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Lysozyme
Kinds of Lysozyme Terms modified by Lysozyme Selected AbstractsGold nanoparticles induce protein crystallizationCRYSTAL RESEARCH AND TECHNOLOGY, Issue 6 2008F. Hodzhaoglu Abstract Nucleation of protein crystals by gold nanoparticles was observed. Lysozyme and ferritin were used as model proteins. The effect was established with uncoated gold nanoparticles and with gold nanoparticles coated by 16-mercaptodecanoic acid. (© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Lysozyme as Pathogen-Recognition Protein in the Hemolymph of Galleria mellonellaENTOMOLOGICAL RESEARCH, Issue 3 2003In Hee LEE ABSTRACT Recognition of invading micro-organisms into hemolymph is a pivotal event for triggering diverse immune mechanisms in insects. It has been known that this recognition was mediated by the binding of hemolymph proteins to pattern-molecules on the cell surface of microbes. Recently, I found that the lysozyme in the G. mellonella hemolymph has binding affinity to cell-walls of Gram (-), (±) bacteria and fungus (Candida albicans). After the hemolymph was incubated with heat-killed microbes and treated with acidic buffer containing high concentration of NaCl, several plasma proteins detached from microbes were detected by reverse phase HPLC and SDS-PAGE analyses. Of binding proteins, it was assumed that the major one might be a lysozyme, which was previously characterized in the G. mellonella hemolymph. Furthermore immunoblot analysis performed with antiserum to G. mellonella lysozyme revealed that it was a lysozyme. [source] Time Controlled Protein Release from Layer-by-Layer Assembled Multilayer Functionalized Agarose HydrogelsADVANCED FUNCTIONAL MATERIALS, Issue 2 2010Sumit Mehrotra Abstract Axons of the adult central nervous system exhibit an extremely limited ability to regenerate after spinal cord injury. Experimentally generated patterns of axon growth are typically disorganized and randomly oriented. Support of linear axonal growth into spinal cord lesion sites has been demonstrated using arrays of uniaxial channels, templated with agarose hydrogel, and containing genetically engineered cells that secrete brain-derived neurotrophic factor (BDNF). However, immobilizing neurotrophic factors secreting cells within a scaffold is relatively cumbersome, and alternative strategies are needed to provide sustained release of BDNF from templated agarose scaffolds. Existing methods of loading the drug or protein into hydrogels cannot provide sustained release from templated agarose hydrogels. Alternatively, here it is shown that pH-responsive H-bonded poly(ethylene glycol)(PEG)/poly(acrylic acid)(PAA)/protein hybrid layer-by-layer (LbL) thin films, when prepared over agarose, provided sustained release of protein under physiological conditions for more than four weeks. Lysozyme, a protein similar in size and isoelectric point to BDNF, is released from the multilayers on the agarose and is biologically active during the earlier time points, with decreasing activity at later time points. This is the first demonstration of month-long sustained protein release from an agarose hydrogel, whereby the drug/protein is loaded separately from the agarose hydrogel fabrication process. [source] A precise boundary element method for macromolecular transport propertiesJOURNAL OF COMPUTATIONAL CHEMISTRY, Issue 9 2004Sergio Aragon Abstract A very precise boundary element numerical solution of the exact formulation of the hydrodynamic resistance problem with stick boundary conditions is presented. BEST, the Fortran 77 program developed for this purpose, computes the full transport tensors in the center of resistance or the center of diffusion for an arbitrarily shaped rigid body, including rotation-translation coupling. The input for this program is a triangulation of the solvent-defined surface of the molecule of interest, given by Connolly's MSROLL or other suitable triangulator. The triangulation is prepared for BEST by COALESCE, a program that allows user control over the quality and number of triangles to describe the surface. High numerical precision is assured by effectively exact integration of the Oseen tensor over triangular surface elements, and by scaling the hydrodynamic computation to the precise surface area of the molecule. Efficiency of computation is achieved by the use of public domain LAPACK routines that call BLAS Level 3 hardware-optimized subroutines available for most processors. A protein computation can be done in less than 10 min of CPU time in a modern Pentium IV processor. The present work includes a complete analysis of the sources of error in the numerical work and techniques to eliminate these errors. The operation of BEST is illustrated with applications to ellipsoids of revolution, and Lysozyme, a small protein. The typical numerical accuracy achieved is 0.05% compared to analytical theory. The numerical precision for a protein is better than 1%, much better than experimental errors in these quantities, and more than 10 times better than traditional bead-based methods. © 2004 Wiley Periodicals, Inc. J Comput Chem 9: 1191,1205, 2004 [source] Use of Lysozyme, Nisin, and EDTA Combined Treatments for Maintaining Quality of Packed Ostrich PattiesJOURNAL OF FOOD SCIENCE, Issue 3 2010Marianna Mastromatteo ABSTRACT:, The antimicrobial effectiveness of lysozyme, nisin, and ethylene diamine tetraacetic acid (EDTA) combination treatments (Mix1: 250 ppm lysozyme, 250 ppm nisin, 5 mM EDTA; Mix2: 500 ppm lysozyme, 500 ppm nisin, 5 mM EDTA) on bacterial growth of ostrich patties packaged in air, vacuum, and 2 different modified atmospheres (MAP1: 80% O2, 20% CO2; MAP2: 5% O2, 30% CO2, 65% N2) was evaluated. Moreover, the lipid oxidation was evaluated as well as color and sensory characteristics. The growth of total viable counts and lactic acid bacteria were strongly inhibited by the antimicrobial treatments in all the running time (Inhibition Index >97%) whereas for Enterobacteriaceae,and Pseudomonas,spp. lower inhibition indices from 12% to about 28% were observed. The lipid oxidation was more pronounced in the control respect to the treated meat patties. Moreover, the mixture at low concentration of lysozyme and nisin showed the best antioxidative effect. High concentrations of lysozyme and nisin showed the greatest color loss. Also, off-odors for the untreated patties developed faster than the treated samples. Practical Application: Great interest is developing in food bio-preservation, because of the ever-increasing needs to protect consumers' health and to valorize the naturalness and safety of food products. [source] Qualification and Quantification of Fish Protein in Prepared Surimi CrabstickJOURNAL OF FOOD SCIENCE, Issue 5 2008Z.H. Reed ABSTRACT:, Species identification and protein quantification in surimi crabstick were achieved using sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). When the Lowry and Kjeldahl protein determination methods were compared, the former showed more consistent results. Densitometric scanning of the gels was used for quantification of total fish protein as well as total egg white protein. The lower molecular weight proteins, 30 kDa and lower, proved to be the most useful in fish species identification as well as egg white protein addition. Using a combination of the myosin heavy chain band and the species-specific myosin light chain (Alaska pollock: 22.5 kDa; Pacific whiting: 24.4 kDa) proved the most accurate in calculating fish protein content of the crabstick sample, while for those samples that contained egg white, quantification was accomplished from the densitometric analysis of the overlapping bands of actin (45 kDa) from fish and ovalbumin from egg white. Lysozyme (14.3 kDa) proved to be a unique protein band in determining the presence of egg white when the content of dried egg white was equal to or exceeded 0.5% of the total weight of the final crabstick. [source] Characterization of Fish-Skin Gelatin Gels and Films Containing the Antimicrobial Enzyme LysozymeJOURNAL OF FOOD SCIENCE, Issue 5 2006C.K. Bower ABSTRACT:, Fish skins are rich in collagen and can be used to produce food-grade gelatin. Films cast from fish-skin gelatins are stable at room temperature and can act as a barrier when applied to foods. Lysozyme is a food-safe, antimicrobial enzyme that can also produce gels and films. When cold-water, fish-skin gelatin is enhanced with lysozyme, the resulting film has antimicrobial properties. The objective of this study was to characterize the effect on strength and barrier properties of lysozyme-enhanced fish-skin gelatin gels and films, and evaluate their activity against potential spoilage bacteria. Solutions containing 6.67% fish-skin gelatin were formulated to contain varying levels of hen-egg-white lysozyme. Gels were evaluated for strength, clarity, and viscoelastic properties. Films were evaluated for water activity, water vapor permeability, and antimicrobial barrier capabilities. Fish-skin gels containing 0.1% and 0.01% lysozyme had pH (4.8) and gelling-temperatures (2.1 °C) similar to lysozyme-free fish-skin gelatin controls. However, gel strength decreased (up to 20%). Turbidities of gels, with or without lysozyme, were comparable at all concentrations. Films cast with gelatin containing lysozyme demonstrated similar water vapor permeabilities and water activities. Lysozyme was still detectable in most fish gelatin films. More antimicrobial activity was retained in films cast with higher lysozyme concentrations and in films where lysozyme was added after the gelatin had been initially heated. These results suggest that fish-skin gelatin gels and films, when formulated with lysozyme, may provide a unique, functional barrier to increase the shelf life of food products. [source] Antimicrobial Effects of Lactoferrin, Lysozyme, and the Lactoperoxidase System and Edible Whey Protein Films Incorporating the Lactoperoxidase System Against Salmonella enterica and Escherichia coli O157:H7JOURNAL OF FOOD SCIENCE, Issue 7 2005Seacheol Min ABSTRACT: Lactoferrin (LF), lysozyme (LZ), the lactoperoxidase system (LPOS), and edible whey protein isolate (WPI) films incorporating LPOS were studied for inhibition of Salmonella enterica and Escherichia coli O157:H7. Antimicrobial effects of LF (5 to 40 mg/mL), LZ (1 to 20 mg/mL), and LPOS (0.5% to 5.0% [w/v] [0.03,.25 g/g, dry basis]) were examined by measuring turbidity of antimicrobial-containing media after inoculation and by examining cell inhibition by WPI films incorporating LPOS (LPOS-WPI films) on an agar recovery medium. Elastic modulus (EM), tensile strength (TS), percent elongation (%E), oxygen permeability (OP), and Hunter L, a and b of WPI films incorporating 0.03 to 0.25 g/g of LPOS were compared with those of plain WPI films without LPOS. The growth of S. enterica and E. coli O157:H7 (4 log colony-forming units [CFU]/mL) in tryptic soy broth (TSB) was not prevented by LF at ,20 and ,40 mg/mL, respectively. S. enterica and E. coli O157:H7 in TSB were not inhibited by LZ at , 6 and , 20 mg/mL, respectively. LPOS at concentrations of 2.75% (w/v) and 1.0% (w/v) reduced S. enterica and E. coli O157:H7 to below the limit of detection (1 CFU/mL) in TSB, respectively. LPOS-WPI films (0.15 g/g) completely inhibited S. enterica and E. coli O157:H7 (4 log CFU/cm2), inoculated either onto agar before placing the film disc or onto top of the film disc. Incorporation of 0.25 g/g of LPOS decreased EM, TS, and %E. The oxygen barrier property of WPI films was improved with the incorporation of LPOS at 0.15 to 0.25 g/g. [source] The defensive role of lysozyme in human gingiva in inflammatory periodontal diseaseJOURNAL OF PERIODONTAL RESEARCH, Issue 5 2009R. Younes Background and Objective:, The presence of lysozyme in human gingiva has not previously been demonstrated. In this study, we looked for evidence for the potential role of lysozyme as a protector of gingival elastic fibres. The objective of this study was also to determine the ex vivo susceptibility to hydrolysis of gingival elastic fibres from patients with or without periodontal disease by human leukocyte elastase and by human cathepsin G. Materials and Methods:, Using gingival tissue sections from eight control, 10 gingivitis and 10 periodontitis patients, we evaluated the area fraction occupied by gingival elastic fibres (after selective staining) by the use of automated image analysis. In the ex vivo experiments, serial tissue sections from four control, four gingivitis, four young periodontitis and four aged periodontitis patients were submitted to the action of human leukocyte elastase and cathepsin G, after which enzymatic activities were determined by image analysis. Indirect immunodetection of lysozyme was also done on tissue sections for all patients included in this study. Results:, Large variations of the area fraction occupied by elastic fibres were observed in human gingiva from young and aged patients with and without periodontal disease. In control and gingivitis patients, leukocyte elastase and cathepsin G had high comparable elastin solubilizing activities. With young and aged periodontitis patients, the two serine proteinases had weak elastin solubilizing activities. Lysozyme appeared to be present at the periphery of gingival elastic fibres in periodontitis patients. Conclusion:, Lysozyme can be considered an important natural protector of elastic fibres in pathological gingiva. [source] Effect of starch on the thermal kinetics and transmittance properties of lysozyme,JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 3 2005Abdellatif A Mohamed Abstract Lysozyme is being tested for use as a preservative in the food industry. The interaction between starch and lysozyme will help in recommending starch or starch fractions as carriers for lysozyme. The effect of starch fractions on the folding and unfolding of lysozyme was estimated by differential scanning calorimetry (DSC), kinetics and transmittance turbidometry. Lysozyme was unfolded (heated to 90 °C) and folded (cooled to 20 °C) five times in the presence of starch fractions. Starch was added at 1 and 2%. Overall, a trend of higher onset temperature (To) values occurred at 2% addition of all starch fractions except amylose. The increase in the number of cycles influenced the effect of starch on lysozyme denaturation. The percentage of lysozyme's ,H values decreased as a new heating and cooling cycle was performed (ie 74.4% of the ,H remained from the first cycle). The effect of amylose (AM) and amylopectin (AP) on the kinetics of lysozyme unfolding and folding was found to be different based on the assumption that the peak DSC temperature is the fastest step of the reaction. The unfolding showed higher activation energy (Ea) in the presence of both AM and AP, while the folding was not significantly changed. The turbidity of the solution containing lysozyme and potato starch showed transmittance in between that of lysozyme and starch. Stirring of the blend kept the transmittance unchanged while an increase in the transmittance was noticed when stirring ceased. Copyright © 2004 Society of Chemical Industry [source] Sustained and Extended Release with Structural and Activity Recovery of Lysozyme from Complexes with Sodium (Sulfamate Carboxylate) Isoprene/Ethylene Oxide Block CopolymerMACROMOLECULAR BIOSCIENCE, Issue 2 2010Gao Gao Abstract The complexation of lysozyme and sodium (sulfamate carboxylate) isoprene/ethylene oxide (SCIEO) at pH,=,7.4 and the release of lysozyme from the complexes in the presence of NaCl were investigated. Through electrostatic and hydrophobic interactions, lysozyme and SCIEO form stable complex nanoparticles. The complexation partially disturbs the structure of lysozyme. Some of the hydrophobic residues of lysozyme are exposed to bind with SCIEO. The complexation leads to loss of most of the lysozyme activity. In the presence of NaCl, lysozyme can be released from the complexes. The released lysozyme molecules recover their native structure and activity completely. In the condition of physiological pH and ionic strength, a sustained and extended release of lysozyme was achieved. [source] Flavin-sensitized Photo-oxidation of Lysozyme and Serum AlbuminPHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 4 2009Yazhou Zhang The excited state processes of riboflavin, flavin mononucleotide and flavin adenine dinucleotide in argon-saturated aqueous solution were studied in the presence of lysozyme or bovine serum albumin (BSA). UV,Vis absorption and fluorescence spectroscopy indicates that the noncovalent flavin-protein binding is relatively weak. Quenching of the flavin triplet state by BSA, observed by time-resolved photolysis, is less efficient than by lysozyme. Light-induced oxidation of the two proteins and reduction of the three flavins were studied. The quantum yields of the former and latter in the absence of oxygen are up to 0.1 and 0.04, respectively. The effects of pH and sensitizer and protein concentrations were examined in greater detail. The proposed reaction is electron transfer from the tryptophan moiety to the flavin triplet rather than excited singlet state. [source] Boron Mimetics: 1,2-Dihydro-1,2-azaborines Bind inside a Nonpolar Cavity of T4 Lysozyme,ANGEWANDTE CHEMIE, Issue 37 2009Lijun Liu Enzym ausgetrickst: Hochauflösende Proteinkristallographie zeigt, dass nichtnatürliche 1,2-Dihydro-1,2-azaborine in die unpolare Tasche des T4-Lysozyms L99A (siehe Bild) in einer Weise binden, die der Bindung ihrer ,natürlichen" All-Kohlenstoff-Isostere stark ähnelt. Die Studie belegt, dass 1,2-Azaborine in der biomedizinischen Forschung als Bor-haltige, hydrophobe Arenmimetika fungieren können. [source] Micronization of lysozyme by supercritical assisted atomizationBIOTECHNOLOGY & BIOENGINEERING, Issue 6 2009Renata Adami Abstract Supercritical Assisted Atomization (SAA) has been used to produce lysozyme microparticles. Lysozyme has been micronized using water, buffered water at pH 6.2 and water,ethanol mixtures at different volume percentages. Precipitated lysozyme particles were spherical, with a narrow particle size distribution (PSD) ranging from 0.1 to 4,µm. The concentration of lysozyme in the liquid solvent mixture had a nonlinear effect on the particle distribution, with an increase of the X0.9 from about 1 to 3,µm varying the enzyme concentration from 5 to 20,mg/mL. Precipitation temperature was set as low as possible to avoid enzyme degradation. High-performance liquid chromatography analysis showed no degradation of lysozyme and the enzyme activity, measured by turbidimetric enzymatic assay, only slightly decreased after SAA processing. Depending on the process conditions lysozyme retained from 95% to 100% of the biological activity compared to the untreated enzyme. Biotechnol. Bioeng. 2009; 104: 1162,1170. © 2009 Wiley Periodicals, Inc. [source] Elucidation of the mechanism and end products of glutaraldehyde crosslinking reaction by X-ray structure analysisBIOTECHNOLOGY & BIOENGINEERING, Issue 3 2007Yariv Wine Abstract Glutaraldehyde has been used for several decades as an effective crosslinking agent for many applications including sample fixation for microscopy, enzyme and cell immobilization, and stabilization of protein crystals. Despite of its common use as a crosslinking agent, the mechanism and chemistry involved in glutaraldehyde crosslinking reaction is not yet fully understood. Here we describe feasibility study and results obtained from a new approach to investigate the process of protein crystals stabilization by glutaraldehyde crosslinking. It involves exposure of a model protein crystal (Lysozyme) to glutaraldehyde in alkaline or acidic pH for different incubation periods and reaction arrest by medium exchange with crystallization medium to remove unbound glutaraldehyde. The crystals were subsequently incubated in diluted buffer affecting dissolution of un-crosslinked crystals. Samples from the resulting solution were subjected to protein composition analysis by gel electrophoresis and mass spectroscopy while crosslinked, dissolution resistant crystals were subjected to high resolution X-ray structural analysis. Data from gel electrophoresis indicated that the crosslinking process starts at specific preferable crosslinking site by lysozyme dimer formation, for both acidic and alkaline pH values. These dimer formations were followed by trimer and tetramer formations leading eventually to dissolution resistant crystals. The crosslinking initiation site and the end products obtained from glutaraldehyde crosslinking in both pH ranges resulted from reactions between lysine residues of neighboring protein molecules and the polymeric form of glutaraldehyde. Reaction rate was much faster at alkaline pH. Different reaction end products, indicating different reaction mechanisms, were identified for crosslinking taking place under alkaline or acidic conditions. Biotechnol. Bioeng. 2007;98:711,718. © 2007 Wiley Periodicals, Inc. [source] Molecularly Imprinted Polymer-Assisted Refolding of LysozymeBIOTECHNOLOGY PROGRESS, Issue 5 2007Mitsuru Haruki For production of active proteins using heterologous expression systems, refolding of proteins from inclusion bodies often creates a bottleneck due to its poor yield. In this study, we show that molecularly imprinted polymer (MIP) toward native lysozyme promotes the folding of chemically denatured lysozyme. The MIP, which was prepared with 1 M acrylamide, 1 M methacrylic acid, 1 M 2-(dimethylamino)ethyl methacrylate, and 5 mg/mL lysozyme, successfully promoted the refolding of lysozyme, whereas the non-imprinted polymer did not. The refolding yield of 90% was achieved when 15 mg of the MIP was added to 0.3 mg of the unfolded lysozyme. The parallel relationship between the refolding yield and the binding capacity of the MIP suggests that MIP promotes refolding through shifting the folding equilibrium toward the native form by binding the refolded protein. [source] A Novel Magnetic Affinity Support for Protein Adsorption and PurificationBIOTECHNOLOGY PROGRESS, Issue 1 2001Xiao-Dong Tong A novel magnetic support was prepared by an oxidization-precipitation method with poly(vinyl alcohol) (PVA) as the entrapment material. Transmission electron microscopy indicated that the magnetic particles had a core-shell structure, containing many nanometer-sized magnetic cores stabilized by the cross-linked PVA. The particles showed a high magnetic responsiveness in magnetic field, and no aggregation of the particles was observed after the particles had been treated in the magnetic field. These facts indicated that the particles were superparamagnetic. Cibacron blue 3GA (CB) was coupled to the particles to prepare a magnetic affinity support (MAS) for protein adsorption. Lysozyme was used as a model protein to test the adsorption properties of the MAS. The adsorption equilibrium of lysozyme to the MAS was described by the Langmuir-type isotherm. The capacity for lysozyme adsorption was more than 70 mg/g MAS (wet weight) at a relatively low CB coupling density (3,5 ,mol/g). In addition, 1.0 M NaCl solution could be used to dissociate the adsorbed lysozyme. Finally, the MAS was recycled for the purification of alcohol dehydrogenase (ADH) from clarified yeast homogenates. Under proper conditions, the magnetic separation yielded over 5-fold purification of the enzyme with 60% recovery of the enzyme activity. [source] [Ru(bpy)2(dcbpy)NHS] Labeling/Aptamer-Based Biosensor for the Detection of Lysozyme by Increasing Sensitivity with Gold Nanoparticle AmplificationCHEMISTRY - AN ASIAN JOURNAL, Issue 11 2008Jianguo Bai Abstract A novel [Ru(bpy)2(dcbpy)NHS] labeling/aptamer-based biosensor combined with gold nanoparticle amplification for the determination of lysozyme with an electrochemiluminescence (ECL) method is presented. In this work, an aptamer, an ECL probe, gold nanoparticle amplification, and competition assay are the main protocols employed in ECL detection. With all the protocols used, an original biosensor coupled with an aptamer and [Ru(bpy)2(dcbpy)NHS] has been prepared. Its high selectivity and sensitivity are the main advantages over other traditional [Ru(bpy)3]2+ biosensors. The electrochemical impedance spectroscopy (EIS) and atomic force microscopy (AFM) characterization illustrate that this biosensor is fabricated successfully. Finally, the biosensor was applied to a displacement assay in different concentrations of lysozyme solution, and an ultrasensitive ECL signal was obtained. The ECL intensity decreased proportionally to the lysozyme concentration over the range 1.0×10,13,1.0×10,8,mol,L,1 with a detection limit of 1.0×10,13,mol,L,1. This strategy for the aptasensor opens a rapid, selective, and sensitive route for the detection of lysozyme and potentially other proteins. [source] Synthesis of Novel Phosphorylated Daidzein Derivatives and ESI Investigation on Their Non-Covalent Complexes with LysozymeCHINESE JOURNAL OF CHEMISTRY, Issue 7 2007Xiao-Lan Chen Abstract Daidzein (7,4,-dihydroxyisoflavone) was phosphorylated by a modified Atherton-Todd reaction. The structures of the five target product, were determined by X-ray, IR, NMR and ESI-MS. Electrospray ionization results show that in the gas phase all the phosphorylated daidzein derivatives could form non-covalent complexes with the protein lysozyme, while non-covalent complexes were not detected in the mixed solution of daidzein with lysozyme. Relative affinity of every non-covalent complex was obtained according to its different decomposition orifice voltage. [source] Phagocyte activation in preterm infants following premature rupture of the membranes or chorioamnionitisACTA PAEDIATRICA, Issue 10 2000I Nupponen Phagocyte activation was studied in 48 preterm infants, gestational age 27.3 ± 0.3 wk, birthweight 968 ± 40 g, during the first postnatal week. Human neutrophil lipocalin as a marker of neutrophil activation was measured in plasma and tracheal aspirate fractions; and lysozyme, as a marker of monocyte and macrophage activation, in plasma. The concentration of plasma human neutrophil lipocalin was 69 (46,126) ,g/l (median and quartiles), tracheal aspirate fraction fluid 213 (71,433) (,g/l and plasma lysozyme 1337 (923,1764) ,g/l. Infants born to mothers with premature rupture of the membranes or clinical chorioamnionitis (group A, n 20) had significantly higher plasma [73 (58,151) vs 53 (38,108) ,g/l; p 0.027], and tracheal aspirate fraction human neutrophil lipocalin [319 (129,540) vs 190 (57,324) ,g/l; p= 0.019], and plasma lysozyme [1739 (1356,2021) vs 1140 (739,1557) ,g/l; p 0.0001] than did infants whose mothers had intact membranes and who had no suspicion of infection (Group B, n 28). In infants born to mothers receiving corticosteroids ante partum, correlations existed between time from treatment to delivery and plasma (r 0.322, p 0.0256) and tracheal aspirate fraction human neutrophil lipocalin (r= 0.314, p 0.0096). Infants born to mothers with at risk of infection are exposed to the potentially harmful effects of activated neutrophils. Premature rupture of the membranes, even without signs of clinical infection of the mother or the fetus, is associated with phagocyte activation that may begin already in utero. Corticosteroid treatment of the mother may cause transient inhibition of neutrophil activation in the newborn. [source] Protein separations using polyelectrolyte multilayer coatings with molecular micelles in open tubular capillary electrochromatographyELECTROPHORESIS, Issue 4 2008Candace A. Luces Abstract Novel polyelectrolyte multilayer (PEM) coatings for enhanced protein separations in open tubular CEC (OT-CEC) are reported. Use of four cationic polymers (poly- L -lysine, poly- L -ornithine, poly- L -lysine-serine, and poly- L -glutamic acid-lysine), and three anionic molecular micelles, sodium poly(N -undecanoyl- L -leucyl-alaninate) (poly- L -SULA), sodium poly(N -undecanoyl- L -leucyl-valinate) (poly- L -SULV), and sodium poly(undecylenic sulfate) (poly-SUS) were investigated in PEM coatings for protein separations. The simultaneous effects of cationic polymer concentration, number of bilayers, temperature, applied voltage, and pH of the BGE on the separation of four basic proteins (,-chymotrypsinogen A, lysozyme, ribonuclease A, and cytochrome c) were analyzed using a Box Behnken experimental design. The influence of NaCl on the run-to-run reproducibility was investigated for PEM coatings containing each cationic polymer. All coatings exhibited excellent reproducibilities with a %RSD of the EOF less than 1% in the presence of NaCl. Optimal conditions were dependent on both the cationic and anionic polymers used in the PEM coatings. Poly- L -glutamic acid-lysine produced the highest resolution and longest migration time. The use of molecular micelles to form PEM coatings resulted in better separations than single cationic coatings. Chiral poly- L -SULA and poly- L -SULV resulted in higher protein resolutions as compared to the achiral, poly-SUS. Furthermore, the use of poly- L -SULV reversed the elution order of lysozyme and cytochrome c when compared to poly- L -SULA and poly-SUS. [source] Atmospheric molding of ionic copolymer MALDI-TOF/MS arrays: A new tool for protein identification/profilingELECTROPHORESIS, Issue 24 2006Alexander Muck Abstract An atmospheric molding protocol has been used to prepare an ionic methacrylate-based copolymer sample support chips for MALDI (pMALDI)-MS by targeting selected groups of various monomers copolymerized during molding, namely, carboxy, sulfo, dimethylalkyamino, and trimethylalkylammonium groups. The new disposable array chips provide analyte-oriented enhancement of protein adsorption to the modified substrates without requiring complicated surface coating or derivatization. The MALDI-MS performance of the new ionic copolymer chips was evaluated for lysozyme, ,-lactoglobulin,A, trypsinogen and carbonic anhydrase,I using washing with solutions prepared in pH or ionic strength steps. On cationic chips, the proteins are washed out at pH lower than their pI values, and on anionic chips at pH higher than their pI values. The ability of the microfabricated pMALDI chip set to selectively adsorb different proteins from real samples and to significantly increase their MS-signal was documented for the transmembrane photosystem,I protein complex from the green alga Chlamydomonas reinhardtii. The proteins were almost exclusively adsorbed according to calculated pI values and grand average of hydropathy (GRAVY) indexes. The new disposable chips reduce manipulation times and increase measurement sensitivity for real-world proteomic samples. The simple atmospheric molding procedure enables additional proteomic operations to be incorporated on disposable MALDI-MS integrated platforms. [source] On-line concentration of proteins in pressurized capillary electrochromatography coupled with electrospray ionization-mass spectrometryELECTROPHORESIS, Issue 7-8 2005Zhen Liang Abstract Pressurized capillary electrochromatography (pCEC) and electrospray ionization-mass spectrometry (ESI-MS) have been hyphenated for protein analysis. Taken cytochrome,c, lysozyme, and insulin as samples, the limits of detection (LODs) for absolute concentrations are 10,11,mol (signal-to-noise ratio S/N = 3) with relative standard deviations (RSDs) of retention time and peak area, respectively, of less than 1.7% and 4.8%. In order to improve the detection sensitivity, on-line concentration by field-enhanced sample-stacking effect and chromatographic zone-sharpening effect has been developed, and parameters affecting separation and detection, such as pH and electrolyte concentration in the mobile phase, separation voltage, as well as enrichment voltage and time, have been studied systematically. Under the optimized conditions, the LODs of the three proteins could be decreased up to 100-fold. In addition, the feasibility of such techniques has been further demonstrated by the analysis of modified insulins at a concentration of 20,,g/mL. [source] Application of dodecyldimethyl (2-hydroxy-3-sulfopropyl) ammonium in wall modification for capillary electrophoresis separation of proteinsELECTROPHORESIS, Issue 3 2005Wei Wei Abstract A zwitterionic surfactant, dodecyldimethyl (2-hydroxy-3-sulfopropyl) ammonium (C12H25N+(CH3)2CH2CHOHCH2SO3,), named dodecyl sulfobetaine (DSB), was used as a novel modifier to coat dynamically capillary walls for capillary electrophoresis separation of basic proteins. The DSB coating suppressed the electroosmotic flow (EOF) in the pH range of 3,12. At high DSB concentration, the EOF was suppressed by more than 8.8,times. The DSB coating also prevented successfully the adsorption of cationic proteins on the capillary wall. Anions, such as Cl,, Br,, I,, SO42,, CO32,, and ClO4,, could be used as running buffer modifiers to adjust the EOF for better separation of analytes. Using this dynamically coated capillary, a mixture of eight inorganic anions achieved complete separation within 4.2,min with the efficiencies from 24,000 to 1,310,000,plates/m. In the presence of ClO4, as EOF adjustor, the separation of a mixture containing four basic proteins (lysozyme, cytochrome c, ,-chymotrypsinogen,A, and myoglobin) yielded efficiencies of 204,000,896,000,plates/m and recoveries of 88%,98%. Migration time reproducibility of these proteins was less than 0.5% relative standard deviation (RSD) from run to run and less than 3.1% RSD from day to day, showing promising application of this novel modifier in protein separation. [source] Multilayer poly(vinyl alcohol)-adsorbed coating on poly(dimethylsiloxane) microfluidic chips for biopolymer separationELECTROPHORESIS, Issue 1 2005Dapeng Wu Abstract A poly(dimethylsiloxane) (PDMS) microfluidic chip surface was modified by multilayer-adsorbed and heat-immobilized poly(vinyl alcohol) (PVA) after oxygen plasma treatment. The reflection absorption infrared spectrum (RAIRS) showed that 88% hydrolyzed PVA adsorbed more strongly than 100% hydrolyzed one on the oxygen plasma-pretreated PDMS surface, and they all had little adsorption on original PDMS surface. Repeating the coating procedure three times was found to produce the most robust and effective coating. PVA coating converted the original PDMS surface from a hydrophobic one into a hydrophilic surface, and suppressed electroosmotic flow (EOF) in the range of pH 3,11. More than 1 000,000 plates/m and baseline resolution were obtained for separation of fluorescently labeled basic proteins (lysozyme, ribonuclease B). Fluorescently labeled acidic proteins (bovine serum albumin, ,-lactoglobulin) and fragments of dsDNA ,X174 RF/HaeIII were also separated satisfactorily in the three-layer 88% PVA-coated PDMS microchip. Good separation of basic proteins was obtained for about 70 consecutive runs. [source] Analysis of the sinusitis nasal lavage fluid proteome using capillary liquid chromatography interfaced to electrospray ionization-quadrupole time of flight- tandem mass spectrometryELECTROPHORESIS, Issue 9 2004Begona Casado Abstract The nasal lavage fluids (NLFs) from four subjects with acute sinusitis were analyzed to investigate the amount of proteins expressed in this pathology at the beginning of the event (day 1) and after 6 days of treatment with antibiotics and a nasal steroid spray. The protein identification was performed with capillary liquid chromatography-electrospray-quadrupole time of flight-(LC-ESI-Q-TOF)-mass spectrometry. The samples collected on the first day contained high-abundant plasma proteins, such as albumin and immunoglobulins, glandular serous cell proteins (lysozyme, lactoferrin, and polymeric immunoglobulin receptor), epithelial keratins, and inflammatory cell proteins (myeloperoxidase, IL-16, and IL-17E). After six days of therapy, the complexity of the proteome was reduced to plasma proteins and lysozyme with no inflammatory markers. The presence of hemoglobin, however, suggested that significant squamous metaplasia with breaches in the epithelial barrier, or nasal steroid-related bleeding, had occurred. The proteomic approach presented here allowed us to identify, in the high complexity of acute sinusitis nasal secretions, the proteins that respond to a pharmacological treatment and that could be suitable as markers of this pathology. [source] Purification and cDNA Cloning of Lysozyme II from Cabbage Butterfly, Artogeia rapae LarvaeENTOMOLOGICAL RESEARCH, Issue 4 2005BANG In Seok ABSTRACT Last instar larvae of cabbage butterfly Artogeia rapae respond to injection of bacteria with a set of inducible antibacterial peptides/proteins. The inducible peptides/proteins are related to the known hinnavins (I and II) and lysozymes (I and II). The lysozyme II has been isolated by heat treatment, cation exchange, and reversed-phase chromatography from immunized hemolymph of last instar larvae. The lysozyme II gene of A. rapae was isolated and its nucleotide sequence was determined by the RACE-PCR from immunized fat body with E. coli. It has an open reading frame of 414 bp nucleotide corresponding to 138 amino acids including an 18 amino acid signal sequence. The molecular weight and the isoelectric point of Artogeia lysozyme II without a signal peptide were 13,649.38 Da and 9.11, respectively. It is great similarity with Manduca lysozyme among other lepidopteran. [source] Lysozyme as Pathogen-Recognition Protein in the Hemolymph of Galleria mellonellaENTOMOLOGICAL RESEARCH, Issue 3 2003In Hee LEE ABSTRACT Recognition of invading micro-organisms into hemolymph is a pivotal event for triggering diverse immune mechanisms in insects. It has been known that this recognition was mediated by the binding of hemolymph proteins to pattern-molecules on the cell surface of microbes. Recently, I found that the lysozyme in the G. mellonella hemolymph has binding affinity to cell-walls of Gram (-), (±) bacteria and fungus (Candida albicans). After the hemolymph was incubated with heat-killed microbes and treated with acidic buffer containing high concentration of NaCl, several plasma proteins detached from microbes were detected by reverse phase HPLC and SDS-PAGE analyses. Of binding proteins, it was assumed that the major one might be a lysozyme, which was previously characterized in the G. mellonella hemolymph. Furthermore immunoblot analysis performed with antiserum to G. mellonella lysozyme revealed that it was a lysozyme. [source] Breakpoints in immunoregulation required for Th1 cells to induce diabetesEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 9 2006Margaret Neighbors Abstract We describe a novel TCR-transgenic mouse line, TCR7, where MHC class,II-restricted, CD4+ T cells are specific for the subdominant H-2b epitope (HEL74,88) of hen egg lysozyme (HEL), and displayed an increased frequency in the thymus and in peripheral lymphoid compartments over that seen in non-transgenic littermate controls. CD4+ T cells responded vigorously to HEL or HEL74,88 epitope presented on APC and could develop into Th1 or Th2 cells under appropriate conditions. Adoptive transfer of TCR7 Ly5.1 T cells into Ly5.2 rat insulin promoter (RIP)-HEL transgenic recipient hosts did not lead to expansion of these cells or result in islet infiltration, although these TCR7 cells could expand upon transfer into mice expressing high levels of HEL in the serum. Islet cell infiltration only occurred when the TCR7 cells had been polarized to either a Th1 or Th2 phenotype prior to transfer, which led to insulitis. Progression from insulitis to autoimmune diabetes only occurred in these recipients when Th1 but not Th2 TCR7 cells were transferred and CTLA-4 signaling was simultaneously blocked. These findings show that regulatory pathways such as CTLA-4 can hold in check already differentiated autoreactive effector Th1 cells, to inhibit the transition from tolerance to autoimmune diabetes. See accompanying commentary at http://dx.doi.org/10.1002/eji.200636591 [source] Enzymes in the acquired enamel pellicleEUROPEAN JOURNAL OF ORAL SCIENCES, Issue 1 2005Christian Hannig The acquired pellicle is a biofilm, free of bacteria, covering oral hard and soft tissues. It is composed of mucins, glycoproteins and proteins, among which are several enzymes. This review summarizes the present state of research on enzymes and their functions in the dental pellicle. Theoretically, all enzymes present in the oral cavity could be incorporated into the pellicle, but apparently enzymes are adsorbed selectively onto dental surfaces. There is clear evidence that enzymes are structural elements of the pellicle. Thereby they exhibit antibacterial properties but also facilitate bacterial colonization of dental hard tissues. Moreover, the immobilized enzymes are involved in modification and in homeostasis of the salivary pellicle. It has been demonstrated that amylase, lysozyme, carbonic anhydrases, glucosyltransferases and fructosyltransferase are immobilized in an active conformation in the pellicle layer formed in vivo. Other enzymes, such as peroxidase or transglutaminase, have been investigated in experimental pellicles. Despite the depicted impact of enzymes on the formation and function of pellicle, broader knowledge on their properties in the in vivo -formed pellicle is required. This might be beneficial in the development of new preventive and diagnostic strategies. [source] | |||||||||||||||||||||||||||||||||||||||||||