-linked bovine β-lactoglobulin

  16. Insights into the swelling process and drug release mechanisms from cross-linked pectin/high amylose starch matrices

    Directory of Open Access Journals (Sweden)

    Fernanda M. Carbinatto

    2014-02-01

    Full Text Available Cross-linked pectin/high amylose mixtures were evaluated as a new excipient for matrix tablets formulations, since the mixing of polymers and cross-linking reaction represent rational tools to reach materials with modulated and specific properties that meet specific therapeutic needs. Objective: In this work the influence of polymer ratio and cross-linking process on the swelling and the mechanism driving the drug release from swellable matrix tablets prepared with this excipient was investigated. Methods: Cross-linked samples were characterized by their micromeritic properties (size and shape, density, angle of repose and flow rate and liquid uptake ability. Matrix tablets were evaluated according their physical properties and the drug release rates and mechanisms were also investigated. Results: Cross-linked samples demonstrated size homogeneity and irregular shape, with liquid uptake ability insensible to pH. Cross-linking process of samples allowed the control of drug release rates and the drug release mechanism was influenced by both polymer ratio and cross-linking process. The drug release of samples with minor proportion of pectin was driven by an anomalous transport and the increase of the pectin proportion contributed to the erosion of the matrix. Conclusion: The cross-linked mixtures of high amylose and pectin showed a suitable excipient for slowing the drug release rates.

  17. Wheat gluten films cross-linked with 1-ethyl-3-(3-demethylaminopropyl) carbodiimide and N-hydroxysuccinimide

    NARCIS (Netherlands)

    Tropini, V.; Lens, J.P.; Mulder, W.J.; Silvestre, F.

    2004-01-01

    Wheat gluten films were cast from aqueous dispersions containing 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) as cross-linking reagents and glycerol as a plasticizer. Cross-linking was carried out to improve film properties such as water sensitivity and tensile

  18. Dually cross-linked single network poly(acrylic acid) hydrogels with superior mechanical properties and water absorbency.

    Science.gov (United States)

    Zhong, Ming; Liu, Yi-Tao; Liu, Xiao-Ying; Shi, Fu-Kuan; Zhang, Li-Qin; Zhu, Mei-Fang; Xie, Xu-Ming

    2016-06-28

    Poly(acrylic acid) (PAA) hydrogels with superior mechanical properties, based on a single network structure with dual cross-linking, are prepared by one-pot free radical polymerization. The network structure of the PAA hydrogels is composed of dual cross-linking: a dynamic and reversible ionic cross-linking among the PAA chains enabled by Fe(3+) ions, and a sparse covalent cross-linking enabled by a covalent cross-linker (Bis). Under deformation, the covalently cross-linked PAA chains remain intact to maintain their original configuration, while the Fe(3+)-enabled ionic cross-linking among the PAA chains is broken to dissipate energy and then recombined. It is found that the mechanical properties of the PAA hydrogels are significantly influenced by the contents of covalent cross-linkers, Fe(3+) ions and water, which can be adjusted within a substantial range and thus broaden the applications of the hydrogels. Meanwhile, the PAA hydrogels have excellent recoverability based on the dynamic and reversible ionic cross-linking enabled by Fe(3+) ions. Moreover, the swelling capacity of the PAA hydrogels is as high as 1800 times in deionized water due to the synergistic effects of ionic and covalent cross-linkings. The combination of balanced mechanical properties, efficient recoverability, high swelling capacity and facile preparation provides a new method to obtain high-performance hydrogels.

  19. Cross-Linking Behavior and Foaming Properties of Bovine a-Lactalbumin after Glycation with Various Saccharides

    NARCIS (Netherlands)

    Haar, ter R.; Westphal, Y.; Wierenga, P.A.; Schols, H.A.; Gruppen, H.

    2011-01-01

    a-Lactalbumin was glycated via the Maillard reaction in the dry state using various mono- and oligosaccharides. The reaction resulted not only in coupling of the saccharides to a-lactalbumin but also in cross-linked proteins. The glycation rate and the extent of cross-link formation were highly

  20. MeV ion beam interaction with polymer films containing cross-linking agents

    International Nuclear Information System (INIS)

    Evelyn, A. L.

    1999-01-01

    Polymer films containing cross linking enhancers were irradiated with MeV alpha particles to determine the effects of MeV ion beam interaction on these materials. The contributed effects from the electronic and nuclear stopping powers were separated by irradiating stacked thin films of polyvinyl chloride (PVC), polystyrene (PS) and polyethersulfone (PES). This layered system allowed most of the effects of the electronic energy deposited to be experienced by the first layers and the last layers to receive most of the effects of the nuclear stopping power. RGA, Raman microprobe analysis, RBS and FTIR measured changes in the chemical structures of the irradiated films. The characterization resolved the effects of the stopping powers on the PVC, PS and PES and the results were compared with those from previously studied polymers that did not contain any cross linking agents

  1. Multi-stimulus-responsive shape-memory polymer nanocomposite network cross-linked by cellulose nanocrystals.

    Science.gov (United States)

    Liu, Ye; Li, Ying; Yang, Guang; Zheng, Xiaotong; Zhou, Shaobing

    2015-02-25

    In this study, we developed a thermoresponsive and water-responsive shape-memory polymer nanocomposite network by chemically cross-linking cellulose nanocrystals (CNCs) with polycaprolactone (PCL) and polyethylene glycol (PEG). The nanocomposite network was fully characterized, including the microstructure, cross-link density, water contact angle, water uptake, crystallinity, thermal properties, and static and dynamic mechanical properties. We found that the PEG[60]-PCL[40]-CNC[10] nanocomposite exhibited excellent thermo-induced and water-induced shape-memory effects in water at 37 °C (close to body temperature), and the introduction of CNC clearly improved the mechanical properties of the mixture of both PEG and PCL polymers with low molecular weights. In addition, Alamar blue assays based on osteoblasts indicated that the nanocomposites possessed good cytocompatibility. Therefore, this thermoresponsive and water-responsive shape-memory nanocomposite could be potentially developed into a new smart biomaterial.

  2. Charge regulation and energy dissipation while compressing and sliding a cross-linked chitosan hydrogel layer

    DEFF Research Database (Denmark)

    Liu, Chao; Thormann, Esben; Tyrode, Eric

    2015-01-01

    , but not the friction coefficient. This suggests that the main energy dissipation mechanism arises from processes occurring within the hydrogel layer, rather than at the silica/hydrogel interface, and we suggest that it is related to stretching of polymer chains between the cross-linking points. We also find...... that an increased cross-linking density, from 40% to 100%, in the hydrogel reduces the friction coefficient....... the concentration of counterions within the hydrogel to keep the interior close to uncharged, which is similar to what has been observed for polyelectrolyte brushes. Changes in pH and ionic strength affect the adhesion force and the friction force between the silica surface and the hydrogel layer...

  3. Cross-linked self-assembled micelle based nanosensor for intracellular pH measurements

    DEFF Research Database (Denmark)

    Ek, Pramod Kumar; Søndergaard, Rikke Vicki; Windschiegl, Barbara

    2014-01-01

    A micelle based nanosensor was synthesized and investigated as a ratiometric pH sensor for use in measurements in living cells by fluorescent microscopy. The nanosensor synthesis was based on self-assembly of an amphiphilic triblock copolymer, which was chemically cross-linked after micelle......-linked by an amidation reaction using 3,6,9-trioxaundecandioic acid cross-linker. The cross-linked micelle was functionalized with two pH sensitive fluorophores and one reference fluorophore, which resulted in a highly uniform ratiometric pH nanosensor with a diameter of 29 nm. The use of two sensor fluorophores...... provided a sensor with a very broad measurement range that seems to be influenced by the chemical design of the sensor. Cell experiments show that the sensor is capable of monitoring the pH distributions in HeLa cells....

  4. Adsorption of hexavalent chromium on modified corn stalk using different cross-linking agents

    Science.gov (United States)

    Chen, Suhong; Zhu, Yi; Han, Zhijun; Feng, Gao; Jia, Yuling; Fu, Kaifang; Yue, Qinyan

    2017-12-01

    In this study, four different types of adsorbents modified from corn stalk were synthesized after the reaction with epichlorohydrin, N,N-dimethylformamide, triethylamine and different cross-linking agents. The surface functional groups and thermal stability of modified corn stalk (MCSs) were characterized using FTIR and TG analysis, respectively. The feasibility of using MCSs to remove Cr(VI) were evaluated. Adsorption isotherms were determined and modeled with Langmuir, Freundlich and Temkin equations. The experimental results showed that MCS modified using diethylenetriamine (DETA) had the best modification effect, and the adsorption capacity of Cr(VI) reached as high as 227.27 mg/g at 323 K. Thermodynamic study showed that the Cr(VI) adsorption onto MCSs was endothermic processes. As a result, MCS by using DETA as cross-linking agent has good potential for the removal of Cr(VI) from aqueous solutions.

  5. Mechanical properties and collagen cross-linking of the patellar tendon in old and young men

    DEFF Research Database (Denmark)

    Couppé, C; Hansen, P; Kongsgaard, M

    2009-01-01

    were higher in OM than in YM (73 +/- 13 vs. 11 +/- 2 mmol/mol; P appreciably influence the dimensions or mechanical properties of the human patellar tendon in vivo. Collagen concentration was reduced, whereas both enzymatic......Age-related loss in muscle mass and strength impairs daily life function in the elderly. However, it remains unknown whether tendon properties also deteriorate with age. Cross-linking of collagen molecules provides structural integrity to the tendon fibrils and has been shown to change with age...... in animals but has never been examined in humans in vivo. In this study, we examined the mechanical properties and pyridinoline and pentosidine cross-link and collagen concentrations of the patellar tendon in vivo in old (OM) and young men (YM). Seven OM (67 +/- 3 years, 86 +/- 10 kg) and 10 YM (27 +/- 2...

  6. The cross-linking influence of electromagnetic radiation on water-soluble polyacrylan compositions with biopolymers

    Directory of Open Access Journals (Sweden)

    B. Grabowska

    2009-01-01

    Full Text Available The results of examinations of the cross-linking influence of electromagnetic radiation - in a microwave range – on polyacrylancompositions with biopolymers, are presented in the hereby paper. The cross-linking process of the tested compositions was determined on the basis of the FT-IR spectroscopic methods. It was shown that microwave operations can lead to the formation of new cross-linkedstructures with strong covalent bonds. The adsorption process and formation of active centres in polymer molecules as well as in highsilica sand were found due to microwave radiations. In this process hydroxyl groups (-OH - present in a polymer - and silane groups (Si- O-H - present in a matrix - are mainly taking part. Spectroscopic and strength tests performed for the system: biopolymer binding agent – matrix indicate that the microwave radiation can be applied for hardening moulding sands with biopolymer binders.

  7. Development and Evaluation of Dual Cross-Linked Pulsatile Beads for Chronotherapy of Rheumatoid Arthritis

    Directory of Open Access Journals (Sweden)

    Abanesh kumar Bansal

    2013-01-01

    Full Text Available In the present investigation, pulsatile release beads were prepared by ionic gelation technique. Lornoxicam dual cross-linked beads were prepared by dropping dispersed phase of lornoxicam, pectin, and sodium alginate into the dispersion phase of different concentrations of calcium chloride solution followed by aluminium chloride solution. The formulated beads were further coated by Eudragit L & S 100 in the ratio 1 : 2 w/w in order to achieve desired lag time. In vitro release study showed lag time of 5–8 h before release of lornoxicam from the formulated beads. Thus, formulated dual cross-linked beads when administered at bed time may release lornoxicam when needed most for chronotherapeutics of early morning rheumatoid arthritis attacks in chronic patients.

  8. Water dispersible, non-cytotoxic, cross-linked luminescent AIE dots: Facile preparation and bioimaging applications

    Science.gov (United States)

    Liu, Meiying; Zhang, Xiqi; Yang, Bin; Deng, Fengjie; Li, Zhen; Wei, Junchao; Zhang, Xiaoyong; Wei, Yen

    2014-12-01

    Fluorescent organic nanoparticles have attracted great current research interest due to their superior properties as compared with small organic dyes and fluorescent inorganic nanoparticles. However, fluorescent organic nanoparticles based on conventional organic dyes often result in significant fluorescence decrease due to the notorious aggregation-caused quenching effect. On the other hand, these fluorescent organic nanoparticles obtained from self-assembly are normally not stable in diluted solution. Therefore, the development of novel fluorescent organic nanoparticles which could overcome these limitations is highly desirable for their practical biomedical applications. In this work, water dispersible, non-cytotoxic and cross-linked luminescent polymeric nanoparticles based on aggregation induced emission dyes were prepared via one pot emulsion polymerization. These cross-linked luminescent polymeric nanoparticles emitted strong red fluorescence and were highly stable in diluted aqueous solution, making them highly potential for various biomedical applications.

  9. Adsorption equilibrium studies of uranium (VI) onto cross-linked chitosan-citric acid

    International Nuclear Information System (INIS)

    Ho Thi Yeu Ly; Nguyen Van Suc; Vo Quang Mai; Nguyen Mong Sinh

    2011-01-01

    Investigation of U(VI) adsorption by the cross- linked chitosan with citric acid was conduced by bath method. Effect of parameters such as pH, contact time, adsorbent dosage and other metal cations was determined. The maximum adsorption capacity of U(VI) at pH 4 was found to be 71.43 mg U(VI) / g cross-linked chitosan - citric acid after 300 min of contact time. The Langmuir and Freundlich isotherm models were used to describe adsorption equilibrium. The correction values, R 2 of two models were found to be 0.991 and 0.997, respectively. Therefore, it could be concluded that the adsorption equilibrium for U(VI) was followed the Langmuir and the Freundlich isotherm models. (author)

  10. Polymeric Nanocapsule from Silica Nanoparticle@Cross-linked Polymer Nanoparticles via One-Pot Approach

    Directory of Open Access Journals (Sweden)

    Shen Ruoping

    2009-01-01

    Full Text Available Abstract A facile strategy was developed here to prepare cross-linked polymeric nanocapsules (CP nanocapsules with silica nanoparticles as templates. The silica nanoparticle@cross-linked polymer nanoparticles were prepared by the encapsulation of the silica nanoparticles by the one-pot approach via surface-initiated atom transfer radical polymerization of hydroxyethyl acrylate in the presence ofN,N′-methylenebisacrylamide as a cross-linker from the initiator-modified silica nanoparticles. After the silica nanoparticle templates were etched with hydrofluoric acid, the CP nanocapsules with particle size of about 100 nm were obtained. The strategy developed was confirmed with Fourier transform infrared, thermogravimetric analysis and transmission electron microscopy.

  11. Bio-inspired borate cross-linking in ultra-stiff graphene oxide thin films.

    Science.gov (United States)

    An, Zhi; Compton, Owen C; Putz, Karl W; Brinson, L Catherine; Nguyen, SonBinh T

    2011-09-01

    Adjacent graphene oxide nanosheets in a thin-film structure have been covalently cross-linked in a fashion similar to the cell walls of higher-order plants. The resulting ultra-stiff structure exhibits a maximum storage modulus of 127 GPa that can be tuned by varying borate concentration. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Vision Restoration with a Collagen Cross-linked Boston Keratoprosthesis Unit

    Science.gov (United States)

    2017-09-01

    tissue by collagen-crosslinking the cornea graft ex vivo using vitamin B2 (riboflavin) and ultraviolet light . The overall objective of this study is...cross-linked using vitamin B2 (riboflavin) and ultraviolet light prior to prosthesis assembly. This study’s population will include patients who are... reflect the company’s specialty of providing ophthalmic tissue. The company goals and their roll in this study was not affected. The name change

  13. Procedure for the fabrication of a cross-linked polyester material

    International Nuclear Information System (INIS)

    D'Alelio, G.F.

    1972-01-01

    The procedures are described for the production of a cross-linked polyester material by means of the irradiation of a radiosensitive polyester with a dose of over 0.5 megarad and under 8 megarads high energy, ionising radiation, corresponding to at least 100,000 ev. The polyester is of the telomerised diacrylpolyester type, and may be in a mixture containing about 1% of a coplymerisable aliphatic monomer, or about 30-90% of an unsaturated aliphatic alkyd resin. (JIW)

  14. Long-term results of cornea collagen cross-linking with riboflavin for keratoconus

    Directory of Open Access Journals (Sweden)

    Vinay Agrawal

    2013-01-01

    Full Text Available Corneal collagen cross-linking with riboflavin and UVA light (CXL is the only method designed to arrest the progression of keratoconus. Visual improvement generally starts 3 months after treatment. Reduction is coma seen on aberrometry in early postoperative phase is also responsible for the improvement in visual acuity. In the light of currently available data we can thus say that CXL is a safe procedure that is successful in arresting keratoconus.

  15. Progress of research on corneal collagen cross-linking for corneal melting

    Directory of Open Access Journals (Sweden)

    Ke-Ren Xiao

    2016-06-01

    Full Text Available Corneal collagen cross-linking(CXLcould increase the mechanical strength, biological stability and halt ectasia progression due to covalent bond formed by photochemical reaction between ultraviolet-A and emulsion of riboflavin between collagen fibers in corneal stroma. Corneal melting is an autoimmune related noninfectious corneal ulcer. The mechanism of corneal melting, major treatment, the basic fundamental of ultraviolet-A riboflavin induced CXL and the clinical researches status and experiment in CXL were summarized in the study.

  16. Corneal Tomographic Changes After UV Cross-Linking for Corneal Ectasia (1-Year Results).

    Science.gov (United States)

    Baksoellah, Zainab; Lavy, Itay; Baydoun, Lamis; Hooijmaijers, Hilde C M; van Dijk, Korine; Melles, Gerrit R J

    2017-12-01

    To evaluate changes in maximum keratometry (Kmax), corneal higher-order aberrations (HOAs), and densitometry (backscattered light) up to 1 year after UV cross-linking and their possible relation with changes in the visual outcome. Retrospective cohort study on 18 eyes of 16 patients, who underwent UV cross-linking after the Dresden protocol for progressive keratoconus or ectasia after laser-assisted in situ keratomileusis. Corrected distance visual acuity (CDVA), Scheimpflug-based corneal tomography, mean image brightness (corneal densitometry) from the anterior 120 μm of the midcornea, and posterior 60 μm of the central 6 mm of the cornea, and HOAs were evaluated. Kmax at 1 month (59.7 ± 6.0D) after UV cross-linking resembled preoperative Kmax (59.3 ± 6.4D, P = 0.368), decreased until 3 months postoperatively (58.3 ± 6.3D, P = 0.002), and stabilized thereafter (P > 0.227). All postoperative corneal densitometry values were higher than preoperative values in all measured depths (P corneal HOAs (4.28 ± 1.64 μm and 3.87 ± 1.62 μm, respectively) resembled preoperative values (4.10 ± 1.70 μm and 3.67 ± 1.62 μm, respectively; P > 0.221) and then decreased until 12 months postoperatively (3.86 ± 1.84 μm and 3.40 ± 1.80 μm, respectively; P 0.345 and P > 0.257, respectively). No relations were found between CDVA and the evaluated parameters (P > 0.05). One year after UV cross-linking, the observation of stable CDVA and thinnest point thickness, together with reduced Kmax suggests no ectasia progression within the study period in these cases. Although HOAs showed a trend toward improvement, corneal densitometry remained elevated.

  17. Chemical cross-linking and mass spectrometry as structure determination tools

    Czech Academy of Sciences Publication Activity Database

    Novák, Petr; Giannakopulos, A.

    2007-01-01

    Roč. 13, - (2007), s. 105-113 ISSN 1469-0667 R&D Projects: GA AV ČR KJB400200501; GA MŠk LC545 Grant - others:SE(XE) Marie Curie Actions TOK, Contract No. MTKD-CT-2004-014407 Institutional research plan: CEZ:AV0Z50200510 Keywords : cross-linking * protein * high order structure Subject RIV: EE - Microbiology, Virology Impact factor: 1.198, year: 2007

  18. Cross-Linking in Collagen by Nonenzymatic Glycation Increases the Matrix Stiffness in Rabbit Achilles Tendon

    OpenAIRE

    Reddy, G. Kesava

    2004-01-01

    Nonenzymatic glycation of connective tissue matrix proteins is a major contributor to the pathology of diabetes and aging. Previously the author and colleagues have shown that nonenzymatic glycation significantly enhances the matrix stability in the Achilles tendon (Reddy et al., 2002, Arch. Biochem. Biophys., 399, 174–180). The present study was designed to gain further insight into glycation-induced collagen cross-linking and its relationship to matrix stiffness in the rabbit Achilles tendo...

  19. Cross-linking in collagen by nonenzymatic glycation increases the matrix stiffness in rabbit achilles tendon.

    Science.gov (United States)

    Reddy, G Kesava

    2004-01-01

    Nonenzymatic glycation of connective tissue matrix proteins is a major contributor to the pathology of diabetes and aging. Previously the author and colleagues have shown that nonenzymatic glycation significantly enhances the matrix stability in the Achilles tendon (Reddy et al., 2002, Arch. Biochem. Biophys., 399, 174-180). The present study was designed to gain further insight into glycation-induced collagen cross-linking and its relationship to matrix stiffness in the rabbit Achilles tendon. The glycation process was initiated by incubating the Achilles tendons (n = 6) in phosphate-buffered saline containing ribose, whereas control tendons (n = 6) were incubated in phosphate-buffered saline without ribose. Eight weeks following glycation, the biomechanical attributes as well as the degree of collagen cross-linking were determined to examine the potential associations between matrix stiffness and molecular properties of collagen. Compared to nonglycated tendons, the glycated tendons showed increased maximum load, stress, strain, Young's modulus of elasticity, and toughness indicating that glycation increases the matrix stiffness in the tendons. Glycation of tendons resulted in a considerable decrease in soluble collagen content and a significant increase in insoluble collagen and pentosidine. Analysis of potential associations between the matrix stiffness and degree of collagen cross-linking showed that both insoluble collagen and pentosidine exhibited a significant positive correlation with the maximum load, stress, and strain, Young's modulus of elasticity, and toughness (r values ranging from.61 to.94) in the Achilles tendons. However, the soluble collagen content present in neutral salt buffer, acetate buffer, and acetate buffer containing pepsin showed an inverse relation with the various biomechanical attributes tested (r values ranging from.22 to.84) in the Achilles tendons. The results of the study demonstrate that glycation-induced collagen cross-linking

  20. In vitro Antiglycation and Cross-Link Breaking Activities of Sri ...

    African Journals Online (AJOL)

    Purpose: To investigate the antiglycation and cross-link breaking activities of Sri Lankan low-grown orthodox Orange Pekoe grade black tea (Camellia sinensis L) Methods: Five concentrations (6.25, 12.5, 25.0, 50.0 or 100.0 ìg/ml) of Black tea brew (BTB) were made using Sri Lankan low-grown Orange Pekoe (O.P.) grade ...

  1. Production of RNA-protein cross links in γ irradiated E. Coli ribosomes

    International Nuclear Information System (INIS)

    Ekert, Bernard; Giocanti, Nicole

    1976-01-01

    γ irradiation in de-aerated conditions of E. coli MRE 600 ribosomes, labelled with 14 C uracil, leads to a decrease of extractibility of 14 C RNA by lithium chloride 4 M-urea 8 M. On the other hand, the radioactivity of the protein fraction increases with irradiation. These results strongly suggest that RNA-protein cross links are formed in irradiated ribosomes [fr

  2. 3.3. Sorption activity of cross-linked polymers of ethynyl-piperidol

    International Nuclear Information System (INIS)

    Khalikov, D.Kh.

    2012-01-01

    The sorption activity of cross-linked polymers of ethynyl-piperidol was studied. The bilirubin sorption was studied as well. The kinetic of bilirubin sorption and human serum albumin at their joint presence in hydrogel solutions was defined. Bilirubin sorption and change of albumin composition was considered. The sorption of middle molecular peptides was considered as well. The sorption of endogenous toxin by means of ethynyl-piperidol polymers was done.

  3. Studies in cross-linking PVC footwear soling compounds using gamma-irradiation

    International Nuclear Information System (INIS)

    Bloom, L.I.

    1983-01-01

    Irradiation cross-linking of polymeric materials has been known for some time, but it is only in recent years that it has been put to commercial advantage. Well known uses are the modification of PVC for higher temperature applications. Fundamental studies were carried out on amongst other materials, plasticised PVC compounds for use in cable applications. The results of this work, encouraged the author to investigate cross-linkable PVC in areas such as footwear soling

  4. Reaction-Diffusion Degradation Model for Delayed Erosion of Cross-Linked Polyanhydride Biomaterials

    OpenAIRE

    Domanskyi, Sergii; Poetz, Katie L.; Shipp, Devon A.; Privman, Vladimir

    2015-01-01

    We develop a theoretical model to explain the long induction interval of water intake that precedes the onset of erosion due to degradation caused by hydrolysis in the recently synthesized and studied cross-linked polyanhydrides. Various kinetic mechanisms are incorporated in the model in an attempt to explain the experimental data for the mass loss profile. Our key finding is that the observed long induction interval is attributable to the nonlinear dependence of the degradation rate constan...

  5. Corneal collagen cross-linking for treatment of non-healing corneal ulcers

    OpenAIRE

    Mitra Zamani; Mahmoodreza Panahi-Bazaz; Mona Assadi

    2015-01-01

    Purpose: To evaluate the efficacy of corneal collagen cross-linking (CXL) for treatment of corneal ulcers not responding to antimicrobial therapy. Methods: Eight patients with corneal ulcers associated with corneal melting, not responding to conventional antibiotic therapy, were treated with CXL. The procedure was performed according to the standardized protocol for keratoconus. Preoperative medications were continued after CXL in all cases. Microbiological exams revealed Pseudomonas aeru...

  6. Sieving of hot gases by hyper-cross-linked nanoscale-hybrid membranes.

    Science.gov (United States)

    Raaijmakers, Michiel J T; Hempenius, Mark A; Schön, Peter M; Vancso, G Julius; Nijmeijer, Arian; Wessling, Matthias; Benes, Nieck E

    2014-01-08

    Macromolecular networks consisting of homogeneously distributed covalently bonded inorganic and organic precursors are anticipated to show remarkable characteristics, distinct from those of the individual constituents. A novel hyper-cross-linked ultrathin membrane is presented, consisting of a giant molecular network of alternating polyhedral oligomeric silsesquioxanes and aromatic imide bridges. The hybrid characteristics of the membrane are manifested in excellent gas separation performance at elevated temperatures, providing a new and key enabling technology for many important industrial scale applications.

  7. Synthesis and Characterisation of Novel Cross Linked Biopolyesters from Olive Oil as Eco-friendly Biodegradable Materials

    Directory of Open Access Journals (Sweden)

    J. Shakina

    2012-01-01

    Full Text Available Novel cross linked biopolyesters were synthesised from naturally available olive oil, oligomeric polytriglyceride fumarate (o-PTF was prepared by glycerolysis of the olive oil followed by condensation with maleic anhydride. The (o-PTF was characterised by spectral (FTIR, UV, NMR and physicochemical properties (specific gravity, viscosity, saponification value, iodine value. The molecular weight of o-PTF was estimated using gel permeation chromatography (GPC. The cross linked biopolyesters were prepared by treating o-PTF of olive with N-vinyl-2-pyrrolidone and vinyl acetate. The cross linked biopolyesters were evaluated for spectral properties (FT IR, cross linked density, thermal properties, mechanical properties and biodegradation. The potential antifungal and anti bacterial activities of the newly prepared biopolyesters were evaluated. The environment degradation was assessed by soil burial test. The outcome of the studies has revealed that the newly prepared cross linked biopolyesters are potential biodegradable material for various consumer application like package materials and agricultural applications.

  8. Three-Year Outcomes of Cross-Linking PLUS (Combined Cross-Linking with Femtosecond Laser Intracorneal Ring Segments Implantation for Management of Keratoconus

    Directory of Open Access Journals (Sweden)

    Mohammed Iqbal Hafez Saleem

    2018-01-01

    Full Text Available Purpose. To analyze the results of three-year outcomes of combined epithelium-on cross-linking with femtosecond laser ICRS (cross-linking PLUS for keratoconus management. Design. A retrospective multicenter clinical study. Methods. 43 eyes of 38 patients were subjected to preoperative and postoperative UCVA, BCVA, refraction, Pentacam pachymetry, and keratometry examinations at 3-, 6-, 12-, 24-, and 36-month follow-up period. Results. The preoperative and postoperative mean UCVA was 1.30 ± 0.48 (logMAR ± SD and 0.82 ± 0.22 respectively. The preoperative and postoperative mean BCVA was 0.90 ± 0.40 and 0.60 ± 0.30, respectively. The preoperative and postoperative mean K average was 50.63 ± 0.87 (D ± SD and 45.56 ± 0.98, respectively. The preoperative and postoperative mean pachymetry was 471 ± 92.36 (μm ± SD and 423 ± 39.58, respectively. The preoperative and postoperative mean astigmatism was 7.55 ± 1.75 and 3.39 ± 1.26, respectively. One eye showed ICRS edge exposure while 6 eyes showed progression of keratoconus. Conclusion. CXL PLUS was proved to be a successful procedure to halt progression (mainly by CXL and to correct the refractive status of the keratoconic eye (mainly by ICRS. CXL PLUS performed a synergistic action correcting and maintaining the correction of both myopic and astigmatic components of keratoconus.

  9. Conventional Corneal Collagen Cross-Linking Versus Transepithelial Diluted Alcohol and Iontophoresis-Assisted Corneal Cross-Linking in Progressive Keratoconus.

    Science.gov (United States)

    Bilgihan, Kamil; Yesilirmak, Nilufer; Altay, Yesim; Yuvarlak, Armagan; Ozdemir, Huseyin Baran

    2017-12-01

    To compare clinical outcomes of conventional corneal cross-linking (C-CXL) and diluted alcohol and iontophoresis-assisted corneal cross-linking (DAI-CXL) for the treatment of progressive keratoconus (KC). Ninety-three eyes of 80 patients with KC were treated by C-CXL (n = 47) or DAI-CXL (n = 46). Visual acuity, keratometry, KC indexes, pachymetry, and aberrations were recorded before treatment and 1, 3, 6, and 12 months after treatment. The demarcation line was assessed 1 month after treatment. A significant improvement in visual acuity was observed at month 3 and month 6 after DAI-CXL and C-CXL, respectively. A significant decrease in maximum keratometry was observed in both groups at month 6. The front symmetry index significantly improved in both groups after 6 months, whereas the Baiocchi Calossi Versaci index significantly improved only after DAI-CXL at month 12 (P = 0.01). Average keratometry and other KC indexes were stable during 12 months of follow-up. Central corneal thickness decreased by 28.6 and 40.2 μm after DAI-CXL and C-CXL at month 1, respectively (P < 0.01), and it reached baseline at the 12th month (P = 0.14) only in the DAI-CXL group. Higher-order aberrations, coma, and spherical aberration significantly worsened at month 1 (P < 0.01) only after C-CXL; however, they improved significantly at month 12 compared with baseline (P < 0.05) in both groups. The demarcation line was visible in all cases at month 1 at a mean depth of 302 ± 56 μm and 311 ± 57 μm after DAI-CXL and C-CXL, respectively (P = 0.7). The DAI-CXL protocol seems as effective as the C-CXL protocol in halting KC progression after 1 year of follow-up.

  10. Interstrand Coupling Properties of LARP High Gradient Quadrupole Cables in Response to Variations in Cable Design and Heat Treatment Condition

    NARCIS (Netherlands)

    Collings, E.W.; Sumption, M.D.; Majoros, M.; Wang, Xiaorong; Dietderich, D.R.; Yagotyntsev, Kostyantyn; Nijhuis, Arend

    Calorimetric measurement of coupling loss versus frequency has been measured on two sets of cored and uncored large Hadron Collider Accelerator Research Program high gradient quadrupole Nb3Sn Rutherford cables. Studied are the responses of the resulting interstrand contact resistances (ICR) to

  11. Chitosan nanogels by template chemical cross-linking in polyion complex micelle nanoreactors.

    Science.gov (United States)

    Maggi, Flavia; Ciccarelli, Serena; Diociaiuti, Marco; Casciardi, Stefano; Masci, Giancarlo

    2011-10-10

    Chitosan covalent nanogels cross-linked with genipin were prepared by template chemical cross-linking of chitosan in polyion complex micelle (PIC) nanoreactors. By using this method, we were able to prepare chitosan nanogels using only biocompatible materials without organic solvents. PIC were prepared by interaction between chitosan (X(n) = 23, 44, and 130) and block copolymer poly(ethylene oxide)-block-poly[sodium 2-(acrylamido)-2-methylpropanesulfonate] (PEO-b-PAMPS) synthesized by single-electron transfer-living radical polymerization (SET-LRP). PIC with small size (diameter about 50 nm) and low polydispersity were obtained up to 5 mg/mL. After cross-linking of chitosan with genipin, the nanoreactors were dissociated by adding NaCl. The dissociation of the nanoreactors and the formation of the nanogels were confirmed by (1)H NMR, DLS, and TEM. The size of the smallest nanogels was about 50 nm in the swollen state and 20 nm in the dry state. The amount of genipin used during reticulation was an important parameter to modulate the size of the nanogels in solution.

  12. Wood Sawdust/Natural Rubber Ecocomposites Cross-Linked by Electron Beam Irradiation.

    Science.gov (United States)

    Manaila, Elena; Stelescu, Maria Daniela; Craciun, Gabriela; Ighigeanu, Daniel

    2016-06-23

    The obtaining and characterization of some polymeric eco-composites based on wood sawdust and natural rubber is presented. The natural rubber was cross-linked using the electron beam irradiation. The irradiation doses were of 75, 150, 300 and 600 kGy and the concentrations of wood sawdust were of 10 and 20 phr, respectively. As a result of wood sawdust adding, the physical and mechanical properties such as hardness, modulus at 100% elongation and tensile strength, showed significant improvements. The presence of wood sawdust fibers has a reinforcing effect on natural rubber, similar or better than of mineral fillers. An increase in the irradiation dose leads to the increasing of cross-link density, which is reflected in the improvement of hardness, modulus at 100% elongation and tensile strength of blends. The cross-linking rates, appreciated using the Flory-Rehner equation, have increased with the amount of wood sawdust in blends and with the irradiation dose. Even if the gel fraction values have varied irregularly with the amount of wood sawdust and irradiation dose it was over 90% for all blends, except for the samples without wood sawdust irradiated with 75 kGy. The water uptake increased with increasing of fiber content and decreased with the irradiation dose.

  13. A Single Molecular Diels-Alder Crosslinker for Achieving Recyclable Cross-Linked Polymers.

    Science.gov (United States)

    Chen, Shengli; Wang, Fenfen; Peng, Yongjin; Chen, Tiehong; Wu, Qiang; Sun, Pingchuan

    2015-09-01

    A triol-functional crosslinker combining the thermoreversible properties of Diels-Alder (DA) adducts in one molecule is designed, synthesized, and used as an ideal substitute of a traditional crosslinker to prepare thermal recyclable cross-linked polyurethanes with excellent mechanical properties and recyclability in a very simple and efficient way. The recycle property of these materials achieved by the DA/retro-DA reaction at a suitable temperature is verified by differential scanning calorimetry and in situ variable temperature solid-state NMR experiments during the cyclic heating and cooling processes. The thermal recyclability and remending ability of the bulk polyurethanes is demonstrated by three polymer processing methods, including hot-press molding, injection molding, and solution casting. It is notable that all the recycled cross-linked polymers display nearly invariable elongation/stress at break compared to the as-synthesized samples. Further end-group functionalization of this single molecular DA crosslinker provides the potential in preparing a wide range of recyclable cross-linked polymers. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Two-photon induced collagen cross-linking in bioartificial cardiac tissue

    Science.gov (United States)

    Kuetemeyer, Kai; Kensah, George; Heidrich, Marko; Meyer, Heiko; Martin, Ulrich; Gruh, Ina; Heisterkamp, Alexander

    2011-08-01

    Cardiac tissue engineering is a promising strategy for regenerative therapies to overcome the shortage of donor organs for transplantation. Besides contractile function, the stiffness of tissue engineered constructs is crucial to generate transplantable tissue surrogates with sufficient mechanical stability to withstand the high pressure present in the heart. Although several collagen cross-linking techniques have proven to be efficient in stabilizing biomaterials, they cannot be applied to cardiac tissue engineering, as cell death occurs in the treated area. Here, we present a novel method using femtosecond (fs) laser pulses to increase the stiffness of collagen-based tissue constructs without impairing cell viability. Raster scanning of the fs laser beam over riboflavin-treated tissue induced collagen cross-linking by two-photon photosensitized singlet oxygen production. One day post-irradiation, stress-strain measurements revealed increased tissue stiffness by around 40% being dependent on the fibroblast content in the tissue. At the same time, cells remained viable and fully functional as demonstrated by fluorescence imaging of cardiomyocyte mitochondrial activity and preservation of active contraction force. Our results indicate that two-photon induced collagen cross-linking has great potential for studying and improving artificially engineered tissue for regenerative therapies.

  15. Application of cross-linked and hydrolyzed arabinoxylans in baking of model rye bread.

    Science.gov (United States)

    Buksa, Krzysztof; Nowotna, Anna; Ziobro, Rafał

    2016-02-01

    The role of water extractable arabinoxylan with varying molar mass and structure (cross-linked vs. hydrolyzed) in the structure formation of rye bread was examined using a model bread. Instead of the normal flour, the dough contained starch, arabinoxylan and protein, which were isolated from rye wholemeal. It was observed that the applied mixes of these constituents result in a product closely resembling typical rye bread, even if arabinoxylan was modified (by cross-linking or hydrolysis). The levels of arabinoxylan required for bread preparation depended on its modification and mix composition. At 3% protein, the maximum applicable level of poorly soluble cross-linked arabinoxylan was 3%, as higher amounts of this preparation resulted in an extensively viscous dough and diminished bread volume. On the other hand highly soluble, hydrolyzed arabinoxylan could be used at a higher level (6%) together with larger amounts of rye protein (3% or 6%). Further addition of arabinoxylan leads to excessive water absorption, resulting in a decreased viscosity of the dough during baking and insufficient gas retention. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Cross-links in carbon nanotube assembly introduced by using polyacrylonitrile as precursor.

    Science.gov (United States)

    Cui, Yanbin; Zhang, Mei

    2013-08-28

    Individual carbon nanotube (CNT) exhibits extraordinary mechanics. However, the properties of the macroscopic CNT-based structure, such as CNT fibers and films, are far lower than that of individual CNT. One of the main reasons is the weak interaction between tubes and bundles in the CNT assemblies. It is understood that the cross-links in CNT assembly play a key role to improve the performance of CNT-based structure. Different approaches have been taken to create CNT joints. Most of these approaches focus on connecting CNTs by generating new covalent bonding between tubes. In this work, we intend to reinforce the CNT network by locking the contacted CNTs. Polyacrylonitrile (PAN) was used as precursor because PAN can form graphitic structures after carbonization. The freestanding superthin CNT sheet and CNT yarn were used to evaluate the effects of the PAN precursor to form cross-links between CNTs. The tensile strength of CNT yarn is improved when the yarn is partially infiltrated with PAN and consequently carbonated. High-resolution transmission electron microscopy observation of the sheets shows that graphite structures are formed and cross-link CNTs in CNT assembly.

  17. Seed-mediated synthesis of cross-linked Pt-NiO nanochains for methanol oxidation

    Energy Technology Data Exchange (ETDEWEB)

    Gu, Zhulan; Bin, Duan; Feng, Yue; Zhang, Ke; Wang, Jin; Yan, Bo; Li, Shumin; Xiong, Zhiping; Wang, Caiqin; Shiraishi, Yukihide; Du, Yukou, E-mail: duyk@suda.edu.cn

    2017-07-31

    Highlights: • Cross-linked Pt-NiO nanochains using seed-mediated growth method are synthesized. • The as-prepared catalysts exhibit higher electrocatalytic activity than Pt/C for MOR. • The Pt-NiO(1:1 by molar) catalyst shows the best electrocatalytic property towards MOR. - Abstract: A simple method was reported for employing NiO nanoparticles act as seeds and then different amounts of Pt{sup 2+} were reduced on the NiO nanoparticles, forming a cross-linked Pt-NiO nanocatalysts. These as-prepared catalysts were characterized using different physical-chemical techniques, including X-ray diffraction (XRD), transmission electron microscopy (TEM) and X-ray photoelectron spectroscopy (XPS). The results indicate that the morphology of the cross-linked Pt-NiO nanochain was successfully produced regardless of the molar ratio of Pt{sup 2+} to NiO precursors. The electrochemical characteristics of Pt-NiO nanochain catalysts were evaluated for the oxidation of methanol as a model reaction, which verify that the Pt-NiO catalysts show enhanced activity and high stability in comparison with the commercial Pt/C catalyst. The optimized ratio of Pt to NiO is 1:1, then tuned by simple adjusting the feed ratio of the precursors as well. The synthesized nanocatalysts will be found the great potential applications as electrocatalysts for fuel cells owe to their enhanced catalytic performance and long-term stability.

  18. Scheimpflug imaged corneal changes on anterior and posterior surfaces after collagen cross-linking

    Directory of Open Access Journals (Sweden)

    Ziad Hassan

    2014-04-01

    Full Text Available AIM: To compare the anterior and posterior corneal parameters before and after collagen cross-linking therapy for keratoconus.METHODS: Collagen cross-linking was performed in 31 eyes of 31 keratoconus patients (mean age 30.6±8.9y. Prior to treatment and an average 7mo after therapy, Scheimpflug analysis was performed using Pentacam HR. In addition to corneal thickness assessments, corneal radius, elevation, and aberrometric measurements were performed both on anterior and posterior corneal surfaces. Data obtained before and after surgery were statistically analyzed.RESULTS: In terms of horizontal and vertical corneal radius, and central corneal thickness no deviations were observed an average 7mo after operation. Corneal higher order aberration showed no difference neither on anterior nor on posterior corneal surfaces. During follow-up period, no significant deviation was detected regarding elevation values obtained by measurement in mm units between the 3.0-8.0 mm-zones.CONCLUSION: Corneal stabilization could be observed in terms of anterior and posterior corneal surfaces, elevation and higher order aberration values 7mo after collagen cross-linking therapy for keratoconus.

  19. Capacity of cognitive radio under imperfect secondary and cross link channel state information

    KAUST Repository

    Sboui, Lokman

    2011-09-01

    In this paper, we study the ergodic capacity of secondary user channel in a spectrum sharing scenario in which the secondary transmitter is instantaneously aware of estimated versions of the cross link (between the secondary transmitter and the primary receiver) and the secondary link Channel State Information (CSI). The secondary link optimal power profile along with the ergodic capacity are derived for a class of fading channels, under an average power constraint and an instantaneous interference outage constraint. We also show that our framework is rather general as it encompasses several previously studied spectrum sharing settings as special cases. In order to gain some insights on the capacity behavior, numerical results are shown for independent Rayleigh fading channels where it is found for instance, that at low SNR regime, only the secondary channel estimation matters and that the cross link CSI has no effect on the ergodic capacity; whereas at high SNR regime, the capacity is rather driven by the cross link CSI. © 2011 IEEE.

  20. Azide-based cross-linking of polymers of intrinsic microporosity (PIMs) for condensable gas separation

    KAUST Repository

    Du, Naiying

    2011-03-11

    Cross-linked polymers of intrinsic microporosity (PIM)s for gas separation membranes, were prepared by a nitrene reaction from a representative PIM in the presence of two different diazide cross-linkers. The reaction temperature was optimized using TGA. The homogenous membranes were cast from THF solutions of different ratios of PIM to azides. The resulting cross-linked structures of the PIMs membranes were formed at 175 °C after 7.5 h and confirmed by TGA, XPS, FT-IR spectroscopy and gel content analysis. These resulting cross-linked polymeric membranes showed excellent gas separation performance and can be used for O 2/N 2 and CO 2/N 2 gas pairs, as well as for condensable gases, such as CO 2/CH 4, propylene/propane separation. Most importantly, and differently from typical gas separation membranes derived from glassy polymers, the crosslinked PIMs showed no obvious CO 2 plasticization up to 20 atm pressure of pure CO 2 and CO 2/CH 4 mixtures. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Cheese whey protein recovery by ultrafiltration through transglutaminase (TG) catalysis whey protein cross-linking.

    Science.gov (United States)

    Wen-Qiong, Wang; Lan-Wei, Zhang; Xue, Han; Yi, Lu

    2017-01-15

    In whey ultrafiltration (UF) production, two main problems are whey protein recovery and membrane fouling. In this study, membrane coupling protein transglutaminase (TG) catalysis protein cross-linking was investigated under different conditions to find out the best treatment. We found that the optimal conditions for protein recovery involved catalyzing whey protein cross-linking with TG (40U/g whey proteins) at 40°C for 60min at pH 5.0. Under these conditions, the recovery rate was increased 15-20%, lactose rejection rate was decreased by 10%, and relative permeate flux was increase 30-40% compared to the sample without enzyme treatment (control). It was noticeable that the total resistance and cake resistance were decreased after enzyme catalysis. This was mainly due to the increased particle size and decreased zeta potential. Therefore, membrane coupling enzyme catalysis protein cross-linking is a potential means for further use. Copyright © 2016. Published by Elsevier Ltd.

  2. Azide-based cross-linking of polymers of intrinsic microporosity (PIMs) for condensable gas separation.

    Science.gov (United States)

    Du, Naiying; Cin, Mauro M Dal-; Pinnau, Ingo; Nicalek, Andrzej; Robertson, Gilles P; Guiver, Michael D

    2011-04-19

    Cross-linked polymers of intrinsic microporosity (PIM)s for gas separation membranes, were prepared by a nitrene reaction from a representative PIM in the presence of two different diazide cross-linkers. The reaction temperature was optimized using TGA. The homogenous membranes were cast from THF solutions of different ratios of PIM to azides. The resulting cross-linked structures of the PIMs membranes were formed at 175 °C after 7.5 h and confirmed by TGA, XPS, FT-IR spectroscopy and gel content analysis. These resulting cross-linked polymeric membranes showed excellent gas separation performance and can be used for O(2) /N(2) and CO(2) /N(2) gas pairs, as well as for condensable gases, such as CO(2) /CH(4) , propylene/propane separation. Most importantly, and differently from typical gas separation membranes derived from glassy polymers, the crosslinked PIMs showed no obvious CO(2) plasticization up to 20 atm pressure of pure CO(2) and CO(2) /CH(4) mixtures. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Seed-mediated synthesis of cross-linked Pt-NiO nanochains for methanol oxidation

    International Nuclear Information System (INIS)

    Gu, Zhulan; Bin, Duan; Feng, Yue; Zhang, Ke; Wang, Jin; Yan, Bo; Li, Shumin; Xiong, Zhiping; Wang, Caiqin; Shiraishi, Yukihide; Du, Yukou

    2017-01-01

    Highlights: • Cross-linked Pt-NiO nanochains using seed-mediated growth method are synthesized. • The as-prepared catalysts exhibit higher electrocatalytic activity than Pt/C for MOR. • The Pt-NiO(1:1 by molar) catalyst shows the best electrocatalytic property towards MOR. - Abstract: A simple method was reported for employing NiO nanoparticles act as seeds and then different amounts of Pt 2+ were reduced on the NiO nanoparticles, forming a cross-linked Pt-NiO nanocatalysts. These as-prepared catalysts were characterized using different physical-chemical techniques, including X-ray diffraction (XRD), transmission electron microscopy (TEM) and X-ray photoelectron spectroscopy (XPS). The results indicate that the morphology of the cross-linked Pt-NiO nanochain was successfully produced regardless of the molar ratio of Pt 2+ to NiO precursors. The electrochemical characteristics of Pt-NiO nanochain catalysts were evaluated for the oxidation of methanol as a model reaction, which verify that the Pt-NiO catalysts show enhanced activity and high stability in comparison with the commercial Pt/C catalyst. The optimized ratio of Pt to NiO is 1:1, then tuned by simple adjusting the feed ratio of the precursors as well. The synthesized nanocatalysts will be found the great potential applications as electrocatalysts for fuel cells owe to their enhanced catalytic performance and long-term stability.

  4. Thermal Conductance in Cross-linked Polymers: Effects of Non-Bonding Interactions.

    Science.gov (United States)

    Rashidi, Vahid; Coyle, Eleanor J; Sebeck, Katherine; Kieffer, John; Pipe, Kevin P

    2017-05-04

    Weak interchain interactions have been considered to be a bottleneck for heat transfer in polymers, while covalent bonds are believed to give a high thermal conductivity to polymer chains. For this reason, cross-linkers have been explored as a means to enhance polymer thermal conductivity; however, results have been inconsistent. Some studies show an enhancement in the thermal conductivity for polymers upon cross-linking, while others show the opposite trend. In this work we study the mechanisms of heat transfer in cross-linked polymers in order to understand the reasons for these discrepancies, in particular examining the relative contributions of covalent (referred to here as "bonding") and nonbonding (e.g., van der Waals and electrostatic) interactions. Our results indicate cross-linkers enhance thermal conductivity primarily when they are short in length and thereby bring polymer chains closer to each other, leading to increased interchain heat transfer by enhanced nonbonding interactions between the chains (nonbonding interactions being highly dependent on interchain distance). This suggests that enhanced nonbonding interactions, rather than thermal pathways through cross-linker covalent bonds, are the primary transport mechanism by which thermal conductivity is increased. We further illustrate this by showing that energy from THz acoustic waves travels significantly faster in polymers when nonbonding interactions are included versus when only covalent interactions are present. These results help to explain prior studies that measure differing trends in thermal conductivity for polymers upon cross-linking with various species.

  5. Structural Analysis and Mechanical Characterization of Hyaluronic Acid-Based Doubly Cross-Linked Networks.

    Science.gov (United States)

    Jha, Amit K; Hule, Rohan A; Jiao, Tong; Teller, Sean S; Clifton, Rodney J; Duncan, Randall L; Pochan, Darrin J; Jia, Xinqiao

    2009-01-01

    We have created a new class of hyaluronic acid (HA)-based hydrogel materials with HA hydrogel particles (HGPs) embedded in and covalently cross-linked to a secondary network. HA HGPs with an average diameter of ∼900 nm and narrow particle size distribution were synthesized using a refined reverse micelle polymerization technique. The average mesh size of the HGPs was estimated to be approximately 5.5 to 7.0 nm by a protein uptake experiment. Sodium periodate oxidation not only introduced aldehyde groups to the particles but also reduced the average particle size. The aldehyde groups generated were used as reactive handles for subsequent cross-linking with an HA derivative containing hydrazide groups. The resulting macroscopic gels contain two distinct hierarchical networks (doubly cross-linked networks, DXNs): one within individual particles and another among different particles. Bulk gels (BGs) formed by direct mixing of HA derivatives with mutually reactive groups were included for comparison. The hydrogel microstructures were collectively characterized by microscopy and neutron scattering techniques. Their viscoelasticity was quantified at low frequencies (0.1-10 Hz) using a controlled stress rheometer and at high frequencies (up to 200 Hz) with a home-built torsional wave apparatus. Both BGs and DXNs are stable elastic gels that become stiffer at higher frequencies. The HA-based DXN offers unique structural hierarchy and mechanical properties that are suitable for soft tissue regeneration.

  6. Human muscle LIM protein dimerizes along the actin cytoskeleton and cross-links actin filaments.

    Science.gov (United States)

    Hoffmann, Céline; Moreau, Flora; Moes, Michèle; Luthold, Carole; Dieterle, Monika; Goretti, Emeline; Neumann, Katrin; Steinmetz, André; Thomas, Clément

    2014-08-01

    The muscle LIM protein (MLP) is a nucleocytoplasmic shuttling protein playing important roles in the regulation of myocyte remodeling and adaptation to hypertrophic stimuli. Missense mutations in human MLP or its ablation in transgenic mice promotes cardiomyopathy and heart failure. The exact function(s) of MLP in the cytoplasmic compartment and the underlying molecular mechanisms remain largely unknown. Here, we provide evidence that MLP autonomously binds to, stabilizes, and bundles actin filaments (AFs) independently of calcium and pH. Using total internal reflection fluorescence microscopy, we have shown how MLP cross-links actin filaments into both unipolar and mixed-polarity bundles. Quantitative analysis of the actin cytoskeleton configuration confirmed that MLP substantially promotes actin bundling in live myoblasts. In addition, bimolecular fluorescence complementation (BiFC) assays revealed MLP self-association. Remarkably, BiFC complexes mostly localize along actin filament-rich structures, such as stress fibers and sarcomeres, supporting a functional link between MLP self-association and actin cross-linking. Finally, we have demonstrated that MLP self-associates through its N-terminal LIM domain, whereas it binds to AFs through its C-terminal LIM domain. Together our data support that MLP contributes to the maintenance of cardiomyocyte cytoarchitecture by a mechanism involving its self-association and actin filament cross-linking. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  7. Self-Healing Gelatin Hydrogels Cross-Linked by Combining Multiple Hydrogen Bonding and Ionic Coordination.

    Science.gov (United States)

    Zhang, Guangzhao; Lv, Lei; Deng, Yonghong; Wang, Chaoyang

    2017-06-01

    Self-healing hydrogels have been studied by many researchers via multiple cross-linking approaches including physical and chemical interactions. It is an interesting project in multifunctional hydrogel exploration that a water soluble polymer matrix is cross-linked by combining the ionic coordination and the multiple hydrogen bonds to fabricate self-healing hydrogels with injectable property. This study introduces a general procedure of preparing the hydrogels (termed gelatin-UPy-Fe) cross-linked by both ionic coordination of Fe 3+ and carboxyl group from the gelatin and the quadruple hydrogen bonding interaction from the ureido-pyrimidinone (UPy) dimers. The gelatin-UPy-Fe hydrogels possess an excellent self-healing property. The effects of the ionic coordination of Fe 3+ and quadruple hydrogen bonding of UPy on the formation and mechanical behavior of the prepared hydrogels are investigated. In vitro drug release of the gelatin-UPy-Fe hydrogels is also observed, giving an intriguing glimpse into possible biological applications. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Cross-Linked Enzyme Aggregates of Naringinase: Novel Biocatalysts for Naringin Hydrolysis

    Directory of Open Access Journals (Sweden)

    Maria H. L. Ribeiro

    2011-01-01

    Full Text Available Cross-linked enzyme aggregates (CLEAs have emerged as interesting biocatalyst design for immobilization. These new generation enzyme biocatalysts, CLEAs, in addition to exhibiting good mechanical stability, can be highly active, since they do not include large amounts of foreign particulate nonenzymatic material and may have increased stability. Naringinase (NGase is an enzyme complex with high potential in pharmaceutical and food industries. In fact, NGase can be used in the biotransformation of steroids, of antibiotics and mainly on glycosides hydrolysis. In this paper, the formation of CLEAs was tried using ammonium sulphate, polyethylene glycol 6000 and tert-butyl alcohol as precipitant agents and glutaraldehyde as cross-linking agent, at different pH, time, and temperature conditions. However, among the precipitant agents tested, only tert-butyl alcohol cross-linked with glutaraldehyde allowed the formation of CLEAs, at pH 4.0 and at temperature between 7 and 10∘C. Different enzyme loadings were tested. The NGase-CLEAs were highly effective in naringin hydrolysis. The operational stability of the NGase-CLEAs aggregates was studied through six successive reutilizations.

  9. A novel sulfonated poly(ether ether ketone) and cross-linked membranes for fuel cells

    Energy Technology Data Exchange (ETDEWEB)

    Li, Hongtao; Zhang, Gang; Wu, Jing; Zhao, Chengji; Zhang, Yang; Shao, Ke; Han, Miaomiao; Lin, Haidan; Zhu, Jing; Na, Hui [Alan G MacDiarmid Institute, College of Chemistry, Jilin University, Qianjin Street 2699, Changchun 130012, Jilin (China)

    2010-10-01

    A novel poly(ether ether ketone) (PEEK) containing pendant carboxyl groups has been synthesized by a nucleophilic polycondensation reaction. Sulfonated polymers (SPEEKs) with different ion exchange capacity are then obtained by post-sulfonation process. The structures of PEEK and SPEEKs are characterized by both FT-IR and {sup 1}H NMR. The properties of SPEEKs as candidates for proton exchange membranes are studied. The cross-linking reaction is performed at 140 C using poly(vinyl alcohol) (PVA) as the cross-linker. In comparison with the non-cross-linked membranes, some properties of the cross-linked membranes are significantly improved, such as water uptake, methanol resistance, mechanical and oxidative stabilities, while the proton conductivity decreases. The effect of PVA content on proton conductivity, water uptake, swelling ratio, and methanol permeability is also investigated. Among all the membranes, SPEEK-C-8 shows the highest selectivity of 50.5 x 10{sup 4} S s cm{sup -3}, which indicates that it is a suitable candidate for applications in direct methanol fuel cells. (author)

  10. Induction of helical structure in a heptapeptide with a metal cross-link: modification of the Lifson-Roig helix-coil theory to account for covalent cross-links.

    Science.gov (United States)

    Kise, Kenneth J; Bowler, Bruce E

    2002-12-31

    A short peptide, acetyl-AHAAAHA-carboxamide, has been synthesized and the histidines cross-linked with a cis-tetraammineruthenium(III) moiety. In the absence of the Ru(III) cross-link, the heptapeptide is essentially structureless, as judged by circular dichroism, NMR chemical shift, and NMR-monitored hydrogen deuterium exchange data. The presence of the cis-Ru(III) cross-link is confirmed by mass spectral data and the characteristic pH dependence of the UV-vis spectrum of the cis-(bis-(imidazole))ruthenium(III) unit. Circular dichroism data indicate that the Ru(III) cross-linked heptapeptide is approximately 37% helical at 0 degrees C. The NMR spectrum of the cross-linked peptide has been fully assigned using TOCSY and ROESY experiments. ROE interactions and J-coupling data provide evidence for helical structure. NMR-monitored hydrogen-deuterium exchange data for the Ru(III)-cross-linked peptide, resolved at the level of the individual amides, give larger protection factors at the ends than in the center of the helix. Steric and polarization effects of the Ru(III) cross-link are proposed to cause this unusual apparent protection pattern. A modification to the Lifson-Roig helix-coil model to account for the effect of the i,i+4 Ru(III) cross-link on the helix-coil transition of a peptide is presented. The model provides an excellent fit to the temperature dependence of the circular dichroism spectrum of the Ru(III)-cross-linked peptide. The modified model indicates that the effect of the cross-link on the nucleation parameter, v(2), is modest (about 7-fold) for residues bounded by the cross-link. Significant increases in the propagation parameter, w, occur for residues within the cross-link. The modification to the Lifson-Roig model accounts for the effect of a Ru(III) cross-link on the circular dichroism spectrum of a previously reported 17 residue peptide.

  11. Impact of styrenic polymer one-step hyper-cross-linking on volatile organic compound adsorption and desorption performance.

    Science.gov (United States)

    Ghafari, Mohsen; Atkinson, John D

    2018-06-05

    A novel one-step hyper-cross-linking method, using 1,2-dichloroethane (DCE) and 1,6-dichlorohexane (DCH) cross-linkers, expands the micropore volume of commercial styrenic polymers. Performance of virgin and modified polymers was evaluated by measuring hexane, toluene, and methyl-ethyl-ketone (MEK) adsorption capacity, adsorption/desorption kinetics, and desorption efficiency. Hyper-cross-linked polymers have up to 128% higher adsorption capacity than virgin polymers at P/P 0  = 0.05 due to micropore volume increases up to 330%. Improvements are most pronounced with the DCE cross-linker. Hyper-cross-linking has minimal impact on hexane adsorption kinetics, but adsorption rates for toluene and MEK decrease by 6-41%. Desorption rates decreased (3-36%) for all materials after hyper-cross-linking, with larger decreases for DCE hyper-cross-linked polymers due to smaller average pore widths. For room temperature desorption, 20-220% more adsorbate remains in hyper-cross-linked polymers after regeneration compared to virgin materials. DCE hyper-cross-linked polymers have 13-92% more residual adsorbate than DCH counterparts. Higher temperatures were required for DCE hyper-cross-linked polymers to completely desorb VOCs compared to the DCH hyper-cross-linked and virgin counterparts. Results show that the one-step hyper-cross-linking method for modifying styrenic polymers improves adsorption capacity because of added micropores, but decreases adsorption/desorption kinetics and desorption efficiency for large VOCs due to a decrease in average pore width. Copyright © 2018 Elsevier B.V. All rights reserved.

  12. Cross-Linking Mast Cell Specific Gangliosides Stimulates the Release of Newly Formed Lipid Mediators and Newly Synthesized Cytokines

    Directory of Open Access Journals (Sweden)

    Edismauro Garcia Freitas Filho

    2016-01-01

    Full Text Available Mast cells are immunoregulatory cells that participate in inflammatory processes. Cross-linking mast cell specific GD1b derived gangliosides by mAbAA4 results in partial activation of mast cells without the release of preformed mediators. The present study examines the release of newly formed and newly synthesized mediators following ganglioside cross-linking. Cross-linking the gangliosides with mAbAA4 released the newly formed lipid mediators, prostaglandins D2 and E2, without release of leukotrienes B4 and C4. The effect of cross-linking these gangliosides on the activation of enzymes in the arachidonate cascade was then investigated. Ganglioside cross-linking resulted in phosphorylation of cytosolic phospholipase A2 and increased expression of cyclooxygenase-2. Translocation of 5-lipoxygenase from the cytosol to the nucleus was not induced by ganglioside cross-linking. Cross-linking of GD1b derived gangliosides also resulted in the release of the newly synthesized mediators, interleukin-4, interleukin-6, and TNF-α. The effect of cross-linking the gangliosides on the MAP kinase pathway was then investigated. Cross-linking the gangliosides induced the phosphorylation of ERK1/2, JNK1/2, and p38 as well as activating both NFκB and NFAT in a Syk-dependent manner. Therefore, cross-linking the mast cell specific GD1b derived gangliosides results in the activation of signaling pathways that culminate with the release of newly formed and newly synthesized mediators.

  13. In Vivo Immune Responses of Cross-Linked Electrospun Tilapia Collagen Membrane.

    Science.gov (United States)

    Hassanbhai, Ammar Mansoor; Lau, Chau Sang; Wen, Feng; Jayaraman, Praveena; Goh, Bee Tin; Yu, Na; Teoh, Swee-Hin

    2017-10-01

    Collagen has been used extensively in tissue engineering applications. However, the source of collagen has been primarily bovine and porcine. In view of the potential risk of zoonotic diseases and religious constraints associated with bovine and porcine collagen, fish collagen was examined as an alternative. The aim of this study is to use tilapia fish collagen to develop a cross-linked electrospun membrane to be used as a barrier membrane in guided bone regeneration. As there is limited data available on the cytotoxicity and immunogenicity of cross-linked tilapia collagen, in vitro and in vivo tests were performed to evaluate this in comparison to the commercially available Bio-Gide ® membrane. In this study, collagen was extracted and purified from tilapia skin and electrospun into a nanofibrous membrane. The resultant membrane was cross-linked to obtain a cross-linked electrospun tilapia collagen (CETC) membrane, which was characterized by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), degradation studies, cytotoxicity studies, and cell proliferation studies. The membranes were also implanted subcutaneously in rats and the host immune responses were examined. The DSC data showed that cross-linking increased the denaturation temperature of tilapia collagen to 58.3°C ± 1.4°C. The in vitro tests showed that CETC exhibited no cytotoxicity toward murine fibroblast L929 cells, and culture of murine preosteoblast MC3T3-E1 cells demonstrated better proliferation on CETC as compared to Bio-Gide. When implanted in rats, CETC caused a higher production of interleukin IL-6 at early time points as compared to Bio-Gide, but there was no long-term inflammatory responses after the acute inflammation phase. This finding was supported with histology data, which clearly illustrated that CETC has a decreased inflammatory response comparable to the benchmark control group. In all, this study demonstrated the viability for the use of CETC as a

  14. Optimization of Formaldehyde Cross-Linking for Protein Interaction Analysis of Non-Tagged Integrin β1

    Directory of Open Access Journals (Sweden)

    Cordula Klockenbusch

    2010-01-01

    Full Text Available Formaldehyde cross-linking of protein complexes combined with immunoprecipitation and mass spectrometry analysis is a promising technique for analysing protein-protein interactions, including those of transient nature. Here we used integrin β1 as a model to describe the application of formaldehyde cross-linking in detail, particularly focusing on the optimal parameters for cross-linking, the detection of formaldehyde cross-linked complexes, the utility of antibodies, and the identification of binding partners. Integrin β1 was found in a high molecular weight complex after formaldehyde cross-linking. Eight different anti-integrin β1 antibodies were used for pull-down experiments and no loss in precipitation efficiency after cross-linking was observed. However, two of the antibodies could not precipitate the complex, probably due to hidden epitopes. Formaldehyde cross-linked complexes, precipitated from Jurkat cells or human platelets and analyzed by mass spectrometry, were found to be composed of integrin β1, α4 and α6 or β1, α6, α2, and α5, respectively.

  15. Nonenzymatic cross-linking pentosidine increase in bone collagen and are associated with disorders of bone mineralization in dialysis patients.

    Science.gov (United States)

    Mitome, Jun; Yamamoto, Hiroyasu; Saito, Mitsuru; Yokoyama, Keitaro; Marumo, Keishi; Hosoya, Tatsuo

    2011-06-01

    Disorders of bone and mineral metabolism are common complications in chronic kidney disease (CKD) patients and lead to significantly increased fracture risk, morbidity, and mortality of cardiovascular disease due to ectopic calcifications, contributing to a worsening prognosis. Bone strength is determined by not only bone mineral density but also bone quality, which is dependent on bone collagen cross-links. Collagen cross-links are classified into enzymatic immature and mature types and nonenzymatic advanced glycation end products (AGEs). Pentosidine is well established as one of the AGEs that accumulates markedly in CKD patients. The chemistry, function, and clinical relevance of cross-links have been revealed, whereas bone quality and the relationship with bone mineralization in CKD patients are not clear. We performed transiliac bone biopsies on 22 dialysis patients (mean age 56 ± 9 years) with severe secondary hyperparathyroidism and measured cross-links by evaluating bone histomorphometry. Cross-links data were compared with age-matched non-CKD subjects (mean age 58 ± 8 years, n = 17). Enzymatic collagen cross-links were formed to a similar extent compared with non-CKD subjects and showed a positive correlation with plasma intact parathyroid hormone. Pentosidine was remarkably increased in dialysis patients and inversely correlated with bone-formation rate/bone volume and mineral apposition rate. This study suggests that AGE collagen cross-links strongly associate with disorders of bone metabolism in dialysis patients.

  16. Characterization of spray-dried microalgal oil encapsulated in cross-linked sodium caseinate matrix induced by microbial transglutaminase.

    Science.gov (United States)

    Bao, Shan-Shan; Hu, Xue-Chao; Zhang, Ka; Xu, Xiao-Kang; Zhang, Hong-Man; Huang, He

    2011-01-01

    Sodium caseinate (SC) cross-linked by microbial transglutaminase (MTGase) for encapsulating microalgal oil was investigated. Protein cross-linking was evidenced in the SDS-PAGE graph. The emulsifying properties of SC depended on the cross-linked time with MTGase. The emulsifying activity and stability indexes of SC increased with the cross-linking time of 30 to 90 min (P30 to P90), and then declined with longer cross-linked time of 180 to 420 min (P180 to P420). The P30 to P90 as wall material for microencapsulation was superior to P180 to P420 and control sample. The microcapsules prepared with P30 to P90 showed more than 97% of microencapsulation efficiency in contrast to about 90% with the P180 to P420. During storage, the microcapsules prepared with P30 to P90 exhibited higher oxidative stability as compared with other microcapsules. A sustained release of microalgal oil was observed, and core release was time dependent and affected by cross-linking degree. Results showed that the powdered microalgal oil prepared with P30 to P90 demonstrated enhanced physicochemical properties and oxidative stability. Practical Application: The novel method using cross-linked proteins as wall material induced by microbial transglutaminase in food industry for sensitive ingredients could convert microcapsules into a stable form, which would lead to its more widespread utilization as a kind of food additive.

  17. Sub- T g Cross-Linking of a Polyimide Membrane for Enhanced CO 2 Plasticization Resistance for Natural Gas Separation

    KAUST Repository

    Qiu, Wulin

    2011-08-09

    Decarboxylation-induced thermal cross-linking occurs at elevated temperatures (∼15 °C above glass transition temperature) for 6FDA-DAM:DABA polyimides, which can stabilize membranes against swelling and plasticization in aggressive feed streams. Despite this advantage, such a high temperature might result in collapse of substructure and transition layers in the asymmetric structure of a hollow fibers based on such a material. In this work, the thermal cross-linking of the 6FDA-DAM:DABA at temperatures much below the glass transition temperature (∼387 °C by DSC) was demonstrated. This sub-Tg cross-linking capability enables extension to asymmetric structures useful for large scale membranes. The resulting polymer membranes were characterized by swelling in known solvents for the un-cross-linked materials, TGA analysis, and permeation tests of aggressive gas feed stream at higher pressure. The annealing temperature and time clearly influence the degree of cross-linking of the membranes, and results in a slight difference in selectivity for membranes under various cross-linking conditions. Results indicate that the sub-Tg thermal cross-linking of 6FDA-DAM:DABA dense film membrane can be carried out completely even at a temperature as low as 330 °C. Permeabilities were tested for the polyimide membranes using both pure gases (He, O2, N2, CH4, CO2) and mixed gases (CO2/CH4). The selectivity of the cross-linked membrane can be maintained even under very aggressive CO2 operating conditions that are not possible without cross-linking. Moreover, the plasticization resistance was demonstrated up to 700 psia for pure CO 2 gas or 1000 psia for 50% CO2 mixed gas feeds. © 2011 American Chemical Society.

  18. CD24 cross-linking induces apoptosis in, and inhibits migration of, MCF-7 breast cancer cells

    Directory of Open Access Journals (Sweden)

    Bae Ji-Yeon

    2008-04-01

    Full Text Available Abstract Background The biological effects of CD24 (FL-80 cross-linking on breast cancer cells have not yet been established. We examined the impact of CD24 cross-linking on human breast cancer cell line MCF-7. Methods MCF-7 and MDA-MB-231 cells were treated with anti-rabbit polyclonal IgG or anti-human CD24 rabbit polyclonal antibodies to induce cross-linking, and then growth was studied. Changes in cell characteristics such as cell cycle modulation, cell death, survival in three-dimensional cultures, adhesion, and migration ability were assayed after CD24 cross-linking in MCF-7. Results Expression of CD24 was analyzed by flow cytometry in MDA-MB-231 and MCF-7 cells where 2% and 66% expression frequencies were observed, respectively. CD24 cross-linking resulted in time-dependent proliferation reduction in MCF-7 cells, but no reduction in MDA-MB-231 cells. MCF-7 cell survival was reduced by 15% in three-dimensional culture after CD24 cross-linking. Increased MCF-7 cell apoptosis was observed after CD24 cross-linking, but no cell cycle arrest was observed in that condition. The migration capacity of MCF-7 cells was diminished by 30% after CD24 cross-linking. Conclusion Our results showed that CD24 cross-linking induced apoptosis and inhibited migration in MCF-7 breast cancer cells. We conclude that CD24 may be considered as a novel therapeutic target for breast cancer.

  19. Structural Architecture of the Nucleosome Remodeler ISWI Determined from Cross-Linking, Mass Spectrometry, SAXS, and Modeling.

    Science.gov (United States)

    Harrer, Nadine; Schindler, Christina E M; Bruetzel, Linda K; Forné, Ignasi; Ludwigsen, Johanna; Imhof, Axel; Zacharias, Martin; Lipfert, Jan; Mueller-Planitz, Felix

    2018-02-06

    Chromatin remodeling factors assume critical roles by regulating access to nucleosomal DNA. To determine the architecture of the Drosophila ISWI remodeling enzyme, we developed an integrative structural approach that combines protein cross-linking, mass spectrometry, small-angle X-ray scattering, and computational modeling. The resulting structural model shows the ATPase module in a resting state with both ATPase lobes twisted against each other, providing support for a conformation that was recently trapped by crystallography. The autoinhibiting NegC region does not protrude from the ATPase module as suggested previously. The regulatory NTR domain is located near both ATPase lobes. The full-length enzyme is flexible and can adopt a compact structure in solution with the C-terminal HSS domain packing against the ATPase module. Our data imply a series of conformational changes upon activation of the enzyme and illustrate how the NTR, NegC, and HSS domains contribute to regulation of the ATPase module. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Affect the Cross Linking Degree and Polymer Composition on the Mechanical Properties of Poly (vinyl alcohol/ Pullu-lan Films

    Directory of Open Access Journals (Sweden)

    Shemaa Abdul Satar Soud

    2018-04-01

    Full Text Available In this study Cross-linked PVA/Pullulan film was prepared. Cross-linking reaction done by addi-tion of gluteraldehyde at different reaction time (10,30and 60 min. Chemical interaction, me-chanical, thermal properties, water solubility and film morphology was studied for cross-linked PVA/Pullulan, PVA and Pullulan only. Thus FTIR investigated formation of hydrogen bonding between pullulan and PVA with (GA. Tensile strength, tensile modulus and elongation (% at break for PVA/Pullulan film was improved with addition of (GA as the reaction time proceed equivalent with increasing PVA content

  1. Effect of the cross-linking agent on performances of NaCS-CS/WSC microcapsules.

    Science.gov (United States)

    Wu, Qing-Xi; Xu, Xin; Wang, Zu-Li; Yao, Shan-Jing; Tong, Wang-Yu; Chen, Yan

    2016-11-01

    Based on the properties of oppositely charged natural polysaccharides, the polyelectrolyte complexes (PECs) prepared with chitosan-related polycationic polyelectrolytes and cellulose-related polyanionic polyelectrolytes have been widely concerned for their potential applications as micro-drug-carriers for colon. However, the poor mechanical property of the PECs becomes the obstacle encountered in practical applications. This study investigated the effect of the cross-linking agent (sodium polyphosphate, PPS) on the performances of sodium cellulose sulfate -chitosan/water soluble chitosan (NaCS-CS/WSC) microcapsules. The results revealed that PPS could penetrate through the PEC film and form tighter interior structures compared with the microcapsules without the addition of cross-linking agent. The NaCS-CS microcapsules and NaCS-WSC microcapsules with or without PPS had distinct microstructures, which could be ascribed to the different physicochemical properties of CS and WSC. During the formation process, CS can be dissolved in water under acidic conditions, while WSC can be directly dissolved and protonated in acid-free aqueous providing NH3(+) groups quickly, which resulted in the microstructure's difference. Further analysis showed the NaCS-CS-PPS microcapsules and NaCS-WSC-PPS microcapsules had lower swelling ratios due to their tighter interior microstructures that formed. The cross-linking agent had important effect on the total mass of PECs that produced; moreover, the decline of zeta potential of NaCS-CS-PPS microcapsules was lower than that of NaCS-CS microcapsules, similar trend was found in the NaCS-WSC-PPS microcapsules compared with NaCS-WSC microcapsules, indicating the PPS participated in the interactions and played a role in the microcapsules' formation process. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Water-resistant, transparent hybrid nanopaper by physical cross-linking with chitosan.

    Science.gov (United States)

    Toivonen, Matti S; Kurki-Suonio, Sauli; Schacher, Felix H; Hietala, Sami; Rojas, Orlando J; Ikkala, Olli

    2015-03-09

    One of the major, but often overlooked, challenges toward high end applications of nanocelluloses is to maintain their high mechanical properties under hydrated or even fully wet conditions. As such, permanent covalent cross-linking or surface hydrophobization are viable approaches, however, the former may hamper processability and the latter may have adverse effect on interfibrillar bonding and resulting material strength. Here we show a concept based on physical cross-linking of cellulose nanofibers (CNF, also denoted as microfibrillated cellulose, MFC, and, nanofibrillated cellulose, NFC) with chitosan for the aqueous preparation of films showing high mechanical strength in the wet state. Also, transparency (∼70-90% in the range 400-800 nm) is achieved by suppressing aggregation and carefully controlling the mixing conditions: Chitosan dissolves in aqueous medium at low pH and under these conditions the CNF/chitosan mixtures form easily processable hydrogels. A simple change in the environmental conditions (i.e., an increase of pH) reduces hydration of chitosan promoting multivalent physical interactions between CNF and chitosan over those with water, resulting effectively in cross-linking. Wet water-soaked films of CNF/chitosan 80/20 w/w show excellent mechanical properties, with an ultimate wet strength of 100 MPa (with corresponding maximum strain of 28%) and a tensile modulus of 4 and 14 GPa at low (0.5%) and large (16%) strains, respectively. More dry films of similar composition display strength of 200 MPa with maximum strain of 8% at 50% air relative humidity. We expect that the proposed, simple concept opens new pathways toward CNF-based material utilization in wet or humid conditions, which has still remained a challenge.

  3. Corneal collagen cross-linking to stop corneal ectasia exacerbated by radial keratotomy.

    Science.gov (United States)

    Mazzotta, Cosimo; Baiocchi, Stefano; Denaro, Rosario; Tosi, Gian Marco; Caporossi, Tomaso

    2011-02-01

    To assess the efficacy of riboflavin ultraviolet A (UV-A) corneal collagen cross-linking in the management of keratoconic corneal ectasia exacerbated by radial keratotomy (RK). A patient with progressive corneal ectasia and hyperopic shift, occurring 10 years after RK performed in the left eye, was treated with riboflavin UV-A corneal collagen cross-linking according to the Siena protocol: Pilocarpin 0.1% drop (1 hour before), lidocaine 4% drops 15 minutes before, mechanical scraping of epithelium (9-mm-diameter area), preirradiation stromal soaking for 10 minutes in riboflavin 0.1%-dextrane 20% (Ricrolin; Sooft Italy) applied every 2 minutes, and 30 minutes of total exposure (6 steps of 5 minutes) to solid-state UV-A illuminator (Caporossi, Baiocchi, Mazzotta Vega X linker; CSO Opthalmics, Florence, Italy), energy delivered 3 mW/cm, and irradiated area 9 mm in diameter. After the operation, uncorrected visual acuity and best spectacle-corrected visual acuity improved from 0.2 to 0.6 and from 0.3 to 0.8 Snellen lines, respectively, in a 12-month follow-up. Improved topographical K readings and corneal symmetry index were also recorded starting from the first postoperative month and continuing thereafter. No adverse effects were recorded after treatment. Riboflavin UV-A-induced corneal cross-linking seems to be a promising surgical option in the management of unstable corneal ectasia exacerbated by RK, particularly in eyes with preexisting keratoconus. A large cohort and longer follow-up are needed to determine its long-term efficacy in this clinical setting.

  4. Accelerated Corneal Collagen Cross-linking for Postoperative LASIK Ectasia: Two-Year Outcomes.

    Science.gov (United States)

    Marino, Gustavo K; Torricelli, Andre A M; Giacomin, Natalia; Santhiago, Marcony R; Espindola, Rodrigo; Netto, Marcelo V

    2015-06-01

    To evaluate the effectiveness and safety of accelerated corneal collagen cross-linking for postoperative LASIK ectasia after 2 years. A prospective, single-center case series was performed with patients treated for postoperative LASIK ectasia. All eyes underwent accelerated corneal collagen cross-linking (CCL-Vario Crosslinking; Peschke Meditrade GmbH, Zurich, Switzerland) at 9 mW/cm(2) for 10 minutes. The main outcome measures were changes in uncorrected distance visual acuity, corrected distance visual acuity, central corneal thickness, corneal topography, and endothelial cell density. These parameters were assessed at baseline and at the 6-month and 1- and 2-year follow-up visit. The study enrolled 40 eyes of 24 patients (15 male and 9 female) with a mean age of 33.8 ± 7.5 years (range: 24 to 52 years) that attained at least 2 years of follow-up. The surgical procedure was uneventful in all cases. All eyes stabilized after treatment without any further signs of progression and no statistically significant changes in the mean uncorrected distance visual acuity (P = .649), corrected distance visual acuity (P = .616), mean keratometry (P =.837), steep keratometry (P = .956), ultrasonic pachymetry (P = .135), slit-scanning pachymetry (P = .276), and endothelial cell density (P = .523). In addition, 72.5% of the patients presented stable or gains of Snellen lines over time. Accelerated corneal collagen cross-linking seems to be safe and effective in halting postoperative LASIK ectasia progression after 2 years of follow-up. However, a longer follow-up period with a larger cohort is needed to validate these findings. Copyright 2015, SLACK Incorporated.

  5. High-performance liquid chromatography estimation of cross-linking of dihydroxyindole moiety in eumelanin.

    Science.gov (United States)

    Ito, Shosuke; Wakamatsu, Kazumasa; Glass, Keely; Simon, John D

    2013-03-15

    Eumelanin pigments consist of various ratios of 5,6-dihydroxyindole-2-carboxylic acid (DHICA) and 5,6-dihydroxyindole (DHI). On alkaline hydrogen peroxide oxidation, these indole moieties give rise to pyrrole-2,3,5-tricarboxylic acid (PTCA) and pyrrole-2,3-dicarboxylic acid (PDCA), respectively. In a recent study, we detected considerable amounts of other pyrrole acids, pyrrole-2,3,4,5-tetracarboxylic acid (PTeCA) and pyrrole-2,3,4-tricarboxylic acid (isoPTCA), among the oxidation products of fossil ink sacs more than 160 million years old. PTeCA and isoPTCA arise from the cross-linking of the DHI moiety of eumelanin at the C2 and/or C3 positions. We mimicked the process of cross-linking by heating synthetic eumelanins prepared from various ratios of DHICA and DHI at 100 °C for 18 days (or at 40 °C for 180 days). The heated eumelanins were analyzed after alkaline peroxide oxidation as PTCA, PDCA, PTeCA, and isoPTCA by high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection. On heating, PTCA decreased rapidly due to decarboxylation, whereas PDCA decreased gradually. Concurrently, PTeCA increased gradually to levels close to PTCA. IsoPTCA also increased gradually at lower levels. Similar changes were observed at 40°C at a much slower rate. These findings suggest that the PTeCA/PTCA ratio may serve as a good indicator of aging (cross-linking) of eumelanin. Copyright © 2012 Elsevier Inc. All rights reserved.

  6. Impact of corneal cross-linking combined with photorefractive keratectomy on blurring strength

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    Labiris G

    2016-04-01

    Full Text Available Georgios Labiris,1,2 Haris Sideroudi,2 Dimitris Angelonias,2 Kimonas Georgantzoglou,2 Vassilios P Kozobolis1,21Department of Ophthalmology, University Hospital of Alexandroupolis, 2Eye Institute of Thrace, Alexandroupolis, GreecePurpose: The aim of this study was to evaluate the impact of corneal cross-linking combined with photorefractive keratectomy (PRK on blurring strength.Methods: A total of 63 patients with keratoconus were recruited for this study, and two study groups were formed according to the therapeutic intervention: corneal collagen cross-linking (CxL group (33 patients received corneal cross-linking according to the Dresden protocol, while the rest additionally received topography-guided photorefractive keratectomy (tCxL. The impact of surgical procedure on blurring strength was assessed by power vector analysis. Potential association between blurring strength and vision-specific quality of life was assessed using the National Eye Institute Visual Function Questionnaire (NEI-VFQ 25 instrument.Results: Blurring strength presented excellent correlation with NEI-VFQ scores both preoperatively and postoperatively (all P<0.01. Both groups demonstrated nonsignificant changes in best-corrected visual acuity; however, only the tCxL group had significant reduction in blurring strength (13.48+10.86 [preoperative], 4.26+7.99 [postoperative], P=0.042.Conclusion: Only the combined treatment (tCxL resulted in significant reduction in blurring strength. Moreover, the excellent correlation of blurring strength with NEI-VFQ scores indicates its reliability as an index of self-reported quality of life in keratoconus, since it seems to address the nonsignificant changes in best-corrected visual acuity following CxL treatments that are conceived as subjective improvement by the patient.Keywords: keratoconus, cornea crosslinking, blurring strength, tCxL

  7. Effect of Age Factor on Corneal Cross-linking Treatment of Keratoconus Patients

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    Lokman Aslan

    2012-12-01

    Full Text Available Pur po se: To evaluate the effect of age on corneal cross-linking treatment in keratoconus patients. Ma te ri al and Met hod: We retrospectively evaluated forty-nine eyes of 45 patients with keratoconus that were applied corneal cross-linking treatment. The patients were divided into three groups; pediatric, adolescent and adult. The first group (25 years old consisted of 17 eyes. Preoperative best-corrected visual acuity (BCVA log-MAR, central corneal thickness (CCT, and maximum corneal steepness (MCS were compared with postoperative 13.8±3.83-month follow-up values. Re sults: After the treatment, a statistically significant improvement in visual acuity was found in group 1 (p<0.50.Visual acuity improvement was not statistically significant in group 2 and group 3 (p=0.434, p=0.21, respectively. Log-MAR-visual acuity differences were not statistically significant among the groups (p=0.182. No statistically significant change was found in postoperative CCT values in all groups (p=0.353, p=0.308, p=0.692, respectively. MCS values did not change statistically significantly at the postoperative evaluation (p=0.605, p=0.574, p=0.266, respectively. There was no statistically significant difference when comparing the three groups (p=0.856. Maximum corneal flattening was observed in group 3, but it was not statistically significant (p=0.282. Dis cus si on: It was determined that the results of corneal cross-linking treatment in keratoconus did not demonstrate any statistically significant changes among the age groups with respect to visual acuity, CCT and MCS values. (Turk J Ophthalmol 2012; 42: 450-3

  8. Maternal obesity enhances collagen accumulation and cross-linking in skeletal muscle of ovine offspring.

    Directory of Open Access Journals (Sweden)

    Yan Huang

    Full Text Available Maternal obesity (MO has harmful effects on both fetal development and subsequent offspring health. We previously demonstrated that MO enhances collagen accumulation in fetal skeletal muscle, but its impact on mature offspring muscle collagen accumulation is unknown. Ewes were fed either a control diet (Con, fed 100% of NRC nutrient recommendations or obesogenic diet (OB, fed 150% of NRC nutrient recommendations from 60 days before conception to birth. All ewes received the Con diet during lactation. Male offspring were euthanized at 2.5 years (mean and the left Longissimus dorsi (LD muscle and semitendinosus (ST muscle were sampled. Collagen concentration increased by 37.8 ± 19.0% (P<0.05 in LD and 31.2 ± 16.0% (P<0.05 in ST muscle of OB compared to Con offspring muscle. Mature collagen cross-linking (pyridinoline concentration was increased for 22.3 ± 7.4% and 36.3 ± 9.9% (P<0.05 in LD and ST muscle of OB group respectively. Expression of lysyl oxidase, lysyl hydroxylase-2b (LH2b and prolyl 4-hydroxylase (P4HA was higher in OB LD and ST muscle. In addition, the expression of metalloproteinases (MMPs was lower but tissue inhibitor of metalloproteinases (TIMPs was higher in OB offspring muscle, indicating reduced collagen remodeling. MO enhanced collagen content and cross-linking in offspring muscle, which might be partially due to reduced collagen remodeling. Our observation that the collagen content and cross-linking are enhanced in MO offspring muscle is significant, because fibrosis is known to impair muscle functions and is a hallmark of muscle aging.

  9. Superior Catalytic Performance of Gold Nanoparticles Within Small Cross-Linked Lysozyme Crystals.

    Science.gov (United States)

    Liu, Mingyue; Wang, Libing; Huang, Renliang; Yu, Yanjun; Su, Rongxin; Qi, Wei; He, Zhimin

    2016-10-08

    Bionanomaterials synthesized by bio-inspired templating methods have emerged as a novel class of composite materials with varied applications in catalysis, detection, drug delivery, and biomedicine. In this study, two kinds of cross-linked lysozyme crystals (CLLCs) of different sizes were applied for the in situ growth of Au nanoparticles (AuNPs). The resulting composite materials were characterized by light microscopy, scanning electron microscopy, transmission electron microscopy, thermogravimetric analysis, and X-ray photoelectron spectroscopy. The catalytic properties of the prepared materials were examined in the catalytic reduction of 4-nitrophenol (4-NP) to 4-aminophenol (4-AP). It was found that the size of the AuNPs increased with an increase in Au loading for both small and large crystals. In addition, small crystals favored homogeneous adsorption and distribution of the metal precursors. And the size of the AuNPs within small crystals could be maintained below 2.5 nm by managing the HAuCl4/lysozyme molar ratio. Furthermore, the lysozyme functional groups blocked the AuNP activity sites, therefore reducing their catalytic activity. This effect was more pronounced for small AuNPs. Moreover, the mass transfer of reactants (4-NP) from solution to AuNPs within the crystals restricted their catalytic reduction, leading to superior catalytic performance of the AuNPs within small cross-linked lysozyme crystals (Au@S-CLLCs) compared to those within large cross-linked lysozyme crystals (Au@L-CLLCs) at similar Au loadings. Finally, an increase in Au loading clogged the crystal channels with increased quantities of larger aggregated AuNPs, thus impeding the catalytic performance of Au@S-CLLCs.

  10. Effect of microbial transglutaminase on the natural actomyosin cross-linking in chicken and beef.

    Science.gov (United States)

    Ahhmed, Abdulatef Mrghni; Nasu, Tetsuo; Huy, Dang Quang; Tomisaka, Yoshito; Kawahara, Satoshi; Muguruma, Michio

    2009-06-01

    The objective of this research was to investigate the difference between chicken and beef in the interaction of actomyosin (myosin B) with microbial transglutaminase (MTG). The gel strength of myosin B was improved in both species and was significantly greater in beef than in chicken (Pactivity. SEMI also suggested that all MTG-dependent mega-structures of protein molecules generated in chicken and beef may vary greatly in size, configuration and complexity after treatment with MTG. We concluded that the optimal cross-links in myosin B induced by MTG are heterogeneous in chicken and beef.

  11. Towards aging mechanisms of cross-linked polyethylene (XLPE) cable insulation materials in nuclear power plants

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Shuaishuai; Fifield, Leonard S.; Bowler, Nicola

    2016-12-19

    Cross-linked polyethylene (XLPE) cable insulation material undergoes simultaneous, accelerated thermal and gamma-radiation aging to simulate the long-term aging environment within nuclear power plants (NPPs). A variety of materials characterization tests, including scanning electron microscopy, thermo-gravimetric analysis, differential scanning calorimetry, oxidation induction time, gel-fraction and dielectric properties measurement, are conducted on pristine and differently aged XLPE samples. A preliminary model of one possible aging mechanism of XLPE cable insulation material under gamma radiation at elevated temperature of 115 °C is suggested.

  12. Oxidation of myosin by haem proteins generates myosin radicals and protein cross-links

    DEFF Research Database (Denmark)

    Lametsch, Marianne Lund; Luxford, Catherine; Skibsted, Leif Horsfelt

    2008-01-01

    as a result of the reaction with activated haem proteins (horseradish peroxidase/H2O2) and met-myoglobin/H2O2) has been investigated by EPR spectroscopy and amino-acid consumption, product formation has been characterized by HPLC, and changes in protein integrity have been determined by SDS/PAGE. Multiple...... of thiyl and tyrosyl radicals is consistent with the observed consumption of cysteine and tyrosine residues, the detection of di-tyrosine by HPLC and the detection of both reducible (disulfide bond) and non-reducible cross-links between myosin molecules by SDS/PAGE. The time course of radical formation...

  13. Corneal cross-linking in a child with osteogenesis imperfecta syndrome and keratoconus

    Directory of Open Access Journals (Sweden)

    Sergio Kwitko

    2017-07-01

    Full Text Available Cross-linking (CXL is a well-established procedure in children with keratoconus (KC, but cases of CXL and osteogenesis imperfecta (OI have not been published in the literature, despite the association between physiopathology of these diseases. This is the first case, to the best of our knowledge, of a young girl with both OI and KC that underwent a CXL treatment. In this case, CXL was performed at 6-years-old prior to an expected progression, without complications and probably stopped further keratoconus progression.

  14. Adsorption properties of cross-linked cellulose-epichlorohydrin polymers in aqueous solution.

    Science.gov (United States)

    Udoetok, Inimfon A; Dimmick, Raquel M; Wilson, Lee D; Headley, John V

    2016-01-20

    Cellulose was cross-linked with epichlorohydrin (EP) at variable levels (CLE-0.5, CLE-2 and CLE-4), where CLE-i denotes the cellulose to EP mole ratios. The cross-linked products were characterized by TGA and FT-IR spectroscopy, pH at the point of zero charge (pHpzc), water swelling, and dye-adsorption methods employing two types of dyes [phenolphthalein (phth) and p-nitrophenol (PNP)]. The characterization methods provide evidence of cross-linking of cellulose in accordance with variations in surface area, PZC, available surface hydroxyl groups, and thermal stability when compared against pristine cellulose. The pHpzc of the sorbent materials was ∼ 6.5 indicating a negative surface charge occurs above pHpzc. The cross-linked polymers possess greater swelling properties relative to pristine cellulose. Detailed adsorption studies were carried out at pH 9 for cellulose and CLE-i with five types single component carboxylate anions [2-hexyldecanoic acid (S1), trans-4-pentylcyclohexanecarboxylic acid (S2), 2-dicyclohexylacetic acid (S3), adamantane carboxylic acid (S4), and cyclohexane carboxylic acid (S5)] at 295 K. The uptake properties of PNP with cellulose and CLE-i were also compared at pH 5 and 9, respectively. CLE-2 had the highest uptake of PNP (Qm=1.22 × 10(-1)mmol/g, pH 9) and S1 (Qm=4.27 mg/g) while cellulose and CLE-4 had the strongest binding affinity (1.43 L/mmol and 5.90 × 10(-2)L/mg), respectively. Uptake of PNP by CLE-0.5 at pH 5 (Q m=5.30 × 10(-2)mmol/g) was higher than uptake at pH 9 (Qm=3.11 × 10(-2)mmol/g). Sorption of CLE-4 with S1, S2 and S3 showed that relative uptake of the surrogates had the following order: S3>S2>S1, where S2 had the strongest binding affinity to CLE-i. CLE-2 had the highest sorption capacity towards Si in an equimolar mixture with evidence of molecular selective uptake. At pH 9, low uptake was mainly related to electrostatic repulsion between the negatively charged sorbent surface and the carboxylate head groups of Si

  15. Scleral wound healing with cross-link technique using riboflavin and ultraviolet A on rabbit eyes

    Directory of Open Access Journals (Sweden)

    Damasceno NA

    2017-07-01

    Full Text Available Nadyr A Damasceno,1 Nadia C Miguel,2 Marcelo Palis Ventura,3 Miguel Burnier Jr,4 Marcos P Avila,5 Eduardo F Damasceno3 1Ophthalmology Department, Hospital Naval Marcílio Dias, 2Laboratory of Neurohistology and Cell Ultrastructure, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, 3Ophthalmology Department, Universidade Federal Fluminense, Niterói, Brazil; 4Ophthalmology Department, McGill University, Montreal, QC, Canada; 5Ophthalmology Department, Universidade Federal de Goiás, Goiania, Brazil Purpose: The aim of study was to evaluate the cross-link using riboflavin and ultraviolet A (UVA for improving scleral wound healing.Materials and methods: This was an experimental study involving four New Zealand rabbits (eight eyes. Therapy procedure was chosen for the right eye and control procedure for the left one. UVA irradiation of 365 nm with a surface irradiance of 3 mW/cm2 and a photosensitizer of riboflavin drops were applied for 30 minutes on the right eye at 2 mm from the limbus. Sclerotomy incision was performed at 2 mm from the limbus in both right (on the cross-link-treated area and left eye. Then, 30 days after surgery, a morphological analysis and histological staining with hematoxylin–eosin and picrosirius red were performed, and the sclerotomy cicatrization of right and left eyes was compared. The variables investigated were as follows: sclerotomy incision pictures and measurements were made using the ImageJ Software. Scleral thickness was measured (employing the anterior optical coherence tomography and the digital caliper. Collagen fiber density stained with picrosirius red staining was measured using the Image Pro Plus software.Results: The morphological analysis showed that in all samples, the right eye presented sclerotomy closure, and in two eyes, among them, there were no visible edges of the sclerotomies incision. The left eye presented sclerotomy closure and incision edges

  16. Laser-induced corneal cross-linking upon photorefractive ablation with riboflavin

    Directory of Open Access Journals (Sweden)

    Kornilovskiy IM

    2016-04-01

    Full Text Available Igor M Kornilovskiy,1 Elmar M Kasimov,2 Ayten I Sultanova,2 Alexander A Burtsev1 1Department of Eye Diseases, Federal State Budgetary Institution “National Pirogov Medical Surgical Centre”, Ministry of Health, Moscow, Russia; 2Department of Eye Diseases, Zarifa Aliyeva National Ophthalmology Center, Ministry of Health, Baku, Azerbaijan Aim: To estimate the biomechanical effect of the laser-induced cross-linking resulting from photorefractive ablation of the cornea with riboflavin.Methods: Excimer laser ablation studies were performed ex vivo (32 eyes of 16 rabbits by phototherapeutic keratectomy (PTK and in vivo (24 eyes of 12 rabbits by transepithelial photorefractive keratectomy (TransPRK, with and without riboflavin saturation of the stroma. Then, we performed corneal optical coherence tomography on 36 eyes of 18 patients with varying degrees of myopia at different times after the TransPRK was performed with riboflavin saturation of the stroma.Results: Biomechanical testing of corneal samples saturated with riboflavin revealed cross-linking effect accompanied by the increase in tensile strength and maximum strength. PTK showed increase in tensile strength from 5.1±1.4 to 7.2±1.6 MPa (P=0.001, while TransPRK showed increase in tensile strength from 8.8±0.9 to 12.8±1.3 MPa (P=0.0004. Maximum strength increased from 8.7±2.5 to 12.0±2.8 N (P=0.005 in PTK and from 12.8±1.6 to 18.3±1.2 N (P=0.0004 in TransPRK. Clinical optical coherence tomography studies of the biomicroscopic transparent cornea at different times after TransPRK showed increased density in the surface layers of the stroma and membrane-like structure beneath the epithelium.Conclusion: Photorefractive ablation of the preliminary corneal stroma saturation with riboflavin causes the effect of laser-induced cross-linking, which is attended with an increase in corneal tensile strength, maximum strength, increased density in the surface layers of the stroma, and formation of

  17. Evaluation of epigallocatechin-3-gallate (EGCG) cross-linked collagen membranes and concerns on osteoblasts

    Energy Technology Data Exchange (ETDEWEB)

    Chu, Chenyu; Deng, Jia [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Xiang, Lin; Wu, Yingying [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Department of Oral Implantology, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Wei, Xiawei [State Key Laboratory of Biotherapy and Laboratory for Aging Research, West China Hospital, Sichuan University, and Collaborative Innovation Center for Biotherapy, Chengdu, Sichuan 610041 (China); Qu, Yili [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Department of Oral Implantology, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Man, Yi, E-mail: manyi780203@126.com [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Department of Oral Implantology, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China)

    2016-10-01

    Collagen membranes have ideal biological and mechanical properties for supporting infiltration and proliferation of osteoblasts and play a vital role in guided bone regeneration (GBR). However, pure collagen can lead to inflammation, resulting in progressive bone resorption. Therefore, a method for regulating the level of inflammatory cytokines at surgical sites is paramount for the healing process. Epigallocatechin-3-gallate (EGCG) is a component extracted from green tea with numerous biological activities including an anti-inflammatory effect. Herein, we present a novel cross-linked collagen membrane containing different concentrations of EGCG (0.0064%, 0.064%, and 0.64%) to regulate the level of inflammatory factors secreted by pre-osteoblast cells; improve cell proliferation; and increase the tensile strength, wettability, and thermal stability of collagen membranes. Scanning electron microscope images show that the surfaces of collagen membranes became smoother and the collagen fiber diameters became larger with EGCG treatment. Measurement of the water contact angle demonstrated that introducing EGCG improved membrane wettability. Fourier transform infrared spectroscopy analyses indicated that the backbone of collagen was intact, and the thermal stability was significant improved in differential scanning calorimetry. The mechanical properties of 0.064% and 0.64% EGCG-treated collagen membranes were 1.5-fold greater than those of the control. The extent of cross-linking was significantly increased, as determined by a 2,4,6-trinitrobenzenesulfonic acid solution assay. The Cell Counting Kit-8 (CCK-8) and live/dead assays revealed that collagen membrane cross-linked by 0.0064% EGCG induced greater cell proliferation than pure collagen membranes. Additionally, real-time polymerase chain reaction and enzyme-linked immunosorbent assay results showed that EGCG significantly affected the production of inflammatory factors secreted by MC3T3-E1 cells. Taken together, our

  18. Progress of corneal collagen cross-linking combined with refractive surgery

    Directory of Open Access Journals (Sweden)

    Na Li

    2014-02-01

    Full Text Available As a photochemical reaction that can stiffen the cornea, corneal collagen cross-linking (CXL is the only promising method of preventing the progress of keratectasia, such as keratoconus and secondary ectasia following refractive surgery. The aim of CXL is to stabilize the underlying condition, with a small chance of visual improvement. Combining CXL with refractive surgery targeting both stabilization and reshaping of the corneal tissue for visual function improvement is a good treatment option. This review aims to provide a comprehensive and unbiased summary of the published research regarding combined CXL and refractive surgery, including measures and results, to help elucidate the future direction of CXL.

  19. Chemical Cross-Linking and H/D Exchange for Fast Refinement of Protein Crystal Structure

    Czech Academy of Sciences Publication Activity Database

    Rozbeský, Daniel; Man, Petr; Kavan, Daniel; Chmelík, Josef; Černý, Jiří; Bezouška, Karel; Novák, Petr

    2012-01-01

    Roč. 84, č. 2 (2012), s. 867-870 ISSN 0003-2700 R&D Projects: GA ČR(CZ) GAP207/10/1040; GA ČR GA303/09/0477; GA ČR GD305/09/H008; GA MŠk LC545 Institutional research plan: CEZ:AV0Z50520701; CEZ:AV0Z50200510 Keywords : MASS-SPECTROMETRY * chemical cross-linking * NKR-P1A receptor Subject RIV: EE - Microbiology, Virology Impact factor: 5.695, year: 2012

  20. Theoretical Basis, Laboratory Evidence, and Clinical Research of Chemical Surgery of the Cornea: Cross-Linking

    Directory of Open Access Journals (Sweden)

    Amanda C. da Paz

    2014-01-01

    Full Text Available Corneal cross-linking (CXL is increasingly performed in ophthalmology with high success rates for progressive keratoconus and other types of ectasia. Despite being an established procedure, some molecular and clinical aspects still require additional studies. This review presents a critical analysis of some established topics and others that are still controversial. In addition, this review examines new technologies and techniques (transepithelial and ultrafast CXL, uses of corneal CXL including natural products and biomolecules as CXL promoters, and evidence for in vitro and in vivo indirect effectiveness.

  1. Evaluation of cross-linked chitosan microparticles containing acyclovir obtained by spray-drying

    International Nuclear Information System (INIS)

    Stulzer, Hellen Karine; Tagliari, Monika Piazzon; Parize, Alexandre Luis; Silva, Marcos Antonio Segatto; Laranjeira, Mauro Cesar Marghetti

    2009-01-01

    The aim of this study was to obtain microparticles containing acyclovir (ACV) and chitosan cross-linked with tripolyphosphate using the spray-drying technique. The resultant system was evaluated through loading efficiency, differential scanning calorimetry (DSC), thermogravimetric analysis (TG), X-ray powder diffraction (XRPD), scanning electron microscopy (SEM), in vitro release and stability studies. The results obtained indicated that the polymer/ACV ratio influenced the final properties of the microparticles, with higher ratios giving the best encapsulation efficiency, dissolution profiles and stability. The DSC and XRPD analyses indicated that the ACV was transformed into amorphous form during the spray-drying process

  2. Effects of Composition of Iron-Cross-Linked Alginate Hydrogels for Cultivation of Human Dermal Fibroblasts

    OpenAIRE

    Machida-Sano, Ikuko; Ogawa, Sakito; Ueda, Hiroyuki; Kimura, Yoshitaka; Satoh, Nao; Namiki, Hideo

    2012-01-01

    We investigated the suitability of ferric-ion-cross-linked alginates (Fe-alginate) with various proportions of L-guluronic acid (G) and D-mannuronic acid (M) residues as a culture substrate for human dermal fibroblasts. High-G and high-M Fe-alginate gels showed comparable efficacy in promoting initial cell adhesion and similar protein adsorption capacities, but superior cell proliferation was observed on high-G than on high-M Fe-alginate as culture time progressed. During immersion in culture...

  3. Evaluation of nanohydroxyapaptite (nano-HA) coated epigallocatechin-3-gallate (EGCG) cross-linked collagen membranes.

    Science.gov (United States)

    Chu, Chenyu; Deng, Jia; Man, Yi; Qu, Yili

    2017-09-01

    Collagen is the main component of extracellular matrix (ECM) with desirable biological activities and low antigenicity. Collagen materials have been widely utilized in guided bone regeneration (GBR) surgery due to its abilities to maintain space for hard tissue growth. However, pure collagen lacks optimal mechanical properties. In our previous study, epigallocatechin-3-gallate (EGCG) cross-linked collagen membranes, with better biological activities and enhanced mechanical properties, may promote osteoblast proliferation, but their effect on osteoblast differentiation is not very significant. Nanohydroxyapatite (nano-HA) is the main component of mineral bone, which possesses exceptional bioactivity properties including good biocompatibility, high osteoconductivity and osteoinductivity, non-immunogenicity and non-inflammatory behavior. Herein, by analyzing the physical and chemical properties as well as the effects on promoting bone regeneration, we have attempted to present a novel EGCG-modified collagen membrane with nano-HA coating, and have found evidence that the novel collagen membrane may promote bone regeneration with a better surface morphology, without destroying collagen backbone. To evaluate the surface morphologies, chemical and mechanical properties of pure collagen membranes, epigallocatechin-3-gallate (EGCG) cross-linked collagen membranes, nano-HA coated collagen membranes, nano-HA coated EGCG-collagen membranes, (ii) to evaluate the bone regeneration promoted by theses membranes. In the present study, collagen membranes were divided into 4 groups: (1) untreated collagen membranes (2) EGCG cross-linked collagen membranes (3) nano-HA modified collagen membranes (4) nano-HA modified EGCG-collagen membranes. Scanning electron microscope (SEM) and Fourier transform infrared spectroscopy (FTIR) were used to evaluate surface morphologies and chemical properties, respectively. Mechanical properties were determined by differential scanning calorimeter (DSC

  4. Irradiation effects on properties of reverse osmosis membrane based on cross-linked aromatic polyamide

    International Nuclear Information System (INIS)

    Nakase, Yoshiaki; Yanagi, Tadashi; Uemura, Tadahiro.

    1994-01-01

    In order to develop a membrane suitable for reverse osmotic condensation of radioactive liquid wastes, a new cross-linked aromatic polyamide composite reverse osmosis membrane (ROM) was irradiated in water or in wet system, and its mechanical and some thermal properties, and the separation performance for inorganic salt were investigated. A membrane was degraded by irradiation more severely in wet system than in dry system, probably due to the reaction with OH-radicals. In the separation performance for NaCl, the salt rejection of the membrane was kept over 88% until irradiation reached 2MGy, maintaining about 90% of its original water flux. (author)

  5. Modulation by propranolol of the lysyl cross-links in aortic elastin and collagen of the aneurysm-prone turkey.

    Science.gov (United States)

    Boucek, R J; Gunja-Smith, Z; Noble, N L; Simpson, C F

    1983-01-15

    dl-Propranolol (propranolol) fed to immature and mature aneurysm-prone turkeys (Broad-Breasted White, BBW) for 6 weeks significantly raised the tensile strength of tissue rings from the abdominal aorta. The drug-mediated increase in tensile strength values was dose-related and independent of its heart rate- and arterial pressure-lowering effects. Propranolol acts, in part, by (a) stimulating lysyl oxidase to produce greater amounts of reactive aldehydes for intermolecular cross-links, (b) enhancing the progression of chemically unstable to stable forms of intermolecular elastin cross-links (lysinonorleucine and the desmosines), and (c) reducing the density of the age-related intermolecular cross-linking of collagen (pyridinoline). These propranolol effects on the lysyl cross-links were demonstrated in both the immature and mature animals and suggest a heretofore unrecognized potential for this widely used cardiovascular drug.

  6. Mechanical properties of the equine superficial digital flexor tendon relate to specific collagen cross-link levels.

    Science.gov (United States)

    Thorpe, C T; Stark, R J F; Goodship, A E; Birch, H L

    2010-11-01

    Damage to the flexor tendons, particularly the superficial digital flexor tendon (SDFT), is one of the most common musculoskeletal injuries sustained by horses competing in all disciplines. Our previous work has shown that SDFTs from different individuals show a wide variation in mechanical strengths; this is important clinically as it may relate to predisposition to injury. The high mechanical strength of tendon relies on the correct orientation of collagen molecules within fibrils and stabilisation by the formation of chemical cross-links between collagen molecules. It is not known whether the variation in SDFT mechanical properties between individuals relates to differences in collagen cross-link levels. Enzyme-derived, intermolecular cross-linking of tendon collagen correlates with mechanical properties of the SDFT. SDFTs were collected from 38 horses and mechanically tested to failure. Structural and material properties were calculated from the load/deformation plot and cross-sectional area for each tendon. Following mechanical testing, pyrrolic cross-link levels were measured using a spectrophotometric assay for Ehrlich's reactivity and pyridinoline levels were quantified by HPLC. Cross-link levels were correlated with mechanical properties and statistical significance tested using a Pearson's correlation test. Pyrrole cross-link levels showed a significant positive correlation with ultimate stress (P = 0.004), yield stress (P = 0.003) and elastic modulus (P = 0.018) of the tendons, despite being a minor cross-link in these tendons. There was no significant correlation of mechanical properties with either hydroxylysyl- or lysyl-pyridinoline levels. Given the low absolute levels of pyrrole, we suggest that the correlation with high mechanical strength is through an indirect mechanism. Understanding the nature of the relationships between pyrrole cross-links, other matrix characteristics and tendon material properties may allow development of strategies to

  7. Effect of pore size and cross-linking of a novel collagen-elastin dermal substitute on wound healing.

    Science.gov (United States)

    Boekema, Bouke K H L; Vlig, Marcel; Olde Damink, Leon; Middelkoop, Esther; Eummelen, Lizette; Bühren, Anne V; Ulrich, Magda M W

    2014-02-01

    Collagen-elastin (CE) scaffolds are frequently used for dermal replacement in the treatment of full-thickness skin defects such as burn wounds. But little is known about the optimal pore size and level of cross-linking. Different formulations of dermal substitutes with unidirectional pores were tested in porcine full-thickness wounds in combination with autologous split skin mesh grafts (SSG). Effect on wound healing was evaluated both macro- and microscopically. CE scaffolds with a pore size of 80 or 100 μm resulted in good wound healing after one-stage grafting. Application of scaffolds with a larger average pore size (120 μm) resulted in more myofibroblasts and more foreign body giant cells (FBGC). Moderate crosslinking impaired wound healing as it resulted in more wound contraction, more FBGC and increased epidermal thickness compared to no cross-linking. In addition, take rate and redness were negatively affected compared to SSG only. Vascularization and the number of myofibroblasts were not affected by cross-linking. Surprisingly, stability of cross-linked scaffolds was not increased in the wound environment, in contrast to in vitro results. Cross-linking reduced the proliferation of fibroblasts in vitro, which might explain the reduced clinical outcome. The non-cross-linked CE substitute with unidirectional pores allowed one-stage grafting of SSG, resulting in good wound healing. In addition, only a very mild foreign body reaction was observed. Cross-linking of CE scaffolds negatively affected wound healing on several important parameters. The optimal non-cross-linked CE substitute is a promising candidate for future clinical evaluation.

  8. Removal of textile dye, direct red 23, with glutaraldehyde cross-linked magnetic chitosan beads.

    Science.gov (United States)

    Sanlier, Senay Hamarat; Ak, Güliz; Yilmaz, Habibe; Ozbakir, Gizem; Cagliyan, Oguzhan

    2013-01-01

    One of the most important classes of pollutants is dyes, and today there are more than 100,000 commercial dyes. Conventional treatment processes are very expensive, so it is essential to develop low-cost sorbent materials with high adsorption capacities. The aim of this study is to prepare magnetic microsized adsorbents that have high adsorption capacity for removal of direct red 23. Through this objective, glutaraldehyde cross-linked magnetic chitosan beads were formed in order to remove the textile dye direct red 23. Barium ferrite was used to give a magnetic property so that the beads could easily be separated from the water after treatment. The effects of barium ferrite, pH, incubation time, dye concentration, and glutaraldehyde amounts were investigated. The Langmuir and Freundlich adsorption models were applied to describe the equilibrium isotherms. The adsorption capacity had a very large value: 1250 mg/g at pH 4.0, at room temperature. Compared with activated carbon, magnetic cross-linked chitosan exhibits excellent performance in the adsorption of anionic dyes and the magnetic properties of beads enable us to remove the beads from the water after treatment. Pseudo-second-order and intraparticle diffusion kinetic models were applied.

  9. Angiotensin I-Converting Enzyme Inhibitor Derived from Cross-Linked Oyster Protein

    Directory of Open Access Journals (Sweden)

    Cheng-Liang Xie

    2014-01-01

    Full Text Available Following cross-linking by microbial transglutaminase, modified oyster proteins were hydrolyzed to improve inhibitory activity against angiotensin-converting enzyme (ACE inhibitory activity with the use of a single protease, or a combination of six proteases. The oyster hydrolysate with the lowest 50% ACE inhibitory concentration (IC50 of 0.40 mg/mL was obtained by two-step hydrolysis of the cross-linked oyster protein using Protamex and Neutrase. Five ACE inhibitory peptides were purified from the oyster hydrolysate using a multistep chromatographic procedure comprised of ion-exchange, size exclusion, and reversed-phase liquid chromatography. Their sequences were identified as TAY, VK, KY, FYN, and YA, using automated Edman degradation and mass spectrometry. These peptides were synthesized, and their IC50 values were measured to be 16.7, 29.0, 51.5, 68.2, and 93.9 μM, respectively. Toxicity of the peptides on the HepG2 cell line was not detected. The oyster hydrolysate also significantly decreased the systolic blood pressure of spontaneously hypertensive rats (SHR. The antihypertensive effect of the oyster hydrolysate on SHR was rapid and long-lasting, compared to commercially obtained sardine hydrolysate. These results suggest that the oyster hydrolysate could be a source of effective nutraceuticals against hypertension.

  10. Localized changes in stromal reflectivity after corneal collagen cross-linking observed with different imaging techniques.

    Science.gov (United States)

    Wittig-Silva, Christine; Chan, Elsie; Pollock, Graeme; Snibson, Grant R

    2013-06-01

    To report changes observed in the corneal stroma using confocal microscopy and Scheimpflug imaging following epithelium-off collagen cross-linking (CXL) in cases of progressive keratoconus. Fifteen eyes of 14 patients were examined before and after CXL using slit-lamp biomicroscopy, Scheimpflug imaging, and confocal microscopy. A subset of patients also had optical coherence tomography imaging performed. After CXL, confocal microscopy revealed a discrete layer of increased reflectivity in the mid to deep stroma. This layer was visible from as early as 1 week following treatment with patchy changes persisting until 12 months. This posterior reflective zone was found to consist of numerous linear or striate reflective structures (intrastromal striate reflections [ISRs]). ISRs were not observed in any of the eyes before the CXL procedure. High-resolution Scheimpflug and optical coherence tomography images also demonstrated a narrow zone of increased reflectivity at a similar depth. A layer-by-layer match was possible between the confocal and Scheimpflug images. The location of the ISR layer appears to correlate with a zone of increased reflectivity visible on postoperative slit-lamp examinations. Altered stromal reflectivity after CXL can be observed with modern diagnostic imaging technologies. These findings seem to correlate not only among the different devices but also with biomicroscopic observations and could potentially provide a non-invasive tool to monitor the cross-linking effect in individual corneas. Copyright 2013, SLACK Incorporated.

  11. Encapsulated eucalyptus oil in ionically cross-linked alginate microcapsules and its controlled release.

    Science.gov (United States)

    Noppakundilograt, Supaporn; Piboon, Phianghathai; Graisuwan, Wilaiporn; Nuisin, Roongkan; Kiatkamjornwong, Suda

    2015-10-20

    Sodium alginate microcapsules containing eucalyptus oil were prepared by oil-in-water emulsification via Shirasu porous glass (SPG) membrane and cross-linked by calcium chloride (CaCl2). SPG membrane pore size of 5.2μm was used to control the size of eucalyptus oil microdroplets. Effects of sodium alginate, having a mannuronic acid/guluronic acid (M/G) ratio of 1.13, eucalyptus oil and CaCl2 amounts on microdroplet sizes and size distribution were elucidated. Increasing sodium alginate amounts from 0.1 to 0.5% (wv(-1)) sodium alginate, the average droplets size increased from 42.2±2.0 to 48.5±0.6μm, with CVs of 16.5±2.2 and 30.2±4.5%, respectively. CaCl2 successfully gave narrower size distribution of cross-linked eucalyptus oil microcapsules. The optimum conditions for preparing the microcapsules, oil loading efficiency, and controlled release of the encapsulated eucalyptus oil from the microcapsules as a function of time at 40°C were investigated. Release model for the oil from microcapsules fitted Ritger-Peppas model with non-Fickian transport mechanism. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Patterning the Stiffness of Elastomeric Nanocomposites by Magnetophoretic Control of Cross-linking Impeder Distribution

    Directory of Open Access Journals (Sweden)

    Suvojit Ghosh

    2015-01-01

    Full Text Available We report a novel method to pattern the stiffness of an elastomeric nanocomposite by selectively impeding the cross-linking reactions at desired locations while curing. This is accomplished by using a magnetic field to enforce a desired concentration distribution of colloidal magnetite nanoparticles (MNPs in the liquid precursor of polydimethysiloxane (PDMS elastomer. MNPs impede the cross-linking of PDMS; when they are dispersed in liquid PDMS, the cured elastomer exhibits lower stiffness in portions containing a higher nanoparticle concentration. Consequently, a desired stiffness pattern is produced by selecting the required magnetic field distribution a priori. Up to 200% variation in the reduced modulus is observed over a 2 mm length, and gradients of up to 12.6 MPa·mm−1 are obtained. This is a significant improvement over conventional nanocomposite systems where only small unidirectional variations can be achieved by varying nanoparticle concentration. The method has promising prospects in additive manufacturing; it can be integrated with existing systems thereby adding the capability to produce microscale heterogeneities in mechanical properties.

  13. Preliminary evaluation of glipizide spheres and compacts from spheres prepared by cross-linking technique.

    Science.gov (United States)

    Garcia, J G; Ghaly, E S

    2001-03-01

    The objective of this research was to use the natural polymer Carrageenan to obtain controlled release spheres loaded with glipizide using the cross-linking technique. The effect of polymer level and drug load were investigated. The drug was dispersed in Carrageenan solution and the dispersion was dropped by a device containing 3 disposable syringes into cross-linking solution containing 3% calcium chloride. After 15 minutes residence time, the spheres were collected by decantation and dried in hot air oven at 38 degrees C +/- 2 degrees C for 24 hours. The dried spheres were successfully compacted into tablets using rotary Manesty B-3B machine equipped with 12/32 inches round flat face punches, target tablet weight was 400 mg +/- 5%. As the polymer level was increased in the sphere formulation, the drug release rate was increased. However, as the drug level was increased in the sphere formulation, the release rate was decreased. This trend was also true for tablets compacted from spheres. The scanning electron microscope photographs supported the dissolution data. More cracks and rough surface were observed in tablets compacted from spheres containing high polymer level and low drug level.

  14. Removal of anionic azo dyes from aqueous solution by functional ionic liquid cross-linked polymer

    International Nuclear Information System (INIS)

    Gao, Hejun; Kan, Taotao; Zhao, Siyuan; Qian, Yixia; Cheng, Xiyuan; Wu, Wenli; Wang, Xiaodong; Zheng, Liqiang

    2013-01-01

    Highlights: • Equilibrium, kinetic and thermodynamic of adsorption of dyes onto PDVB-IL was investigated. • PDVB-IL has a high adsorption capacity to treat dyes solution. • Higher adsorption capacity is due to the functional groups of PDVB-IL. • Molecular structure of dyes influences the adsorption capacity. -- Abstract: A novel functional ionic liquid based cross-linked polymer (PDVB-IL) was synthesized from 1-aminoethyl-3-vinylimidazolium chloride and divinylbenzene for use as an adsorbent. The physicochemical properties of PDVB-IL were investigated by Fourier transform infrared spectroscopy, scanning electron microscopy and thermogravimetric analysis. The adsorptive capacity was investigated using anionic azo dyes of orange II, sunset yellow FCF, and amaranth as adsorbates. The maximum adsorption capacity could reach 925.09, 734.62, and 547.17 mg/g for orange II, sunset yellow FCF and amaranth at 25 °C, respectively, which are much better than most of the other adsorbents reported earlier. The effect of pH value was investigated in the range of 1–8. The result shows that a low pH value is found to favor the adsorption of those anionic azo dyes. The adsorption kinetics and isotherms are well fitted by a pseudo second-order model and Langmuir model, respectively. The adsorption process is found to be dominated by physisorption. The introduction of functional ionic liquid moieties into cross-linked poly(divinylbenzene) polymer constitutes a new and efficient kind of adsorbent

  15. Preparation and biological activity studies of resveratrol loaded ionically cross-linked chitosan-TPP nanoparticles.

    Science.gov (United States)

    Wu, Jie; Wang, Yaping; Yang, Hao; Liu, Xiangyu; Lu, Zhong

    2017-11-01

    Nanoparticles with size range of 10-500nm can be efficiently delivered into cancer cells by the Enhanced Permeability and Retention (EPR) effect. Here, we prepared resveratrol (Res) loaded chitosan (CS) nanoparticles with the size of 172-217nm by an ionic cross-linking method, with sodium tripolyphosphate (TPP) as the cross-linking agent, to improve the stability, solubility and tumors targeting of the natural anti-cancer drug Res. The prepared Res loaded CS-TPP nanoparticles presented long-term storage stability and UV light stability. The cumulative drug release from nanoparticles in mimetic tumor tissue condition (pH 6.5) was higher than that in physiological condition (pH 7.4). Further, Res-loaded CS-TPP nanoparticles maintained the antioxidant activity of Res even after UV light irradiation. Cell viability study shows that the as prepared drug loaded nanoparticles had similar antiproliferative activity on hepatocellular carcinoma cells SMMC 7721 and lower cytotoxicity on normal hepatocyte cells L02 compared with free Res. Fluorescence microscopy observation revealed that the nanoparticles were efficiently taken in by SMMC 7721 cells. This work indicates the potential use of drug loaded CS-TPP nanoparticles for the efficient delivery of bioactive Res for chemotherapy. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Viability, Apoptosis, Proliferation, Activation, and Cytokine Secretion of Human Keratoconus Keratocytes after Cross-Linking

    Directory of Open Access Journals (Sweden)

    Xuefei Song

    2015-01-01

    Full Text Available Purpose. The purpose of this study was to determine the impact of cross-linking (CXL on viability, apoptosis, proliferation, activation, and cytokine secretion of human keratoconus (KC keratocytes, in vitro. Methods. Primary KC keratocytes were cultured in DMEM/Ham’s F12 medium supplemented with 10% FCS and underwent UVA illumination (370 nm, 2 J/cm2 during exposure to 0.1% riboflavin and 20% Dextran in PBS. Twenty-four hours after CXL, viability was assessed using Alamar blue assay; apoptosis using APO-DIRECT Kit; proliferation using ELISA-BrdU kit; and CD34 and alpha-smooth muscle actin (α-SMA expression using flow cytometry. Five and 24 hours after CXL, FGFb, HGF, TGFβ1, VEGF, KGF, IL-1β, IL-6, and IL-8 secretion was measured using enzyme-linked-immunoabsorbent assay (ELISA. Results. Following CXL, cell viability and proliferation decreased (P0.06. Five hours after CXL, FGFb secretion increased significantly (P=0.037; however no other cytokine secretion differed significantly from controls after 5 or 24 hours (P>0.12. Conclusions. Cross-linking decreases viability, triggers apoptosis, and inhibits proliferation, without an impact on multipotent hematopoietic stem cell transformation and myofibroblastic transformation of KC keratocytes. CXL triggers FGFb secretion of KC keratocytes transiently (5 hours, normalizing after 24 hours.

  17. Efficacy of iontophoresis-assisted epithelium-on corneal cross-linking for keratoconus

    Directory of Open Access Journals (Sweden)

    Hong-Zhen Jia

    2018-04-01

    Full Text Available Corneal cross-linking (CXL is a noninvasive therapeutic procedure for keratoconus that is aimed at improving corneal biomechanical properties by induction of covalent cross-links between stromal proteins. It is accomplished by ultraviolet A (UVA radiation of the cornea, which is first saturated with photosensitizing riboflavin. It has been shown that standard epithelium-off CXL (S-CXL is efficacious, and it has been recommended as the standard of care procedure for keratoconus. However, epithelial removal leads to pain, transient vision loss, and a higher risk of corneal infection. To avoid these disadvantages, transepithelial CXL was developed. Recently, iontophoresis has been adopted to increase riboflavin penetration through the epithelium. Several clinical observations have demonstrated the safety and efficacy of iontophoresis-assisted epithelium-on CXL (I-CXL for keratoconus. This review aimed to provide a comprehensive summary of the published studies regarding I-CXL and a comparison between I-CXL and S-CXL. All articles used in this review were mainly retrieved from the PubMed database. Original articles and reviews were selected if they were related to the I-CXL technique or related to the comparison between I-CXL and S-CXL.

  18. In Situ Dual Cross-Linking of Neat Biogel with Controlled Mechanical and Delivery Properties.

    Science.gov (United States)

    Kelmansky, Regina; Shagan, Alona; Rozenblit, Boris; Omar, Rawan; Lufton, Maayan; Mizrahi, Boaz

    2017-10-02

    Injectable biomaterials play a critical role in many biomedical applications. These materials, however, often have limitations in mechanical and drug-eluting properties attributed to their high water content and the weak secondary forces holding them together. Here we describe a new injectable material based on two complementary water-free, prepolymers modified with succinimidyl carbonate (SC) or with NH 2 end groups that form a stiff matrix upon mixing. Cross-linking involves an immediate reaction between PEG 4 -SC and PEG 4 -NH 2 that forms carbamate bonds and a delayed reaction of PEG 4 -SC with hydroxyl functional groups that forms carbonate bonds. The mechanical properties, swelling, and erosion kinetics of this biomaterial can be fine-tuned by varying the ratio between the two prepolymers. Bovine serum albumin and poorly water-soluble free base doxorubicin were readily loaded into this system, resulting in a high drug loading content attributed to the absence of water in the formulation. Controlled release over a period of 1 to 30 days was observed, depending on mixture composition and drug properties. The injectable nature of the formulation, its tailored mechanical properties, the fact that it can be cross-linked by two separate mechanisms, and its ability to incorporate and release hydrophilic and hydrophobic drugs make it very attractive as a drug delivery system.

  19. Hydroxyhomocitrulline Is a Collagen-Specific Carbamylation Mark that Affects Cross-link Formation.

    Science.gov (United States)

    Taga, Yuki; Tanaka, Keisuke; Hamada, Chieko; Kusubata, Masashi; Ogawa-Goto, Kiyoko; Hattori, Shunji

    2017-10-19

    Carbamylation is a non-enzymatic post-translational modification that physiologically occurs during aging and is a risk factor for various diseases. The most common product of carbamylation is homocitrulline (HCit), where a lysine (Lys) amino group has reacted with urea-derived cyanate. HCit has recently been detected in collagen; however, given that 15%-90% of total Lys in collagen is hydroxylated, it is unclear how hydroxylation affects collagen carbamylation. Here, we identified a collagen-specific carbamylation product, hydroxyhomocitrulline (HHCit), and showed that high levels of HHCit are correlated with age in rat tissue collagen and in vivo carbamylation in mice, as well as with the decline of kidney function in the serum of dialysis patients. Proteomic analysis of the carbamylated collagens identified α2(I) Lys 933 , a major cross-linking site, as a preferential HHCit site. Furthermore, our results suggest that hydroxylysine carbamylation affects the mechanical properties of connective tissue by competitively inhibiting collagen cross-link formation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Identification of PDGF-BB binding to thymosin β4 by chemical cross-linking.

    Science.gov (United States)

    Knop, Jana; App, Christine; Huff, Thomas; Iavarone, Federica; Castagnola, Massimo; Hannappel, Ewald

    2015-01-01

    The purpose of our work was to identify unknown interaction partners of thymosin β4 (Tβ4). It was suggested that Tβ4 could be an antifibrotic drug for treatment of liver fibrogenesis, because Tβ4 prevents the platelet-derived growth factor-BB (PDGF-BB)-induced activation of hepatic stellate cells (HSCs). Very little information is available how Tβ4 counteracts the PDGF-BB-induced activation of HSCs. We propose the hypothesis that Tβ4 could bind directly to PDGF-BB and thereby reduce the concentration of free PDGF-BB available for binding to the PDGF-β receptor. To prove our suggestion of a direct interaction between Tβ4 and PDGF-BB, we carried out chemical as well as photochemical cross-linking experiments between the two pure proteins in vitro. We identified an interaction between Tβ4 and PDGF-BB by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) cross-linking as well as through biotin label transfer using a bifunctional photoactivatable derivative of Tβ4. In an in vitro system, PDGF-BB was identified as the first extracellular partner interacting with Tβ4. This interaction could influence PDGF-BB binding to its receptor and abolish PDGF-BB-related effects. Direct interaction of Tβ4 with extracellular factors should be considered as a potential mechanism to explain the pleiotropic effects of β-thymosins.

  1. Evaluation of Epithelial Integrity with Various Transepithelial Corneal Cross-Linking Protocols for Treatment of Keratoconus

    Directory of Open Access Journals (Sweden)

    Suphi Taneri

    2014-01-01

    Full Text Available Purpose. Corneal collagen cross-linking (CXL has been demonstrated to stiffen cornea and halt progression of ectasia. The original protocol requires debridement of central corneal epithelium to facilitate diffusion of a riboflavin solution to stroma. Recently, transepithelial CXL has been proposed to reduce risk of complications associated with epithelial removal. Aim of the study is to evaluate the impact of various transepithelial riboflavin delivery protocols on corneal epithelium in regard to pain and epithelial integrity in the early postoperative period. Methods. One hundred and sixty six eyes of 104 subjects affected by progressive keratoconus underwent transepithelial CXL using 6 different riboflavin application protocols. Postoperatively, epithelial integrity was evaluated at slit lamp and patients were queried regarding their ocular pain level. Results. One eye had a corneal infection associated with an epithelial defect. No other adverse event including endothelial decompensation or endothelial damage was observed, except for epithelial damages. Incidence of epithelial defects varied from 0 to 63%. Incidence of reported pain varied from 0 to 83%. Conclusion. Different transepithelial cross-linking protocols have varying impacts on epithelial integrity. At present, it seems impossible to have sufficient riboflavin penetration without any epithelial disruption. A compromise between efficacy and epithelial integrity has to be found.

  2. Dual Cross-Linked Carboxymethyl Sago Pulp-Gelatine Complex Coacervates for Sustained Drug Delivery

    Directory of Open Access Journals (Sweden)

    Saravanan Muniyandy

    2015-06-01

    Full Text Available In the present work, we report for the first time the complex coacervation of carboxymethyl sago pulp (CMSP with gelatine for sustained drug delivery. Toluene saturated with glutaraldehyde and aqueous aluminum chloride was employed as cross-linkers. Measurements of zeta potential confirm neutralization of two oppositely charged colloids due to complexation, which was further supported by infrared spectroscopy. The coacervates encapsulated a model drug ibuprofen and formed microcapsules with a loading of 29%–56% w/w and an entrapment efficiency of 85%–93% w/w. Fresh coacervates loaded with drug had an average diameter of 10.8 ± 1.93 µm (n = 3 ± s.d.. The coacervates could encapsulate only the micronized form of ibuprofen in the absence of surfactant. Analysis through an optical microscope evidenced the encapsulation of the drug in wet spherical coacervates. Scanning electron microscopy revealed the non-spherical geometry and surface roughness of dried drug-loaded microcapsules. X-ray diffraction, differential scanning calorimetry and thermal analysis confirmed intact and crystalline ibuprofen in the coacervates. Gas chromatography indicated the absence of residual glutaraldehyde in the microcapsules. Dual cross-linked microcapsules exhibited a slower release than mono-cross-linked microcapsules and could sustain the drug release over the period of 6 h following Fickian diffusion.

  3. Probing Rubber Cross-Linking Generation of Industrial Polymer Networks at Nanometer Scale.

    Science.gov (United States)

    Gabrielle, Brice; Gomez, Emmanuel; Korb, Jean-Pierre

    2016-06-23

    We present improved analyses of rheometric torque measurements as well as (1)H double-quantum (DQ) nuclear magnetic resonance (NMR) buildup data on polymer networks of industrial compounds. This latter DQ NMR analysis allows finding the distribution of an orientation order parameter (Dres) resulting from the noncomplete averaging of proton dipole-dipole couplings within the cross-linked polymer chains. We investigate the influence of the formulation (filler and vulcanization systems) as well as the process (curing temperature) ending to the final polymer network. We show that DQ NMR follows the generation of the polymer network during the vulcanization process from a heterogeneous network to a very homogeneous one. The time variations of microscopic Dres and macroscopic rheometric torques present power-law behaviors above a threshold time scale with characteristic exponents of the percolation theory. We observe also a very good linear correlation between the kinetics of Dres and rheometric data routinely performed in industry. All these observations confirm the description of the polymer network generation as a critical phenomenon. On the basis of all these results, we believe that DQ NMR could become a valuable tool for investigating in situ the cross-linking of industrial polymer networks at the nanometer scale.

  4. Borax cross-linked guar gum hydrogels as potential adsorbents for water purification.

    Science.gov (United States)

    Thombare, Nandkishore; Jha, Usha; Mishra, Sumit; Siddiqui, M Z

    2017-07-15

    With the aim to explore new adsorbents for water purification, guar gum based hydrogels were synthesized by cross-linking with borax at different percentage. The cross-linking was confirmed through characterization by FTIR spectroscopy, SEM morphology, thermal studies and water absorption capacity. To examine the adsorption/absorption performance of different grades of hydrogels, their flocculation efficiency was studied in kaolin suspension at different pH by standard jar test procedure. The flocculation efficiency of the test materials was compared with the commercially used coagulant, alum and also residues of Al and K left in the treated water were comparatively studied. The synthesized hydrogels were also tested for their efficiency of removing Aniline Blue dye by UV-vis spectrophotometer study. The best grade hydrogel outperformed alum, at extremely low concentration and also showed dye removing efficiency up to 94%. The single step synthesized green products thus exhibited great potential as water purifying agents. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Study for HDR cross-link for formation flight satellite systems

    Science.gov (United States)

    Nishinaga, Nozomu; Takayama, Yoshihisa; Takahashi, Takashi; Ogawa, Yasuo; Kubooka, Toshihiro; Umehara, Hiroaki

    2003-07-01

    In this paper we studied about optical and millimeter radio wave cross-link for formation flight communication satellite system in a geostationary orbit. In formation flight system, since the distance among the satellites becomes short compared with the conventional inter satellite comunication link, the size and the weight of the communication system can be reduced. If the high data rate cross-link among the satellites which is equal to the whole transponder bandwidth can be established, the functionally distributed communications satellite system can be constructed. Then, by exchanging a part of the satellite system without the physical contact, in other words, by exchange some old satellites and new ones, the fully reconfigurable and long lifetime (from the point of the function) satellite communication system which can follow the paradigm shift in the terrestrial communications technology can be realize. On the other hand, however, since the maximum of relative angle error among two satellites is enlarged, the tracking becomes difficult. In this study, it turns the electric power which is gotten from the shortening the distance to making beam width large. Here, we examine communication among two satellites (10Km distance) where the data rate is 1Gbps.

  6. Analysis of ligand-receptor cross-linked fragments by mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Son, C.D. [University of Tennessee, Knoxville (UTK); Sargsyan, H. [City University of New York (CUNY); Hurst, Gregory {Greg} B [ORNL; Naider, F. [City University of New York (CUNY); Becker, J.M. [University of Tennessee, Knoxville (UTK)

    2005-01-01

    G-protein coupled receptors (GPCRs) are a class of integral membrane receptor proteins that are characterized by a signature seven-transmembrane (7-TM) configuration. The a-factor receptor (Ste2p) from Saccharomyces cerevisiae is a GPCR that, upon binding of a peptide ligand, transduces a signal to initiate a cascade of events leading to the mating of haploid yeast cells. This study summarizes the application of affinity purification and of matrix-assisted laser-desorption ionization time-of-flight (MALDI-TOF) experiments using biotinylated photoactivatable a-factor analogs. Affinity purification and enrichment of biotinylated peptides by monomeric avidin beads resulted in mass spectrometric detection of specific signals corresponding to crosslinked fragments of Ste2p. Data obtained from cyanogen bromide (CNBr) fragments of receptor cross-linked to an a-factor analog with the photoaffinity group p-benzoyl-L-phenylalanine on position 1 were in agreement with the previous results reported by our laboratory suggesting the cross-linking between position 1 of a-factor and a region of Ste2p covering residues 251 294.

  7. Laser cross-linking protein captures for living cells on a biochip

    Science.gov (United States)

    Lin, Chih-Lang; Pan, Ming-Jeng; Chen, Hai-Wen; Lin, Che-Kuan; Lin, Chuen-Fu; Baldeck, Patrice L.

    2014-03-01

    In this study, bio-sensing pads are proposed to capture living cells, which are fabricated on cover glasses by cross-linking proteins/antibodies using laser induced photochemistry. The biological functions of the cross-linked protein/antibody were verified by capturing Staphylococcus aureus (S. aureus), Leptospira, and red blood cells (RBCs), separately, with associated protein/antibody sensing pads. The experimental results show that S. aureus were bound on GFP-AcmA' pad after minutes of incubation and phosphate buffered saline (PBS) rinsing. No binding was observed with reference pad made of neutral bovine serum albumin (BSA). Second, A-type RBCs were chosen as the model cell to demonstrate the blood typing feasibility of the anti-A pad in microchannel. The A-type RBCs were captured only by the anti-A pad, but not the reference pad made of BSA. The same experimental model was carried out on the Leptospira, which stuck on the blood serum pad after PBS rinsing, but not BSA pad. This study provides a potential platform for simple and direct detection of living full cells without culture that could be used in point-of-care settings.

  8. Transparent, Superflexible Doubly Cross-Linked Polyvinylpolymethylsiloxane Aerogel Superinsulators via Ambient Pressure Drying.

    Science.gov (United States)

    Zu, Guoqing; Shimizu, Taiyo; Kanamori, Kazuyoshi; Zhu, Yang; Maeno, Ayaka; Kaji, Hironori; Shen, Jun; Nakanishi, Kazuki

    2018-01-23

    Aerogels have many attractive properties but are usually costly and mechanically brittle, which always limit their practical applications. While many efforts have been made to reinforce the aerogels, most of the reinforcement efforts sacrifice the transparency or superinsulating properties. Here we report superflexible polyvinylpolymethylsiloxane, (CH 2 CH(Si(CH 3 )O 2/2 )) n , aerogels that are facilely prepared from a single precursor vinylmethyldimethoxysilane or vinylmethyldiethoxysilane without organic cross-linkers. The method is based on consecutive processes involving radical polymerization and hydrolytic polycondensation, followed by ultralow-cost, highly scalable, ambient-pressure drying directly from alcohol as a drying medium without any modification or additional solvent exchange. The resulting aerogels and xerogels show a homogeneous, tunable, highly porous, doubly cross-linked nanostructure with the elastic polymethylsiloxane network cross-linked with flexible hydrocarbon chains. An outstanding combination of ultralow cost, high scalability, uniform pore size, high surface area, high transparency, high hydrophobicity, excellent machinability, superflexibility in compression, superflexibility in bending, and superinsulating properties has been achieved in a single aerogel or xerogel. This study represents a significant progress of porous materials and makes the practical applications of transparent flexible aerogel-based superinsulators realistic.

  9. Degradability of cross-linked polyurethanes based on synthetic polyhydroxybutyrate and modified with polylactide.

    Science.gov (United States)

    Brzeska, Joanna; Morawska, Magda; Sikorska, Wanda; Tercjak, Agnieszka; Kowalczuk, Marek; Rutkowska, Maria

    2017-01-01

    In many areas of application of conventional non-degradable cross-linked polyurethanes (PUR), there is a need for their degradation under the influence of specific environmental factors. It is practiced by incorporation of sensitive to degradation compounds (usually of natural origin) into the polyurethane structure, or by mixing them with polyurethanes. Cross-linked polyurethanes (with 10 and 30%wt amount of synthetic poly([ R,S ]-3-hydroxybutyrate) (R,S-PHB) in soft segments) and their physical blends with poly([d,l]-lactide) (PDLLA) were investigated and then degraded under hydrolytic (phosphate buffer solution) and oxidative (CoCl 2 /H 2 O 2 ) conditions. The rate of degradation was monitored by changes of samples mass, morphology of surface and their thermal properties. Despite the small weight losses of samples, the changes of thermal properties of polymers and topography of their surface indicated that they were susceptible to gradual degradation under oxidative and hydrolytic conditions. Blends of PDLLA and polyurethane with 30 wt% of R,S -PHB in soft segments and PUR/PDLLA blends absorbed more water and degraded faster than polyurethane with low amount of R,S -PHB.

  10. Enzymatically Cross-linked Alginic-Hyaluronic acid Composite Hydrogels As Cell Delivery Vehicles

    Science.gov (United States)

    Ganesh, Nitya; Hanna, Craig; Nair, Shantikumar V.; Nair, Lakshmi S.

    2013-01-01

    An injectable composite gel was developed from alginic and hyaluronic acid. The ezymatically cross-linked injectable gels were prepared via the oxidative coupling of tyramine modified sodium algiante and sodium hyaluronate in the presence of horse radish peroxidase (HRP) and hydrogen peroxide (H2O2). The composite gels were prepared by mixing equal parts of the two tryaminated polymer solutions in 10U HRP and treating with 1.0% H2O2. The properties of the alginate gels were significanly affected by the addition of hyaluronic acid. The percentage water absorption and storage modulus of the composite gels were found to be lower than the alginate gels. The alginate and composite gels showed lower protein release compared to hyaluronate gels in the absence of hyaluronidase. Even hyaluronate gels showed only approximately 10% protein release after 14 days incubation in phosphate buffer solution. ATDC-5 cells encapsulated in the injectable gels showed high cell viability. The composite gels showed the presence of enlarged spherical cells with significantly higher metabolic activity compared to cells in hyaluronic and alginic acid gels. The results suggest the potential of the composite approach to develop covalently cross-linked hydrogels with tuneable physical, mechanical, and biological properties. PMID:23357799

  11. Bioinspired composites from cross-linked galactoglucomannan and microfibrillated cellulose: Thermal, mechanical and oxygen barrier properties.

    Science.gov (United States)

    Oinonen, Petri; Krawczyk, Holger; Ek, Monica; Henriksson, Gunnar; Moriana, Rosana

    2016-01-20

    In this study, new wood-inspired films were developed from microfibrillated cellulose and galactoglucomannan-lignin networks isolated from chemothermomechanical pulping side streams and cross-linked using laccase enzymes. To the best of our knowledge, this is the first time that cross-linked galactoglucomannan-lignin networks have been used for the potential development of composite films inspired by woody-cell wall formation. Their capability as polymeric matrices was assessed based on thermal, structural, mechanical and oxygen permeability analyses. The addition of different amounts of microfibrillated cellulose as a reinforcing agent and glycerol as a plasticizer on the film performances was evaluated. In general, an increase in microfibrillated cellulose resulted in a film with better thermal, mechanical and oxygen barrier performance. However, the presence of glycerol decreased the thermal stability, stiffness and oxygen barrier properties of the films but improved their elongation. Therefore, depending on the application, the film properties can be tailored by adjusting the amounts of reinforcing agent and plasticizer in the film formulation. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Comparative Study of One-Step Cross-Linked Electrospun Chitosan-Based Membranes

    Directory of Open Access Journals (Sweden)

    Yanet E. Aguirre-Chagala

    2017-01-01

    Full Text Available Chitosan membranes are widely applied for tissue engineering; however, a major drawback is their low resistance in aqueous phases and therefore the structure collapses impeding their long-term use. Although there is extensive research, because of chitosan’s importance as a biomaterial, studies involving chitosan-based membranes are still needed. Herein, a detailed investigation of diverse chemical routes to cross-link fibers in situ by electrospinning process is described. In case of using genipin as cross-linker, a close relationship with the content and the mean diameter values is reported, suggesting a crucial effect over the design of nanostructures. Also, the physical resistance is enhanced for the combination of two types of methods, such as chemical and physical methods. Cross-linked fibers upon exposure to long wave ultraviolet A (UVA light change their morphology, but not their chemical composition. When they are incubated in aqueous phase for 70 days, they show an extensive improvement of their macrostructural integrity which makes them attractive candidates for tissue engineering application. As a result, the thermal properties of these materials reveal less crystallinity and higher temperature of degradation.

  13. Polymeric redox-responsive delivery systems bearing ammonium salts cross-linked via disulfides

    Directory of Open Access Journals (Sweden)

    Christian Dollendorf

    2013-08-01

    Full Text Available A redox-responsive polycationic system was synthesized via copolymerization of N,N-diethylacrylamide (DEAAm and 2-(dimethylaminoethyl methacrylate (DMAEMA. N,N’-bis(4-chlorobutanoylcystamine was used as disulfide-containing cross-linker to form networks by the quaternization of tertiary amine groups. The insoluble cationic hydrogels become soluble by reduction of disulfide to mercaptanes by use of dithiothreitol (DTT, tris(2-carboxyethylphosphine (TCEP or cysteamine, respectively. The soluble polymeric system can be cross-linked again by using oxygen or hydrogen peroxide under basic conditions. The redox-responsive polymer networks can be used for molecular inclusion and controlled release. As an example, phenolphthalein, methylene blue and reactive orange 16 were included into the network. After treatment with DTT a release of the dye could be recognized. Physical properties of the cross-linked materials, e.g., glass transition temperature (Tg, swelling behavior and cloud points (Tc were investigated. Redox-responsive behavior was further analyzed by rheological measurements.

  14. Effects of Composition of Iron-Cross-Linked Alginate Hydrogels for Cultivation of Human Dermal Fibroblasts

    Directory of Open Access Journals (Sweden)

    Ikuko Machida-Sano

    2012-01-01

    Full Text Available We investigated the suitability of ferric-ion-cross-linked alginates (Fe-alginate with various proportions of L-guluronic acid (G and D-mannuronic acid (M residues as a culture substrate for human dermal fibroblasts. High-G and high-M Fe-alginate gels showed comparable efficacy in promoting initial cell adhesion and similar protein adsorption capacities, but superior cell proliferation was observed on high-G than on high-M Fe-alginate as culture time progressed. During immersion in culture medium, high-G Fe-alginate showed little change in gel properties in terms of swelling and polymer content, but the properties of high-M Fe-alginate gel were altered due to loss of ion cross-linking. However, the degree of cell proliferation on high-M Fe-alginate gel was improved after it had been stabilized by immersion in culture medium until no further changes occurred. These results suggest that the mode of cross-linkage between ferric ions and alginate differs depending on alginate composition and that the major factor giving rise to differences in cell growth on the two types of Fe-alginate films is gel stability during culture, rather than swelling of the original gel, polymer content, or protein adsorption ability. Our findings may be useful for extending the application of Fe-alginate to diverse biomedical fields.

  15. Seed-mediated synthesis of cross-linked Pt-NiO nanochains for methanol oxidation

    Science.gov (United States)

    Gu, Zhulan; Bin, Duan; Feng, Yue; Zhang, Ke; Wang, Jin; Yan, Bo; Li, Shumin; Xiong, Zhiping; Wang, Caiqin; Shiraishi, Yukihide; Du, Yukou

    2017-07-01

    A simple method was reported for employing NiO nanoparticles act as seeds and then different amounts of Pt2+ were reduced on the NiO nanoparticles, forming a cross-linked Pt-NiO nanocatalysts. These as-prepared catalysts were characterized using different physical-chemical techniques, including X-ray diffraction (XRD), transmission electron microscopy (TEM) and X-ray photoelectron spectroscopy (XPS). The results indicate that the morphology of the cross-linked Pt-NiO nanochain was successfully produced regardless of the molar ratio of Pt2+ to NiO precursors. The electrochemical characteristics of Pt-NiO nanochain catalysts were evaluated for the oxidation of methanol as a model reaction, which verify that the Pt-NiO catalysts show enhanced activity and high stability in comparison with the commercial Pt/C catalyst. The optimized ratio of Pt to NiO is 1:1, then tuned by simple adjusting the feed ratio of the precursors as well. The synthesized nanocatalysts will be found the great potential applications as electrocatalysts for fuel cells owe to their enhanced catalytic performance and long-term stability.

  16. Genipin-cross-linked poly(L-lysine)-based hydrogels: synthesis, characterization, and drug encapsulation.

    Science.gov (United States)

    Wang, Steven S S; Hsieh, Ping-Lun; Chen, Pei-Shan; Chen, Yu-Tien; Jan, Jeng-Shiung

    2013-11-01

    Genipin-cross-linked hydrogels composed of biodegradable and pH-sensitive cationic poly(L-lysine) (PLL), poly(L-lysine)-block-poly(L-alanine) (PLL-b-PLAla), and poly(L-lysine)-block-polyglycine (PLL-b-PGly) polypeptides were synthesized, characterized, and used as carriers for drug delivery. These polypeptide hydrogels can respond to pH-stimulus and their gelling and mechanical properties, degradation rate, and drug release behavior can be tuned by varying polypeptide composition and cross-linking degree. Comparing with natural polymers, the synthetic polypeptides with well-defined chain length and composition can warrant the preparation of the hydrogels with tunable properties to meet the criteria for specific biomedical applications. These hydrogels composed of natural building blocks exhibited good cell compatibility and enzyme degradability and can support cell attachment/proliferation. The evaluation of these hydrogels for in vitro drug release revealed that the controlled release profile was a biphasic pattern with a mild burst release and a moderate release rate thereafter, suggesting the drug molecules were encapsulated inside the gel matrix. With the versatility of polymer chemistry and conjugation of functional moieties, it is expected these hydrogels can be useful for biomedical applications such as polymer therapeutics and tissue engineering. Copyright © 2013 Elsevier B.V. All rights reserved.

  17. DFT Modeling of Cross-Linked Polyethylene: Role of Gold Atoms and Dispersion Interactions.

    Science.gov (United States)

    Blaško, Martin; Mach, Pavel; Antušek, Andrej; Urban, Miroslav

    2018-02-08

    Using DFT modeling, we analyze the concerted action of gold atoms and dispersion interactions in cross-linked polyethylene. Our model consists of two oligomer chains (PEn) with 7, 11, 15, 19, or 23 carbon atoms in each oligomer cross-linked with one to three Au atoms through C-Au-C bonds. In structures with a single gold atom the C-Au-C bond is located in the central position of the oligomer. Binding energies (BEs) with respect to two oligomer radical fragments and Au are as high as 362-489 kJ/mol depending on the length of the oligomer chain. When the dispersion contribution in PEn-Au-PEn oligomers is omitted, BE is almost independent of the number of carbon atoms, lying between 293 and 296 kJ/mol. The dispersion energy contributions to BEs in PEn-Au-PEn rise nearly linearly with the number of carbon atoms in the PEn chain. The carbon-carbon distance in the C-Au-C moiety is around 4.1 Å, similar to the bond distance between saturated closed shell chains in the polyethylene crystal. BEs of pure saturated closed shell PEn-PEn oligomers are 51-187 kJ/mol. Both Au atoms and dispersion interactions contribute considerably to the creation of nearly parallel chains of oligomers with reasonably high binding energies.

  18. Multivalent anchoring and cross-linking of mussel-inspired antifouling surface coatings.

    Science.gov (United States)

    Wei, Qiang; Becherer, Tobias; Mutihac, Radu-Cristian; Noeske, Paul-Ludwig Michael; Paulus, Florian; Haag, Rainer; Grunwald, Ingo

    2014-08-11

    In this work, we combine nature's amazing bioadhesive catechol with the excellent bioinert synthetic macromolecule hyperbranched polyglycerol (hPG) to prepare antifouling surfaces. hPG can be functionalized by different amounts of catechol groups for multivalent anchoring and cross-linking because of its highly branched architecture. The catecholic hPGs can be immobilized on various surfaces including metal oxides, noble metals, ceramics, and polymers via simple incubation procedures. The effect of the catechol amount on the immobilization, surface morphology, stability, and antifouling performance of the coatings was studied. Both anchoring and cross-linking interactions provided by catechols can enhance the stability of the coatings. When the catechol groups on the hPG are underrepresented, the tethering of the coating is not effective; while an overrepresentation of catechol groups leads to protein adsorption and cell adhesion. Thus, only a well-balanced amount of catechols as optimized and described in this work can supply the coatings with both good stability and antifouling ability.

  19. Low-Temperature Cross-Linking of PEDOT:PSS Films Using Divinylsulfone.

    Science.gov (United States)

    Mantione, Daniele; Del Agua, Isabel; Schaafsma, Wandert; ElMahmoudy, Mohammed; Uguz, Ilke; Sanchez-Sanchez, Ana; Sardon, Haritz; Castro, Begoña; Malliaras, George G; Mecerreyes, David

    2017-05-31

    Recent interest in bioelectronics has prompted the exploration of properties of conducting polymer films at the interface with biological milieus. Poly(3,4-ethylenedioxythiophene) doped with poly(styrenesulfonate) (PEDOT:PSS) from a commercially available source has been used as a model system for these studies. Different cross-linking schemes have been used to stabilize films of this material against delamination and redispersion, but the cost is a decrease in the electrical conductivity and/or additional heat treatment. Here we introduce divinylsulfone (DVS) as a new cross-linker for PEDOT:PSS. Thanks to the higher reactiveness of the vinyl groups of DVS, the cross-linking can be performed at room temperature. In addition, DVS does not reduce electronic conductivity of PEDOT:PSS but rather increases it by acting as a secondary dopant. Cell culture studies show that PEDOT:PSS:DVS films are cytocompatible and support neuroregeneration. As an example, we showed that this material improved the transconductance value and stability of an organic electrochemical transistor (OECT) device. These results open the way for the utilization of DVS as an effective cross-linker for PEDOT:PSS in bioelectronics applications.

  20. Synthesis of Covalently Cross-Linked Colloidosomes from Peroxidized Pickering Emulsions

    Directory of Open Access Journals (Sweden)

    Nadiya Popadyuk

    2016-10-01

    Full Text Available A new approach to the formation of cross-linked colloidosomes was developed on the basis of Pickering emulsions that were stabilized exclusively by peroxidized colloidal particles. Free radical polymerization and a soft template technique were used to convert droplets of a Pickering emulsion into colloidosomes. The peroxidized latex particles were synthesized in the emulsion polymerization process using amphiphilic polyperoxide copolymers poly(2-tert-butylperoxy-2-methyl-5-hexen-3-ine-co-maleic acid (PM-1-MAc or poly[N-(tert-butylperoxymethylacrylamide]-co-maleic acid (PM-2-MAc, which were applied as both initiators and surfactants (inisurfs. The polymerization in the presence of the inisurfs results in latexes with a controllable amount of peroxide and carboxyl groups at the particle surface. Peroxidized polystyrene latex particles with a covalently grafted layer of inisurf PM-1-MAc or PM-2-MAc were used as Pickering stabilizers to form Pickering emulsions. A mixture of styrene and/or butyl acrylate with divinylbenzene and hexadecane was applied as a template for the synthesis of colloidosomes. Peroxidized latex particles located at the interface are involved in the radical reactions of colloidosomes formation. As a result, covalently cross-linked colloidosomes were obtained. It was demonstrated that the structure of the synthesized (using peroxidized latex particles colloidosomes depends on the amount of functional groups and pH during the synthesis. Therefore, the size and morphology of colloidosomes can be controlled by latex particle surface properties.

  1. Adsorption of Pb{sup 2+} on chitosan cross-linked with triethylene-tetramine

    Energy Technology Data Exchange (ETDEWEB)

    Tang, X.H. [College of Chemistry and Chemical Engineering, Hunan University, Changsha (China); College of Environmental and Chemical Engineering, Nanchang Institute of Aeronautical Technology (China); Zhang, X.M.; Zhou, A.L. [College of Environmental and Chemical Engineering, Nanchang Institute of Aeronautical Technology (China); Guo, C.C. [College of Chemistry and Chemical Engineering, Hunan University, Changsha (China)

    2007-07-15

    A novel triethylene-tetramine cross-linked chitosan (CCTS) was synthesized via the cross-linking of triethylene-tetramine and epichlorohydrin activated chitosan. Its structure was characterized by elemental analysis, infrared spectroscopy and X-ray diffraction analysis, and the surface topography was determined with ESEM. The results were in agreement with expectations. The capacity of CCTS to adsorb Pb{sup 2+} ions from aqueous solutions was examined, and equilibrium and kinetic investigations were undertaken. The adsorption isotherms were fitted well by the Langmuir equation (R > 0.999). The maximum adsorbed amount, at pH 5.5, with an initial concentration of 3 mmol/L (621 ppm), was 378.8 mg/g. The adsorption process could be best described by a second-order equation (R = 1). This suggests that the rate-limiting step may be the chemical adsorption (chemisorption) step and not the mass transport. The separation factor used was 0 < R{sub L} < 1. Therefore, it can be concluded that CCTS is an effective adsorbent for the collection of Pb{sup 2+}. (Abstract Copyright [2007], Wiley Periodicals, Inc.)

  2. Effect of nordihydroguaiaretic acid cross-linking on fibrillar collagen: in vitro evaluation of fibroblast adhesion strength and migration

    Directory of Open Access Journals (Sweden)

    Ana Y. Rioja

    2017-04-01

    Full Text Available Fixation is required to reinforce reconstituted collagen for orthopedic bioprostheses such as tendon or ligament replacements. Previous studies have demonstrated that collagen fibers cross-linked by the biocompatible dicatechol nordihydroguaiaretic acid (NDGA have mechanical strength comparable to native tendons. This work focuses on investigating fibroblast behavior on fibrillar and NDGA cross-linked type I collagen to determine if NDGA modulates cell adhesion, morphology, and migration. A spinning disk device that applies a range of hydrodynamic forces under uniform chemical conditions was employed to sensitively quantify cell adhesion strength, and a radial barrier removal assay was used to measure cell migration on films suitable for these quantitative in vitro assays. The compaction of collagen films, mediated by the drying and cross-linking fabrication process, suggests a less open organization compared to native fibrillar collagen that likely allowed the collagen to form more inter-chain bonds and chemical links with NDGA polymers. Fibroblasts strongly adhered to and migrated on native and NDGA cross-linked fibrillar collagen; however, NDGA modestly reduced cell spreading, adhesion strength and migration rate. Thus, it is hypothesized that NDGA cross-linking masked some adhesion receptor binding sites either physically, chemically, or both, thereby modulating adhesion and migration. This alteration in the cell-material interface is considered a minimal trade-off for the superior mechanical and compatibility properties of NDGA cross-linked collagen compared to other fixation approaches.

  3. The effect of pH on hydrolysis, cross-linking and barrier properties of starch barriers containing citric acid.

    Science.gov (United States)

    Olsson, Erik; Menzel, Carolin; Johansson, Caisa; Andersson, Roger; Koch, Kristine; Järnström, Lars

    2013-11-06

    Citric acid cross-linking of starch for e.g. food packaging applications has been intensely studied during the last decade as a method of producing water-insensitive renewable barrier coatings. We managed to improve a starch formulation containing citric acid as cross-linking agent for industrial paper coating applications by adjusting the pH of the starch solution. The described starch formulations exhibited both cross-linking of starch by citric acid as well as satisfactory barrier properties, e.g. fairly low OTR values at 50% RH that are comparable with EVOH. Furthermore, it has been shown that barrier properties of coated papers with different solution pH were correlated to molecular changes in starch showing both hydrolysis and cross-linking of starch molecules in the presence of citric acid. Hydrolysis was shown to be almost completely hindered at solution pH≥4 at curing temperatures≤105 °C and at pH≥5 at curing temperatures≤150 °C, whereas cross-linking still occurred to some extent at pH≤6.5 and drying temperatures as low as 70 °C. Coated papers showed a minimum in water vapor transmission rate at pH 4 of the starch coating solution, corresponding to the point where hydrolysis was effectively hindered but where a significant degree of cross-linking still occurred. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. Bone fracture toughness and strength correlate with collagen cross-link maturity in a dose-controlled lathyrism mouse model.

    Science.gov (United States)

    McNerny, Erin M B; Gong, Bo; Morris, Michael D; Kohn, David H

    2015-03-01

    Collagen cross-linking is altered in many diseases of bone, and enzymatic collagen cross-links are important to bone quality, as evidenced by losses of strength after lysyl oxidase inhibition (lathyrism). We hypothesized that cross-links also contribute directly to bone fracture toughness. A mouse model of lathyrism using subcutaneous injection of up to 500 mg/kg β-aminopropionitrile (BAPN) was developed and characterized (60 animals across 4 dosage groups). Three weeks of 150 or 350 mg/kg BAPN treatment in young, growing mice significantly reduced cortical bone fracture toughness, strength, and pyridinoline cross-link content. Ratios reflecting relative cross-link maturity were positive regressors of fracture toughness (HP/[DHLNL + HLNL] r(2)  = 0.208, p toughness and strength. Thus, cross-link profile perturbations associated with bone disease may provide insight into bone mechanical quality and fracture risk. © 2014 American Society for Bone and Mineral Research.

  5. Cross-linked poly(arylene ether ketone) proton exchange membranes with high ion exchange capacity for fuel cells

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, S.; Hai, S.D.; Kim, D. [School of Chemical Engineering, Sungkyunkwan University, Suwon, Kyunggi (Korea, Republic of)

    2012-08-15

    Sulfonated poly(arylene ether ketone) (SPAEK) possessing the pendant carboxylic acid groups was synthesized. The carboxylic acid groups of SPAEK were reacted with a cross-linking reagent to prepare a cross-linked membrane with a high ion exchange capacity (IEC), a high oxidative stability, and an excellent mechanical strength. The cross-linking hindered the mobility of the polymer chains and thus strongly affected the water uptake and the methanol permeability of the membranes. Also, as the cross-linker used in this study bore sulfonic acid groups, cross-linking did not lead to a noticeable loss of the proton conductivity. The cross-linked SPAEK membrane with 20% cross-linking density, CSPAEK-20% membrane, exhibited a high proton conductivity of 0.045 S cm{sup -1} associated with a high IEC value of 1.78 mmol g{sup -1} but a low methanol permeability of 4.3 x 10{sup -7} cm{sup 2} s{sup -1}. The CSPAEK-20% membrane also showed excellent cell performance and oxidation resistance. (Copyright copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  6. Cross-linking staphylococcal enterotoxin A bound to major histocompatibility complex class I is required for TNF-alpha secretion

    Science.gov (United States)

    Wright, A. D.; Chapes, S. K.

    1999-01-01

    The mechanism of how superantigens function to activate cells has been linked to their ability to bind and cross-link the major histocompatibility complex class II (MHCII) molecule. Cells that lack the MHCII molecule also respond to superantigens, however, with much less efficiency. Therefore, the purpose of this study was to confirm that staphylococcal enterotoxin A (SEA) could bind the MHCI molecule and to test the hypothesis that cross-linking SEA bound to MHCII-deficient macrophages would induce a more robust cytokine response than without cross-linking. We used a capture enzyme-linked immunosorbent assay and an immunprecipitation assay to directly demonstrate that MHCI molecules bind SEA. Directly cross-linking MHCI using monoclonal antibodies or cross-linking bound SEA with an anti-SEA antibody or biotinylated SEA with avidin increased TNF-alpha and IL-6 secretion by MHCII(-/-) macrophages. The induction of a vigorous macrophage cytokine response by SEA/anti-SEA cross-linking of MHCI offers a mechanism to explain how MHCI could play an important role in superantigen-mediated pathogenesis. Copyright 1999 Academic Press.

  7. Collagen Cross-Linking Using Riboflavin and Ultraviolet-A for Corneal Thinning Disorders

    Science.gov (United States)

    Pron, G; Ieraci, L; Kaulback, K

    2011-01-01

    Executive Summary Objective The main objectives for this evidence-based analysis were to determine the safety and effectiveness of photochemical corneal collagen cross-linking with riboflavin (vitamin B2) and ultraviolet-A radiation, referred to as CXL, for the management of corneal thinning disease conditions. The comparative safety and effectiveness of corneal cross-linking with other minimally invasive treatments such as intrastromal corneal rings was also reviewed. The Medical Advisory Secretariat (MAS) evidence-based analysis was performed to support public financing decisions. Subject of the Evidence-Based Analysis The primary treatment objective for corneal cross-linking is to increase the strength of the corneal stroma, thereby stabilizing the underlying disease process. At the present time, it is the only procedure that treats the underlying disease condition. The proposed advantages for corneal cross-linking are that the procedure is minimally invasive, safe and effective, and it can potentially delay or defer the need for a corneal transplant. In addition, corneal cross-linking does not adversely affect subsequent surgical approaches, if they are necessary, or interfere with corneal transplants. The evidence for these claims for corneal cross-linking in the management of corneal thinning disorders such as keratoconus will be the focus of this review. The specific research questions for the evidence review were as follows: Technical: How technically demanding is corneal cross-linking and what are the operative risks? Safety: What is known about the broader safety profile of corneal cross-linking? Effectiveness - Corneal Surface Topographic Affects: What are the corneal surface remodeling effects of corneal cross-linking? Do these changes interfere with subsequent interventions, particularly corneal transplant known as penetrating keratoplasty (PKP)? Effectiveness -Visual Acuity: What impacts does the remodeling have on visual acuity? Are these impacts

  8. Collagen cross-linking using riboflavin and ultraviolet-a for corneal thinning disorders: an evidence-based analysis.

    Science.gov (United States)

    Pron, G; Ieraci, L; Kaulback, K

    2011-01-01

    The main objectives for this evidence-based analysis were to determine the safety and effectiveness of photochemical corneal collagen cross-linking with riboflavin (vitamin B(2)) and ultraviolet-A radiation, referred to as CXL, for the management of corneal thinning disease conditions. The comparative safety and effectiveness of corneal cross-linking with other minimally invasive treatments such as intrastromal corneal rings was also reviewed. The Medical Advisory Secretariat (MAS) evidence-based analysis was performed to support public financing decisions. SUBJECT OF THE EVIDENCE-BASED ANALYSIS: The primary treatment objective for corneal cross-linking is to increase the strength of the corneal stroma, thereby stabilizing the underlying disease process. At the present time, it is the only procedure that treats the underlying disease condition. The proposed advantages for corneal cross-linking are that the procedure is minimally invasive, safe and effective, and it can potentially delay or defer the need for a corneal transplant. In addition, corneal cross-linking does not adversely affect subsequent surgical approaches, if they are necessary, or interfere with corneal transplants. The evidence for these claims for corneal cross-linking in the management of corneal thinning disorders such as keratoconus will be the focus of this review. The specific research questions for the evidence review were as follows: TECHNICAL: How technically demanding is corneal cross-linking and what are the operative risks? What is known about the broader safety profile of corneal cross-linking?Effectiveness - Corneal Surface Topographic Affects:What are the corneal surface remodeling effects of corneal cross-linking?Do these changes interfere with subsequent interventions, particularly corneal transplant known as penetrating keratoplasty (PKP)?Effectiveness -Visual Acuity:What impacts does the remodeling have on visual acuity?Are these impacts predictable, stable, adjustable and

  9. A gas phase cleavage reaction of cross-linked peptides for protein complex topology studies by peptide fragment fingerprinting from large sequence database

    NARCIS (Netherlands)

    Buncherd, H.; Roseboom, W.; de Koning, L.J.; de Koster, C.G.; de Jong, L.

    2014-01-01

    A high molecular weight fraction of a HeLa cell nuclear extract containing nearly 1100 identified proteins was cross-linked with bis(succinimidyl)-3-azidomethyl glutarate (BAMG). The azido group in cross-linked peptides can be reduced to an amine group. Reduction enables isolation of cross-linked

  10. [Riboflavin photoprotection with cross-linking effect in photorefractive ablation of the cornea].

    Science.gov (United States)

    Kornilovskiy, I M; Sultanova, A I; Burtsev, A A

    2016-01-01

    Photorefractive ablation is inevitably accompanied by oxidative stress of the cornea and weakening of its biomechanical and photoprotective properties. To validate the expediency of riboflavin use in photorefractive ablation for photoprotection of the cornea and cross-linking. The effects of riboflavin use in photorefractive ablation was first studied in a series of in vitro and in vivo experiments performed on 56 eyes of 28 rabbits, and then on 232 eyes of 142 patients with different degrees of myopia. Biomechanical testing of corneal samples was performed with Zwick/RoellВZ 2.5/TN1S tensile-testing machine. Transepithelial photorefractive keratectomy (TransPRK) and femtosecond laser-assisted in situ keratomileusis (Femto-LASIK) were performed on Wavelight-Allegretto200, MEL-80, and WaveLight-EX500 excimer laser systems and also VisuMax and WaveLight-FS200 femtosecond lasers. For preliminary examinations, an appropriate set of diagnostic tools was used. In vivo experiments have proved that, in the absence of conservative therapy, riboflavin is able to produce both photoprotective and cross-linking effects to the cornea. Corneal syndrome was thus reduced and re-epithelialization after TransPRK accelerated. Biomechanical testing of corneal samples revealed an increase in tolerated load from 12.9±1.4 N to 18.3±1.2 N (p=0.0002) and tensile strength from 8.6±1.7 MPa to 12.4±1.7 MPa (p=0.007). Clinical studies conducted in a group of patients with mild to moderate myopia have also confirmed the photoprotective effect of riboflavin at months 1-12 after TransPRK. There were no significant differences in uncorrected visual acuity (ranged from 0.80±0.16 to 0.85±0.15) and corrected visual acuity at baseline (0.83±0.14). Evaluation of the optical and refractive effect achieved after Femto-LASIK with riboflavin photoprotection in the fellow eye has shown that this technique is not inferior to the traditional one as to its refractive accuracy, but provides better

  11. Use of cross-linked carboxymethyl cellulose for soft-tissue augmentation: preliminary clinical studies

    Directory of Open Access Journals (Sweden)

    Mauro Leonardis

    2010-11-01

    Full Text Available Mauro Leonardis1, Andrea Palange2, Rodrigo FV Dornelles3, Felipe Hund41Department of Plastic Surgery, Salvator Mundi International Hospital, Roma, Italy; 2Department of Aesthetic Medicine, Fisiobios, Roma, Italy; 3Department of Plastic Surgery, Núcleo de Plástica Avançada, São Paulo, SP, Brazil; 4Department of Plastic Surgery, Consultorio de Cirurgia Plastica, Criciuma, SC, BrazilPurpose: The continual search for new products for soft-tissue augmentation has in recent years led to the introduction of long lasting alternatives to hyaluronic acids and collagen that are composed of other polymers able to improve clinical persistence over time. This is the first report in which sodium carboxymethyl cellulose (CMC has been chemically treated by the cross-linking process and thus used as a hydrogel for soft-tissue augmentation through injection with thin needles. The study evaluates, from a clinical point of view, the behavior of cross-linked carboxymethyl cellulose hydrogel used in the aesthetic field and its side effects so as to check the safety and performance of the polymer following intradermal injections.Patients and methods: This work shows the preliminary results of an ongoing clinical study conducted between 2006 and 2009, performed on 84 healthy volunteers (62 females, 22 males aged between 18 and 72 years, for the treatment of 168 nasolabial folds, 45 perioral wrinkles, and 39 lip volume.Results: Study results show an excellent correction of facial defects. Tolerance and aesthetic quality of the correction obtained indicate considerable safety features and absence of side effects. From a clinical point of view, hydrogel is gradually absorbed into the injection site without migration issues.Conclusion: Cross-linked CMC hydrogel proves to be an ideal agent for soft tissue augmentation with regard to safety and ease of application. It did not cause infection, extrusion, migration, or adverse reactions in the patients who have been

  12. Characterization of the somatogenic receptor in rat liver. Hydrodynamic properties and affinity cross-linking

    International Nuclear Information System (INIS)

    Husman, B.; Haldosen, L.A.; Andersson, G.; Gustafsson, J.A.

    1988-01-01

    Rat liver somatogenic receptors have been characterized by gel permeation chromatography, sucrose density gradients in H 2 O and D 2 O, and affinity cross-linking using 125 I-bovine growth hormone (bGH) as a specific somatogenic receptor ligand. Cross-linking of 125 I-bovine growth hormone to a Triton X-100-treated low density fraction isolated from livers of late pregnant rats followed by sodium dodecylsulfate-polyacrylamide gel electrophoresis under reducing conditions showed three major binders with Mr 95,000, 86,000, and 43,000 and a minor binder of Mr 55,000, after correction for bound ligand assuming a 1:1 binding ratio of ligand-receptor. The Mr 86,000, 55,000, and 43,000 species were recovered in the detergent-soluble supernatant after high-speed centrifugation, whereas the Mr 95,000 species remained Triton X-100 insoluble. Detergent-soluble 125 I-bGH-receptor complexes were further analyzed by sedimentation into sucrose density gradients. The sedimentation coefficient was S20,w = 5.2 S and the partial specific volume v = 0.72 ml/g. Gel permeation chromatography on a Sepharose S-400 column indicated a Stokes radius of 61 A for the 125 I-bGH-receptor-Triton X-100 complex. Based on these figures, the molecular weight of the complex was calculated as 131,100. The molecular weight of the ligand-free receptor-Triton X-100 complex was calculated as Mr 109,100. Affinity cross-linking and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the 61 A peak from Sephacryl S-400 chromatography (cf. above) showed two binding entities, one major and one minor with Mr values 86,000 and 43,000, respectively, in the absence of reductant. When electrophoresis was run in the presence of reductant the Mr 43,000 species was the major binding entity

  13. Evaluation of cross-linked aggregates from purified Bacillus subtilis levansucrase mutants for transfructosylation reactions

    Directory of Open Access Journals (Sweden)

    Munguia Agustin

    2009-07-01

    Full Text Available Abstract Background Increasing attention has been focused on inulin and levan-type oligosaccharides, including fructosyl-xylosides and other fructosides due to their nutraceutical properties. Bacillus subtilis levansucrase (LS catalyzes the synthesis of levan from sucrose, but it may also transfer the fructosyl moiety from sucrose to acceptor molecules included in the reaction medium. To study transfructosylation reactions with highly active and robust derivatives, cross-linked enzyme aggregates (CLEAs were prepared from wild LS and two mutants. CLEAs combine the catalytic features of pure protein preparations in terms of specific activity with the mechanical behavior of industrial biocatalysts. Results Two types of procedures were used for the preparation of biocatalysts from purified wild type LS (WT LS B. subtilis and the R360K and Y429N LS mutants: purified enzymes aggregated with glutaraldehyde (cross-linked enzyme aggregates: CLEAs, and covalently immobilized enzymes in Eupergit C®. The biocatalysts were characterized and used for fructoside synthesis using xylose as an acceptor model. CLEAs were able to catalyze the synthesis of fructosides as efficiently as soluble enzymes. The specific activity of CLEAs prepared from wild type LS (44.9 U/mg of CLEA, R360K (56.5 U/mg of CLEA and Y429N (1.2 U/mg of CLEA mutants were approximately 70, 40 and 200-fold higher, respectively, than equivalent Eupergit C® immobilized enzyme preparations (U/mg of Eupergit, where units refer to global LS activity. In contrast, the specific activity of the free enzymes was 160, 171.2 and 1.5 U/mg of protein, respectively. Moreover, all CLEAs had higher thermal stability than corresponding soluble enzymes. In the long term, the operational stability was affected by levan synthesis. Conclusion This is the first report of cross-linked transglycosidases aggregates. CLEAs prepared from purified LS and mutants have the highest specific activity for immobilized

  14. Genetically encoded releasable photo-cross-linking strategies for studying protein-protein interactions in living cells.

    Science.gov (United States)

    Yang, Yi; Song, Haiping; He, Dan; Zhang, Shuai; Dai, Shizhong; Xie, Xiao; Lin, Shixian; Hao, Ziyang; Zheng, Huangtao; Chen, Peng R

    2017-10-01

    Although protein-protein interactions (PPIs) have crucial roles in virtually all cellular processes, the identification of more transient interactions in their biological context remains challenging. Conventional photo-cross-linking strategies can be used to identify transient interactions, but these approaches often suffer from high background due to the cross-linked bait proteins. To solve the problem, we have developed membrane-permeable releasable photo-cross-linkers that allow for prey-bait separation after protein complex isolation and can be installed in proteins of interest (POIs) as unnatural amino acids. Here we describe the procedures for using two releasable photo-cross-linkers, DiZSeK and DiZHSeC, in both living Escherichia coli and mammalian cells. A cleavage after protein photo-cross-linking (CAPP ) strategy based on the photo-cross-linker DiZSeK is described, in which the prey protein pool is released from a POI after affinity purification. Prey proteins are analyzed using mass spectrometry or 2D gel electrophoresis for global comparison of interactomes from different experimental conditions. An in situ cleavage and mass spectrometry (MS)-label transfer after protein photo-cross-linking (IMAPP) strategy based on the photo-cross-linker DiZHSeC is also described. This strategy can be used for the identification of cross-linking sites to allow detailed characterization of PPI interfaces. The procedures for photo-cross-linker incorporation, photo-cross-linking of interaction partners and affinity purification of cross-linked complexes are similar for the two photo-cross-linkers. The final section of the protocol describes prey-bait separation (for CAPP) and MS-label transfer and identification (for IMAPP). After plasmid construction, the CAPP and IMAPP strategies can be completed within 6 and 7 d, respectively.

  15. Wear of PEEK-OPTIMA® and PEEK-OPTIMA®-Wear Performance articulating against highly cross-linked polyethylene.

    Science.gov (United States)

    East, Rebecca H; Briscoe, Adam; Unsworth, Anthony

    2015-03-01

    The idea of all polymer artificial joints, particularly for the knee and finger, has been raised several times in the past 20 years. This is partly because of weight but also to reduce stress shielding in the bone when stiffer materials such as metals or ceramics are used. With this in mind, pin-on-plate studies of various polyetheretherketone preparations against highly cross-linked polyethylene were conducted to investigate the possibility of using such a combination in the design of a new generation of artificial joints. PEEK-OPTIMA(®) (no fibre) against highly cross-linked polyethylene gave very low wear factors of 0.0384 × 10(-6) mm(3)/N m for the polyetheretherketone pins and -0.025 × 10(-6) mm(3)/N m for the highly cross-linked polyethylene plates. The carbon-fibre-reinforced polyetheretherketone (PEEK-OPTIMA(®)-Wear Performance) also produced very low wear rates in the polyetheretherketone pins but produced very high wear in the highly cross-linked polyethylene, as might have been predicted since the carbon fibres are quite abrasive. When the fibres were predominantly tangential to the sliding plane, the mean wear factor was 0.052 × 10(-6) mm(3)/N m for the pins and 49.3 × 10(-6) mm(3)/N m for the highly cross-linked polyethylene plates; a half of that when the fibres ran axially in the pins (0.138 × 10(-6) mm(3)/N m for the pins and 97.5 × 10(-6) mm/ N m for the cross-linked polyethylene plates). PEEK-OPTIMA(®) against highly cross-linked polyethylene merits further investigation. © IMechE 2015.

  16. Investigation of cross-linking characteristics of novel hole-transporting materials for solution-processed phosphorescent OLEDs

    Science.gov (United States)

    Lee, Jaemin; Ameen, Shahid; Lee, Changjin

    2016-04-01

    After the success of commercialization of the vacuum-evaporated organic light-emitting diodes (OLEDs), solutionprocessing or printing of OLEDs are currently attracting much research interests. However, contrary to various kinds of readily available vacuum-evaporable OLED materials, the solution-processable OLED materials are still relatively rare. Hole-transporting layer (HTL) materials for solution-processed OLEDs are especially limited, because they need additional characteristics such as cross-linking to realize multilayer structures in solution-processed OLEDs, as well as their own electrically hole-transporting characteristics. The presence of such cross-linking characteristics of solutionprocessable HTL materials therefore makes them more challenging in the development stage, and also makes them essence of solution-processable OLED materials. In this work, the structure-property relationships of thermally crosslinkable HTL materials were systematically investigated by changing styrene-based cross-linking functionalities and modifying the carbazole-based hole-transporting core structures. The temperature dependency of the cross-linking characteristics of the HTL materials was systematically investigated by the UV-vis. absorption spectroscopy. The new HTL materials were also applied to green phosphorescent OLEDs, and their device characteristics were also investigated based on the chemical structures of the HTL materials. The device configuration was [ITO / PEDOT:PSS / HTL / EML / ETL / CsF / Al]. We found out that the chemical structures of the cross-linking functionalities greatly affect not only the cross-linking characteristics of the resultant HTL materials, but also the resultant OLED device characteristics. The increase of the maximum luminance and efficiency of OLEDs was evident as the cross-linking temperature decreases from higher than 200°C to at around 150°C.

  17. Latent heat storage in a fixed-bed packed with cross-linked polymer particles

    Science.gov (United States)

    Morita, Hiroki; Miyatake, Osamu

    Experimental and numerical analyses were carried out to investigate the discharge characteristics of a fixed-bed, using form-stable cross-linked cylindrical polymer particles as a phase change type of heat storage material and ethylene glycol as heat transfer fluid. Firstly, particle-to-fluid heat transfer coefficient in a fixed-bed, essential for analyzing the discharge characteristics of the latent heat storage column, were evaluated experimentally using Schumann's extended theory, and an empirical formula for estimating the heat transfer coefficient was presented. Secondly, by using the empirical formula, the transient temperature distribution in the column and the transient response of the outlet temperature of the heat transfer fluid were calculated numerically, and the latter was compared with measurements of the outlet temperature of the fluid. The numerical results were found to be in good agreement with the experimental results.

  18. Removal of hexavalent chromium from aqueous solution by barium ion cross-linked alginate beads

    Directory of Open Access Journals (Sweden)

    Uzaşçı Sesil

    2014-07-01

    Full Text Available Barium ion cross-linked alginate beads have shown great affinity to toxic hexavalent chromium ions in aqueous solution, contrary to the traditional calcium alginate beads. The adsorption experiments were carried out by the batch contact method. The optimal pH for removal was found to be pH 4. The equilibrium was established in 4 h and the removal efficiency of chromium (VI was found as 95%. The adsorption data fit well with Langmuir and Freundlich isotherms. The maximum chromium (VI adsorption capacity determined from Langmuir isotherm was 36.5 mg/g dry alginate beads. Our study suggests that barium alginate beads can be used as cost-effective and efficient adsorbents for the removal of chromium (VI from contaminated waters.

  19. Star polymers as unit cells for coarse-graining cross-linked networks

    Science.gov (United States)

    Molotilin, Taras Y.; Maduar, Salim R.; Vinogradova, Olga I.

    2018-03-01

    Reducing the complexity of cross-linked polymer networks by preserving their main macroscale properties is key to understanding them, and a crucial issue is to relate individual properties of the polymer constituents to those of the reduced network. Here we study polymer networks in a good solvent, by considering star polymers as their unit elements, and first quantify the interaction between their centers of masses. We then reduce the complexity of a network by replacing sets of its bridged star polymers by equivalent effective soft particles with dense cores. Our coarse graining allows us to approximate complex polymer networks by much simpler ones, keeping their relevant mechanical properties, as illustrated in computer experiments.

  20. Glucose-sensitive microcapsules from glutaraldehyde cross-linked hemoglobin and glucose oxidase.

    Science.gov (United States)

    Qi, Wei; Yan, Xuehai; Duan, Li; Cui, Yue; Yang, Yang; Li, Junbai

    2009-05-11

    We have fabricated the glucose-sensitive protein hollow microcapsules from hemoglobin (Hb) and glucose oxidase (GOx) via layer-by-layer technique with glutaraldehyde (GA) as a cross-linking agent. We demonstrated the coupled enzymatic reactions between Hb and GOx within capsules by means of CLSM. The real-time monitoring of the consumption of glucose may be useful for the design of a glucose sensor. Furthermore, to further study the effect of glucose on the capsules, we investigated the wall permeability of (Hb/GOx)(5) capsules. It was found that the glucose consumption could enhance the permeability of (Hb/GOx)(5) capsules. This is significant for the development of advanced drug delivery devices. Therefore, the fabricated Hb/GOx capsules may be a potential candidate for the creation of multifunctional devices.