Laser ablation of the protein lysozyme

DEFF Research Database (Denmark)

Schou, Jørgen; Canulescu, Stela; Amoruso, Salvatore

produced thin films of average thickness up to 300 nm, which not only contained a significant amount of intact molecules, but also maintained the bioactivity. These films were produced by a nanosecond laser in the UV regime at 355 nm with 2 J/cm2. The surprising fact that these molecules can be transferred......Lysozyme is a well-known protein, which is used in food processing because of its bactericidal properties. The mass (14307 amu) is in the range in which it easily can be monitored by mass spectrometric methods, for example by MALDI (Matrix assisted laser desorption ionization). We have recently...... to a substrate as intact molecules by the violent laser impact ( up to 50 mJ/pulse) has not yet been understood. One issue is that up to 150 ng/pulse is removed by the laser, and much of the material is ejected from the target in relatively large chunks. We have explored as well the excitation mechanics by laser...

Lysozymes in the animal kingdom.

Science.gov (United States)

Callewaert, Lien; Michiels, Chris W

2010-03-01

Lysozymes (EC 3.2.1.17) are hydrolytic enzymes, characterized by their ability to cleave the beta-(1,4)-glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine in peptidoglycan, the major bacterial cell wall polymer. In the animal kingdom, three major distinct lysozyme types have been identified--the c-type (chicken or conventional type), the g-type (goose-type) and the i-type (invertebrate type) lysozyme. Examination of the phylogenetic distribution of these lysozymes reveals that c-type lysozymes are predominantly present in the phylum of the Chordata and in different classes of the Arthropoda. Moreover, g-type lysozymes (or at least their corresponding genes) are found in members of the Chordata, as well as in some bivalve mollusks belonging to the invertebrates. In general, the latter animals are known to produce i-type lysozymes. Although the homology in primary structure for representatives of these three lysozyme types is limited, their three-dimensional structures show striking similarities. Nevertheless, some variation exists in their catalytic mechanisms and the genomic organization of their genes. Regarding their biological role, the widely recognized function of lysozymes is their contribution to antibacterial defence but, additionally, some lysozymes (belonging to different types) are known to function as digestive enzymes.

Chitosan and Cystatin/Lysozyme Preparation as Protective Edible Films Components

Directory of Open Access Journals (Sweden)

Anna Zimoch-Korzycka

2015-01-01

Full Text Available This work characterizes biological, physical, and chemical properties of films formed from an aqueous solution of hydroxypropyl methylcellulose (HPMC, with different concentrations of chitosan (CH and bioactive cystatin/lysozyme preparation (C/L. The properties of biocomposites were examined by Dynamic Mechanical Analysis (DMA, Fourier’s transfer infrared spectroscopy (FTIR, water vapour permeability (WVP, and tensile testing. Antimicrobial activity against Micrococcus flavus, Bacillus cereus, Escherichia coli, Pseudomonas fluorescens, and Candida famata was conducted. Films glass transition and storage modulus were dependent on the C/L and CH concentration. Modulus values decreased during the temperature scan and with higher reagents levels. An increase of CH and C/L concentrations in the films resulted in a decrease in tensile strength from 2.62 to 1.08 MPa. It suggests the hydrolyzing influence of C/L, also observed in smaller peak size of α relaxation. C/L addition caused shifting Tg to higher temperature. DMA and FTIR analysis proved that HPMC and CH are compatible polymers. Water resistance was improved with rising CH concentration from 1.08E-09 to 7.71E−10 g/m∗s∗Pa. The highest inhibition zone in M. flavus and C. famata was recorded at the highest concentration of CH and C/L.

Laser ablation of the lysozyme protein: a model system for soft materials

DEFF Research Database (Denmark)

Schou, Jørgen; Matei, Andreea; Constantinescu, Catalin

.3 1015 molecules per pulse. This is perhaps one of the highest ablation yields ever measured. Films with a significant number of intact lysozyme molecules have been produced by PLD (pulsed laser deposition) and MAPLE (Matrix-assisted pulsed laser evaporation). The deposition of intact molecules...... is expected in MAPLE, but is surprising in PLD, where a high degree of thermal fragmentation is typically required for generation of a sufficient amount of volatile decomposition products that drive the transfer of molecules to the film substrate. The experimental results will be discussed based...

Pulsed laser deposition of the lysozyme protein: an unexpected “Inverse MAPLE” process

DEFF Research Database (Denmark)

Schou, Jørgen; Matei, Andreea; Constantinescu, Catalin

2012-01-01

Films of organic materials are commonly deposited by laser assisted methods, such as MAPLE (matrix-assisted pulsed laser evaporation), where a few percent of the film material in the target is protected by a light-absorbing volatile matrix. Another possibility is to irradiate the dry organic...... the ejection and deposition of lysozyme. This can be called an “inverse MAPLE” process, since the ratio of “matrix” to film material in the target is 10:90, which is inverse of the typical MAPLE process where the film material is dissolved in the matrix down to several wt.%. Lysozyme is a well-known protein...

Evaluating the antimicrobial activity of Nisin, Lysozyme and Ethylenediaminetetraacetate incorporated in starch based active food packaging film.

Science.gov (United States)

Bhatia, Sugandha; Bharti, Anoop

2015-06-01

The pleothera of micro organisms obtained from contaminated food cultured in a starch broth was effectively tested against antibacterial agents, i.e. nisin, lysozyme and chelating agent EDTA. A variety of combination treatments of these antimicrobial agents and their incorporation in Starch based active packaging film according to their permissibility standards was done. 4 variables of Nisin concentration (ranging from 0 to 750 IU/ml), 3 variables of lysozyme concentration (ranging from 0 to 500 IU/ml) and 3 variables of EDTA concentration from (0 to 20 μM) were chosen. Bacterial inhibition by combination of different levels of different factors without antimicrobial films was evaluated using a liquid incubation method. The samples were assayed for turbidity at interval of 2, 4 and 24 h to check effectiveness of combined effects of antimicrobial agents which proved a transitory bactericidal effect for short incubation times. Zone of Inhibition was observed in the antimicrobial films prepared by agar diffusion method. Statistical analysis of experimental data for their antimicrobial spectrum was carried out by multi regression analysis and ANOVA using Design-Expert software to plot the final equation in terms of coded factors as antimicrobial agents. The experimental data indicated that the model was highly significant. Results were also evaluated graphically using response surface showing interactions between two factors, keeping other factor fixed at values at the center of domain. Synergy was also determined among antibacterial agents using the fractional inhibitory concentration (FIC) index which was observed to be 0.56 supporting the hypothesis that nisin and EDTA function as partial synergistically. The presented work aimed to screen in quick fashion the combinatorial effect of three antimicrobial agents and evaluating their efficacy in anti microbial film development.

Genetic control of the humoral immune response to avian egg white lysozymes in the chicken

International Nuclear Information System (INIS)

Flanagan, M.P.

1987-01-01

Chickens from two closely related sublines, GHs-B6 and GHs-B13, differing serologically at the major histocompatibility complex, were significantly different in their humoral response to three avian egg white lysozymes. Specific antisera levels were measured by radioimmunoassay using 125 I-labeled lysozymes. Antibodies elicited in response to these lysozymes are assumed to be directed against sites on these lysozymes where their amino acid sequence differs from that of the recipient G. domesticus egg white lysozyme (HEL). GHs-B6 birds produced a high level of antibody in response to immunization of turkey (TEL), pheasant (PhL) and guinea hen (GHL) lysozymes. GHs-B13 birds produced no detectable antibody to TEL, were intermediate in their response to PhL and equaled the antibody production of GHs-B6 birds in response to GHL. Antisera to each lysozyme were examined for crossreactivity with all other lysozymes by use of a competitive binding assay

[Lysozyme in the treatment of juvenile laryngeal papillomatosis. A new concept in its etiopathogenesis].

Science.gov (United States)

Altamar-Ríos, J

1990-01-01

The A. inform about the results achieved with lysozyme chlorhydrate in the treatment of 15 patients with juvenile laryngeal papillomatosis. The lysozyme is an electropositive enzyme which synthesis is related to the degree of proteins and vitamin B complex ingestion. Lysozyme is a component of the immunitary inespecific system, serving to prevent against HPV-DNA at the level of the secretory film of the mucociliary apparatus of the respiratory mucous membrane. Furthermore, lysozyme hydrolyzes the mucopolysaccharide of the connective tissue and inhibits the virus-DNA replication. 100-300 mgr daily during 30-60 days simultaneously with hyperproteic diet and vitamin B complex (after correction of the nutrimental deficiencies) brought about the evanishment of papillomatosis. The A. suggest that the predisposition to infection by virus DNA is primarily of immunitary origin, because of lysozyme deficiency, and secondary due to a low intake of proteins and vitamin B complex.

Pulsed laser deposition of lysozyme: the dependence on shot numbers and the angular distribution

DEFF Research Database (Denmark)

Constantinescu, C.; Matei, A.; Schou, Jørgen

2013-01-01

The ejection of molecules from a pressed solid target of lysozyme induced by laser ablation in the UV-regime at a wavelength of 355 nm was investigated. The ablation studies were carried out in vacuum at a laser fluence of 2 J/cm2 for which a significant fraction of proteins remains intact....... This was verified by matrix-assisted laser desorption ionization (MALDI) spectrometry of thin films deposited on silicon substrates. The deposition rate of lysozyme was found to decrease with the number of shots and was correlated with increasing thermal damage of the lysozyme. This was monitored by measurements...... of the optical reflectivity of dry lysozyme. The angular distribution of the mass deposition can be fitted well by Anisimov’s hydrodynamic model. The total deposited yield over the entire hemisphere from direct laser ablation of lysozyme was estimated from this model and found to be three orders of magnitude...

Electrophoretic Approach for the Simultaneous Deposition and Functionalization of Reduced Graphene Oxide Nanosheets with Diazonium Compounds: Application for Lysozyme Sensing in Serum.

Science.gov (United States)

Wang, Qian; Vasilescu, Alina; Wang, Qi; Coffinier, Yannick; Li, Musen; Boukherroub, Rabah; Szunerits, Sabine

2017-04-12

Electrophoretic deposition (EPD) of reduced graphene oxide nanosheets (rGO) offers several advantages over other surface coating approaches, including process simplicity, uniformity of the deposited films, and good control of the film thickness. The EPD conditions might also be of interest for the reduction of diazonium salts, which upon the release of N 2 molecules and generation of radicals, can form covalent bonds with the sp 2 hybridized carbon lattice atoms of rGO films. In this work, we report on the coating of gold electrodes in one step with rGO/polyethylenimine (PEI) thin films and their simultaneous modification using different phenyl (Ph) diazonium salt precursors bearing various functionalities such as -B(OH) 2 , -COOH, and -C≡CH. We show further the interest of such interfaces for designing highly sensitive sensing platforms. Azide-terminated lysozyme aptamers were clicked onto the rGO/PEI/Ph-alkynyl matrix and used for the sensing of lysozyme levels in patients suffering from inflammatory bowel disease (IBD), where lysozyme levels are up-regulated. The approach attained the required demand for the determination of lysozyme level in patients suffering from IBD with a 200 fM detection limit and a linear range up to 20 pM without signal amplification.

Modified lysozymes as novel broad spectrum natural antimicrobial agents in foods.

Science.gov (United States)

Aminlari, Ladan; Hashemi, Marjan Mohammadi; Aminlari, Mahmoud

2014-06-01

In recent years much attention and interest have been directed toward application of natural antimicrobial agents in foods. Some naturally occurring proteins such as lactoperoxidase, lactoferrin, and lysozyme have received considerable attention and are being considered as potential antimicrobial agents in foods. Lysozyme kills bacteria by hydrolyzing the peptidoglycan layer of the cell wall of certain bacterial species, hence its application as a natural antimicrobial agent has been suggested. However, limitations in the action of lysozyme against only Gram-positive bacteria have prompted scientists to extend the antimicrobial effects of lysozyme by several types of chemical modifications. During the last 2 decades extensive research has been directed toward modification of lysozyme in order to improve its antimicrobial properties. This review will report on the latest information available on lysozyme modifications and examine the applicability of the modified lysozymes in controlling growth of Gram-positive and Gram-negative bacteria in foods. The results of modifications of lysozyme using its conjugation with different small molecule, polysaccharides, as well as modifications using proteolytic enzymes will be reviewed. These types of modifications have not only increased the functional properties of lysozyme (such as solubility and heat stability) but also extended the antimicrobial activity of lysozyme. Many examples will be given to show that modification can decrease the count of Gram-negative bacteria in bacterial culture and in foods by as much as 5 log CFU/mL and in some cases essentially eliminated Escherichia coli. In conclusion this review demonstrates that modified lysozymes are excellent natural food preservatives, which can be used in food industry. The subject described in this review article can lead to the development of methods to produce new broad-spectrum natural antimicrobial agents, based on modification of chicken egg white lysozyme, which

Solvent freeze out crystallization of lysozyme from a lysozyme-ovalbumin mixture

Energy Technology Data Exchange (ETDEWEB)

Diaz Borbon, V.; Ulrich, J. [Martin-Luther-Universitaet Halle-Wittenberg, Zentrum fuer Ingenieurwissenschaft, Verfahrenstechnik/TVT, 06099 Halle Saale (Germany)

2012-05-15

Hen egg white lysozyme (HEWL) crystallization conditions from an ovalbumin-lysozyme mixture were found by screening tests and further located in pseudo-phase diagrams. This information was used to set up the initial conditions for the solvent freeze out (SFO) process. The process uses the freezing of ice to create the supersaturation for the proteins to crystallize out of the solution. The crystallization of HEWL (15 mg/mL) out of a lysozyme-ovalbumin mixture (1.7 mg/mL) is carried out by SFO. Under the reported conditions, a crystallization yield of 69 % was obtained. A mean crystal size of 77.8 {mu}m was enhanced in a crystallization time of 15.1 h. The lysozyme nature of the crystals is proven by SDS PAGE and enzymatic activity tests. (copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

Antimicrobial lysozyme-containing starch microgel to target and inhibit amylase-producing microorganisms

NARCIS (Netherlands)

Li, Y.; Kadam, S.; Abee, T.; Slaghek, T.M.; Timmermans, J.W.; Cohen Stuart, M.A.; Norde, W.; Kleijn, M.J.

2012-01-01

The aim of this study is to determine the release of lysozyme from oxidized starch microgels and subsequently test its antimicrobial activity. The gels are made of oxidized potato starch polymers, which are chemically cross-linked by sodium trimetaphosphate (STMP). The microgel is negatively charged

Zero-order release of lysozyme from (poly)ethylene glycol)/poly(butylene terephthalate) matrices

NARCIS (Netherlands)

Bezemer, J.M.; Radersma, R.; Grijpma, Dirk W.; Dijkstra, Pieter J.; Feijen, Jan; van Blitterswijk, Clemens

2000-01-01

Protein release from a series of biodegradable poly(ether ester) multiblock copolymers, based on poly(ethylene glycol) (PEG) and poly(butylene terephthalate) (PBT) was investigated. Lysozyme-containing PEG/PBT films and microspheres were prepared using an emulsion technique. Proteins were

Electrostatic interactions in protein adsorption probed by comparing lysozyme and succinylated lysozyme

NARCIS (Netherlands)

Veen, van der M.; Norde, W.; Cohen Stuart, M.A.

2004-01-01

The influence of electrostatic interactions on protein adsorption was studied by comparing the adsorption of lysozyme and succinylated lysozyme at silica surfaces. The succinylation affects the charge of the protein, but also the stability. Although changes in stability can have an influence on

Coassembly of Lysozyme and Amphiphilic Biomolecules Driven by Unimer-Aggregate Equilibrium.

Science.gov (United States)

Tao, Yuanyuan; Ma, Xiaoteng; Cai, Yaqian; Liu, Li; Zhao, Hanying

2018-04-12

Synthesis and self-assembly of bioconjugates composed of proteins and synthetic molecules have been widely studied because of the potential applications in medicine, biotechnology, and nanotechnology. One of the challenging research studies in this area is to develop organic solvent-free approaches to the synthesis and self-assembly of amphiphilic bioconjugates. In this research, dialysis-assisted approach, a method based on unimer-aggregate equilibrium, was applied in the coassembly of lysozyme and conjugate of cholesterol and glutathione (Ch-GSH). In phosphate buffer solution, amphiphilic Ch-GSH conjugate self-assembles into vesicles, and the vesicle solution is dialyzed against lysozyme solution. Negatively charged Ch-GSH unimers produced in the unimer-vesicle exchange equilibrium, diffuse across the dialysis membrane and have electrostatic interaction with positively charged lysozyme, resulting in the formation of Ch-GSH-lysozyme bioconjugate. Above a critical concentration, the three-component bioconjugate molecules self-assemble into bioactive vesicles.

Serum lysozyme determinations, April-June 1961

Energy Technology Data Exchange (ETDEWEB)

Finch, S C; Lamphere, J P; Jablon, S

1963-04-18

Serum lysozyme levels were determined on 670 consecutive subjects seen for regularly scheduled clinic examinations of the Adult Health Study in Hiroshima. Serum lysozyme levels were found to vary significantly with the absolute peripheral granulocyte count, age, sex, and month of study. A high level of correlation also was noted between serum lysozyme and diabetes mellitus. This was at least in part attributable to greater average age in patients with diabetes. Suggestive relationship was established between serum lysozyme levels, respiratory diseases, and tuberculosis. These changes are believed to reflect active inflammation with excessive destruction of granulocytes and parenchymal tissues in those patients with the more acute processes. No relationship was found between serum lysozyme and previous exposure to ionizing radiation. These studies indicate that the serum lysozyme level may be useful in the study of the kinetics of leukopoiesis, the aging process, and in the detection of subtle inflammatory processes. 21 references, 5 tables.

1H NMR studies of human lysozyme: Spectral assignment and comparison with hen lysozyme

International Nuclear Information System (INIS)

Redfield, C.; Dobson, C.M.

1990-01-01

Complete main-chain (NH and αCH) 1 H NMR assignments are reported for the 130 residues of human lysozyme, along with extensive assignments for side-chain protons. Analysis of 2-D NOESY experiments shows that the regions of secondary structure for human lysozyme in solution are essentially identical with those found previously in a similar study of hen lysozyme and are in close accord with the structure of the protein reported previously from x-ray diffraction studies in the crystalline state. Comparison of the chemical shifts, spin-spin coupling constants, and hydrogen exchange behavior are also consistent with closely similar structures for the two proteins in solution. In a number of cases specific differences in the NMR parameters between hen and human lysozymes can be correlated with specific differences observed in the crystal structures

Complex coacervates of hyaluronic acid and lysozyme

DEFF Research Database (Denmark)

Water, Jorrit J.; Schack, Malthe M.; Velazquez-Campoy, Adrian

2014-01-01

stoichiometry was determined using solution depletion and isothermal titration calorimetry. The binding stoichiometry of lysozyme to hyaluronic acid (870 kDa) determined by solution depletion was found to be 225.9 ± 6.6 mol, or 0.1 bound lysozyme molecules per hyaluronic acid monomer. This corresponded well...... with that obtained by isothermal titration calorimetry of 0.09 bound lysozyme molecules per hyaluronic acid monomer. The complexation did not alter the secondary structure of lysozyme measured by Fourier-transform infrared spectroscopy overlap analysis and had no significant impact on the Tm of lysozyme determined...

Lysozymes in the animal kingdom

Indian Academy of Sciences (India)

In the animal kingdom, three major distinct lysozyme types have been ... reveals that c-type lysozymes are predominantly present in the phylum of the Chordata and in different classes of the Arthropoda. ... Journal of Biosciences | News ...

Study of a photo-induced lysozyme-riboflavin bond

Energy Technology Data Exchange (ETDEWEB)

Ferrer, I; Silva, E

1985-01-01

Irradiation of lysozyme in the presence of riboflavin results in the formation of a lysozyme-riboflavin adduct. Reduction and carboxymethylation of the four disulfide bonds as well as the chemical modification of the Tyr residues and the photochemical alteration of the His residue in lysozyme, do not affect the formation of the photo-induced lysozyme-riboflavin bond. When the lysozyme-riboflavin adduct was subjected to mild acid hydrolysis and ion exchange chromatography, the retention of a compound containing /sup 14/C-riboflavin was observed. Free /sup 14/C-riboflavin, on the contrary is not retained by the column. The photo-oxidation of free Trp in the presence of /sup 14/C-riboflavin, gave a compound which bound to the ion exchange resin like the above-mentioned derivative. The photo-oxidation of the Trp residues in lysozyme and in peptides obtained from lysozyme showed very high quantum yields, and these values were directly related to the incorporation of /sup 14/C-riboflavin in these samples.

Study of a photo-induced lysozyme-riboflavin bond

International Nuclear Information System (INIS)

Ferrer, I.; Silva, E.

1985-01-01

Irradiation of lysozyme in the presence of riboflavin results in the formation of a lysozyme-riboflavin adduct. Reduction and carboxymethylation of the four disulfide bonds as well as the chemical modification of the Tyr residues and the photochemical alteration of the His residue in lysozyme, do not affect the formation of the photo-induced lysozyme-riboflavin bond. When the lysozyme-riboflavin adduct was subjected to mild acid hydrolysis and ion exchange chromatography, the retention of a compound containing 14 C-riboflavin was observed. Free 14 C-ribboflavin, on the contrary is not retained by the column. The photo-oxidation of free Trp in the presence of 14 C-riboflavin, gave a compound which bound to the ion exchange resin like the above-mentioned derivative. The photo-oxidation of the Trp residues in lysozyme and in peptides obtained from lysozyme showed very high quantum yields, and these values were directly related to the incorporation of 14 C-riboflavin in these samples. (orig.)

The Antimicrobial Peptide Lysozyme Is Induced after Multiple Trauma

Directory of Open Access Journals (Sweden)

Tim Klüter

2014-01-01

Full Text Available The antimicrobial peptide lysozyme is an important factor of innate immunity and exerts high potential of antibacterial activity. In the present study we evaluated the lysozyme expression in serum of multiple injured patients and subsequently analyzed their possible sources and signaling pathways. Expression of lysozyme was examined in blood samples of multiple trauma patients from the day of trauma until 14 days after trauma by ELISA. To investigate major sources of lysozyme, its expression and regulation in serum samples, different blood cells, and tissue samples were analysed by ELISA and real-time PCR. Neutrophils and hepatocytes were stimulated with cytokines and supernatant of Staphylococcus aureus. The present study demonstrates the induction and release of lysozyme in serum of multiple injured patients. The highest lysozyme expression of all tested cells and tissues was detected in neutrophils. Stimulation with trauma-related factors such as interleukin-6 and S. aureus induced lysozyme expression. Liver tissue samples of patients without trauma show little lysozyme expression compared to neutrophils. After stimulation with bacterial fragments, lysozyme expression of hepatocytes is upregulated significantly. Toll-like receptor 2, a classic receptor of Gram-positive bacterial protein, was detected as a possible target for lysozyme induction.

Interaction of magnetic nanoparticles with lysozyme amyloid fibrils

Energy Technology Data Exchange (ETDEWEB)

Gdovinová, Veronika [Institute of Experimental Physics SAS, Watsonova 47, 040 01 Košice (Slovakia); Frank Laboratory of Neutron Physics, Joint Institute for Nuclear Research, Joliot-Curie 6, 141980 Dubna, Moscow Region (Russian Federation); Tomašovičová, Natália, E-mail: nhudak@saske.sk [Institute of Experimental Physics SAS, Watsonova 47, 040 01 Košice (Slovakia); Frank Laboratory of Neutron Physics, Joint Institute for Nuclear Research, Joliot-Curie 6, 141980 Dubna, Moscow Region (Russian Federation); Batko, Ivan; Batková, Marianna; Balejčíková, Lucia [Institute of Experimental Physics SAS, Watsonova 47, 040 01 Košice (Slovakia); Garamus, Vasyl M. [Helmholtz-Zentrum Geesthacht: Zentrum fr Material, und Kstenforschung GmbH, Max-Plank-Strae 1, Geesthacht 216502 (Germany); Petrenko, Viktor I. [Frank Laboratory of Neutron Physics, Joint Institute for Nuclear Research, Joliot-Curie 6, 141980 Dubna, Moscow Region (Russian Federation); Physics Department, Taras Shevchenko Kyiv National University, Volodymyrska Street 64, 01601 Kyiv (Ukraine); Avdeev, Mikhail V. [Frank Laboratory of Neutron Physics, Joint Institute for Nuclear Research, Joliot-Curie 6, 141980 Dubna, Moscow Region (Russian Federation); Kopčanský, Peter [Institute of Experimental Physics SAS, Watsonova 47, 040 01 Košice (Slovakia)

2017-06-01

This work is devoted to the structural study of complex solutions of magnetic nanoparticles with lysozyme amyloid fibrils due to possible ordering of such system by applying the external magnetic field. The interaction of magnetic nanoparticles with amyloid fibrils has been followed by atomic force microscopy and small-angle X-ray scattering. It has been observed that magnetic nanoparticles (MNPs) adsorb to lysozyme amyloid fibrils. It was found that MNPs alter amyloids structures, namely the diameter of lysozyme amyloid fibrils is increased whereas the length of fibrils is decreased. In the same time MNPs do not change the helical pitch significantly. - Highlights: • Solution of MNPs with lysozyme amyloid fibrils was characterized by AFM and SAXS. • MNPs adsorb to lysozyme amyloid fibrils. • Diameter and size of lysozyme amyloid fibrils change due to doping with MNPs.

Recognition of lysozyme using surface imprinted bacterial cellulose nanofibers.

Science.gov (United States)

Saylan, Yeşeren; Tamahkar, Emel; Denizli, Adil

2017-11-01

Here, we developed the lysozyme imprinted bacterial cellulose (Lyz-MIP/BC) nanofibers via the surface imprinting strategy that was designed to recognize lysozyme. This study includes the molecular imprinting method onto the surface of bacterial cellulose nanofibers in the presence of lysozyme by metal ion coordination, as well as further characterizations methods FTIR, SEM and contact angle measurements. The maximum lysozyme adsorption capacity of Lyz-MIP/BC nanofibers was found to be 71 mg/g. The Lyz-MIP/BC nanofibers showed high selectivity for lysozyme towards bovine serum albumin and cytochrome c. Overall, the Lyz-MIP/BC nanofibers hold great potential for lysozyme recognition due to the high binding capacity, significant selectivity and excellent reusability.

A more clear insight of the lysozyme crystal composition

Energy Technology Data Exchange (ETDEWEB)

Mueller, C.; Ulrich, J. [Martin-Luther-Universitaet Halle-Wittenberg, Zentrum fuer Ingenieurswissenschaften, Verfahrenstechnik/TVT, 06099 Halle Saale (Germany)

2011-07-15

Crystallization can be used as a purification method for proteins. Lysozyme was chosen as a model substance. Changing crystallization conditions will lead as shown to different lysozyme crystal morphologies with different properties. Beside others, lysozyme crystals can show a Tetragonal, High Temperature and Low Temperature Orthorhombic crystal morphology. Experiments such as conductivity measurements, pH tests, chloride detection tests, experiments using methylene blue as a dye and dissolution experiments were carried out to investigate the composition of the lysozyme crystals. It is proven that lysozyme crystals are made up of the initial buffer solution components: lysozyme (the protein), water which is part of the crystal lattice, salt ions which are attached to the protein molecule and voids filled with the buffer solution containing the crystallization agent (e.g. salt). Interesting dissolution behaviours of the lysozyme crystals were observed which are not described so far elsewhere (copyright 2011 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

The association of lysozyme with casein

NARCIS (Netherlands)

Roos, de A.L.; Walstra, P.; Geurts, T.J.

1998-01-01

The association of hen eggs’ lysozyme with caseins was studied by using three casein substrates: (I) solutions of the various caseins, (II) artificially made casein micelles of various compositions and (III) caseins adsorbed onto soya-oil emulsion droplets. In solution, lysozyme associated most

Molecular Cloning and Characterization of a New C-type Lysozyme Gene from Yak Mammary Tissue

Directory of Open Access Journals (Sweden)

Ming Feng Jiang

2015-12-01

Full Text Available Milk lysozyme is the ubiquitous enzyme in milk of mammals. In this study, the cDNA sequence of a new chicken-type (c-type milk lysozyme gene (YML, was cloned from yak mammary gland tissue. A 444 bp open reading frames, which encodes 148 amino acids (16.54 kDa with a signal peptide of 18 amino acids, was sequenced. Further analysis indicated that the nucleic acid and amino acid sequences identities between yak and cow milk lysozyme were 89.04% and 80.41%, respectively. Recombinant yak milk lysozyme (rYML was produced by Escherichia coli BL21 and Pichia pastoris X33. The highest lysozyme activity was detected for heterologous protein rYML5 (M = 1,864.24 U/mg, SD = 25.75 which was expressed in P. pastoris with expression vector pPICZαA and it clearly inhibited growth of Staphylococcus aureus. Result of the YML gene expression using quantitative polymerase chain reaction showed that the YML gene was up-regulated to maximum at 30 day postpartum, that is, comparatively high YML can be found in initial milk production. The phylogenetic tree indicated that the amino acid sequence was similar to cow kidney lysozyme, which implied that the YML may have diverged from a different ancestor gene such as cow mammary glands. In our study, we suggest that YML be a new c-type lysozyme expressed in yak mammary glands that plays a role as host immunity.

Aptamer-Based Electrochemical Sensing of Lysozyme

Directory of Open Access Journals (Sweden)

Alina Vasilescu

2016-06-01

Full Text Available Protein analysis and quantification are required daily by thousands of laboratories worldwide for activities ranging from protein characterization to clinical diagnostics. Multiple factors have to be considered when selecting the best detection and quantification assay, including the amount of protein available, its concentration, the presence of interfering molecules, as well as costs and rapidity. This is also the case for lysozyme, a 14.3-kDa protein ubiquitously present in many organisms, that has been identified with a variety of functions: antibacterial activity, a biomarker of several serious medical conditions, a potential allergen in foods or a model of amyloid-type protein aggregation. Since the design of the first lysozyme aptamer in 2001, lysozyme became one of the most intensively-investigated biological target analytes for the design of novel biosensing concepts, particularly with regards to electrochemical aptasensors. In this review, we discuss the state of the art of aptamer-based electrochemical sensing of lysozyme, with emphasis on sensing in serum and real samples.

Technology Optimization of Lysozyme's Fresh Maintaining Effect on Apple.

Science.gov (United States)

Jun-Hong, Liu; Kun-Yu, Wang

2016-10-03

Lysozyme is a kind of alkaline globin, which functions well in the degradation of the cell wall of microbe. Currently, lysozyme is widely used in various fields, such as medicine, fruit, and vegetable industry, dairy industry, and so on. Therefore, the exploitation and utilization of lysozyme is of significant economic benefit. Taking apple as material, weight loss ratio and reducing sugar content as objectives, this paper studied the fresh-keeping effect of lysozyme. Influential factors, lysozyme concentration, soaking time, modified temperature, and reaction time were discussed in detail. The results showed that reducing sugar content was 2.043% and the weight loss ratio was the minimum in the presence of soaking time of 2 min, modified temperature of 65 °C, reaction time of 4 d, and lysozyme concentration of 0.5 g/L. © 2016 Institute of Food Technologists®.

The Antimicrobial Peptide Lysozyme Is Induced after Multiple Trauma

OpenAIRE

Klüter, Tim; Fitschen-Oestern, Stefanie; Lippross, Sebastian; Weuster, Matthias; Mentlein, Rolf; Steubesand, Nadine; Neunaber, Claudia; Hildebrand, Frank; Pufe, Thomas; Tohidnezhad, Mersedeh; Beyer, Andreas; Seekamp, Andreas; Varoga, Deike

2014-01-01

The antimicrobial peptide lysozyme is an important factor of innate immunity and exerts high potential of antibacterial activity. In the present study we evaluated the lysozyme expression in serum of multiple injured patients and subsequently analyzed their possible sources and signaling pathways. Expression of lysozyme was examined in blood samples of multiple trauma patients from the day of trauma until 14 days after trauma by ELISA. To investigate major sources of lysozyme, its expression ...

A novel electrochemical aptamer-antibody sandwich assay for lysozyme detection.

Science.gov (United States)

Ocaña, Cristina; Hayat, Akhtar; Mishra, Rupesh; Vasilescu, Alina; del Valle, Manel; Marty, Jean-Louis

2015-06-21

In this paper, we have reported a novel electrochemical aptamer-antibody based sandwich biosensor for the detection of lysozyme. In the sensing strategy, an anti-lysozyme aptamer was immobilized onto the carbon electrode surface by covalent binding via diazonium salt chemistry. After incubating with a target protein (lysozyme), a biotinylated antibody was used to complete the sandwich format. The subsequent additions of avidin-alkaline phosphatase as an enzyme label, and a 1-naphthyl phosphate substrate (1-NPP) allowed us to determine the concentration of lysozyme (Lys) via Differential Pulse Voltammetry (DPV) of the generated enzyme reaction product, 1-naphthol. Using this strategy, a wide detection range from 5 fM to 5 nM was obtained for a target lysozyme, with a detection limit of 4.3 fM. The control experiments were carried out by using bovine serum albumin (BSA), cytochrome c and casein. The results showed that the proposed biosensor had good specificity, stability and reproducibility for lysozyme analysis. In addition, the biosensor was applied for detecting lysozyme in spiked wine samples, and very good recovery rates were obtained in the range from 95.2 to 102.0% for lysozyme detection. This implies that the proposed sandwich biosensor is a promising analytical tool for the analysis of lysozyme in real samples.

An improved 96-well turbidity assay for T4 lysozyme activity.

Science.gov (United States)

Toro, Tasha B; Nguyen, Thao P; Watt, Terry J

2015-01-01

T4 lysozyme (T4L) is an important model system for investigating the relationship between protein structure and function. Despite being extensively studied, a reliable, quantitative activity assay for T4L has not been developed. Here, we present an improved T4L turbidity assay as well as an affinity-based T4L expression and purification protocol. This assay is designed for 96-well format and utilizes conditions amenable for both T4L and other lysozymes. This protocol enables easy, efficient, and quantitative characterization of T4L variants and allows comparison between different lysozymes. Our method: •Is applicable for all lysozymes, with enhanced sensitivity for T4 lysozyme compared to other 96-well plate turbidity assays;•Utilizes standardized conditions for comparing T4 lysozyme variants and other lysozymes; and•Incorporates a simplified expression and purification protocol for T4 lysozyme.

Inheritance of the lysozyme inhibitor Ivy was an important evolutionary step by Yersinia pestis to avoid the host innate immune response.

Science.gov (United States)

Derbise, Anne; Pierre, François; Merchez, Maud; Pradel, Elizabeth; Laouami, Sabrina; Ricard, Isabelle; Sirard, Jean-Claude; Fritz, Jill; Lemaître, Nadine; Akinbi, Henry; Boneca, Ivo G; Sebbane, Florent

2013-05-15

Yersinia pestis (the plague bacillus) and its ancestor, Yersinia pseudotuberculosis (which causes self-limited bowel disease), encode putative homologues of the periplasmic lysozyme inhibitor Ivy and the membrane-bound lysozyme inhibitor MliC. The involvement of both inhibitors in virulence remains subject to debate. Mutants lacking ivy and/or mliC were generated. We evaluated the mutants' ability to counter lysozyme, grow in serum, and/or counter leukocytes; to produce disease in wild-type, neutropenic, or lysozyme-deficient rodents; and to induce host inflammation. MliC was not required for lysozyme resistance and the development of plague. Deletion of ivy decreased Y. pestis' ability to counter lysozyme and polymorphonuclear neutrophils, but it did not affect the bacterium's ability to grow in serum or resist macrophages. Y. pestis lacking Ivy had attenuated virulence, unless animals were neutropenic or lysozyme deficient. The Ivy mutant induced inflammation to a degree similar to that of the parental strain. Last, Y. pseudotuberculosis did not require Ivy to counter lysozyme and for virulence. Ivy is required to counter lysozyme during infection, but its role as a virulence factor is species dependent. Our study also shows that a gene that is not necessary for the virulence of an ancestral bacterium may become essential in the emergence of a new pathogen.

Study of ethanol-lysozyme interactions using neutron diffraction

International Nuclear Information System (INIS)

Lehmann, M.S.; Mason, S.A.; McIntyre, G.J.

1985-01-01

Single-crystal neutron diffraction has been used to observe the interactions between deuterated ethanol (CD3CD2OH) and lysozyme in triclinic crystals of hen egg white lysozyme soaked in 25% (v/v) ethanol solutions. A total of 6047 observed reflections to a resolution of 2 A were used, and 13 possible ethanol sites were identified. The three highest occupied sites are close to locations for bromoethanol found in an earlier study by Yonath et al. [Yonath, A., Podjarny, A., Honig, B., Traub, W., Sielecki, A., Herzberg, O., and Moult, J. (1978) Biophys. Struct. Mech. 4, 27-36]. Structure refinements including a model for the flat solvent lead to a final crystallographic agreement factor of 0.097. Comparison with earlier neutron studies on triclinic lysozyme showed that neither the molecular structure nor the thermal motions were affected significantly by the ethanol. A detailed analysis of the ethanol-lysozyme contacts showed 61% of these to be with hydrophobic sites, in agreement with the dominant hydrophobic nature of ethanol. This, together with the fact that the molecular structure of lysozyme is not perturbed, suggests a model for denaturation of lysozyme by alcohol, which proceeds via a dehydration of the protein at high alcohol concentration

The interaction of lysozyme with caffeine, theophylline and theobromine in solution.

Science.gov (United States)

Zhang, Hong-Mei; Tang, Bo-Ping; Wang, Yan-Qing

2010-10-01

The interactions of lysozyme with caffeine (Caf), theophylline (Tph) and theobromine (Tbr) were investigated using UV-Vis absorption, fluorescence, synchronous fluorescence, and three-dimensional fluorescence spectra techniques. The results revealed that Caf (Tph or Tbr) caused the fluorescence quenching of lysozyme by the formation of Caf (Tph or Tbr)-lysozyme complex. The binding constants (K(A)) and thermodynamic parameters (ΔG°, ΔH°, ΔS°) at two different temperatures, the binding locality, and the binding power were obtained. The results showed that the process of binding Caf (Tph or Tbr) to lysozyme was a spontaneous molecular interaction procedure and the hydrophobic and electrostatic interactions play a major role in stabilizing the complex; The distance r between donor (lysozyme) and acceptor (Caf, Tph or Tbr) was obtained according to fluorescence resonance energy transfer. The effect of Caf (Tph or Tbr) on the conformation of lysozyme was analyzed using synchronous fluorescence and three-dimensional fluorescence spectra techniques. The results showed that the binding of Caf (Tph or Tbr) to lysozyme induced some micro-environmental and conformational changes in lysozyme and disturbed the environment of the polypeptide of lysozyme.

Suppressive effects of lysozyme on polyphosphate-mediated vascular inflammatory responses

Energy Technology Data Exchange (ETDEWEB)

Chung, Jiwoo [College of Pharmacy, CMRI, Research Institute of Pharmaceutical Sciences, BK21 Plus KNU Multi-Omics Based Creative Drug Research Team, Kyungpook National University, Daegu 41566 (Korea, Republic of); Ku, Sae-Kwang [Department of Anatomy and Histology, College of Korean Medicine, Daegu Haany University, Gyeongsan 38610 (Korea, Republic of); Lee, Suyeon [College of Pharmacy, CMRI, Research Institute of Pharmaceutical Sciences, BK21 Plus KNU Multi-Omics Based Creative Drug Research Team, Kyungpook National University, Daegu 41566 (Korea, Republic of); Bae, Jong-Sup, E-mail: baejs@knu.ac.kr [College of Pharmacy, CMRI, Research Institute of Pharmaceutical Sciences, BK21 Plus KNU Multi-Omics Based Creative Drug Research Team, Kyungpook National University, Daegu 41566 (Korea, Republic of)

2016-06-10

Lysozyme, found in relatively high concentration in blood, saliva, tears, and milk, protects us from the ever-present danger of bacterial infection. Previous studies have reported proinflammatory responses of endothelial cells to the release of polyphosphate(PolyP). In this study, we examined the anti-inflammatory responses and mechanisms of lysozyme and its effects on PolyP-induced septic activities in human umbilical vein endothelial cells (HUVECs) and mice. The survival rates, septic biomarker levels, behavior of human neutrophils, and vascular permeability were determined in PolyP-activated HUVECs and mice. Lysozyme suppressed the PolyP-mediated vascular barrier permeability, upregulation of inflammatory biomarkers, adhesion/migration of leukocytes, and activation and/or production of nuclear factor-κB, tumor necrosis factor-α, and interleukin-6. Furthermore, lysozyme demonstrated protective effects on PolyP-mediated lethal death and the levels of the related septic biomarkers. Therefore, these results indicated the therapeutic potential of lysozyme on various systemic inflammatory diseases, such as sepsis or septic shock. -- Highlights: •PolyP is shown to be an important mediator of vascular inflammation. •Lysozyme inhibited PolyP-mediated hyperpermeability. •Lysozyme inhibited PolyP-mediated septic response. •Lysozyme reduced PolyP-induced septic mortality.

Suppressive effects of lysozyme on polyphosphate-mediated vascular inflammatory responses

International Nuclear Information System (INIS)

Chung, Jiwoo; Ku, Sae-Kwang; Lee, Suyeon; Bae, Jong-Sup

2016-01-01

Lysozyme, found in relatively high concentration in blood, saliva, tears, and milk, protects us from the ever-present danger of bacterial infection. Previous studies have reported proinflammatory responses of endothelial cells to the release of polyphosphate(PolyP). In this study, we examined the anti-inflammatory responses and mechanisms of lysozyme and its effects on PolyP-induced septic activities in human umbilical vein endothelial cells (HUVECs) and mice. The survival rates, septic biomarker levels, behavior of human neutrophils, and vascular permeability were determined in PolyP-activated HUVECs and mice. Lysozyme suppressed the PolyP-mediated vascular barrier permeability, upregulation of inflammatory biomarkers, adhesion/migration of leukocytes, and activation and/or production of nuclear factor-κB, tumor necrosis factor-α, and interleukin-6. Furthermore, lysozyme demonstrated protective effects on PolyP-mediated lethal death and the levels of the related septic biomarkers. Therefore, these results indicated the therapeutic potential of lysozyme on various systemic inflammatory diseases, such as sepsis or septic shock. -- Highlights: •PolyP is shown to be an important mediator of vascular inflammation. •Lysozyme inhibited PolyP-mediated hyperpermeability. •Lysozyme inhibited PolyP-mediated septic response. •Lysozyme reduced PolyP-induced septic mortality.

MUCOADHESIVE GEL WITH IMMOBILIZED LYSOZYME: PREPARATION AND PROPERTIES

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Dekina S. S.

2015-08-01

Full Text Available The study of non-covalent immobilized lysozyme, as well as physico-chemical and biochemical properties of obtained mucoadhesive gel was the aim of the research. Lysozyme activity was determined by bacteriolytic method (Micrococcus lysodeikticus cells acetone powder was a substrate. Lysozyme immobilization was conducted by the method of entrapment in gel. Enzyme carrier interaction was studied by viscometric, spectrophotometric and spectrofluorimetric methods. Mucoadhesive gel with immobilized lysozyme, possessing antiinflammatory and antimicrobial activities, was prepared. Due to immobilization, protein-polymer complex with the original enzymatic activity was formed. The product is characterized by high mucoadhesive properties, quantitative retaining of protein and bacteriolytic activity, prolonged release of the enzyme, improved biochemical characteristics (extended pH-activity profile, stability in acidic medium and during storage for 2 years, and it is perspective for further studies. The proposed method for lysozyme immobilization in the carboxymethyl cellulose sodium salt gel allows to obtain a stable, highly efficient product, with high adhesive properties for attachment to the mucous membranes, that is promising for use in biomedicine.

Phase behavior of mixtures of oppositely charged nanoparticles: Heterogeneous Poisson-Boltzmann cell model applied to lysozyme and succinylated lysozyme

NARCIS (Netherlands)

Biesheuvel, P.M.; Lindhoud, S.; Vries, de R.J.; Stuart, M.A.C.

2006-01-01

We study the phase behavior of mixtures of oppositely charged nanoparticles, both theoretically and experimentally. As an experimental model system we consider mixtures of lysozyme and lysozyme that has been chemically modified in such a way that its charge is nearly equal in magnitude but opposite

Expression and prognostic significance of lysozyme in male breast cancer

International Nuclear Information System (INIS)

Serra, Carlos; Baltasar, Aniceto; Medrano, Justo; Vizoso, Francisco; Alonso, Lorena; Rodríguez, Juan C; González, Luis O; Fernández, María; Lamelas, María L; Sánchez, Luis M; García-Muñiz, José L

2002-01-01

Lysozyme, one of the major protein components of human milk that is also synthesized by a significant percentage of breast carcinomas, is associated with lesions that have a favorable outcome in female breast cancer. Here we evaluate the expression and prognostic value of lysozyme in male breast cancer (MBC). Lysozyme expression was examined by immunohistochemical methods in a series of 60 MBC tissue sections and in 15 patients with gynecomastia. Staining was quantified using the HSCORE (histological score) system, which considers both the intensity and the percentage of cells staining at each intensity. Prognostic value of lysozyme was retrospectively evaluated by multivariate analysis taking into account conventional prognostic factors. Lysozyme immunostaining was negative in all cases of gynecomastia. A total of 27 of 60 MBC sections (45%) stained positively for this protein, but there were clear differences among them with regard to the intensity and percentage of stained cells. Statistical analysis showed that lysozyme HSCORE values in relation to age, tumor size, nodal status, histological grade, estrogen receptor status, metastasis and histological type did not increase the statistical significance. Univariate analysis confirmed that both nodal involvement and lysozyme values were significant predictors of short-term relapse-free survival. Multivariate analysis, according to Cox's regression model, also showed that nodal status and lysozyme levels were significant independent indicators of short-term relapse-free survival. Tumor expression of lysozyme is associated with lesions that have an unfavorable outcome in male breast cancer. This milk protein may be a new prognostic factor in patients with breast cancer

Biomimetic mineralization of calcium carbonate/carboxymethylcellulose microspheres for lysozyme immobilization

International Nuclear Information System (INIS)

Lu Zheng; Zhang Juan; Ma Yunzi; Song Siyue; Gu Wei

2012-01-01

Porous calcium carbonate/carboxymethylcellulose (CaCO 3 /CMC) microspheres were prepared by the biomimetic mineralization method for lysozyme immobilization via adsorption. The size and morphology of CaCO 3 /CMC microspheres were characterized by transmitted electron microscopy (TEM) and zeta potential measurement. The lysozyme immobilization was verified by Fourier transform infrared (FTIR) spectroscopy. The effects of pHs and temperatures on lysozyme adsorption were investigated as well. It was revealed that CaCO 3 /CMC microspheres could immobilize lysozyme efficiently via electrostatic interactions and a maximum adsorption capacity of 450 mg/g was achieved at pH 9.2 and 25 °C. Moreover, it was found that the adsorption process fitted well with the Langmuir isothermal model. In addition, UV, fluorescence, and circular dichroism (CD) spectroscopic studies showed that lysozyme maintained its original secondary structure during the adsorption/desorption process. Our study therefore demonstrated that CaCO 3 /CMC microsphere can be used as a cost-effective and efficient support for lysozyme immobilization. - Graphical abstract: CaCO 3 /CMC microsphere was prepared by a facile biomimetic mineralization method and can be used as an efficient and cost-effective support for lysozyme immobilization. Highlights: ► CaCO 3 /CMC microspheres were prepared by the biomimetic mineralization method. ► Lysozyme was efficiently immobilized to CaCO 3 /CMC microspheres via adsorption. ► A maximum adsorption capacity of 450 mg/g was obtained at pH 9.2 and 25 °C. ► The original secondary structure of lysozyme was maintained upon immobilization.

Antimicrobial Membranes of Bio-Based PA 11 and HNTs Filled with Lysozyme Obtained by an Electrospinning Process

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Valeria Bugatti

2018-03-01

Full Text Available Bio-based membranes were obtained using Polyamide 11 (PA11 from renewable sources and a nano-hybrid composed of halloysite nanotubes (HNTs filled with lysozyme (50 wt % of lysozyme, as a natural antimicrobial molecule. Composites were prepared using an electrospinning process, varying the nano-hybrid loading (i.e., 1.0, 2.5, 5.0 wt %. The morphology of the membranes was investigated through SEM analysis and there was found to be a narrow average fiber diameter (0.3–0.5 μm. The mechanical properties were analyzed and correlated to the nano-hybrid content. Controlled release of lysozyme was followed using UV spectrophotometry and the release kinetics were found to be dependent on HNTs–lysozyme loading. The experimental results were analyzed by a modified Gallagher–Corrigan model. The application of the produced membranes, as bio-based pads, for extending the shelf life of chicken slices has been tested and evaluated.

Penetration and fusion of phospholipid vesicles by lysozyme

International Nuclear Information System (INIS)

Kim, J.; Kim, H.

1989-01-01

The lysozyme-induced fusion of phosphatidylserine/phosphatidylethanolamine vesicles as studied at a wide range of pH is found to correlate well with the binding of this protein to the vesicles. An identical 6000 molecular weight segment of lysozyme at the N-terminal region is found to be protected from tryptic digestion when initially incubated with vesicles at several pH values. Only this segment is labeled by dansyl chloride, which is partitioned into the bilayer. These results suggest the penetration of one segment of lysozyme into the bilayer. Photoactivated labeling of the membrane-penetrating segment of lysozyme with 3-(trifluoromethyl)-3-([ 125 I]iodophenyl)diazirine ([ 125 I]TID) and subsequent identification of the labeled residues by Edman degradation and gamma-ray counting indicate that four amino acids from the N-terminal are located outside the hydrophobic core of the bilayer. Although treatment of the membrane-embedded segment with aminopeptidase failed to cleave any amino acids from the N-terminal, it appears that a loop of lysozyme segment near the N-terminal penetrates into the bilayer at acidic pH. A helical wheel diagram shows that the labeling is done mainly on one surface of the alpha-helix. The penetration kinetics as studied by time-dependent [ 125 I]TID labeling coincide with the fusion kinetics, strongly suggesting that the penetration of the lysozyme segment into the vesicles is the cause of the fusion

Penetration and fusion of phospholipid vesicles by lysozyme

Energy Technology Data Exchange (ETDEWEB)

Kim, J.; Kim, H. (Korea Advanced Institute of Science and Technology, Seoul)

1989-10-01

The lysozyme-induced fusion of phosphatidylserine/phosphatidylethanolamine vesicles as studied at a wide range of pH is found to correlate well with the binding of this protein to the vesicles. An identical 6000 molecular weight segment of lysozyme at the N-terminal region is found to be protected from tryptic digestion when initially incubated with vesicles at several pH values. Only this segment is labeled by dansyl chloride, which is partitioned into the bilayer. These results suggest the penetration of one segment of lysozyme into the bilayer. Photoactivated labeling of the membrane-penetrating segment of lysozyme with 3-(trifluoromethyl)-3-(({sup 125}I)iodophenyl)diazirine (({sup 125}I)TID) and subsequent identification of the labeled residues by Edman degradation and gamma-ray counting indicate that four amino acids from the N-terminal are located outside the hydrophobic core of the bilayer. Although treatment of the membrane-embedded segment with aminopeptidase failed to cleave any amino acids from the N-terminal, it appears that a loop of lysozyme segment near the N-terminal penetrates into the bilayer at acidic pH. A helical wheel diagram shows that the labeling is done mainly on one surface of the alpha-helix. The penetration kinetics as studied by time-dependent ({sup 125}I)TID labeling coincide with the fusion kinetics, strongly suggesting that the penetration of the lysozyme segment into the vesicles is the cause of the fusion.

The Anti-sigma Factor RsiV Is a Bacterial Receptor for Lysozyme: Co-crystal Structure Determination and Demonstration That Binding of Lysozyme to RsiV Is Required for σV Activation.

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Jessica L Hastie

2016-09-01

Full Text Available σ factors provide RNA polymerase with promoter specificity in bacteria. Some σ factors require activation in order to interact with RNA polymerase and transcribe target genes. The Extra-Cytoplasmic Function (ECF σ factor, σV, is encoded by several Gram-positive bacteria and is specifically activated by lysozyme. This activation requires the proteolytic destruction of the anti-σ factor RsiV via a process of regulated intramembrane proteolysis (RIP. In many cases proteases that cleave at site-1 are thought to directly sense a signal and initiate the RIP process. We previously suggested binding of lysozyme to RsiV initiated the proteolytic destruction of RsiV and activation of σV. Here we determined the X-ray crystal structure of the RsiV-lysozyme complex at 2.3 Å which revealed that RsiV and lysozyme make extensive contacts. We constructed RsiV mutants with altered abilities to bind lysozyme. We find that mutants that are unable to bind lysozyme block site-1 cleavage of RsiV and σV activation in response to lysozyme. Taken together these data demonstrate that RsiV is a receptor for lysozyme and binding of RsiV to lysozyme is required for σV activation. In addition, the co-structure revealed that RsiV binds to the lysozyme active site pocket. We provide evidence that in addition to acting as a sensor for the presence of lysozyme, RsiV also inhibits lysozyme activity. Thus we have demonstrated that RsiV is a protein with multiple functions. RsiV inhibits σV activity in the absence of lysozyme, RsiV binds lysozyme triggering σV activation and RsiV inhibits the enzymatic activity of lysozyme.

Effects of Purification on the Crystallization of Lysozyme

Science.gov (United States)

Ewing, Felecia L.; Forsythe, Elizabeth L.; Van Der Woerd, Mark; Pusey, Marc L.

1996-01-01

We have additionally purified a commercial lysozyme preparation by cation exchange chromatography, followed by recrystallization. This material is 99.96% pure with respect to macromolecular impurities. At basic pH, the purified lysozyme gave only tetragonal crystals at 20 C. Protein used directly from the bottle, prepared by dialysis against distilled water, or which did not bind to the cation exchange column had considerably altered crystallization behavior. Lysozyme which did not bind to the cation exchange column was subsequently purified by size exclusion chromatography. This material gave predominately bundles of rod-shaped crystals with some small tetragonal crystals at lower pHs. The origin of the bundled rod habit was postulated to be a thermally dependent tetragonal- orthorhombic change in the protein structure. This was subsequently ruled out on the basis of crystallization behavior and growth rate experiments. This suggests that heterogeneous forms of lysozyme may be responsible. These results demonstrate three classes of impurities: (1) small molecules, which may be removed by dialysis; (2) macromolecules, which are removable by chromatographic techniques; and (3) heterogeneous forms of the protein, which can be removed in this case by cation exchange chromatography. Of these, heterogeneous forms of the lysozyme apparently have the greatest affect on its crystallization behavior.

Production of human lactoferrin and lysozyme in the milk of transgenic dairy animals: past, present, and future.

Science.gov (United States)

Cooper, Caitlin A; Maga, Elizabeth A; Murray, James D

2015-08-01

Genetic engineering, which was first developed in the 1980s, allows for specific additions to animals' genomes that are not possible through conventional breeding. Using genetic engineering to improve agricultural animals was first suggested when the technology was in the early stages of development by Palmiter et al. (Nature 300:611-615, 1982). One of the first agricultural applications identified was generating transgenic dairy animals that could produce altered or novel proteins in their milk. Human milk contains high levels of antimicrobial proteins that are found in low concentrations in the milk of ruminants, including the antimicrobial proteins lactoferrin and lysozyme. Lactoferrin and lysozyme are both part of the innate immune system and are secreted in tears, mucus, and throughout the gastrointestinal (GI) tract. Due to their antimicrobial properties and abundance in human milk, multiple lines of transgenic dairy animals that produce either human lactoferrin or human lysozyme have been developed. The focus of this review is to catalogue the different lines of genetically engineered dairy animals that produce either recombinant lactoferrin or lysozyme that have been generated over the years as well as compare the wealth of research that has been done on the in vitro and in vivo effects of the milk they produce. While recent advances including the development of CRISPRs and TALENs have removed many of the technical barriers to predictable and efficient genetic engineering in agricultural species, there are still many political and regulatory hurdles before genetic engineering can be used in agriculture. It is important to consider the substantial amount of work that has been done thus far on well established lines of genetically engineered animals evaluating both the animals themselves and the products they yield to identify the most effective path forward for future research and acceptance of this technology.

Use of Lysozyme as a Feed Additive on Rumen Fermentation and Methane Emission

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Ashraf A. Biswas

2016-11-01

Full Text Available This study was conducted to determine the effect of lysozyme addition on in vitro rumen fermentation and to identify the lysozyme inclusion rate for abating methane (CH4 production. An in vitro ruminal fermentation technique was done using a commercial concentrate to rice straw ratio of 8:2 as substrate. The following treatments were applied wherein lysozyme was added into 1 mg dry matter substrate at different levels of inclusion: Without lysozyme, 2,000, 4,000, and 8,000 U lysozyme. Results revealed that, lysozyme addition had a significant effect on pH after 24 h of incubation, with the highest pH (p<0.01 observed in 8,000 U lysozyme, followed by the 4,000 U, 2,000 U, and without lysozyme. The highest amounts of acetic acid, propionic acid (p<0.01 and total volatile fatty acid (TVFA (p<0.05 were found in 8,000 U after 24 h of incubation. The CH4 concentration was the lowest in the 8,000 U and the highest in the without lysozyme addition after 24 h of incubation. There was no significant differences in general bacteria, methanogen, or protozoan DNA copy number. So far, addition of lysozyme increased the acetate, propionate, TVFA, and decreased CH4 concentration. These results suggest that lysozyme supplementation may improve in vitro rumen fermentation and reduce CH4 emission.

Complex coacervate core micelles with a lysozyme-modified corona.

Science.gov (United States)

Danial, Maarten; Klok, Harm-Anton; Norde, Willem; Stuart, Martien A Cohen

2007-07-17

This paper describes the preparation, characterization, and enzymatic activity of complex coacervate core micelles (C3Ms) composed of poly(acrylic acid) (PAA) and poly(N-methyl-2-vinyl pyridinium iodide)-b-poly(ethylene oxide) (PQ2VP-PEO) to which the antibacterial enzyme lysozyme is end-attached. C3Ms were prepared by polyelectrolyte complex formation between PAA and mixtures containing different ratios of aldehyde and hydroxyl end-functionalized PQ2VP-PEO. This resulted in the formation of C3Ms containing 0-40% (w/w) of the aldehyde end-functionalized PQ2VP-PEO block copolymer (PQ2VP-PEO-CHO). Chemical conjugation of lysozyme was achieved via reductive amination of the aldehyde groups, which are exposed at the surface of the C3M, with the amine groups present in the side chains of the lysine residues of the protein. Dynamic and static light scattering indicated that the conjugation of lysozyme to C3Ms prepared using 10 and 20% (w/w) PQ2VP-PEO-CHO resulted in the formation of unimicellar particles. Multimicellar aggregates, in contrast, were obtained when lysozyme was conjugated to C3Ms prepared using 30 or 40% (w/w) PQ2VP-PEO-CHO. The enzymatic activity of the unimicellar lysozyme-C3M conjugates toward the hydrolysis of the bacterial substrate Micrococcus lysodeikticus was comparable to that of free lysozyme. For the multimicellar particles, in contrast, significantly reduced enzymatic rates of hydrolysis, altered circular dichroism, and red-shifted tryptophan fluorescence spectra were measured. These results are attributed to the occlusion of lysozyme in the interior of the multimicellar conjugates.

Potential toxicity and affinity of triphenylmethane dye malachite green to lysozyme.

Science.gov (United States)

Ding, Fei; Li, Xiu-Nan; Diao, Jian-Xiong; Sun, Ye; Zhang, Li; Ma, Lin; Yang, Xin-Ling; Zhang, Li; Sun, Ying

2012-04-01

Malachite green is a triphenylmethane dye that is used extensively in many industrial and aquacultural processes, generating environmental concerns and health problems to human being. In this contribution, the complexation between lysozyme and malachite green was verified by means of computer-aided molecular modeling, steady state and time-resolved fluorescence, and circular dichroism (CD) approaches. The precise binding patch of malachite green in lysozyme has been identified from molecular modeling and ANS displacement, Trp-62, Trp-63, and Trp-108 residues of lysozyme were earmarked to possess high-affinity for this dye, the principal forces in the lysozyme-malachite green adduct are hydrophobic and π-π interactions. Steady state fluorescence proclaimed the complex of malachite green with lysozyme yields quenching through static type, which substantiates time-resolved fluorescence measurements that lysozyme-malachite green conjugation formation has an affinity of 10(3)M(-1). Moreover, via molecular modeling and also CD data, we can safely arrive at a conclusion that the polypeptide chain of lysozyme partially destabilized upon complexation with malachite green. The data emerged here will help to further understand the toxicological action of malachite green in human body. Copyright © 2012 Elsevier Inc. All rights reserved.

TEACHER-PRODUCED INSTRUCTIONAL FILMS IN CHEMISTRY, 8MM AND SUPER 8.

Science.gov (United States)

O'CONNOR, ROD; SLABAUGH, WENDELL

TECHNIQUES FOR PRODUCING 8MM INSTRUCTIONAL FILMS IN CHEMISTRY ARE PRESENTED. IN PART I A PHILOSOPHY OF TEACHER-PRODUCED FILMS IS DEVELOPED, EMPHASIZING THE VALUE OF THE LOCAL SETTING, AND CUSTOM-MADE CONTENTS. APPLICATIONS SUGGESTED ARE (1) TECHNIQUE INSTRUCTION, (2) FILMED EXPERIMENTS, (3) INSTRUMENT FAMILIARIZATION, (4) LECTURE AIDS, AND (5)…

Deuterium retention properties of co-deposited carbon films produced at wall gaps

International Nuclear Information System (INIS)

Nobuta, Yuji; Kanazawa, Jun; Yamauchi, Yuji; Hino, Tomoaki; Yokoyama, Kenji; Suzuki, Satoshi; Ezato, Koichiro; Enoeda, Mikio; Akiba, Masato; Akamaru, Satoshi; Hatano, Yuji

2013-01-01

Deuterium retention properties in co-deposited carbon film produced in gap and the relationship between this retention behavior and the crystal structure of carbon film were investigated. In the case of a wide gap, the atomic ratio of deuterium to carbon (D/C) in the film was almost constant at any depth in the gap, while in the case of a narrow gap the D/C ratio decreased with increasing distance from the gap entrance. The micro structure of carbon film tended to be more amorphous for the film produced at locations deeper in the gap. Thermal desorption spectra of D 2 in the film produced near the gap entrance showed one broad main peak at around 1100 K, while that in the film produced near the bottom showed very sharp peaks at around 950 K. This difference in desorption behavior was related with the differences of micro structure. (author)

Nanocomposites Based on PCL and Halloysite Nanotubes Filled with Lysozyme: Effect of Draw Ratio on the Physical Properties and Release Analysis

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Valeria Bugatti

2017-08-01

Full Text Available Halloysite nanotubes (HNTs were loaded with lsozyme, as antimicrobial molecule, at a HNTs/lysozyme ratio of 1:1. Such a nano-hybrid was incorporated into a poly (ε-caprolactone (PCL matrix at 10 wt % and films were obtained. The nano-composites were submitted to a cold drawn process at three different draw ratios, λ = 3, 4, and 5, where λ is l(final length/l0(initial length. Morphology, physical, and barrier properties of the starting nanocomposite and drawn samples were studied, and correlated to the release of the lysozyme molecule. It was demonstrated that with a simple mechanical treatment it is possible to obtain controlled release systems for specific active packaging requirements.

Plasticized chitosan/polyolefin films produced by extrusion.

Science.gov (United States)

Matet, Marie; Heuzey, Marie-Claude; Ajji, Abdellah; Sarazin, Pierre

2015-03-06

Plasticized chitosan and polyethylene blends were produced through a single-pass extrusion process. Using a twin-screw extruder, chitosan plasticization was achieved in the presence of an acetic acid solution and glycerol, and directly mixed with metallocene polyethylene, mPE, to produce a masterbatch. Different dilutions of the masterbatch (2, 5 and 10 wt% of plasticized chitosan), in the presence of ethylene vinyl acetate, EVA, were subsequently achieved in single screw film extrusion. Very small plasticized chitosan domains (number average diameter <5 μm) were visible in the polymeric matrix. The resulting films presented a brown color and increasing haze with chitosan plasticized content. Mechanical properties of the mPE films were affected by the presence of plasticized chitosan, but improvement was observed as a result of some compatibility between mPE and chitosan in the presence of EVA. Finally the incorporation of plasticized chitosan affected mPE water vapor permeability while oxygen permeability remained constant. Copyright © 2014 Elsevier Ltd. All rights reserved.

Methods for producing thin film charge selective transport layers

Science.gov (United States)

Hammond, Scott Ryan; Olson, Dana C.; van Hest, Marinus Franciscus Antonius Maria

2018-01-02

Methods for producing thin film charge selective transport layers are provided. In one embodiment, a method for forming a thin film charge selective transport layer comprises: providing a precursor solution comprising a metal containing reactive precursor material dissolved into a complexing solvent; depositing the precursor solution onto a surface of a substrate to form a film; and forming a charge selective transport layer on the substrate by annealing the film.

A population-based study of salivary lysozyme concentrations and candidal counts.

Science.gov (United States)

Yeh, C K; Dodds, M W; Zuo, P; Johnson, D A

1997-01-01

The relationship between salivary lysozyme concentration and oral candida load was examined in 595 adults. Unstimulated whole saliva, and citrate-stimulated parotid and submandibular/sublingual saliva were collected from each participant. Candida colony-forming units (c.f.u.) in unstimulated whole saliva were determined. An enzyme-linked immunosorbent assay for lysozyme using commercially available antibodies was developed. This assay showed a linear relation of salivary lysozyme concentrations from 0.5 to 4.0 ng/ml. Significant negative relations were observed between lysozyme concentration and flow rate: r = -0.16 (p candida counts (r = 0.18: p candida in whole saliva revealed that lysozyme concentrations were higher in the high candida (> or = 1000 c.f.u./ml) group than in the zero and moderate candida categories in stimulated parotid saliva (p candida load increases.

The antibacterial protein lysozyme identified as the termite egg recognition pheromone.

Directory of Open Access Journals (Sweden)

Kenji Matsuura

Full Text Available Social insects rely heavily on pheromone communication to maintain their sociality. Egg protection is one of the most fundamental social behaviours in social insects. The recent discovery of the termite-egg mimicking fungus 'termite-ball' and subsequent studies on termite egg protection behaviour have shown that termites can be manipulated by using the termite egg recognition pheromone (TERP, which strongly evokes the egg-carrying and -grooming behaviours of workers. Despite the great scientific and economic importance, TERP has not been identified because of practical difficulties. Herein we identified the antibacterial protein lysozyme as the TERP. We isolated the target protein using ion-exchange and hydrophobic interaction chromatography, and the MALDI-TOF MS analysis showed a molecular size of 14.5 kDa. We found that the TERP provided antibacterial activity against a gram-positive bacterium. Among the currently known antimicrobial proteins, the molecular size of 14.5 kDa limits the target to lysozyme. Termite lysozymes obtained from eggs and salivary glands, and even hen egg lysozyme, showed a strong termite egg recognition activity. Besides eggs themselves, workers also supply lysozyme to eggs through frequent egg-grooming, by which egg surfaces are coated with saliva containing lysozyme. Reverse transcript PCR analysis showed that mRNA of termite lysozyme was expressed in both salivary glands and eggs. Western blot analysis confirmed that lysozyme production begins in immature eggs in queen ovaries. This is the first identification of proteinaceous pheromone in social insects. Researchers have focused almost exclusively on hydrocarbons when searching for recognition pheromones in social insects. The present finding of a proteinaceous pheromone represents a major step forward in, and result in the broadening of, the search for recognition pheromones. This novel function of lysozyme as a termite pheromone illuminates the profound influence

Molecular dynamics simulations of lysozyme in water/sugar solutions

Energy Technology Data Exchange (ETDEWEB)

Lerbret, A. [Department of Food Science, Cornell University, 101 Stocking Hall, Ithaca, NY 14853 (United States); Affouard, F. [Laboratoire de Dynamique et Structure des Materiaux Moleculaires, UMR CNRS 8024, Universite Lille I, 59655 Villeneuve d' Ascq Cedex (France)], E-mail: frederic.affouard@univ-lille1.fr; Bordat, P. [Laboratoire de Chimie Theorique et de Physico-Chimie Moleculaire, UMR 5624, Universite de Pau et des Pays de l' Adour, 64000 Pau (France); Hedoux, A.; Guinet, Y.; Descamps, M. [Laboratoire de Dynamique et Structure des Materiaux Moleculaires, UMR CNRS 8024, Universite Lille I, 59655 Villeneuve d' Ascq Cedex (France)

2008-04-18

Structural and dynamical properties of the solvent at the protein/solvent interface have been investigated by molecular dynamics simulations of lysozyme in trehalose, maltose and sucrose solutions. Results are discussed in the framework of the bioprotection phenomena. The analysis of the relative concentration of water oxygen atoms around lysozyme suggests that lysozyme is preferentially hydrated. When comparing the three sugars, trehalose is seen more excluded than maltose and sucrose. The preferential exclusion of sugars from the protein surface induces some differences in the behavior of trehalose and maltose, particularly at 50 and 60 wt% concentrations, that are not observed experimentally in binary sugar/mixtures. The dynamical slowing down of the solvent is suggested to mainly arise from the homogeneity of the water/sugar matrices controlled by the percolation of the sugar hydrogen bonds networks. Furthermore, lysozyme strongly increases relaxation times of solvent molecules at the protein/solvent interface.

Interaction of anthraquinone dyes with lysozyme: Evidences from spectroscopic and docking studies

International Nuclear Information System (INIS)

Paramaguru, G.; Kathiravan, A.; Selvaraj, S.; Venuvanalingam, P.; Renganathan, R.

2010-01-01

The interaction between lysozyme and anthraquinone dyes such as Alizarin Red S, Acid blue 129 and Uniblue was studied using steady state, time resolved fluorescence measurements and docking studies. Addition of anthraquinone dyes effectively quenched the intrinsic fluorescence of lysozyme. Fluorescence quenching of lysozyme by dyes has revealed the formation of complex. The number of binding sites (n) and binding constant (K) for all the three dyes was calculated by relevant fluorescence quenching data. Based on Foerster's non-radiative energy transfer theory, distance (r 0 ) between the donor (lysozyme) and acceptor (dyes) as well as the critical energy transfer distance (R 0 ) has also been calculated. The interaction between dyes and lysozyme occurs through static quenching mechanism as confirmed by time resolved spectroscopy. The conformational change of lysozyme has been analyzed using synchronous fluorescence measurement. Finally, docking studies revealed that specific interactions were observed with the residue of Trp 62.

ABCC-JNIH adult health study: Hiroshima. Serum lysozyme determinations, April-June 1961

Energy Technology Data Exchange (ETDEWEB)

Finch, S C; Lamphere, J P; Jablon, S

1961-01-01

Serum lysozyme levels were determined on 670 consecutive subjects seen for regularly scheduled clinic examinations of the Adult Health Study in Hiroshima. Serum lysozyme levels were found to vary significantly with the absolute peripheral granulocyte count, age, sex, and month of study. A high level of correlation also was noted between serum lysozyme and diabetes mellitus. This was at least in part attributable to greater average age in patients with diabetes. A suggestive relationship was established between serum lysozyme levels, respiratory diseases, and tuberculosis. These changes are believed to reflect active inflammation with excessive destruction of granulocytes and parenchymal tissues in those patients with the more acute processes. No relationship was found between serum lysozyme and previous exposure to ionizing radiation. These studies indicate that the serum lysozyme level may be useful in the study of the kinetics of leukopoiesis, the aging process, and in the detection of subtle inflammatory processes.

Influence of lysozyme complexation with purified Aldrich humic acid on lysozyme activity

NARCIS (Netherlands)

Li, Y.; Tan, W.F.; Wang, M.X.; Liu, F.; Weng, L.P.; Norde, W.; Koopal, L.K.

2012-01-01

Humic acid is an important component of dissolved organic matter and in two previous papers it has been shown that purified Aldrich humic acid (PAHA) forms strong complexes with the oppositely charged protein lysozyme (LSZ). The complexation and aggregation of enzymes with humic acids may lead to

Growth and optical microscopy observation of the lysozyme crystals

OpenAIRE

R.Vlokh; L.Marsel; I.Teslyuk; O.G.Vlokh

2001-01-01

The little single lysozyme crystals in the capillary after 15 days of growth process with average size 0.1´0.1´0.16mm3 were obtained. It was shown that lysozyme crystals are optically anisotropical and birefringence along a axis is Dn=(2.2±0.5)´10-3 in visible spectrum region. From the measurements of crystallographic angles follows that on the {001} faces angles equal a=81o, b=99o. On the sexangle faces angles equal e=100o, f=140o and g=120o. On the base of obtained results the lysozyme crys...

Microstructure and thermal and functional properties of biodegradable films produced using zein

Directory of Open Access Journals (Sweden)

Crislene Barbosa de Almeida

2018-03-01

Full Text Available Abstract Research is being conducted in an attempt to produce biodegradable packaging to replace plastic products, thereby reducing solid waste disposal. In this work, zein films were produced from vegetable oils (macadamia, olive and buriti and from pure oleic acid. The surface of zein-based films made using oleic acid has a good lipid distribution. The high content of oleic acid produced a film with the greatest elongation at break (8.08 ± 2.71% due to the greater homogeneity of the protein matrix. The different oils did not affect the glass transition temperature (Tg. Tg curves of films with fatty acids showed a reduction in mass at between 50 and 120 °C due to water evaporation. At 120 °C the weight loss was 3-5% and above this temperature further weight loss was observed with the highest loss being seen in the film made using pure oleic acid. In conclusion, although biodegradable films were produced using the four different oils, the film made from pure oleic acid has the best characteristics.

Computational study of aggregation mechanism in human lysozyme[D67H].

Directory of Open Access Journals (Sweden)

Dharmeshkumar Patel

Full Text Available Aggregation of proteins is an undesired phenomena that affects both human health and bioengineered products such as therapeutic proteins. Finding preventative measures could be facilitated by a molecular-level understanding of dimer formation, which is the first step in aggregation. Here we present a molecular dynamics (MD study of dimer formation propensity in human lysozyme and its D67H variant. Because the latter protein aggregates while the former does not, they offer an ideal system for testing the feasibility of the proposed MD approach which comprises three stages: i partially unfolded conformers involved in dimer formation are generated via high-temperature MD simulations, ii potential dimer structures are searched using docking and refined with MD, iii free energy calculations are performed to find the most stable dimer structure. Our results provide a detailed explanation for how a single mutation (D67H turns human lysozyme from non-aggregating to an aggregating protein. Conversely, the proposed method can be used to identify the residues causing aggregation in a protein, which can be mutated to prevent it.

Lysozyme-immobilized electrospun PAMA/PVA and PSSA-MA/PVA ion-exchange nanofiber for wound healing.

Science.gov (United States)

Tonglairoum, Prasopchai; Ngawhirunpat, Tanasait; Rojanarata, Theerasak; Opanasopit, Praneet

2014-08-27

Abstract This research was aimed to develop the lysozyme immobilized ion-exchange nanofiber mats for wound healing. To promote the healing process, the PSSA-MA/PVA and PAMA ion-exchange nanofiber mats were fabricated to mimic the extracellular matrix structure using electrospinning process followed by thermally crosslinked. Lysozyme was immobilized on the ion-exchane nanofibers by an adsorption method. The ion-exchange nanofibers were investigated using SEM, FTIR and XRPD. Moreover, the lysozyme-immobilized ion-exchange nanofibers were further investigated for lysozyme content and activity, lysozyme release and wound healing activity. The fiber diameters of the mats were in the nanometer range. Lysozyme was gradually absorbed into the PSSA-MA/PVA nanofiber with higher extend than that is absorbed on the PAMA/PVA nanofiber and exhibited higher activity than lysozyme-immobilized PAMA/PVA nanofiber. The total contents of lysozyme on the PSSA-MA/PVA and PAMA/PVA nanofiber were 648 and 166 µg/g, respectively. FTIR and lysozyme activity results confirmed the presence of lysozyme on the nanofiber mats. The lysozyme was released from the PSSA-MA/PVA and PAMA/PVA nanofiber in the same manner. The lysozyme-immobilized PSSA-MA/PVA nanofiber mats and lysozyme-immobilized PAMA/PVA nanofiber mats exhibited significantly faster healing rate than gauze and similar to the commercial antibacterial gauze dressing. These results suggest that these nanofiber mats could provide the promising candidate for wound healing application.

Interaction of anthraquinone dyes with lysozyme: Evidences from spectroscopic and docking studies

Energy Technology Data Exchange (ETDEWEB)

Paramaguru, G.; Kathiravan, A.; Selvaraj, S.; Venuvanalingam, P. [School of Chemistry, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu (India); Renganathan, R., E-mail: rrengas@gmail.com [School of Chemistry, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu (India)

2010-03-15

The interaction between lysozyme and anthraquinone dyes such as Alizarin Red S, Acid blue 129 and Uniblue was studied using steady state, time resolved fluorescence measurements and docking studies. Addition of anthraquinone dyes effectively quenched the intrinsic fluorescence of lysozyme. Fluorescence quenching of lysozyme by dyes has revealed the formation of complex. The number of binding sites (n) and binding constant (K) for all the three dyes was calculated by relevant fluorescence quenching data. Based on Foerster's non-radiative energy transfer theory, distance (r{sub 0}) between the donor (lysozyme) and acceptor (dyes) as well as the critical energy transfer distance (R{sub 0}) has also been calculated. The interaction between dyes and lysozyme occurs through static quenching mechanism as confirmed by time resolved spectroscopy. The conformational change of lysozyme has been analyzed using synchronous fluorescence measurement. Finally, docking studies revealed that specific interactions were observed with the residue of Trp 62.

Immunological response in egg-sensitive adults challenged with cheese containing or not containing lysozyme.

Science.gov (United States)

Rossi, Filippo; Iaconelli, Amerigo; Fiorentini, Lucia; Zito, Francesco; Donati, Maria Benedett; De Cristofaro, Maria Laura; Piva, Gianfranco; Mingrone, Geltrude

2012-12-01

Lysozyme is an enzyme that hydrolyzes bacterial peptidoglicans. For this reason, it is used in cheese manufacturing in order to prevent a defect of long-ripened hard cheese called "late blowing" due to the outgrowth of spores of Clostridium tyrobutyricum and Clostridium butyricum. Moreover, germination of Listeria monocytogenes spores into vegetative cells is also sensitive to lysozyme. The enzyme can be an allergenic molecule, and for this reason there are concerns about its use in food industry. The immunological and clinical response of consumption of lysozyme-containing cheese has been evaluated in 25 egg-sensitive subjects with or without lysozyme sensitization. A total of 25 egg-sensitive subjects were enrolled in this study. All the subjects were already treated for egg-sensitization and presented a positive skin prick test. All the subjects had a body mass index ≤ 25 kg/m(2) and were in the age range of 20-50 years. Each subject was studied twice and received randomly 30 g of Grana Padano (containing lysozyme) or TrentinGrana cheese (lysozyme-free) of two different aging periods: 16 or 24 months. A washout period of 1 week between each cheese intake was adopted. Blood samples were taken in fasting conditions and 1 hour after cheese intake and IgA, total IgE, and lysozyme-, ovomucoid-, and ovalbumin-specific IgE were measured. No adverse reactions were observed in both groups of patients after cheese samples were given. Lysozyme did not determine any variation of specific IgE compared with basal level. In lysozyme-sensitive patients a significant relationship between IgA and lysozyme-specific IgE was observed when lysozyme-containing cheese was given, confirming that lysozyme can pass the gut barrier. Neither adverse events nor immunological responses were observed after ingestion of cheese containing lysozyme. However, the immunological properties of peptides deriving from cheese protein hydrolysis need to be clarified, as does the effect of lysozyme on

Immobilization of lysozyme-cellulose amide-linked conjugates on cellulose i and ii cotton nanocrystalline preparations

Science.gov (United States)

Lysozyme was attached through an amide linkage between some of the protein’s aspartate and glutamate residues to amino-glycine-cellulose (AGC), which was prepared by esterification of glycine to preparations of cotton nanocrystals (CNC). The nanocrystalline preparations were produced through acid h...

Adsorption of lysozyme unto silica and polystyrene surfaces in ...

African Journals Online (AJOL)

The adsorption capacity of lysozyme (chicken egg white) from aqueous solutions unto silica and polystyrene interfaces was studied at varying lysozyme concentrations and ionic strength. The studies revealed an increase in adsorption capacity with increase in concentration and with maximum adsorption densities of 1.34 ...

Antimicrobial activity of lysozyme with special relevance to milk

African Journals Online (AJOL)

STORAGESEVER

2008-12-29

Dec 29, 2008 ... Lysozyme is among the minor milk proteins that has attracted increased .... while there is a general agreement that surface attachment polymers and ..... and form aggregates as a result of electrostatic and hydrophobic ...... conformational changes and antimicrobial activity of lysozyme upon reduction of its ...

Dielectric and gravimetric studies of water binding to lysozyme

International Nuclear Information System (INIS)

Bone, S.

1996-01-01

Time domain dielectric spectroscopy and hydration isotherm measurements as a function of temperature have been applied to hydrated lysozyme powder. Two dielectric dispersions were identified, the first centred at approximately 8 MHz and a second above 1 GHz. The higher dispersion is considered to be the result of rotational relaxation of water molecules bound to the enzyme. In this case the results indicate the existence of a population of 32 water molecules per lysozyme molecule which are irrotationally bound to the lysozyme structure. A larger population of water molecules is relatively free to respond to the electric field and exhibits a dipole moment close to that of vapour phase water molecules. Multi-temperature hydration isotherm measurements are used to calculate enthalpies and entropies associated with the binding of water to lysozyme. Discontinuities both in dielectric and in thermodynamic characteristics in the range 10-14% hydration are interpreted as a re-ordering of the water structure on the enzyme surface

Antimicrobial activity of lysozyme with special relevance to milk ...

African Journals Online (AJOL)

This review discusses the antimicrobial activity of lysozyme with special emphasis on milk's lysozyme, and attempts to shed some light on the recent advances elucidating the mechanism of its antimicrobial activity against sensitive microorganisms as well as the means used by some bacteria to resist such an activity.

Complex nature of serum lysozyme activity: evidence of thermolability in inflammatory bowel disease

Energy Technology Data Exchange (ETDEWEB)

Ward, M; Mitchell, W D; Eastwood, M [Western General Hospital, Edinburgh (UK)

1978-01-01

In patients with Crohn's disease and ulcerative colitis, alterations in serum storage temperature produced significant changes in serum lysozyme activity (SLA) as measured by the lysoplate method. This was not the case in healthy controls or in a group with other gastrointestinal disorders. Electrophoretic separation of serum revealed two components of lysozyme-type lytic activity but only one in extracts of gut mucosa, leucocytes, and egg white. The major lytic component of serum migrated towards the cathode and reacted with specific antilysozyme serum, but the minor component which migrated towards the anode did not. Although the cause of this anionic lytic activity is uncertain, it contributes to total serum activity as estimated by any method utilizing the lysis of Micrococcus lysodeikticus, and may possibly be related to the observed thermolability.

Method for producing fluorinated diamond-like carbon films

Science.gov (United States)

Hakovirta, Marko J.; Nastasi, Michael A.; Lee, Deok-Hyung; He, Xiao-Ming

2003-06-03

Fluorinated, diamond-like carbon (F-DLC) films are produced by a pulsed, glow-discharge plasma immersion ion processing procedure. The pulsed, glow-discharge plasma was generated at a pressure of 1 Pa from an acetylene (C.sub.2 H.sub.2) and hexafluoroethane (C.sub.2 F.sub.6) gas mixture, and the fluorinated, diamond-like carbon films were deposited on silicon substrates. The film hardness and wear resistance were found to be strongly dependent on the fluorine content incorporated into the coatings. The hardness of the F-DLC films was found to decrease considerably when the fluorine content in the coatings reached about 20%. The contact angle of water on the F-DLC coatings was found to increase with increasing film fluorine content and to saturate at a level characteristic of polytetrafluoroethylene.

Growth of thin films of low molecular weight proteins by matrix assisted pulsed laser evaporation (MAPLE)

DEFF Research Database (Denmark)

Matei, Andreea; Schou, Jørgen; Constantinescu, C.

2011-01-01

Thin films of lysozyme and myoglobin grown by matrix assisted pulsed laser evaporation (MAPLE) from a water ice matrix have been investigated. The deposition rate of these two low molecular weight proteins (lysozyme: 14307 amu and myoglobin: 17083 amu) exhibits a maximum of about 1–2 ng/cm2 per....... The results for lysozyme demonstrate that the fragmentation rate of the proteins during the MAPLE process is not influenced by the pH of the water solution prior to freezing....

Optical and magnetic properties of porous graphene films produced by electrospraying

International Nuclear Information System (INIS)

Zhao, Jun; Yang, Shan-Shan; Chen, Li-Qing; Zhang, Zhao-Chun; Zheng, Hou-Li

2013-01-01

Graphene films have been produced by electrospraying on SiO 2 -coated silicon substrate and subsequent heat treatment, offering a simple and typical method to produce porous graphene films and exhibiting a good adhesion to silicon substrate. The microstructures of as-prepared graphene films were characterized by field emission scanning electron microscopy, transmission electron microscopy, selected area electron diffraction and atomic force microscopy. X-ray photoelectron spectroscopy, infrared spectroscopy and Raman spectroscopy further confirmed the formation of porous graphene films. Moreover, the reflection spectrum of as-prepared graphene films was studied by ultraviolet–visible spectroscopy, revealing that light absorption played dominant roles at 375 and 635 nm, respectively. Finally, the resistance and magnetoresistance were measured, and some preliminary theoretical explanations were proposed. - Highlights: ► Porous graphene films were produced by electrospraying. ► Light absorption plays dominant roles at 375 and 635 nm. ► A negative magnetoresistance is emerged at low temperature. ► A 2D weak localization effect arises from random stacking of graphene

ISOLATION AND PURIFICATION OF LYSOZYME FROM THE HEN EGG WHITE

Directory of Open Access Journals (Sweden)

S. S.

2015-12-01

Full Text Available The aim of the research was the development of the method of lysozyme isolation from hen egg proteins. Lysozyme was isolated by differential heat denaturation of proteins with changing of the medium pH value, followed by neutralization, dialysis and additional purification by gel chromatography on Sephadex G-50. Activity was determined by bacteriolytic method (with Micrococcus lysodeikticus 4698 as a substrate. The enzyme purity and molecular mass were determined using SDS-electrophoresis and massspectrometry. The method of lysozyme isolation from hen egg proteins with the enzyme yield of 3.2 ± 0.2% and bacteriolytic activity of 22 025 ± 1 500 U/mg is modified. According to electrophoresis data, the isolated enzyme is characterized by high degree of purity (~95–98% and is comparable with lysozyme of AppliChem company by main physical and chemical characteristics. The obtaining product is stored in a crystalline form at low temperature (–24 оC for 9 months. The proposed method allows obtaining active and stable lysozyme with high purity from hen egg protein in laboratory conditions for the usage in biotechnology.

A study of the interaction between malachite green and lysozyme by steady-state fluorescence.

Science.gov (United States)

Ding, Fei; Liu, Wei; Liu, Feng; Li, Zhi-Yuan; Sun, Ying

2009-09-01

The interaction of a N-methylated diaminotriphenylmethane dye, malachite green, with lysozyme was investigated by fluorescence spectroscopic techniques under physiological conditions. The binding parameters have been evaluated by fluorescence quenching methods. The results revealed that malachite green caused the fluorescence quenching of lysozyme through a static quenching procedure. The thermodynamic parameters like DeltaH and DeltaS were calculated to be -15.33 kJ mol(-1) and 19.47 J mol(-1) K(-1) according to van't Hoff equation, respectively, which proves main interaction between malachite green and lysozyme is hydrophobic forces and hydrogen bond contact. The distance r between donor (lysozyme) and acceptor (malachite green) was obtained to be 3.82 nm according to Frster's theory. The results of synchronous fluorescence, UV/vis and three-dimensional fluorescence spectra showed that binding of malachite green with lysozyme can induce conformational changes in lysozyme. In addition, the effects of common ions on the constants of lysozyme-malachite green complex were also discussed.

Immobilization of lysozyme on cotton fabrics; synthesis, characterication, and activity

Science.gov (United States)

The antimicrobial activity of lysozyme derives from the hydrolysis of the bacterial cell wall polysaccharide at the glycosidic bond that links N-acetyl-glucosamine and N-acetyl-muramic acid. Maintaining the activity of lysozyme while bound to a cellulose substrate is a goal toward developing enzyme...

Probabilistic approach to lysozyme crystal nucleation kinetics.

Science.gov (United States)

Dimitrov, Ivaylo L; Hodzhaoglu, Feyzim V; Koleva, Dobryana P

2015-09-01

Nucleation of lysozyme crystals in quiescent solutions at a regime of progressive nucleation is investigated under an optical microscope at conditions of constant supersaturation. A method based on the stochastic nature of crystal nucleation and using discrete time sampling of small solution volumes for the presence or absence of detectable crystals is developed. It allows probabilities for crystal detection to be experimentally estimated. One hundred single samplings were used for each probability determination for 18 time intervals and six lysozyme concentrations. Fitting of a particular probability function to experimentally obtained data made possible the direct evaluation of stationary rates for lysozyme crystal nucleation, the time for growth of supernuclei to a detectable size and probability distribution of nucleation times. Obtained stationary nucleation rates were then used for the calculation of other nucleation parameters, such as the kinetic nucleation factor, nucleus size, work for nucleus formation and effective specific surface energy of the nucleus. The experimental method itself is simple and adaptable and can be used for crystal nucleation studies of arbitrary soluble substances with known solubility at particular solution conditions.

Analysis of two lysozyme genes and antimicrobial functions of their recombinant proteins in Asian seabass.

Directory of Open Access Journals (Sweden)

Gui Hong Fu

Full Text Available Lysozymes are important proteins of the innate immune system for the defense against bacterial infection. We cloned and analyzed chicken-type (c-type and goose-type (g-type lysozymes from Asian seabass (Lates calcarifer. The deduced amino acid sequence of the c-type lysozyme contained 144 residues and possessed typical structure residues, conserved catalytic residues (Glu(50 and Asp(67 and a "GSTDYGIFQINS" motif. The deduced g-type lysozyme contained 187 residues and possessed a goose egg white lysozyme (GEWL domain containing three conserved catalytic residues (Glu(71, Asp(84, Asp(95 essential for catalytic activity. Real time quantitative PCR (qRT-PCR revealed that the two lysozyme genes were constitutively expressed in all the examined tissues. The c-type lysozyme was most abundant in liver, while the g-type lysozyme was predominantly expressed in intestine and weakly expressed in muscle. The c-type and g-type transcripts were up-regulated in the kidney, spleen and liver in response to a challenge with Vibrio harveyi. The up-regulation of the c-type lysozyme was much stronger than that of the g-type lysozyme in kidney and spleen. The recombinant proteins of the c-type and g-type lysozymes showed lytic activities against the bacterial pathogens Vibrio harveyi and Photobacterium damselae in a dosage-dependent manner. We identified single nucleotide polymorphisms (SNPs in the two lysozyme genes. There were significant associations of these polymorphisms with resistance to the big belly disease. These results suggest that the c- and g-type genes play an important role in resistance to bacterial pathogens in fish. The SNP markers in the two genes associated with the resistance to bacterial pathogens may facilitate the selection of Asian seabass resistant to bacterial diseases.

Carbon films produced from ionic liquid carbon precursors

Science.gov (United States)

Dai, Sheng; Luo, Huimin; Lee, Je Seung

2013-11-05

The invention is directed to a method for producing a film of porous carbon, the method comprising carbonizing a film of an ionic liquid, wherein the ionic liquid has the general formula (X.sup.+a).sub.x(Y.sup.-b).sub.y, wherein the variables a and b are, independently, non-zero integers, and the subscript variables x and y are, independently, non-zero integers, such that ax=by, and at least one of X.sup.+ and Y.sup.- possesses at least one carbon-nitrogen unsaturated bond. The invention is also directed to a composition comprising a porous carbon film possessing a nitrogen content of at least 10 atom %.

Binding of Lysozyme to Spherical Poly(styrenesulfonate Gels

Directory of Open Access Journals (Sweden)

Martin Andersson

2018-01-01

Full Text Available Polyelectrolyte gels are useful as carriers of proteins and other biomacromolecules in, e.g., drug delivery. The rational design of such systems requires knowledge about how the binding and release are affected by electrostatic and hydrophobic interactions between the components. To this end we have investigated the uptake of lysozyme by weakly crosslinked spherical poly(styrenesulfonate (PSS microgels and macrogels by means of micromanipulator assisted light microscopy and small angle X-ray scattering (SAXS in an aqueous environment. The results show that the binding process is an order of magnitude slower than for cytochrome c and for lysozyme binding to sodium polyacrylate gels under the same conditions. This is attributed to the formation of very dense protein-rich shells in the outer layers of the microgels with low permeability to the protein. The shells in macrogels contain 60 wt % water and nearly charge stoichiometric amounts of lysozyme and PSS in the form of dense complexes of radius 8 nm comprising 30–60 lysozyme molecules. With support from kinetic modelling results we propose that the rate of protein binding and the relaxation rate of the microgel are controlled by the protein mass transport through the shell, which is strongly affected by hydrophobic and electrostatic interactions. The mechanism explains, in turn, an observed dependence of the diffusion rate on the apparent degree of crosslinking of the networks.

Renaturation of reduced hen egg white lysozyme containing two blocked sulfhydryl groups

Energy Technology Data Exchange (ETDEWEB)

Acharya, A.S.; Taniuchi, H.

1980-10-01

Formation of native lysozyme from the reduced form involves many pathways in two processes: incorrect pairing of half-cystine residues by oxidation and rearrangement of disulfide (SS) bonds. The energy barrier against suflhydryl (SH)-disulfide interchange of the native or nativelike species thus formed causes accumulation of these species. For example, the enzymatically active isomers containing three presumably native SS bonds and one open SS bond may be thermodynamically favorable over the nonnative isomers and can be formed from reduced lysozyme or lysozyme containing scrambled SS bonds by nonobligatory and flexible pathways. As an extension of these observations formation of nativelike species from reduced lysozyme containing the average of two carboxymethyl (CM)-cysteine was investigated.

New techniques for producing thin boron films

International Nuclear Information System (INIS)

Thomas, G.E.

1988-01-01

A review will be presented of methods for producing thin boron films using an electron gun. Previous papers have had the problem of spattering of the boron source during the evaporation. Methods for reducing this problem will also be presented. 12 refs., 4 figs

Isolation and characterization of a c-type lysozyme from the nurse shark.

Science.gov (United States)

Hinds Vaughan, Nichole; Smith, Sylvia L

2013-12-01

Lysozyme is a ubiquitous antibacterial enzyme that occurs in numerous invertebrate and vertebrate species. Three forms have been described c-type, g-type and i-type which differ in primary structure. Shark lysozyme has not been characterized; here we report on the isolation and characterization of lysozyme from unstimulated shark (Ginglymostoma cirratum) leukocytes and provide amino acid sequence data across the highly conserved active site of the molecule identifying it to be a c-type lysozyme. A leukocyte lysate was applied either (a) to the first of two sequential DE-52 cellulose columns or alternatively, (b) to a DEAE-Sepharose column. Lysozyme activity in lysate and active fractions was identified by zones of lysis of Micrococcus lysodeikticus cell walls on lysoplates and zones of growth inhibition in agar diffusion assays using Planococcus citreus as the target organism. SDS-PAGE analysis revealed a 14 kDa protein which was identified as lysozyme by mass spectroscopic analysis of peptides, reactivity against anti-HEWL antibodies on a Western blot, hydrolysis of M. lysodeikticus cell walls, and inhibition of growth of P. citreus on AU-gel blots in which the area of growth inhibition correlated to a 14 kDa protein. Copyright © 2013 Elsevier Ltd. All rights reserved.

A novel C-type lysozyme from Mytilus galloprovincialis: insight into innate immunity and molecular evolution of invertebrate C-type lysozymes.

Directory of Open Access Journals (Sweden)

Qing Wang

Full Text Available A c-type lysozyme (named as MgCLYZ gene was cloned from the mussel Mytilus galloprovincialis. Blast analysis indicated that MgCLYZ was a salivary c-type lysozyme which was mainly found in insects. The nucleotide sequence of MgCLYZ was predicted to encode a polypeptide of 154 amino acid residues with the signal peptide comprising the first 24 residues. The deduced mature peptide of MgCLYZ was of a calculated molecular weight of 14.4 kD and a theoretical isoelectric point (pI of 8.08. Evolution analysis suggested that bivalve branch of the invertebrate c-type lysozymes phylogeny tree underwent positive selection during evolution. By quantitative real-time RT-PCR (qRT-PCR analysis, MgCLYZ transcript was widely detected in all examined tissues and responded sensitively to bacterial challenge in hemocytes and hepatopancreas. The optimal temperature and pH of recombinant MgCLYZ (rMgCLYZ were 20°C and 4, respectively. The rMgCLYZ displayed lytic activities against Gram-positive bacteria including Micrococcus luteus and Staphyloccocus aureus, and Gram-negative bacteria including Vibrio anguillarum, Enterobacter cloacae, Pseudomonas putida, Proteus mirabilis and Bacillus aquimaris. These results suggest that MgCLYZ perhaps play an important role in innate immunity of M. galloprovincialis, and invertebrate c-type lysozymes might be under positive selection in a species-specific manner during evolution for undergoing adaptation to different environment and diverse pathogens.

Anisotropy of the Coulomb Interaction between Folded Proteins: Consequences for Mesoscopic Aggregation of Lysozyme

Science.gov (United States)

Chan, Ho Yin; Lankevich, Vladimir; Vekilov, Peter G.; Lubchenko, Vassiliy

2012-01-01

Toward quantitative description of protein aggregation, we develop a computationally efficient method to evaluate the potential of mean force between two folded protein molecules that allows for complete sampling of their mutual orientation. Our model is valid at moderate ionic strengths and accounts for the actual charge distribution on the surface of the molecules, the dielectric discontinuity at the protein-solvent interface, and the possibility of protonation or deprotonation of surface residues induced by the electric field due to the other protein molecule. We apply the model to the protein lysozyme, whose solutions exhibit both mesoscopic clusters of protein-rich liquid and liquid-liquid separation; the former requires that protein form complexes with typical lifetimes of approximately milliseconds. We find the electrostatic repulsion is typically lower than the prediction of the Derjaguin-Landau-Verwey-Overbeek theory. The Coulomb interaction in the lowest-energy docking configuration is nonrepulsive, despite the high positive charge on the molecules. Typical docking configurations barely involve protonation or deprotonation of surface residues. The obtained potential of mean force between folded lysozyme molecules is consistent with the location of the liquid-liquid coexistence, but produces dimers that are too short-lived for clusters to exist, suggesting lysozyme undergoes conformational changes during cluster formation. PMID:22768950

The Preventive Effect of L-Lysine on Lysozyme Glycation in Type 2 Diabetes

Directory of Open Access Journals (Sweden)

Hossein Mirmiranpour

2016-01-01

Full Text Available Lysozyme is a bactericidal enzyme whose structure and functions change in diabetes. Chemical chaperones are small molecules including polyamines (e.g. spermine, amino acids (e.g. L-lysine and polyols (e.g. glycerol. They can improve protein conformation in several stressful conditions such as glycation. In this study, the authors aimed to observe the effect of L-lysine as a chemical chaperone on structure and function of glycated lysozyme. In this study, in vitro and in vivo effects of L-lysine on lysozyme glycation were investigated. Lysozyme was incubated with glucose and/or L-lysine, followed by an investigation of its structure by electrophoresis, fluorescence spectroscopy, and circular dichroism spectroscopy and also assessment of its bactericidal activity against M. lysodeikticus. In the clinical trial, patients with type 2 diabetes mellitus (T2DM were randomly divided into two groups of 25 (test and control. All patients received metformin and glibenclamide for a three months period. The test group was supplemented with 3 g/day of L-lysine. The quantity and activity of lysozyme and other parameters were then measured. Among the test group, L-lysine was found to reduce the advanced glycation end products (AGEs in the sera of patients with T2DM and in vitro condition. This chemical chaperone reversed the alteration in lysozyme structure and function due to glycation and resulted in increased lysozyme activity. Structure and function of glycated lysozyme are significantly improved by l-lysine; therefore it can be considered an effective therapeutic supplementation in T2DM, decreasing the risk of infection in these patients.

Crystallization, data collection and phasing of two digestive lysozymes from Musca domestica

Energy Technology Data Exchange (ETDEWEB)

Marana, S. R.; Cançado, F. C. [Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo (Brazil); Valério, A. A. [Centro de Biologia Molecular e Estrutural (CeBiMe), Laboratório Nacional de Luz Síncrotron (LNLS), CP 6192, Campinas, SP 13084-971 (Brazil); Departamento de Bioquímica, Universidade Estadual de Campinas, Campinas (Brazil); Ferreira, C.; Terra, W. R. [Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo (Brazil); Barbosa, J. A. R. G., E-mail: joao@lnls.br [Centro de Biologia Molecular e Estrutural (CeBiMe), Laboratório Nacional de Luz Síncrotron (LNLS), CP 6192, Campinas, SP 13084-971 (Brazil); Departamento de Bioquímica, Universidade Estadual de Campinas, Campinas (Brazil); Departamento de Genética e Evolução, Universidade Estadual de Campinas, Campinas (Brazil); Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo (Brazil)

2006-08-01

The digestive lysozymes 1 and 2 from M. domestica were crystallized by vapour diffusion. The crystallographic data were processed to a maximum resolution of 1.9 Å in both cases. Lysozymes are mostly known for their defensive role against bacteria, but in several animals lysozymes have a digestive function. Here, the initial crystallographic characterization of two digestive lysozymes from Musca domestica are presented. The proteins were crystallized using the sitting-drop vapour-diffusion method in the presence of ammonium sulfate or PEG/2-propanol as the precipitant. X-ray diffraction data were collected to a maximum resolution of 1.9 Å using synchrotron radiation. The lysozyme 1 and 2 crystals belong to the monoclinic space group P2{sub 1} (unit-cell parameters a = 36.52, b = 79.44, c = 45.20 Å, β = 102.97°) and the orthorhombic space group P2{sub 1}2{sub 1}2 (unit-cell parameters a = 73.90, b = 96.40, c = 33.27 Å), respectively. The crystal structures were solved by molecular replacement and structure refinement is in progress.

A zinc complex of heparan sulfate destabilises lysozyme and alters its conformation

International Nuclear Information System (INIS)

Hughes, Ashley J.; Hussain, Rohanah; Cosentino, Cesare; Guerrini, Marco; Siligardi, Giuliano; Yates, Edwin A.; Rudd, Timothy R.

2012-01-01

Highlights: ► Zinc–heparan sulfate complex destabilises lysozyme, a model amyloid protein. ► Addition of zinc, without heparan sulfate, stabilises lysozyme. ► Heparan sulfate cation complexes provide alternative protein folding routes. -- Abstract: The naturally occurring anionic cell surface polysaccharide heparan sulfate is involved in key biological activities and is implicated in amyloid formation. Following addition of Zn–heparan sulfate, hen lysozyme, a model amyloid forming protein, resembled β-rich amyloid by far UV circular dichroism (increased β-sheet: +25%), with a significantly reduced melting temperature (from 68 to 58 °C) by fluorescence shift assay. Secondary structure stability of the Zn–heparan sulfate complex with lysozyme was also distinct from that with heparan sulfate, under stronger denaturation conditions using synchrotron radiation circular dichroism. Changing the cation associated with heparan sulfate is sufficient to alter the conformation and stability of complexes formed between heparan sulfate and lysozyme, substantially reducing the stability of the protein. Complexes of heparan sulfate and cations, such as Zn, which are abundant in the brain, may provide alternative folding routes for proteins.

Control of Electrostatic Interactions Between F-Actin And Genetically Modified Lysozyme in Aqueous Media

International Nuclear Information System (INIS)

Sanders, L.K.; Xian, W.; Guaqueta, C.; Strohman, M.; Vrasich, C.R.; Luijten, E.; Wong, G.C.L.

2009-01-01

The aim for deterministic control of the interactions between macroions in aqueous media has motivated widespread experimental and theoretical work. Although it has been well established that like-charged macromolecules can aggregate under the influence of oppositely charged condensing agents, the specific conditions for the stability of such aggregates can only be determined empirically. We examine these conditions, which involve an interplay of electrostatic and osmotic effects, by using a well defined model system composed of F-actin, an anionic rod-like polyelectrolyte, and lysozyme, a cationic globular protein with a charge that can be genetically modified. The structure and stability of actin-lysozyme complexes for different lysozyme charge mutants and salt concentrations are examined by using synchrotron x-ray scattering and molecular dynamics simulations. We provide evidence that supports a structural transition from columnar arrangements of F-actin held together by arrays of lysozyme at the threefold interstitial sites of the actin sublattice to marginally stable complexes in which lysozyme resides at twofold bridging sites between actin. The reduced stability arises from strongly reduced partitioning of salt between the complex and the surrounding solution. Changes in the stability of actin-lysozyme complexes are of biomedical interest because their formation has been reported to contribute to the persistence of airway infections in cystic fibrosis by sequestering antimicrobials such as lysozyme. We present x-ray microscopy results that argue for the existence of actin-lysozyme complexes in cystic fibrosis sputum and demonstrate that, for a wide range of salt conditions, charge-reduced lysozyme is not sequestered in ordered complexes while retaining its bacterial killing activity.

The interaction of the protein lysozyme with bacteria E. coli observed using nanodiamond labelling

International Nuclear Information System (INIS)

Perevedentseva, Elena; Cheng, C-Y; Chung, P-H; Tu, J-S; Hsieh, Y-H; Cheng, C-L

2007-01-01

The application of a nanometre-sized diamond in Raman-detectable biolabelling is demonstrated in this study. The interaction of a lysozyme-nanodiamond complex with bacteria E. coli was observed via Raman mapping using the diamond Raman signal as the labelling marker. The results are compared with scanning electron microscope observations, and the adsorbed lysozyme's functionality is analysed. High antibacterial activity of lysozyme-nanodiamond complex was observed, equivalent to active lysozyme in solution. The results suggest that nanodiamond labelling can be effective and that it can be applied in ambient conditions without complicated sample pre-treatments

Surface Plasmon Resonance based sensing of lysozyme in serum on Micrococcus lysodeikticus-modified graphene oxide surfaces.

Science.gov (United States)

Vasilescu, Alina; Gáspár, Szilveszter; Gheorghiu, Mihaela; David, Sorin; Dinca, Valentina; Peteu, Serban; Wang, Qian; Li, Musen; Boukherroub, Rabah; Szunerits, Sabine

2017-03-15

Lysozyme is an enzyme found in biological fluids, which is upregulated in leukemia, renal diseases as well as in a number of inflammatory gastrointestinal diseases. We present here the development of a novel lysozyme sensing concept based on the use of Micrococcus lysodeikticus whole cells adsorbed on graphene oxide (GO)-coated Surface Plasmon Resonance (SPR) interfaces. M. lysodeikticus is a typical enzymatic substrate for lysozyme. Unlike previously reported sensors which are based on the detection of lysozyme through bioaffinity interactions, the bioactivity of lysozyme will be used here for sensing purposes. Upon exposure to lysozyme containing serum, the integrity of the bacterial cell wall is affected and the cells detach from the GO based interfaces, causing a characteristic decrease in the SPR signal. This allows sensing the presence of clinically relevant concentrations of lysozyme in undiluted serum samples. Copyright © 2016 Elsevier B.V. All rights reserved.

Interaction of Lysozyme with Rhodamine B: A combined analysis of spectroscopic & molecular docking.

Science.gov (United States)

Millan, Sabera; Satish, Lakkoji; Kesh, Sandeep; Chaudhary, Yatendra S; Sahoo, Harekrushna

2016-09-01

The interaction of Rhodamine B (RB) with Lysozyme (Lys) was investigated by different optical spectroscopic techniques such as absorption, fluorescence, and circular-dichroism (CD), along with molecular docking studies. The fluorescence results (including steady-state and time-resolved mode) revealed that the addition of RB effectively causes strong quenching of intrinsic fluorescence in Lysozyme and mostly, by the static quenching mechanism. Different binding and thermodynamic parameters were calculated at different temperatures and the binding constant value was found to be 2963.54Lmol(-1) at 25°C. The average distance (r0) was found to be 3.31nm according to Förster's theory of non-radiative energy transfer between Lysozyme and RB. The conformational change in Lysozyme during interaction with RB was confirmed from absorbance, synchronous fluorescence, and circular dichroism measurements. Finally, molecular docking studies were done to confirm that the dye binds with Lysozyme. Copyright © 2016 Elsevier B.V. All rights reserved.

The interaction of the protein lysozyme with bacteria E. coli observed using nanodiamond labelling

Energy Technology Data Exchange (ETDEWEB)

Perevedentseva, Elena; Cheng, C-Y; Chung, P-H; Tu, J-S; Hsieh, Y-H; Cheng, C-L [Department of Physics, National Dong Hwa University, Hualien 97401, Taiwan (China)

2007-08-08

The application of a nanometre-sized diamond in Raman-detectable biolabelling is demonstrated in this study. The interaction of a lysozyme-nanodiamond complex with bacteria E. coli was observed via Raman mapping using the diamond Raman signal as the labelling marker. The results are compared with scanning electron microscope observations, and the adsorbed lysozyme's functionality is analysed. High antibacterial activity of lysozyme-nanodiamond complex was observed, equivalent to active lysozyme in solution. The results suggest that nanodiamond labelling can be effective and that it can be applied in ambient conditions without complicated sample pre-treatments.

The influence of different pathogens on the lysozyme activity into tissues of rat oral cavity

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A. P. Levitsky

2017-08-01

Full Text Available Aim: To determine action of the different pathogens on the lysozyme activity into tissues of oral cavity and serum. Methods: The lysozyme activities was determined into oral mucosa cheek, tongue gum and serum of 158 white rats (11 series experiments. The pathogens were used: atropine, protamine sulfat, indometacyn, bee poison, hydrasine sulfat, cytostatic cyclofosfan, lincomycin, lipopolysaccharide, composition of antibiotic and omeprasol for ACBT Results: The  whole of pathogens decreased lysozyme activity (mean in 1,6-2,5 times into oral tissues and on 16 % into serum. The specific lowering of lysozyme activities (Δ%/mg pathogen was low most for lipopolysaccharide, especially after oral application usage (exceeding was in tens times. Conclusion: The lysozyme activity lowering may play significant role in pathogenesis of stomatologic diseases/ Lipopolysaccharide (LPS send lysozyme activity lowering most especially after oral application. Probably, the antilysozyme action of pathogens realize by LPS. The stomatogenic factor in pathogenesis and profilactic of noninfection diseases is important.

Impact of lysozyme on stability mechanism of nanozirconia aqueous suspension

Energy Technology Data Exchange (ETDEWEB)

Szewczuk-Karpisz, Katarzyna, E-mail: k.szewczuk-karpisz@wp.pl; Wiśniewska, Małgorzata

2016-08-30

Highlights: • Adsorption and stabilization-destabilization properties of lysozyme (LSZ) in the nanozirconia-biopolymer solution system were determined. • The stability measurements were performed using turbidimetric method. • Lysozyme macromolecules undergo adsorption on the ZrO{sub 2} surface under electrostatic adsorbent-adsorbate attraction, i.e. at pH 6 and 9. • The biopolymer adsorption impact on the zirconia stability varies at different pH values. - Abstract: The effect of lysozyme (LSZ) presence on the zirconium(IV) oxide (ZrO{sub 2}) aqueous suspension stability was examined. The applied zirconia contains mesopores (with a diameter about 30 nm) and its mean particle size is about 100 nm. To determine the stability mechanism of ZrO{sub 2} suspension in the biopolymer presence, the adsorption and electrokinetic (surface charge density and zeta potential) measurements were performed in the pH range 3–10. The lysozyme adsorption on the nanozirconia surface proceeds mainly through electrostatic forces. Under solid-polymer repulsion conditions, there is no adsorption of lysozyme (pH < 6, C{sub NaCl} 0.01 mol/dm{sup 3}). The increase of solution ionic strength to 0.2 mol/dm{sup 3} causes screening of unfavourable forces and biopolymer adsorption becomes possible. The LSZ addition to the ZrO{sub 2} suspension influences its stability. At pH 3, 4.6 and 7.6, slight improvement of the system stability was obtained. In turn, at pH 9 considerable destabilization of nanozirconia particles covered by polymeric layers occurs.

The dissolution phenomenon of lysozyme crystals

Energy Technology Data Exchange (ETDEWEB)

Mueller, C.; Ulrich, J. [Martin Luther University Halle-Wittenberg, Department of Thermal Separation Processes, Centre of Engineering Science, Halle/Saale (Germany)

2012-02-15

Dissolution studies on lysozyme crystals were carried out since the observed dissolution pattern look different from non-protein dissolved crystals. The Tetragonal, High Temperature and Low Temperature Orthorhombic morphologies, crystallized using sodium chloride, were chosen and the influence of different pH, salt and protein concentration on their dissolution was investigated. An increase in pH and/or salt concentration can modify the dissolution behaviour. The pattern of the crystals during the dissolution process will, therefore, develop differently. Frequently a skeleton like crystal pattern followed by a falling apart of the crystals is observed. The multi-component character of the lysozyme crystal (protein, water, buffer, salt) as well as ''solvatomorphism'' gives first insights in the phenomena happening in the dissolution process. (copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

Pure and Sn-doped ZnO films produced by pulsed laser deposition

DEFF Research Database (Denmark)

Holmelund, E.; Schou, Jørgen; Tougaard, S.

2002-01-01

A new technique, metronome doping, has been used for doping of films during pulsed laser deposition (PLD). This technique makes it possible to dope continuously during film growth with different concentrations of a dopant in one deposition sequence. Films of pure and doped ZnO have been produced...

Hydrophobic nano-carrier for lysozyme adsorption

Indian Academy of Sciences (India)

1 polymer in pH 7.0 phosphate buffer at ... lysozyme concentration, temperature and ionic strength are varied and .... tions at initial and final adsorption medium were measured ... ties such as very high specific surface area, low mass transfer.

Influence of lysozyme on the precipitation of calcium carbonate: a kinetic and morphologic study

Science.gov (United States)

Jimenez-Lopez, Concepcion; Rodriguez-Navarro, Alejandro; Dominguez-Vera, Jose M.; Garcia-Ruiz, Juan M.

2003-05-01

Several mechanisms have been proposed to explain the interactions between proteins and mineral surfaces, among them a combination of electrostatic, stereochemical interactions and molecular recognition between the protein and the crystal surface. To identify the mechanisms of interaction in the lysozyme-calcium carbonate model system, the effect of this protein on the precipitation kinetics and morphology of calcite crystals was examined. The solution chemistry and morphology of the solid were monitored over time in a set of time-series free-drift experiments in which CaCO 3 was precipitated from solution in a closed system at 25°C and 1 atm total pressure, in the presence and absence of lysozyme. The precipitation of calcite was preceded by the precipitation of a metastable phase that later dissolved and gave rise to calcite as the sole phase. With increasing lysozyme concentration, the nucleation of both the metastable phase and calcite occurred at lower Ω calcite, indicating that lysozyme favored the nucleation of both phases. Calcite growth rate was not affected by the presence of lysozyme, at least at protein concentrations ranging from 0 mg/mL to 10 mg/mL. Lysozyme modified the habit of calcite crystals. The degree of habit modification changed with protein concentration. At lower concentrations of lysozyme, the typical rhombohedral habit of calcite crystals was modified by the expression of {110} faces, which resulted from the preferential adsorption of protein on these faces. With increasing lysozyme concentration, the growth of {110}, {100}, and finally {001} faces was sequentially inhibited. This adsorption sequence may be explained by an electrostatic interaction between lysozyme and calcite, in which the inhibition of the growth of {110}, {100}, and {001} faces could be explained by a combined effect of the density of carbonate groups in the calcite face and the specific orientation (perpendicular) of these carbonate groups with respect to the calcite

Fabrication of Sericin/Agrose Gel Loaded Lysozyme and Its Potential in Wound Dressing Application

Directory of Open Access Journals (Sweden)

Meirong Yang

2018-04-01

Full Text Available Sericin is a biomaterial resource for its significant biodegradability, biocompatibility, hydrophilicity, and reactivity. Designing a material with superabsorbent, antiseptic, and non-cytotoxic wound dressing properties is advantageous to reduce wound infection and promote wound healing. Herein, we propose an environment-friendly strategy to obtain an interpenetrating polymer network gel through blending sericin and agarose and freeze-drying. The physicochemical characterizations of the sericin/agarose gel including morphology, porosity, swelling behavior, crystallinity, secondary structure, and thermal property were well characterized. Subsequently, the lysozyme loaded sericin/agarose composite gel was successfully prepared by the solution impregnation method. To evaluate the potential of the lysozyme loaded sericin/agarose gel in wound dressing application, we analyzed the lysozyme loading and release, antimicrobial activity, and cytocompatibility of the resulting gel. The results showed the lysozyme loaded composite gel had high porosity, excellent water absorption property, and good antimicrobial activities against Escherichia coli and Staphylococcus aureus. Also, the lysozyme loaded gel showed excellent cytocompatibility on NIH3T3 and HEK293 cells. So, the lysozyme loaded sericin/agarose gel is a potential alternative biomaterial for wound dressing.

The effects of xylitol and sorbitol on lysozyme- and peroxidase-related enzymatic and candidacidal activities.

Science.gov (United States)

Kim, Bum-Soo; Chang, Ji-Youn; Kim, Yoon-Young; Kho, Hong-Seop

2015-07-01

To investigate whether xylitol and sorbitol affect enzymatic and candidacidal activities of lysozyme, the peroxidase system, and the glucose oxidase-mediated peroxidase system. Xylitol and sorbitol were added to hen egg-white lysozyme, bovine lactoperoxidase, glucose oxidase-mediated peroxidase, and whole saliva in solution and on hydroxyapatite surfaces. The enzymatic activities of lysozyme, peroxidase, and glucose oxidase-mediated peroxidase were determined by the turbidimetric method, the NbsSCN assay, and production of oxidized o-dianisidine, respectively. Candidacidal activities were determined by comparing colony forming units using Candida albicans ATCC strains 10231, 11006, and 18804. While xylitol and sorbitol did not affect the enzymatic activity of hen egg-white lysozyme both in solution and on hydroxyapatite surfaces, they did inhibit the enzymatic activity of salivary lysozyme significantly in solution, but not on the surfaces. Xylitol and sorbitol enhanced the enzymatic activities of both bovine lactoperoxidase and salivary peroxidase significantly in a dose-dependent manner in solution, but not on the surfaces. Sorbitol, but not xylitol, inhibited the enzymatic activity of glucose oxidase-mediated peroxidase significantly. Both xylitol and sorbitol did not affect candidacidal activities of hen egg-white lysozyme, the bovine lactoperoxidase system, or the glucose oxidase-mediated bovine lactoperoxidase system. Xylitol and sorbitol inhibited salivary lysozyme activity, but enhanced both bovine lactoperoxidase and salivary peroxidase activities significantly in solution. Xylitol and sorbitol did not augment lysozyme- and peroxidase-related candidacidal activities. Copyright © 2015 Elsevier Ltd. All rights reserved.

Modified denatured lysozyme effectively solubilizes fullerene c60 nanoparticles in water

Science.gov (United States)

Siepi, Marialuisa; Politi, Jane; Dardano, Principia; Amoresano, Angela; De Stefano, Luca; Monti, Daria Maria; Notomista, Eugenio

2017-08-01

Fullerenes, allotropic forms of carbon, have very interesting pharmacological effects and engineering applications. However, a very low solubility both in organic solvents and water hinders their use. Fullerene C60, the most studied among fullerenes, can be dissolved in water only in the form of nanoparticles of variable dimensions and limited stability. Here the effect on the production of C60 nanoparticles by a native and denatured hen egg white lysozyme, a highly basic protein, has been systematically studied. In order to obtain a denatured, yet soluble, lysozyme derivative, the four disulfides of the native protein were reduced and exposed cysteines were alkylated by 3-bromopropylamine, thus introducing eight additional positive charges. The C60 solubilizing properties of the modified denatured lysozyme proved to be superior to those of the native protein, allowing the preparation of biocompatible highly homogeneous and stable C60 nanoparticles using lower amounts of protein, as demonstrated by dynamic light scattering, transmission electron microscopy and atomic force microscopy studies. This lysozyme derivative could represent an effective tool for the solubilization of other carbon allotropes.

Isolation and partial purification of lysozyme from saliva of Bali cattle ...

African Journals Online (AJOL)

About 100 μg/ml lysozyme of 14.2 kDa, determined through sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), per 4800 μg/ml saliva was obtained. Based on turbidity assay of Staphylococcus aureus, it was revealed that inhibition activities of 40 μg/ml isolated lysozyme were comparable to 800 μg/ml ...

Bio-templated CdSe quantum dots green synthesis in the functional protein, lysozyme, and biological activity investigation

International Nuclear Information System (INIS)

Wang, Qisui; Li, Song; Liu, Peng; Min, Xinmin

2012-01-01

Bifunctional fluorescence (CdSe Quantum Dots) – protein (Lysozyme) nanocomposites were synthesized at room temperature by a protein-directed, solution-phase, green-synthetic method. Fluorescence (FL) and absorption spectra showed that CdSe QDs were prepared successfully with Lyz. The average particle size and crystalline structure of QDs were investigated by high-resolution transmission electron microscopy (HRTEM) and X-ray diffraction (XRD), respectively. With attenuated total reflection-fourier transform infrared (ATR-FTIR) spectra and thermogravimetric (TG) analysis, it was confirmed that there is interaction between QDs and amide I, amide II groups in Lyz. FL polarization was measured and FL imaging was done to monitor whether QDs could be responsible for possible changes in the conformation and activity of Lyz. Interestingly, the results showed Lyz still retain the biological activity after formation of QDs, but the secondary structure of the Lyz was changed. And the advantage of this synthesis method is producing excellent fluorescent QDs with specifically biological function. -- Highlights: ► Lysozyme-directed green synthesis of CdSe quantum dots. ► Lysozyme still retain the biological activity after formation of CdSe. ► The method is the production of fluorescent QDs with highly specific and functions.

Molecular Characterization of a Lysozyme Gene and Its Altered Expression Profile in Crowded Beet Webworm (Loxostege sticticalis.

Directory of Open Access Journals (Sweden)

Hailong Kong

Full Text Available There is growing evidence that insects living in high-density populations exhibit an increase in immune function to counter a higher risk of disease. This phenomenon, known as density-dependent prophylaxis, has been experimentally tested in a number of insect species. Although density-dependent prophylaxis is especially prevalent in insects exhibiting density-dependent phase polyphenism, the molecular mechanism remains unclear. Our previous study demonstrated that the antibacterial activity of lysozyme is important for this process in the beet webworm Loxostege sticticalis. In this study, a lysozyme cDNA from L. sticticalis was cloned and characterized. The full-length cDNA is 1078 bp long and contains an open reading frame of 426 bp that encodes 142 amino acids. The deduced protein possesses structural characteristics of a typical c-type lysozyme and clusters with c-type lysozymes from other Lepidoptera. LsLysozyme was found to be expressed throughout all developmental stages, showing the highest level in pupae. LsLysozyme was also highly expressed in the midgut and fat body. Elevated LsLysozyme expression was observed in L. sticticalis larvae infected by Beauveria bassiana and in larvae reared under crowding conditions. In addition, the expression level of LsLysozyme in infected larvae reared at a density of 10 larvae per jar was significantly higher compared to those reared at a density of l or 30 larvae per jar. These results suggest that larval crowding affects the gene expression profile of this lysozyme. This study provides additional insight into the expression of an immune-associated lysozyme gene and helps us to better understand the immune response of L. sticticalis under crowding conditions.

Formation of Silica-Lysozyme Composites Through Co-Precipitation and Adsorption

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Daniela B. van den Heuvel

2018-04-01

Full Text Available Interactions between silica and proteins are crucial for the formation of biosilica and the production of novel functional hybrid materials for a range of industrial applications. The proteins control both precipitation pathway and the properties of the resulting silica–organic composites. Here, we present data on the formation of silica–lysozyme composites through two different synthesis approaches (co-precipitation vs. adsorption and show that the chemical and structural properties of these composites, when analyzed using a combination of synchrotron-based scattering (total scattering and small-angle X-ray scattering, spectroscopic, electron microscopy, and potentiometric methods vary dramatically. We document that while lysozyme was not incorporated into nor did its presence alter the molecular structure of silica, it strongly enhanced the aggregation of silica particles due to electrostatic and potentially hydrophobic interactions, leading to the formation of composites with characteristics differing from pure silica. The differences increased with increasing lysozyme content for both synthesis approaches. Yet, the absolute changes differ substantially between the two sets of composites, as lysozyme did not just affect aggregation during co-precipitation but also particle growth and likely polymerization during co-precipitation. Our results improve the fundamental understanding of how organic macromolecules interact with dissolved and nanoparticulate silica and how these interactions control the formation pathway of silica–organic composites from sodium silicate solutions, a widely available and cheap starting material.

Formation of Silica-Lysozyme Composites Through Co-Precipitation and Adsorption

Science.gov (United States)

van den Heuvel, Daniela B.; Stawski, Tomasz M.; Tobler, Dominique J.; Wirth, Richard; Peacock, Caroline L.; Benning, Liane G.

2018-04-01

Interactions between silica and proteins are crucial for the formation of biosilica and the production of novel functional hybrid materials for a range of industrial applications. The proteins control both precipitation pathway and the properties of the resulting silica-organic composites. Here we present data on the formation of silica-lysozyme composites through two different synthesis approaches (co-precipitation vs. adsorption) and show that the chemical and structural properties of these composites, when analyzed using a combination of synchrotron-based scattering (total scattering and SAXS), spectroscopic, electron microscopy and potentiometric methods vary dramatically. We document that while lysozyme was not incorporated into nor did its presence alter the molecular structure of silica, it strongly enhanced the aggregation of silica particles due to electrostatic and potentially hydrophobic interactions, leading to the formation of composites with characteristics differing from pure silica. The differences increased with increasing lysozyme content for both synthesis approaches. Yet, the absolute changes differ substantially between the two sets of composites, as lysozyme did not just affect aggregation during co-precipitation but also particle growth and likely polymerization during co-precipitation. Our results improve the fundamental understanding of how organic macromolecules interact with dissolved and nanoparticulate silica and how these interactions control the formation pathway of silica-organic composites from sodium silicate solutions, a widely available and cheap starting material.

Playing Producer: An alternative perspective on video games as film

Directory of Open Access Journals (Sweden)

Olivia Huang

2017-06-01

Full Text Available This paper proposes a perspective on analyzing video games as film by comparing the interactions of the player with those of a producer or other member in the film-making process. This contrasts with existing methodologies which focus on formal characteristics or narratology. This proposal also provides a method for combining the interactivity of games with the storytelling capacity of cinema without encountering the narrative paradox.

Effects of modified β-cyclodextrin on thermal stability and conformation of lysozyme

International Nuclear Information System (INIS)

Kamiyama, Tadashi; Satoh, Megumi; Tateishi, Takahiro; Nojiri, Tomoaki; Takeuchi, Daisuke; Kimura, Takayoshi

2012-01-01

Highlights: ► Effects of cyclodextrin on stability and conformation of lysozyme were clarified. ► The CD influences the hydrophobic interaction of lysozyme by the inclusion. ► The CD relatively destabilized the folded state by stabilizing the unfolded state. ► The destabilization depends on the concentration and the substituent of CD. ► The conformation of lysozyme was more spread at unfolded state by inclusion of CD. - Abstract: Effects of cyclic oligosaccharide cyclodextrin (CD) on stability and conformation of lysozyme were clarified thermodynamically and rheologically by DSC, viscosity, and circular dichroism measurements. The modified β-CD relatively destabilized the folded state of lysozyme by stabilizing the unfolded state due to inclusion of hydrophobic part into the hydrophobic interior of CD. The order of higher destabilization effect was acetyl-β-CD > methyl-β-CD > hydroxypropyl-β-CD. Apparent number of bound CD to unfolded state for methyl-, hydroxypropyl-, and acetyl-β-CD is 6.7 ± 0.7, 4.2 ± 1.1, and 18.6 ± 4.3 and the binding constant is 5.5 ± 0.8, 6.7 ± 2.4, and 4.4 ± 1.2 L mol −1 , respectively. The viscosity for unfolded state was increased with an increase in the each modified β-CD concentration, suggesting that the inclusion of CD on a part of hydrophobic core at unfolded state leads to break the hydrophobic core, then lysozyme would be more spread structure. The substituent of CD can accelerate instability by directly breaking hydrogen bond and/or can restrain instability by increase in hydrophobic interaction. The fact that the each modified CDs has different destabilization effect shows a possibility to control the stability of protein by the substitution of CD.

Characterizing protein activities on the lysozyme and nanodiamond complex prepared for bio applications.

Science.gov (United States)

Perevedentseva, E; Cai, P-J; Chiu, Y-C; Cheng, C-L

2011-02-01

Recently, nanodiamond particles have attracted increasing attention as a promising nanomaterial for its biocompatibility, easy functionalization and conjugation with biomolecules, and its superb physical/chemical properties. Nanodiamonds are mainly used as markers for cell imaging, using its fluorescence or Raman signals for detection, and as carriers for drug delivery. For the success of these applications, the biomolecule associated with the nanodiamond has to retain its functionality. In this work, the protein activities of egg white lysozyme adsorbed on nanodiamond particles of different sizes is investigated. The lysozyme nanodiamond complex is used here as a protein model for analyzing its structural conformation changes and, correspondingly, its enzymatic activity after the adsorption. Fourier-transform infrared spectroscopy (FTIR) is used for the analysis of the sensitive protein secondary structure. To access the activities of the adsorbed lysozyme, a fluorescence-based assay is used. The process of adsorption is also analyzed using UV-visible spectroscopic measurements in combination with analysis of nanodiamond properties with FTIR, Raman spectroscopy, and ζ-potential measurements. It is found that the activity of lysozyme upon adsorption depends on the nanodiamond's size and surface properties, and that the nanodiamond particles can be selected and treated, which do not alter the lysozyme functional properties. Such nanodiamonds can be considered convenient nanoparticles for various bioapplications.

Crystallization of Hevamine, an Enzyme with Lysozyme/Chitinase Activity from Hevea brasiliensis Latex

NARCIS (Netherlands)

ROZEBOOM, HJ; BUDIANI, A; BEINTEMA, JJ

1990-01-01

Hevamine, an enzyme with both lysozyme and chitinase activity, was isolated and purified from Hevea brasiliensis (rubber tree) latex. The enzyme (molecular weight 29,000) is homologous to certain “pathogenesis-related” proteins from plants, but not to hen egg-white or phage T4 lysozyme. To

Regular arrangement of periodates bound to lysozyme

Czech Academy of Sciences Publication Activity Database

Ondráček, Jan; Weiss, M.S.; Brynda, Jiří; Fiala, J.; Jursík, F.; Řezáčová, Pavlína; Jenner, L.B.; Sedláček, Juraj

2005-01-01

Roč. 61, Pt9 (2005), s. 1181-1189 ISSN 0907-4449 Institutional research plan: CEZ:AV0Z50520514 Keywords : hen egg white lysozyme * periodate * epitaxy Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.401, year: 2005

Macromolecular complexes of lysozyme with kappa carrageenan

NARCIS (Netherlands)

Antonov, Y.A.; Zhuravleva, I.L.; Cardinaels, R.; Moldenaers, P.

2018-01-01

We present a structural study of the complexation and binding of lysozyme (Lys) with kappa carrageenan (kCG) by means of turbidity measurements, phase analysis, dynamic and electrophoretic light scattering, differential scanning microcalorimetry (DSMC), confocal laser scanning (CLSM) microscopy,

Interaction of lysozyme protein with different sized silica nanoparticles and their resultant structures

Energy Technology Data Exchange (ETDEWEB)

Yadav, Indresh, E-mail: iykumarindresh288@gmail.com; Aswal, V. K. [Solid State Physics Division, Bhabha Atomic Research Centre, Mumbai 400 085 (India); Kohlbrecher, J. [Laboratory for Neutron Scattering, Paul Scherrer Institut, CH-5232 PSI Villigen (Switzerland)

2016-05-23

The interaction of model protein-lysozyme with three different sized anionic silica nanoparticles has been studied by UV-vis spectroscopy, dynamic light scattering (DLS) and small-angle neutron scattering (SANS). The surface area and curvature of the nanoparticles change with size, which significantly influence their interaction with protein. The lysozyme adsorbs on the surface of the nanoparticles due to electrostatic attraction and leads to the phase transformation from one phase (clear) to two-phase (turbid) of the nanoparticle-protein system. The dominance of lysozyme induced short-range attraction over long-range electrostatic repulsion between nanoparticles is responsible for phase transformation and modeled by the two-Yukawa potential. The magnitude of the attractive interaction increases with the size of the nanoparticles as a result the phase transformation commences relatively at lower concentration of lysozyme. The structure of the nanoparticle-protein system in two-phase is characterized by the diffusion limited aggregate type of mass fractal morphology.

Molecular cloning, sequence analysis and phylogeny of first caudata g-type lysozyme in axolotl (Ambystoma mexicanum).

Science.gov (United States)

Yu, Haining; Gao, Jiuxiang; Lu, Yiling; Guang, Huijuan; Cai, Shasha; Zhang, Songyan; Wang, Yipeng

2013-11-01

Lysozymes are key proteins that play important roles in innate immune defense in many animal phyla by breaking down the bacterial cell-walls. In this study, we report the molecular cloning, sequence analysis and phylogeny of the first caudate amphibian g-lysozyme: a full-length spleen cDNA library from axolotl (Ambystoma mexicanum). A goose-type (g-lysozyme) EST was identified and the full-length cDNA was obtained using RACE-PCR. The axolotl g-lysozyme sequence represents an open reading frame for a putative signal peptide and the mature protein composed of 184 amino acids. The calculated molecular mass and the theoretical isoelectric point (pl) of this mature protein are 21523.0 Da and 4.37, respectively. Expression of g-lysozyme mRNA is predominantly found in skin, with lower levels in spleen, liver, muscle, and lung. Phylogenetic analysis revealed that caudate amphibian g-lysozyme had distinct evolution pattern for being juxtaposed with not only anura amphibian, but also with the fish, bird and mammal. Although the first complete cDNA sequence for caudate amphibian g-lysozyme is reported in the present study, clones encoding axolotl's other functional immune molecules in the full-length cDNA library will have to be further sequenced to gain insight into the fundamental aspects of antibacterial mechanisms in caudate.

Receptor-mediated endocytosis of lysozyme in renal proximal tubules of the frog Rana temporaria

Directory of Open Access Journals (Sweden)

E.V. Seliverstova

2015-04-01

Full Text Available The mechanism of protein reabsorption in the kidney of lower vertebrates remains insufficiently investigated in spite of raising interest to the amphibian and fish kidneys as a useful model for physiological and pathophysiological examinations. In the present study, we examined the renal tubular uptake and the internalization rote of lysozyme after its intravenous injection in the wintering frog Rana temporaria using immunohisto- and immunocytochemistry and specific markers for some endocytic compartments. The distinct expression of megalin and cubilin in the proximal tubule cells of lysozyme-injected frogs was revealed whereas kidney tissue of control animals showed no positive immunoreactivity. Lysozyme was detected in the apical endocytic compartment of the tubular cells and colocalized with clathrin 10 min after injection. After 20 min, lysozyme was located in the subapical compartment negative to clathrin (endosomes, and intracellular trafficking of lysozyme was coincided with the distribution of megalin and cubilin. However, internalized protein was retained in the endosomes and did not reach lysosomes within 30 min after treatment that may indicate the inhibition of intracellular trafficking in hibernating frogs. For the first time, we provided the evidence that lysozyme is filtered through the glomeruli and absorbed by receptor-mediated clathrin-dependent endocytosis in the frog proximal tubule cells. Thus, the protein uptake in the amphibian mesonephros is mediated by megalin and cubilin that confirms a critical role of endocytic receptors in the renal reabsorption of proteins in amphibians as in mammals.

21 CFR 862.1490 - Lysozyme (muramidase) test system.

Science.gov (United States)

2010-04-01

... serum, plasma, leukocytes, and urine. Lysozyme measurements are used in the diagnosis and treatment of monocytic leukemia and kidney disease. (b) Classification. Class I (general controls). The device is exempt...

Synthesis of Ag{sub 2}S nanorods by biomimetic method in the lysozyme matrix

Energy Technology Data Exchange (ETDEWEB)

Qin, Dezhi, E-mail: dezhiqin@163.com; Zhang, Li; He, Guoxu; Zhang, Qiuxia

2013-09-01

Graphical abstract: - Highlights: • Firstly, Ag{sub 2}S nanorods were synthesized by biomimetic method in the lysozyme solutions. • The study of the interaction between Ag{sup +} and the lysozyme. • Discussion of possible formation mechanism of Ag{sub 2}S nanorods. • The synthesis process of lyso-conjugated Ag{sub 2}S nanocrystals is facile, effective and environment friendly. - Abstract: Ag{sub 2}S nanorods were successfully synthesized by biomimetic route in the lysozyme solution at physiological temperature and atmospheric pressure. The transmission electron microscopy (TEM) images revealed that the prepared nanorods are uniform and monodisperse with homogeneous size about 50 nm in diameter and 150 nm in length. The optical property of Ag{sub 2}S nanocrystals was studied by the ultraviolet–visible (UV–vis) and photoluminescence (PL) spectroscopy, the results show that the products exhibit well-defined emission at 471 nm and 496 nm excited by 292 nm. The interaction of Ag{sup +}/Ag{sub 2}S with the lysozyme was investigated through Fourier transform infrared (FT-IR) spectroscopy, which shows that the cooperation effect of the lysozyme and Ag{sup +} could be responsible for the formation of as obtained Ag{sub 2}S nanorods.

Prevention of Bacterial Contamination of a Silica Matrix Containing Entrapped β-Galactosidase through the Action of Covalently Bound Lysozymes

Directory of Open Access Journals (Sweden)

Heng Li

2017-02-01

Full Text Available β-galactosidase was successfully encapsulated within an amino-functionalised silica matrix using a “fish-in-net” approach and molecular imprinting technique followed by covalent binding of lysozyme via a glutaraldehyde-based method. Transmission electron microscopy (TEM, X-ray diffraction (XRD, scanning electron microscopy (SEM, and Fourier transform infrared (FTIR spectroscopy were used to characterise the silica matrix hosting the two enzymes. Both encapsulated β-galactosidase and bound lysozyme exhibited high enzymatic activities and outstanding operational stability in model reactions. Moreover, enzyme activities of the co-immobilised enzymes did not obviously change relative to enzymes immobilised separately. In antibacterial tests, bound lysozyme exhibited 95.5% and 89.6% growth inhibition of Staphylococcus aureus ATCC (American type culture collection 653 and Escherichia coli ATCC 1122, respectively. In milk treated with co-immobilised enzymes, favourable results were obtained regarding reduction of cell viability and high lactose hydrolysis rate. In addition, when both co-immobilised enzymes were employed to treat milk, high operational and storage stabilities were observed. The results demonstrate that the use of co-immobilised enzymes holds promise as an industrial strategy for producing low lactose milk to benefit people with lactose intolerance.

Destruction of Salmonellae on poultry meat with lysozyme, EDTA, x ray, microwave, and chlorine

International Nuclear Information System (INIS)

Teotia, J.S.; Miller, B.F.

1975-01-01

Lysozyme, ethylenediaminetetracetic acid, chlorine, x-irradiation and microwaves were used in experimental attempts to eliminate Salmonella senftenberg 775W or Salmonella typhimurium from turkey drumsticks and whole carcasses. Turkey drumsticks or whole carcasses were artificially contaminated with S. senfenberg 775W or S. typhimurium in concentrations ranging between 5 x 10 5 to 8 x 10 5 viable cells per ml. of contaminating fluid. After each treatment, samples were cultured, plated, and tested according to standard methods to determine the susceptibility of Salmonella organisms to the particular treatment. A 0.1 percent solution of lysozyme eliminated the S. senftenberg 775W at 22 0 C within three hours. A 0.5 percent solution of ethylenediaminetetracetic acid failed to destroy the test organism under the same conditions. Eighty thousand rads of X-ray eliminated the test organism on turkey drumsticks but failed to remove it from whole turkey carcasses. Microwaves eliminated the S. senftenberg 775W in 150 seconds from turkey drumsticks and ten minutes from broiler chicken carcasses. Aqueous solutions containing 3400 and 2125 ppM chlorine failed to destroy the test organism on turkey drumsticks at 21 0 C. in 9 and 24 hours. None of the treatments changed the appearance of the skin or meat, except microwaves produced a partially-cooked appearance. Chlorine produced off-color drumsticks

Serum lysozyme activity in coeliac disease: a possible aid to athe diagnosis of malignant change.

OpenAIRE

Cooper, B T; Ukabam, S O; Barry, R E; Read, A E

1981-01-01

Serum lysozyme activities were measured in 34 control subjects, 13 untreated adult coeliac patients, 21 adult coeliac patients on gluten-free diet, and eight coeliac patients with a histiocytic lymphoma. Serum lysozyme activities were raised in three untreated patients, three patients treated with a gluten-free diet, and in only two patients with coeliac disease and lymphoma. Serum lysozyme estimations cannot be recommended as an aid to the diagnosis of lymphoma in patients with coeliac disease.

[Inactivating Effect of Heat-Denatured Lysozyme on Murine Norovirus in Bread Fillings].

Science.gov (United States)

Takahashi, Michiko; Yasuda, Yuka; Takahashi, Hajime; Takeuchi, Akira; Kuda, Takashi; Kimura, Bon

2018-01-01

In this study, we investigated the viability of murine norovirus strain 1 (MNV-1), a surrogate for human norovirus, in bread fillings used for making stuffed buns and pastries. The inactivating effect of heat-denatured lysozyme, which was recently reported to have an antiviral effect, on MNV-1 contaminating the bread fillings was also examined. MNV-1 was inoculated into two types of fillings (chocolate cream, marmalade jam) at 4.5 log PFU/g, and the bread fillings were stored at 4℃ for 5 days. MNV-1 remained viable in the bread fillings during storage. However, addition of 1% heat-denatured lysozyme to the fillings resulted in a decrease of MNV-1 infectivity immediately after inoculation, in both fillings. On the fifth day of storage, MNV-1 infectivity was decreased by 1.2 log PFU/g in chocolate cream and by 0.9 log PFU/g in marmalade jam. Although the mechanism underlying the anti-norovirus effect of heat-denatured lysozyme has not been clarified, our results suggest that heat-denatured lysozyme can be used as an inactivating agent against norovirus in bread fillings.

Visualization study of film drops produced by bubble bursting

International Nuclear Information System (INIS)

Ma Chao; Bo Hanliang

2012-01-01

The phenomenon that bubble bursting results in drops production is common in the steam generator of the nuclear power plant, and the fine drops generated by this way is one of the most important source of the drop entrainment in the vapor stream. The visualization experiment about the film drops produced by the bursting bubbles at a free water surface was studied using a high-speed video camera. The results show that the bubble cap breaks up in a single point, within the limits of bubble size in the experiment at present. The whole process can be distinguished into four successive stages: A primary inertial drainage, the bubble cap puncture at the foot or on the top, the film rolls-up and the liquid ring appearing with the hole expanding, and fine film drops emission under the effect of destabilization of a Rayleigh-Taylor type. The expression about the bubble radius and the film drops number is obtain by fitting the experiment data at the bubble radius range from 3-25 mm. The result trend agrees well with the previous work. (authors)

An electron microscopy study of the diversity of Streptococcus sanguinis cells induced by lysozyme in vitro.

Science.gov (United States)

Hao, Yuqing; Li, Li; Li, Wei; Zhou, Xuedong; Lu, Junjun

2010-01-01

Bacterial virulence could be altered by the antimicrobial agents of the host. Our aim was to identify the damage and survival of Streptococcus sanguinis induced by lysozymes in vitro and to analyse the potential of oral microorganisms to shirk host defences, which cause infective endocarditis. S. sanguinis ATCC 10556 received lysozyme at concentrations of 12.5, 25, 50 and 100 microg/ml. Cells were examined by electron microscopy. The survival was assessed by colony counting and construction of a growth curve. Challenged by lysozymes, cells mainly exhibited cell wall damage, which seemed to increase with increasing lysozyme concentration and longer incubation period in the presence of ions. Cells with little as well as apparent lesion were observed under the same treatment set, and anomalous stick and huge rotund bodies were occasionally observed. After the removal of the lysozyme, some damaged cells could be reverted to its original form with brain heart infusion (BHI), and their growth curve was similar to the control cells. After further incubation in BHI containing lysozyme, S. sanguinis cell damage stopped progressing, and their growth curve was also similar to the control cells. The results suggested that the S. sanguinis lesions caused by the lysozyme in the oral cavity may be nonhomogeneous and that some damaged cells could self-repair and survive. It also indicated that S. sanguinis with damaged cell walls may survive and be transmitted in the bloodstream.

Thermodynamic Exploration of Eosin-Lysozyme Binding: A Physical Chemistry and Biochemistry Laboratory Experiment

Science.gov (United States)

Huisman, Andrew J.; Hartsell, Lydia R.; Krueger, Brent P.; Pikaart, Michael J.

2010-01-01

We developed a modular pair of experiments for use in the undergraduate physical chemistry and biochemistry laboratories. Both experiments examine the thermodynamics of the binding of a small molecule, eosin Y, to the protein lysozyme. The assay for binding is the quenching of lysozyme fluorescence by eosin through resonant energy transfer. In…

Synergistic cytotoxicity and mechanism of caffeine and lysozyme on hepatoma cell line HepG2

Science.gov (United States)

Yang, Hongchao; Li, Jingjuan; Cui, Lin; Ren, Yanqing; Niu, Liying; Wang, Xinguo; Huang, Yun; Cui, Lijian

2018-03-01

The influences of caffeine, lysozyme and the joint application of them on the hepatoma cell line HepG2 proliferation inhibition and cell apoptosis were observed by 3-(4, 5-dimethyl-2-thiazyl)-2, 5-diphenyl-2H-tetrazolium bromide assay and Hoechst 33342, which showed the proliferation inhibition rate of the joint application on HepG2 cells was 47.21%, significantly higher than caffeine or lysozyme, and the joint application promoted the apoptosis of HepG2 cells obviously. Van't Hoff classical thermodynamics formula, the Föster theory of non-radiation energy transfer and fluorescence phase diagram were used to manifest that the process of lysozyme binding to caffeine followed a two-state model, which was spontaneous at low temperature driven by enthalpy change, and the predominant intermolecular force was hydrogen bonding or Van der Waals force to stabilize caffeine-lysozyme complex with the distance 5.86 nm. The attenuated total reflection-Fourier transform infrared spectra indicated that caffeine decreased the relative contents of α-helix and β-turn, which inferred the structure of lysozyme tended to be "loose". Synchronous fluorescence spectra and ultraviolet spectra supported the above conclusion. The amino acid residues in the cleft of lysozyme were exposed and electropositivity was increased attributing to the loose structure, which were conducive to increasing caffeine concentration on the HepG2 cell surface by electrostatic interaction to show synergistic effect. The great quantities of microvilli on the liver cancer cell membrane surface, is beneficial for the lysozyme-caffeine compound to aggregate on cell surface to increase the concentration of caffeine to play stronger physiological role by electrostatic effect.

Physics properties of TiO_2 films produced by dip-coating technique

International Nuclear Information System (INIS)

Teloeken, A.C.; Alves, A.K.; Berutti, F.A.; Tabarelli, A.; Bergmann, C.P.

2014-01-01

The use of titanium dioxide (TiO_2) as a photocatalyst to produce hydrogen has been of great interest because of their chemical stability, low cost and non-toxicity. TiO_2 occurs in three different crystal forms: rutile, anatase and brokita. Among these, the anatase phase generally exhibits the best photocatalytic behavior, while the rutile phase is the most stable. Among the various techniques of deposition, dip-coating technique produces films with good photocatalytic properties, using simple and inexpensive equipment. In this work TiO_2 films were obtained by dip-coating. The films were characterized using X-ray diffraction, scanning electron microscopy, profilometry, contact angle measurements and photocurrent. The microstructure and physical properties were evaluated in relation of the temperature and the addition of an additive. (author)

In Vitro Effect of Lysozyme on Albumin Deposition to Hydrogel Contact Lens Materials.

Science.gov (United States)

Babaei Omali, Negar; Subbaraman, Lakshman N; Heynen, Miriam; Fadli, Zohra; Coles-Brennan, Chantal; Jones, Lyndon W

2017-11-01

Albumin deposition on contact lenses could be detrimental to contact lens (CL) wear because this may increase the risk of bacterial binding and reduce comfort. Lysozyme deposition on selected lens materials would reduce albumin deposition on lenses. This study aims to determine if lysozyme deposition on CLs could act as a barrier against subsequent albumin adsorption, using an in vitro model. Six hydrogel CL materials (etafilcon A, polymacon, nelfilcon A, omafilcon A, ocufilcon B, and nesofilcon A) were evaluated. Four CLs of each type were soaked in lysozyme solution for 16 hours at 37°C. Lysozyme-coated lenses were then placed in vials with 1.5 mL of artificial tear solution containing I-labeled albumin for 16 hours at 37°C with shaking. Four uncoated lenses of each type were used as controls. Lenses soaked in radiolabeled albumin were rinsed in a phosphate-buffered saline solution, and radioactive counts were measured directly on lenses using a gamma counter. Albumin uptake on lenses was measured using a calibration curve by plotting radioactive counts versus protein concentration. Results are reported as mean ± SD. Lysozyme-coated etafilcon A lenses exhibited lower levels of deposited albumin than uncoated etafilcon A lenses (58 ± 12 vs. 84 ± 5 ng/lens; P albumin adsorption between control (uncoated) and lysozyme-coated polymacon (105 ± 10 vs. 110 ± 34 ng/lens), nelfilcon A (51 ± 7 vs. 42 ± 20 ng/lens), omafilcon A (90 ± 20 vs. 80 ± 38 ng/lens), ocufilcon B (87 ± 20 vs. 115 ± 50 ng/lens), and nesofilcon A (170 ± 29 vs. 161 ± 10 ng/lens) lens materials (P > .05). Uncoated nesofilcon A lenses deposited the highest amount of albumin when compared with other uncoated lenses (P albumin, which may potentially be beneficial to CL wearers.

Effect of Encapsulation on Antimicrobial Activity of Herbal Extracts with Lysozyme

Directory of Open Access Journals (Sweden)

Petra Matouskova

2016-01-01

Full Text Available Resistance of microorganisms to antibiotics has increased. The use of natural components with antimicrobial properties can be of great significance to reduce this problem. The presented work is focused on the study of the effect of encapsulation of selected plant and animal antimicrobial substances (herbs, spices, lysozyme and nisin on their activity and stability. Antimicrobial components were packaged into liposomes and polysaccharide particles (alginate, chitosan and starch. Antimicrobial activity was tested against two Gram-positive (Bacillus subtilis and Micrococcus luteus and two Gram-negative (Escherichia coli and Serratia marcescens bacteria. Encapsulation was successful in all types of polysaccharide particles and liposomes. The prepared particles exhibited very good long-term stability, especially in aqueous conditions. Antimicrobial activity was retained in all types of particles. Liposomes with encapsulated herb and spice extracts exhibited very good inhibitory effect against all tested bacterial strains. Most of herbal extracts had very good antimicrobial effect against the tested Gram-negative bacterial strains, while Gram-positive bacteria were more sensitive to lysozyme particles. Thus, particles with co-encapsulated herbs and lysozyme are more active against different types of bacteria, and more stable and more effective during long-term storage. Particles with encapsulated mixture of selected plant extracts and lysozyme could be used as complex antimicrobial preparation with controlled release in the production of food and food supplements, pharmaceutical and cosmetic industries.

Effect of ethanol as a co-solvent on the aerosol performance and stability of spray-dried lysozyme

DEFF Research Database (Denmark)

Ji, Shuying; Thulstrup, Peter Waaben; Mu, Huiling

2016-01-01

In the spray drying process, organic solvents can be added to facilitate drying, accommodate certain functional excipients, and modify the final particle characteristics. In this study, lysozyme was used as a model pharmaceutical protein to study the effect of ethanol as a co...... the spray drying process. The enzymatic activities of the spray-dried lysozyme showed no significant impact of ethanol; however, the lysozyme enzymatic activity was ca. 25% lower compared to the starting material. In conclusion, the addition of ethanol as a co-solvent in the spray drying feed for lysozyme......-solvent on the stability and aerosol performance of spray-dried protein. Lysozyme was dissolved in solutions with various ratios of ethanol and water, and subsequently spray-dried. A change from spherical particles into wrinkled and folded particles was observed upon increasing the ratio of ethanol in the feed...

Cloning the human lysozyme cDNA: Inverted Alu repeat in the mRNA and in situ hybridization for macrophages and Paneth cells

International Nuclear Information System (INIS)

Chung, L.P.; Keshav, S.; Gordon, S.

1988-01-01

Lysozyme is a major secretory product of human and rodent macrophages and a useful marker for myelomonocytic cells. Based on the known human lysozyme amino acid sequence, oligonucleotides were synthesized and used as probes to screen a phorbol 12-myristate 13-acetate-treated U937 cDNA library. A full-length human lysozyme cDNA clone, pHL-2, was obtained and characterized. Sequence analysis shows that human lysozyme, like chicken lysozyme, has in 18-amino-acid-long signal peptide, but unlike the chicken lysozyme cDNA, the human lysozyme cDNA has a >1-kilobase-long 3' nontranslated sequence. Interestingly, within this 3' region, an inverted repeat of the Alu family of repetitive sequences was discovered. In RNA blot analyses, DNA probes prepared from pHL-2 can be used to detect lysozyme mRNA not only from human but also from mouse and rat. Moreover, by in situ hybridization, complementary RNA transcripts have been used as probes to detect lysozyme mRNA in mouse macrophages and Paneth cells. This human lysozyme cDNA clone is therefore likely to be a useful molecular probe for studying macrophage distribution and gene expression

Controlled Release of Lysozyme from Double-Walled Poly(Lactide-Co-Glycolide (PLGA Microspheres

Directory of Open Access Journals (Sweden)

Rezaul H. Ansary

2017-10-01

Full Text Available Double-walled microspheres based on poly(lactide-co-glycolide (PLGA are potential delivery systems for reducing a very high initial burst release of encapsulated protein and peptide drugs. In this study, double-walled microspheres made of glucose core, hydroxyl-terminated poly(lactide-co-glycolide (Glu-PLGA, and carboxyl-terminated PLGA were fabricated using a modified water-in-oil-in-oil-in-water (w1/o/o/w2 emulsion solvent evaporation technique for the controlled release of a model protein, lysozyme. Microspheres size, morphology, encapsulation efficiency, lysozyme in vitro release profiles, bioactivity, and structural integrity, were evaluated. Scanning electron microscopy (SEM images revealed that double-walled microspheres comprising of Glu-PLGA and PLGA with a mass ratio of 1:1 have a spherical shape and smooth surfaces. A statistically significant increase in the encapsulation efficiency (82.52% ± 3.28% was achieved when 1% (w/v polyvinyl alcohol (PVA and 2.5% (w/v trehalose were incorporated in the internal and external aqueous phase, respectively, during emulsification. Double-walled microspheres prepared together with excipients (PVA and trehalose showed a better control release of lysozyme. The released lysozyme was fully bioactive, and its structural integrity was slightly affected during microspheres fabrication and in vitro release studies. Therefore, double-walled microspheres made of Glu-PLGA and PLGA together with excipients (PVA and trehalose provide a controlled and sustained release for lysozyme.

Biofilm formation and transcriptome analysis of Streptococcus gallolyticus subsp. gallolyticus in response to lysozyme.

Directory of Open Access Journals (Sweden)

Imke Grimm

Full Text Available Streptococcus gallolyticus subsp. gallolyticus is a commensal bacterium of the human gastrointestinal tract, and a pathogen causing infective endocarditis and other biofilm-associated infections via exposed collagen. This study focuses on the characterization of the biofilm formation and collagen adhesion of S. gallolyticus subsp. gallolyticus under different conditions. In this study, it has been observed that the isolate UCN 34 is resistant to 20 mg/ml lysozyme in BHI medium, whereas the strain BAA-2069 builds more biofilm in the presence of lysozyme compared to in a control of BHI without lysozyme. A transcriptome analysis with whole genome microarrays of these two isolates in BHI medium with lysozyme compared to control without lysozyme revealed changes in gene expression levels. In the isolate BAA-2069, 67 genes showed increased expression in the presence of lysozyme, while in the isolate UCN 34, 165 genes showed increased expression and 30 genes showed decreased expression through lysozyme treatment. Products of genes which were higher expressed are in involved in transcription and translation, in cell-wall modification, in hydrogen peroxide resistance and in bacterial immunity. Furthermore, the adhesion ability of different strains of S. gallolyticus subsp. gallolyticus to collagen type I and IV was analyzed. Thereby, we compared the adhesion of 46 human isolates with 23 isolates from animals. It was shown that the adhesion ability depends significantly on whether the isolate was isolated from human or animal. For example, high adhesion ability was observed for strain UCN 34 isolated from an infective endocarditis patient, whereas strain DSM 16831 isolated from koala feces adhered only marginally to collagen. Full genome microarray analysis of these two strains revealed strain-dependent gene expression due to adhesion. The expression of 25 genes of a transposon and 15 genes of a phage region in strain DSM 16831 were increased, which

Production of biofunctionalized MoS2 flakes with rationally modified lysozyme: a biocompatible 2D hybrid material

Science.gov (United States)

Siepi, Marialuisa; Morales-Narváez, Eden; Domingo, Neus; Monti, Daria Maria; Notomista, Eugenio; Merkoçi, Arben

2017-09-01

Bioapplications of 2D materials embrace demanding features in terms of environmental impact, toxicity and biocompatibility. Here we report on the use of a rationally modified lysozyme to assist the exfoliation of MoS2 bulk crystals suspended in water through ultrasonic exfoliation. The design of the proposed lysozyme derivative provides this exfoliated 2D-materail with both, hydrophobic groups that interact with the surface of MoS2 and hydrophilic groups exposed to the aqueous medium, which hinders its re-aggregation. This approach, clarified also by molecular docking studies, leads to a stable material (ζ-potential, 27  ±  1 mV) with a yield of up to 430 µg ml-1. The bio-hybrid material was characterized in terms of number of layers and optical properties according to different slots separated by diverse centrifugal forces. Furthermore the obtained material was proved to be biocompatible using human normal keratinocytes and human cancer epithelial cells, whereas the method was demonstrated to be applicable to produce other 2D materials such as graphene. This approach is appealing for the advantageous production of high quality MoS2 flakes and their application in biomedicine and biosensing. Moreover, this method can be applied to different starting materials, making the denatured lysozyme a promising bio-tool for surface functionalization of 2D materials.

Binding and Inhibitory Effect of the Dyes Amaranth and Tartrazine on Amyloid Fibrillation in Lysozyme.

Science.gov (United States)

Basu, Anirban; Suresh Kumar, Gopinatha

2017-02-16

Interaction of two food colorant dyes, amaranth and tartrazine, with lysozyme was studied employing multiple biophysical techniques. The dyes exhibited hypochromic changes in the presence of lysozyme. The intrinsic fluorescence of lysozyme was quenched by both dyes; amaranth was a more efficient quencher than tartrazine. The equilibrium constant of amaranth was higher than that of tartarzine. From FRET analysis, the binding distances for amaranth and tartrazine were calculated to be 4.51 and 3.93 nm, respectively. The binding was found to be dominated by non-polyelectrolytic forces. Both dyes induced alterations in the microenvironment surrounding the tryptophan and tyrosine residues of the protein, with the alterations being comparatively higher for the tryptophans than the tyrosines. The interaction caused significant loss in the helicity of lysozyme, the change being higher with amaranth. The binding of both dyes was exothermic. The binding of amaranth was enthalpy driven, while that of tartrazine was predominantly entropy driven. Amaranth delayed lysozyme fibrillation at 25 μM, while tartrazine had no effect even at 100 μM. Nevertheless, both dyes had a significant inhibitory effect on fibrillogenesis. The present study explores the potential antiamyloidogenic property of these azo dyes used as food colorants.

SERS activity of silver and gold nanostructured thin films deposited by pulsed laser ablation

Science.gov (United States)

Agarwal, N. R.; Tommasini, M.; Fazio, E.; Neri, F.; Ponterio, R. C.; Trusso, S.; Ossi, P. M.

2014-10-01

Nanostructured Au and Ag thin films were obtained by nanosecond pulsed laser ablation in presence of a controlled Ar atmosphere. Keeping constant other deposition parameters such as target-to-substrate distance, incidence angle, laser wavelength and laser fluence, the film morphology, revealed by SEM, ranges from isolated NPs to island structures and sensibly depends on gas pressure (10-100 Pa) and on the laser pulse number (500-3 × 10). The control of these two parameters allows tailoring the morphology and correspondingly the optical properties of the films. The position and width of the surface plasmon resonance peak, in fact, can be varied with continuity. The films showed remarkable surface-enhanced Raman activity (SERS) that depends on the adopted deposition conditions. Raman maps were acquired on micrometer-sized areas of both silver and gold substrates selected among those with the strongest SERS activity. Organic dyes of interest in cultural heritage studies (alizarin, purpurin) have been also considered for bench marking the substrates produced in this work. Also the ability to detect the presence of biomolecules was tested using lysozyme in a label free configuration.

Starch/poly (butylene adipate-co-terephthalate/montmorillonite films produced by blow extrusion

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Rodrigo A. L. Santos

2014-07-01

Full Text Available This study aims to prepare biodegradable films from cassava starch, poly (butylene adipate-co-terephthalate (PBAT, and montmorillonite (MMT using blow-extrusion process and analyze the effects of different types and concentrations of MMT on the microstructure, physicochemical, and mechanical properties of the resulting films. The films were produced by blending 30% of PBAT with glycerol (17.5%, starch (49.0-52.5%, and four different types of montmorillonite (Cloisite® Na+, 10A, 15A, and 30B at two different concentrations (1.75% and 3.5%. All the films prepared in this study showed an increase in the basal spacing of MMT layers. In particular, the films with 10A and 30B showed the highest increase in intercalation basal spacing, suggesting the formation of intercalated composites. The addition of nanoclays decreased the elongation of films. The addition of Cloisite® 10A resulted in films with the lowest WVP values and the highest stability to water adsorption under different RH conditions.

Isothermal Titration Calorimetry and Macromolecular Visualization for the Interaction of Lysozyme and Its Inhibitors

Science.gov (United States)

Wei, Chin-Chuan; Jensen, Drake; Boyle, Tiffany; O'Brien, Leah C.; De Meo, Cristina; Shabestary, Nahid; Eder, Douglas J.

2015-01-01

To provide a research-like experience to upper-division undergraduate students in a biochemistry teaching laboratory, isothermal titration calorimetry (ITC) is employed to determine the binding constants of lysozyme and its inhibitors, N-acetyl glucosamine trimer (NAG[subscript 3]) and monomer (NAG). The extremely weak binding of lysozyme/NAG is…

Expression and Activity of Lysozyme in Apis Mellifera Carnica Brood Infested with Varroa Destructor

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Zaobidna Ewa A.

2017-12-01

Full Text Available Varroa destructor is a parasitic mite that attacks the honey bee, and previous studies have suggested that parasitosis caused by this mite is accompanied by immunosuppresion in the host. In this study, the effect of mite infestation on the expression of the lysozyme-1 (lys-1 gene and lysozyme activity in Apis mellifera carnica was determined. The experiment was carried out on the five developmental stages of honey bee workers and drones. Developmental and gender-related differences in gene expression and lysozyme activity were observed in a Varroa destructor-infested brood. The relative expression of the lys-1 gene increased in a infested worker brood and decreased in a drone brood except for P3 pupae. In the final stage of development, the lys-1 gene expression was significantly lower in infested newly emerged workers and drones. Changes in the relative expression of the lys-1 gene in infested individuals was poorly manifested at the level of enzyme activity, whereas at the two final stages of development (P5 and I there was a positive correlation between relative lys-1 expression and lysozyme activity in infested bees of both genders (r=0.988, r=0.999, respectively. The results of this study indicate that V. destructor influences the lysozyme-linked immune response in bees.

Size-dependent interaction of silica nanoparticles with lysozyme and bovine serum albumin proteins

Science.gov (United States)

Yadav, Indresh; Aswal, Vinod K.; Kohlbrecher, Joachim

2016-05-01

The interaction of three different sized (diameter 10, 18, and 28 nm) anionic silica nanoparticles with two model proteins—cationic lysozyme [molecular weight (MW) 14.7 kDa)] and anionic bovine serum albumin (BSA) (MW 66.4 kDa) has been studied by UV-vis spectroscopy, dynamic light scattering (DLS), and small-angle neutron scattering (SANS). The adsorption behavior of proteins on the nanoparticles, measured by UV-vis spectroscopy, is found to be very different for lysozyme and BSA. Lysozyme adsorbs strongly on the nanoparticles and shows exponential behavior as a function of lysozyme concentration irrespective of the nanoparticle size. The total amount of adsorbed lysozyme, as governed by the surface-to-volume ratio, increases on lowering the size of the nanoparticles for a fixed volume fraction of the nanoparticles. On the other hand, BSA does not show any adsorption for all the different sizes of the nanoparticles. Despite having different interactions, both proteins induce similar phase behavior where the nanoparticle-protein system transforms from one phase (clear) to two phase (turbid) as a function of protein concentration. The phase behavior is modified towards the lower concentrations for both proteins with increasing the nanoparticle size. DLS suggests that the phase behavior arises as a result of the nanoparticles' aggregation on the addition of proteins. The size-dependent modifications in the interaction potential, responsible for the phase behavior, have been determined by SANS data as modeled using the two-Yukawa potential accounting for the repulsive and attractive interactions in the systems. The protein-induced interaction between the nanoparticles is found to be short-range attraction for lysozyme and long-range attraction for BSA. The magnitude of attractive interaction irrespective of protein type is enhanced with increase in the size of the nanoparticles. The total (attractive+repulsive) potential leading to two-phase formation is found to be

The Natural Antimicrobial Enzyme Lysozyme is Up-Regulated in Gastrointestinal Inflammatory Conditions

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Carlos A. Rubio

2014-01-01

Full Text Available The cells that line the mucosa of the human gastrointestinal tract (GI, that is, oral cavity, oesophagus, stomach, small intestine, large intestine, and rectum are constantly challenged by adverse micro-environmental factors, such as different pH, enzymes, and bacterial flora. With exception of the oral cavity, these microenvironments also contain remnant cocktails of secreted enzymes and bacteria from upper organs along the tract. The density of the GI bacteria varies, from 103/mL near the gastric outlet, to 1010/mL at the ileocecal valve, to 1011 to 1012/mL in the colon. The total microbial population (ca. 1014 exceeds the total number of cells in the tract. It is, therefore, remarkable that despite the prima facie inauspicious mixture of harmful secretions and bacteria, the normal GI mucosa retains a healthy state of cell renewal. To counteract the hostile microenvironment, the GI epithelia react by speeding cell exfoliation (the GI mucosa has a turnover time of two to three days, by increasing peristalsis, by eliminating bacteria through secretion of plasma cell-immunoglobulins and by increasing production of natural antibacterial compounds, such as defensin-5 and lysozyme. Only recently, lysozyme was found up-regulated in Barrett’s oesophagitis, chronic gastritis, gluten-induced atrophic duodenitis (coeliac disease, collagenous colitis, lymphocytic colitis, and Crohn’s colitis. This up-regulation is a response directed to the special types of bacteria recently detected in these diseases. The aim of lysozyme up-regulation is to protect individual mucosal segments to chronic inflammation. The molecular mechanisms connected to the crosstalk between the intraluminal bacterial flora and the production of lysozyme released by the GI mucosae, are discussed. Bacterial resistance continues to exhaust our supply of commercial antibiotics. The potential use of lysozyme to treat infectious diseases is receiving much attention.

A comparative study on the aggregating effects of guanidine thiocyanate, guanidine hydrochloride and urea on lysozyme aggregation

Energy Technology Data Exchange (ETDEWEB)

Emadi, Saeed, E-mail: emadi@iasbs.ac.ir; Behzadi, Maliheh

2014-08-08

Highlights: • Lysozyme aggregated in guanidine thiocyanate (1.0 and 2.0 M). • Lysozyme aggregated in guanidine hydrochloride (4 and 5 M). • Lysozyme did not aggregated at any concentration (0.5–5 M) of urea. • Unfolding pathway is more important than unfolding per se in aggregation. - Abstract: Protein aggregation and its subsequent deposition in different tissues culminate in a diverse range of diseases collectively known as amyloidoses. Aggregation of hen or human lysozyme depends on certain conditions, namely acidic pH or the presence of additives. In the present study, the effects on the aggregation of hen egg-white lysozyme via incubation in concentrated solutions of three different chaotropic agents namely guanidine thiocyanate, guanidine hydrochloride and urea were investigated. Here we used three different methods for the detection of the aggregates, thioflavin T fluorescence, circular dichroism spectroscopy and atomic force microscopy. Our results showed that upon incubation with different concentrations (0.5, 1.0, 2.0, 3.0, 4.0, 5.0 M) of the chemical denaturants, lysozyme was aggregated at low concentrations of guanidine thiocyanate (1.0 and 2.0 M) and at high concentrations of guanidine hydrochloride (4 and 5 M), although no fibril formation was detected. In the case of urea, no aggregation was observed at any concentration.

Obtaining of lysozyme spherulitic forms at ambient temperature using pyrrolidinium octanoat as ionic liquid additive

Directory of Open Access Journals (Sweden)

Claudia Simona ŞTEFAN

2012-12-01

Full Text Available Pyrrolidinium octanoate carboxylate (Py+CnH2n+1COO-; PyO in abbreviation was used as additive for advanced crystallization of Lysozyme protein, to investigate the impact of protic ionic liquid on the protein crystal morphology. The ionic liquid was synthesized by acidic-base Brönsted neutralization, and its purity was checked by HPLC. The protein crystallization was made through the method of vapour diffusion with hanging drops. Crystallization experiments of Lysozyme with the addition of PyO were performed at 0.4 M PyO and respectively 1.6 M. The morphological of spherulitic forms of Lysozyme in aqueous solutions of PyO protic ionic liquid was investigated by optical microscopy after trials were incubated at ambient temperature (18-20°C, in various growth periods (3 days and 1 week. Hanging-drop vapour diffusion crystallization experiments with the addition of 0.4 M of PyO show that Lysozyme crystallized in type I spherulitic form. This is assumed to be a result of heterogeneous nucleation, with thin needles radially growing outwardfrom a more or less spherical particle. Hanging-drop vapour diffusioncrystallization experiments revealed that the addition of 1.6 M of PyO led to a second type of spherulitic form of the Lysozyme.

The disruptive effect of lysozyme on the bacterial cell wall explored by an in-silico structural outlook.

Science.gov (United States)

Primo, Emiliano D; Otero, Lisandro H; Ruiz, Francisco; Klinke, Sebastián; Giordano, Walter

2018-01-01

The bacterial cell wall, a structural unit of peptidoglycan polymer comprised of glycan strands consisting of a repeating disaccharide motif [N-acetylglucosamine (NAG) and N-acetylmuramylpentapeptide (NAM pentapeptide)], encases bacteria and provides structural integrity and protection. Lysozymes are enzymes that break down the bacterial cell wall and disrupt the bacterial life cycle by cleaving the linkage between the NAG and NAM carbohydrates. Lab exercises focused on the effects of lysozyme on the bacterial cell wall are frequently incorporated in biochemistry classes designed for undergraduate students in diverse fields as biology, microbiology, chemistry, agronomy, medicine, and veterinary medicine. Such exercises typically do not include structural data. We describe here a sequence of computer tasks designed to illustrate and reinforce both physiological and structural concepts involved in lysozyme effects on the bacterial cell-wall structure. This lab class usually lasts 3.5 hours. First, the instructor presents introductory concepts of the bacterial cell wall and the effect of lysozyme on its structure. Then, students are taught to use computer modeling to visualize the three-dimensional structure of a lysozyme in complex with bacterial cell-wall fragments. Finally, the lysozyme inhibitory effect on a bacterial culture is optionally proposed as a simple microbiological assay. The computer lab exercises described here give students a realistic understanding of the disruptive effect of lysozymes on the bacterial cell wall, a crucial component in bacterial survival. © 2017 by The International Union of Biochemistry and Molecular Biology, 46(1):83-90, 2018. © 2017 The International Union of Biochemistry and Molecular Biology.

Development of tantalum oxynitride thin films produced by PVD: Study of structural stability

Energy Technology Data Exchange (ETDEWEB)

Cristea, D. [Centro de Física, Universidade do Minho, Campus de Gualtar, 4710-057 Braga (Portugal); Department of Materials Science, Transylvania University, 500036 Brasov (Romania); Crisan, A. [Department of Materials Science, Transylvania University, 500036 Brasov (Romania); Barradas, N.P.; Alves, E. [Instituto Superior Técnico, Universidade Técnica de Lisboa Estrada Nacional 10, ao km 139,7 2695-066, Bobadela LRS (Portugal); Moura, C.; Vaz, F. [Centro de Física, Universidade do Minho, Campus de Gualtar, 4710-057 Braga (Portugal); Cunha, L., E-mail: lcunha@fisica.uminho.pt [Centro de Física, Universidade do Minho, Campus de Gualtar, 4710-057 Braga (Portugal)

2013-11-15

The purpose of this work is to study the evolution of the structure and of the thermal stability of a group of tantalum oxynitride thin films, prepared by magnetron sputtering, under the influence of vacuum annealing, up to a temperature of 800 °C. When varying the partial pressure of the reactive gases (P{sub O{sub 2+N{sub 2}}}), during the deposition process, the films change from a structure with a combination of poorly developed crystallites of the tetragonal β-Ta and of the face centred cubic (fcc) Ta(O,N) phases, for the films deposited with low P{sub O2+N2}, to a quasi-amorphous structure, for the films deposited with highest pressures. For intermediate pressures, the films reveal the presence of the fcc-Ta(O,N) structure. This structure corresponds to O atoms substituting some of the N atoms on the fcc-TaN structure and/or N atoms substituting O atoms of the fcc-γ-TaO structure. When subjected to the thermal annealing at 700 °C or higher, the film produced with lowest partial pressure revealed a remarkable structural change. New diffraction peaks appear and can only be attributed to a sub-stoichiometric hexagonal tantalum nitride structure. The film did not reveal any signs of delamination or cracks after all annealing temperatures. The two films produced with highest partial pressure proved to be the most stable. Structurally, they maintain the amorphous structure after all the annealing treatments and, in addition, no cracks or delamination were detected.

Pulsed electric field (PEF)-induced aggregation between lysozyme, ovalbumin and ovotransferrin in multi-protein system.

Science.gov (United States)

Wu, Li; Zhao, Wei; Yang, Ruijin; Yan, Wenxu

2015-05-15

The aggregation of multi-proteins is of great interest in food processing and a good understanding of the formation of aggregates during PEF processing is needed for the application of the process to pasteurize protein-based foods. The aggregates formation of a multi-protein system (containing ovalbumin, ovotransferrin and lysozyme) was studied through turbidity, size exclusion chromatography and SDS-PAGE patterns for interaction studies and binding forces. Results from size exclusion chromatography indicated that there was no soluble aggregates formed during PEF processing. The existence of lysozyme was important to form insoluble aggregates in the chosen ovalbumin solution. The results of SDS-PAGE patterns indicated that lysozyme was prone to precipitate, and was relatively the higher component of aggregates. Citric acid could be effective in inhibiting lysozyme from interacting with other proteins during PEF processing. Blocking the free sulphydryl by N-ethylmaleimide (NEM) did not affect aggregation inhibition. Copyright © 2014 Elsevier Ltd. All rights reserved.

Ethylenediaminetetraacetate and lysozyme improves antimicrobial activities of ovotransferrin against Escherichia coli O157:H7.

Science.gov (United States)

Ko, K Y; Mendoncam, A F; Ismail, H; Ahn, D U

2009-02-01

The aim of this study was to evaluate the effect of EDTA, lysozyme, or the combination of EDTA and lysozyme on the antibacterial activity of ovotransferrin against Escherichia coli O157:H7. Ovotransferrin solutions (20 mg/mL) containing 100 mM NaHCO3 (OS) with added EDTA (2.0 or 2.5 mg/mL), lysozyme (1.0, 1.5, or 2.0 mg/mL), or both were prepared. The antibacterial activities of OS, OSE (OS+EDTA), or OSL (OS+lysozyme) against E. coli O157:H7 in model systems were investigated by turbidity and viability tests. In addition, OSE, OSL, or OSEL (OS+EDTA+lysozyme) was applied to irradiated pork chops and commercial hams to determine whether the solutions had antibacterial activity on meat products. The effect of the initial cell population on the antibacterial activity of OSE, OSL, and OSEL was determined. Ethylenediaminetetraacetate at 2 mg/mL plus OS induced a reduction of approximately 3 to 4 log in viable E. coli O157:H7 cells in brain heart infusion broth media, and 1 mg/mL of lysozyme plus OS resulted in a reduction of approximately 0.5 to 1.0 log during a 36-h incubation at 35 degrees C. However, neither OSE nor OSEL showed a significant antibacterial effect on pork chops and hams during storage at 10 degrees C. The initial cell number in media did not affect the antibacterial activity of OSE or OSEL against E. coli O157:H7. This study demonstrates that combinations of ovotransferrin, NaHCO3, and EDTA have the potential to control E. coli O157:H7.

Dye-Affinity Nanofibrous Membrane for Adsorption of Lysozyme: Preparation and Performance Evaluation

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Steven Sheng-Shih Wang

2018-01-01

Full Text Available Polyacrylonitrile (PAN nanofibrous membrane was prepared by an electrospinning technique. After heat treatment and alkaline hydrolysis, the weak ion exchange membrane was grafted with chitosan molecule and then covalently immobilized with a Cibacron Blue F3GA (CB. Fibre diameter, porosity and pore size of the membrane and immobilized dye density were characterized. Furthermore, the membrane was applied to evaluate the binding performance of lysozyme under various operating parameters (pH, chitosan mass per volume ratio, dye concentration, ionic strength and temperature in batch mode. The experimental results were directly applied to purify lysozyme from chicken egg white by membrane chromatography. The results showed that the capture efficiency, recovery yield and purification factor were 90 and 87 %, and 47-fold, respectively, in a single step. The binding capacity remained consistent after five repeated cycles of adsorption-desorption operations. This work demonstrates that the dye-affinity nanofibrous membrane holds great potential for purification of lysozyme from real feedstock.

Experimental study of the polymer powder film thickness uniformity produced by the corona discharge

Science.gov (United States)

Fazlyyyakhmatov, Marsel

2017-01-01

The results of an experimental study of the polymer powder film thickness uniformity are presented. Polymer powder films are produced by the electrostatic field of corona discharge. Epoxy and epoxy-polyester powder films with thickness in the range of 30-120 microns are studied. Experimentally confirmed possibility of using these coatings as protective matching layer of piezoceramic transducers at frequencies of 0.5-15 MHz.

Near-surface hydrogen depletion of diamond-like carbon films produced by direct ion deposition

Science.gov (United States)

Markwitz, Andreas; Gupta, Prasanth; Mohr, Berit; Hübner, René; Leveneur, Jerome; Zondervan, Albert; Becker, Hans-Werner

2016-03-01

Amorphous atomically flat diamond-like carbon (DLC) coatings were produced by direct ion deposition using a system based on a Penning ion source, butane precursor gas and post acceleration. Hydrogen depth profiles of the DLC coatings were measured with the 15N R-NRA method using the resonant nuclear reaction 1H(15N, αγ)12C (Eres = 6.385 MeV). The films produced at 3.0-10.5 kV acceleration voltage show two main effects. First, compared to average elemental composition of the film, the near-surface region is hydrogen depleted. The increase of the hydrogen concentration by 3% from the near-surface region towards the bulk is attributed to a growth model which favours the formation of sp2 hybridised carbon rich films in the film formation zone. Secondly, the depth at which the maximum hydrogen concentration is measured increases with acceleration voltage and is proportional to the penetration depth of protons produced by the ion source from the precursor gas. The observed effects are explained by a deposition process that takes into account the contributions of ion species, hydrogen effusion and preferential displacement of atoms during direct ion deposition.

Deposition and characterization of ITO films produced by laser ablation at 355 nm

DEFF Research Database (Denmark)

Holmelund, E.; Thestrup Nielsen, Birgitte; Schou, Jørgen

2002-01-01

Indium tin oxide (ITO) films have been deposited by pulsed laser deposition (PLD) at 355 nm. Even though the absorption of laser light at the wavelength 355 nm is much smaller than that of the standard excimer lasers for PLD at 248 nm and 193 nm, high-quality films can be produced. At high fluence...

Ultra-sensitive quantification of lysozyme based on element chelate labeling and capillary electrophoresis–inductively coupled plasma mass spectrometry

International Nuclear Information System (INIS)

Yang, MingWei; Wu, WeiHua; Ruan, YaJuan; Huang, LiMei; Wu, Zujian; Cai, Yong; Fu, FengFu

2014-01-01

Graphical abstract: An ultra-sensitive method for the determination of lysozyme was developed based on the Gd 3+ chelate labeling and CE–ICP–MS. The proposed method has an extremely low detection limit of 3.89 attomole and has been successfully used to detect lysozyme in saliva sample, showing excellent reliability. The success of the present method provides a new possibility for biological assays and clinical diagnoses. -- Highlights: •An ultra-sensitive method for detecting lysozyme based on CE–ICP–MS was described. •The proposed method has an extremely low detection limit of 3.89 attomole. •It can be used to detect trace lysozyme in saliva sample with a satisfied recovery. •The method provides a new potential for sensitive detection of low-abundant proteins. -- Abstract: In this study, an ultra-sensitive method for the quantification of lysozyme based on the Gd 3+ diethylenetriamine-N,N,N′,N″,N″-pentaacetic acid labeling and capillary electrophoresis–inductively coupled plasma mass spectrometry (CE–ICP–MS) was described. The Gd 3+ -tagged lysozyme was effectively separated by capillary electrophoresis (CE) and sensitively determined by inductively coupled plasma mass spectrometry (ICP–MS). Based on the gadolinium-tagging and CE–ICP–MS, the lysozyme was determined within 12 min with an extremely low detection limit of 3.89 attomole (3.89 × 10 −11 mol L −1 for 100 nL of sample injection) and a RSD < 6% (n = 5). The proposed method has been successfully used to detect lysozyme in saliva samples with a recovery of 91–106%, suggesting that our method is sensitive and reliable. The success of the present method provides a new potential for the biological assays and sensitive detection of low-abundant proteins

Adsorption of lysozyme unto silica and polystyrene surfaces in ...

African Journals Online (AJOL)

user

2011-04-11

Apr 11, 2011 ... surfaces were well fitted by the Langmuir adsorption isotherm model with maximum adsorption .... following reasons: (1) Lysozyme is a globular protein with ... vigorously for 1 h to attain equilibrium adsorption and allowed to.

Functional Micrococcus lysodeikticus layers deposited by laser technique for the optical sensing of lysozyme.

Science.gov (United States)

Dinca, Valentina; Zaharie-Butucel, Diana; Stanica, Luciana; Brajnicov, Simona; Marascu, Valentina; Bonciu, Anca; Cristocea, Andra; Gaman, Laura; Gheorghiu, Mihaela; Astilean, Simion; Vasilescu, Alina

2018-02-01

Whole cell optical biosensors, made by immobilizing whole algal, bacterial or mammalian cells on various supports have found applications in several fields, from ecology and ecotoxicity testing to biopharmaceutical production or medical diagnostics. We hereby report the deposition of functional bacterial layers of Micrococcus lysodeikticus (ML) via Matrix-Assisted Pulsed Laser Evaporation (MAPLE) on poly(diallyldimethylamonium) (PDDA)-coated-glass slides and their application as an optical biosensor for the detection of lysozyme in serum. Lysozyme is an enzyme upregulated in inflammatory diseases and ML is an enzymatic substrate for this enzyme. The MAPLE-deposited bacterial interfaces were characterised by Scanning Electron Microscopy (SEM), Atomic Force Microscopy (AFM), Fourier-Transformed Infrared Spectroscopy (FTIR), Raman and optical microscopy and were compared with control interfaces deposited via layer-by-layer on the same substrate. After MAPLE deposition and coating with graphene oxide (GO), ML-modified interfaces retained their functionality and sensitivity to lysozyme's lytic action. The optical biosensor detected lysozyme in undiluted serum in the clinically relevant range up to 10μgmL -1 , in a fast and simple manner. Copyright © 2017 Elsevier B.V. All rights reserved.

Thickness determination of large-area films of yttria-stabilized zirconia produced by pulsed laser deposition

Energy Technology Data Exchange (ETDEWEB)

Pryds, N. [Materials Research Department, Riso National Laboratory, DK-4000 Roskilde (Denmark)]. E-mail: nini.pryds@risoe.dk; Toftmann, B. [Department of Optics and Plasma Research, Riso National Laboratory, DK-4000 Roskilde (Denmark); Bilde-Sorensen, J.B. [Materials Research Department, Riso National Laboratory, DK-4000 Roskilde (Denmark); Schou, J. [Department of Optics and Plasma Research, Riso National Laboratory, DK-4000 Roskilde (Denmark); Linderoth, S. [Materials Research Department, Riso National Laboratory, DK-4000 Roskilde (Denmark)

2006-04-30

Films of yttria-stabilized zirconia (YSZ) on a polished silicon substrate of diameter up to 125 mm have been produced in a large-area pulsed laser deposition (PLD) setup under typical PLD conditions. The film thickness over the full film area has been determined by energy-dispersive X-ray spectrometry in a scanning electron microscope (SEM) with use of a method similar to one described by Bishop and Poole. The attenuation of the electron-induced X-rays from the Si wafer by the film was monitored at a number of points along a diameter and the thickness was determined by Monte Carlo simulations of the attenuation for various values of film thickness with the program CASINO. These results have been compared with direct measurements in the SEM of the film thickness on a cross-section on one of the wafers. The results of these measurements demonstrate the ability of this technique to accurately determine the thickness of a large film, i.e. up to diameters of 125 mm, in a relatively short time, without destroying the substrate, without the need of a standard sample and without the need of a flat substrate. We have also demonstrated that by controlling the deposition parameters large-area YSZ films with uniform thickness can be produced.

Thickness determination of large-area films of yttria-stabilized zirconia produced by pulsed laser deposition

International Nuclear Information System (INIS)

Pryds, N.; Toftmann, B.; Bilde-Sorensen, J.B.; Schou, J.; Linderoth, S.

2006-01-01

Films of yttria-stabilized zirconia (YSZ) on a polished silicon substrate of diameter up to 125 mm have been produced in a large-area pulsed laser deposition (PLD) setup under typical PLD conditions. The film thickness over the full film area has been determined by energy-dispersive X-ray spectrometry in a scanning electron microscope (SEM) with use of a method similar to one described by Bishop and Poole. The attenuation of the electron-induced X-rays from the Si wafer by the film was monitored at a number of points along a diameter and the thickness was determined by Monte Carlo simulations of the attenuation for various values of film thickness with the program CASINO. These results have been compared with direct measurements in the SEM of the film thickness on a cross-section on one of the wafers. The results of these measurements demonstrate the ability of this technique to accurately determine the thickness of a large film, i.e. up to diameters of 125 mm, in a relatively short time, without destroying the substrate, without the need of a standard sample and without the need of a flat substrate. We have also demonstrated that by controlling the deposition parameters large-area YSZ films with uniform thickness can be produced

Poly lactic acid based injectable delivery systems for controlled release of a model protein, lysozyme.

Science.gov (United States)

Al-Tahami, Khaled; Meyer, Amanda; Singh, Jagdish

2006-02-01

The objective of this study was to evaluate the critical formulation parameters (i.e., polymer concentration, polymer molecular weight, and solvent nature) affecting the controlled delivery of a model protein, lysozyme, from injectable polymeric implants. The conformational stability and biological activity of the released lysozyme were also investigated. Three formulations containing 10%, 20%, and 30% (w/v) poly lactic acid (PLA) in triacetin were investigated. It was found that increasing polymer concentration in the formulations led to a lower burst effect and a slower release rate. Formulation with a high molecular weight polymer showed a greater burst effect as compared to those containing low molecular weight. Conformational stability and biological activity of released samples were studied by differential scanning calorimeter and enzyme activity assay, respectively. The released samples had significantly (P solution kept at same conditions). Increasing polymer concentration increased both the conformational stability and the biological activity of released lysozyme. In conclusion, phase sensitive polymer-based delivery systems were able to deliver a model protein, lysozyme, in a conformationally stable and biologically active form at a controlled rate over an extended period.

Use of lysozyme from chicken egg white as a nitrite replacer in an Italian-type chicken sausage

Directory of Open Access Journals (Sweden)

Nalaka Sandun Abeyrathne

2015-09-01

Full Text Available Background: Sodium or potassium nitrite is widely used as a curing agent in sausages and other cured meat products. Nitrite has strong antimicrobial and antioxidant effects and generates cured meat color. Nitrite, however, can react with secondary or tertiary amines in meat to form carcinogenic, teratogenic and mutagenic N-nitroso compounds. Several findings have been suggested that high consumption of processed meat may increase the risk of cancer, and emphasized that dietary nitrosamines are positively associated with cancer. Lysozyme is one of the major egg proteins that have antimicrobial and antioxidant characteristics. Therefore, lysozyme can be used in meat processing to prevent microbial growth and oxidative degradation in meat products during storage. This study is focused on evaluating the antimicrobial and antioxidant effects of lysozyme extracted from egg white as a replacer of nitrite in a cooked Italian-type chicken sausage. Methods: Four curing treatments including 100% nitrite (control, 100% lysozyme (treatment 1, 25% nitrite + 75% lysozyme (treatment 2 and 50% nitrite + 50% lysozyme (treatment 3 were used to prepare Italian-type chicken sausage samples. Recipe was developed with 64% (w/w meat, 17% (w/w binder (bread crumble, 12% (w/w ice, 4% (w/w vegetable oil, 2% (w/w salt, 1% (w/w spices (chili, black pepper, cardamom. Prepared samples were cooked in an 80 °C smoke house to a core temperature of 65 °C and cooled in cold water to 20-25 °C subsequently packed in polyethylene and stored in a freezer (-18 °C. The antimicrobial effect lysozyme was tested using Escherichia coli and Salmonella. The growth of these pathogens at 0, 3 and 5 days of storage of spore inoculation was determined. The antioxidant activity of lysozyme was determined using the TBARS value during the 25 d storage period. The redness (a*, lightness (L*, and yellowness (b* of sausages were analyzed using a Minolta color meter (CR 410, Konica Minolta Inc

Activity and conformation of lysozyme in molecular solvents, protic ionic liquids (PILs) and salt-water systems.

Science.gov (United States)

Wijaya, Emmy C; Separovic, Frances; Drummond, Calum J; Greaves, Tamar L

2016-09-21

Improving protein stabilisation is important for the further development of many applications in the pharmaceutical, specialty chemical, consumer product and agricultural sectors. However, protein stabilization is highly dependent on the solvent environment and, hence, it is very complex to tailor protein-solvent combinations for stable protein maintenance. Understanding solvent features that govern protein stabilization will enable selection or design of suitable media with favourable solution environments to retain protein native conformation. In this work the structural conformation and activity of lysozyme in 29 solvent systems were investigated to determine the role of various solvent features on the stability of the enzyme. The solvent systems consisted of 19 low molecular weight polar solvents and 4 protic ionic liquids (PILs), both at different water content levels, and 6 aqueous salt solutions. Small angle X-ray scattering, Fourier transform infrared spectroscopy and UV-vis spectroscopy were used to investigate the tertiary and secondary structure of lysozyme along with the corresponding activity in various solvation systems. At low non-aqueous solvent concentrations (high water content), the presence of solvents and salts generally maintained lysozyme in its native structure and enhanced its activity. Due to the presence of a net surface charge on lysozyme, electrostatic interactions in PIL-water systems and salt solutions enhanced lysozyme activity more than the specific hydrogen-bond interactions present in non-ionic molecular solvents. At higher solvent concentrations (lower water content), solvents with a propensity to exhibit the solvophobic effect, analogous to the hydrophobic effect in water, retained lysozyme native conformation and activity. This solvophobic effect was observed particularly for solvents which contained hydroxyl moieties. Preferential solvophobic effects along with bulky chemical structures were postulated to result in less

[The estimation of systemic chemotherapy treatment administered in breast cancer on lysozyme activity in tears--preliminary report].

Science.gov (United States)

Wojciechowska, Katarzyna; Jurowski, Piotr; Wieckowska-Szakiel, Marzena; Rózalska, Barbara

2012-01-01

Estimation of cytostatics influence used in breast cancer treatment on lysozyme activity in human tears depend on time of treatment. 8 women were treated at the base of chemotherapy schema: docetaxel with doxorubicin and 4 women treated with schema CMF: cyclophosphamide, methotrexate, 5-fluorouracil. Lysozyme activity in tears was assessed by measurement of diameter zone of Micrococcus lysodeicticus growth inhibition. It was revealed that both chemotherapy schema caused statistically significant reduction of diameter zone of M. lysodeicticus growth inhibition, after first and second course of chemotherapy treatment. After second chemotherapy course CMF schema induced loss of lysozyme activity in patient's tears (zero mm of M. lysodeicticus diameter zone growth inhibition). Systemic chemotherapy administered in breast cancer induce reduction of lysozyme activity in tears, that may cause higher morbidity of ocular surface infections caused by Gram-positive bacteria.

Near-surface hydrogen depletion of diamond-like carbon films produced by direct ion deposition

Energy Technology Data Exchange (ETDEWEB)

Markwitz, Andreas, E-mail: A.Markwitz@gns.cri.nz [GNS Science, Lower Hutt (New Zealand); The MacDiarmid Institute for Advanced Materials and Nanotechnology (New Zealand); Gupta, Prasanth [GNS Science, Lower Hutt (New Zealand); The MacDiarmid Institute for Advanced Materials and Nanotechnology (New Zealand); Mohr, Berit [GNS Science, Lower Hutt (New Zealand); Hübner, René [Institute of Ion Beam Physics and Materials Research, Helmholtz-Zentrum Dresden-Rossendorf (Germany); Leveneur, Jerome; Zondervan, Albert [GNS Science, Lower Hutt (New Zealand); Becker, Hans-Werner [RUBION, Ruhr-University Bochum (Germany)

2016-03-15

Amorphous atomically flat diamond-like carbon (DLC) coatings were produced by direct ion deposition using a system based on a Penning ion source, butane precursor gas and post acceleration. Hydrogen depth profiles of the DLC coatings were measured with the 15N R-NRA method using the resonant nuclear reaction {sup 1}H({sup 15}N, αγ){sup 12}C (E{sub res} = 6.385 MeV). The films produced at 3.0–10.5 kV acceleration voltage show two main effects. First, compared to average elemental composition of the film, the near-surface region is hydrogen depleted. The increase of the hydrogen concentration by 3% from the near-surface region towards the bulk is attributed to a growth model which favours the formation of sp{sup 2} hybridised carbon rich films in the film formation zone. Secondly, the depth at which the maximum hydrogen concentration is measured increases with acceleration voltage and is proportional to the penetration depth of protons produced by the ion source from the precursor gas. The observed effects are explained by a deposition process that takes into account the contributions of ion species, hydrogen effusion and preferential displacement of atoms during direct ion deposition.

Covalent immobilization of lysozyme onto woven and knitted crimped polyethylene terephthalate grafts to minimize the adhesion of broad spectrum pathogens

International Nuclear Information System (INIS)

Al Meslmani, Bassam M.; Mahmoud, Gihan F.; Leichtweiß, Thomas; Strehlow, Boris; Sommer, Frank O.; Lohoff, Michael D.; Bakowsky, Udo

2016-01-01

Graft-associated infections entirely determine the short-term patency of polyethylene terephthalate PET cardiovascular graft. We attempted to enzymatically inhibit the initial bacterial adhesion to PET grafts using lysozyme. Lysozyme was covalently immobilized onto woven and knitted forms of crimped PET grafts by the end-point method. Our figures of merit revealed lysozyme immobilization yield of 15.7 μg/cm"2, as determined by the Bradford assay. The activity of immobilized lysozyme on woven and knitted PET manifested 58.4% and 55.87% using Micrococcus lysodeikticus cells, respectively. Noteworthy, the adhesion of vein catheter-isolated Staphylococcus epidermidis decreased by 6- to 8-folds and of Staphylococcus aureus by 11- to 12-folds, while the Gram-negative Escherichia coli showed only a decrease by 3- to 4-folds. The anti-adhesion efficiency was specific for bacterial cells and no significant effect was observed on adhesion and growth of L929 cells. In conclusion, immobilization of lysozyme onto PET grafts can inhibit the graft-associated infection. - Highlights: • Lysozyme was covalently immobilized on crimped polyethylene terephthalate (PET). • The activity of immobilized lysozyme was meaningfully reduced. • The maintained activity significantly declined the adhesion of Gram-positive stains. • The enzymatic anti-adhesion efficiency reported lesser extent against Gram-negative. • The anti-bacterial activity displayed no significant effect on cells compatibility.

Covalent immobilization of lysozyme onto woven and knitted crimped polyethylene terephthalate grafts to minimize the adhesion of broad spectrum pathogens

Energy Technology Data Exchange (ETDEWEB)

Al Meslmani, Bassam M., E-mail: almeslmanib@yahoo.com [Department of Pharmaceutical Technology and Biopharmaceutics, Marburg University, Ketzerbach 63, 35037 Marburg (Germany); Mahmoud, Gihan F., E-mail: mahmoudg@staff.uni-marburg.de [Department of Pharmaceutical Technology and Biopharmaceutics, Marburg University, Ketzerbach 63, 35037 Marburg (Germany); Department of Pharmaceutics and Industrial Pharmacy, Helwan University, Ain Helwan, 11795 Cairo (Egypt); Leichtweiß, Thomas, E-mail: Thomas.Leichtweiss@phys.Chemie.uni-giessen.de [Institute of Physical Chemistry, Justus-Liebig-University Giessen, Heinrich-Buff-Ring 58, 35392 Giessen (Germany); Strehlow, Boris, E-mail: strehlo4@staff.uni-marburg.de [Department of Pharmaceutical Technology and Biopharmaceutics, Marburg University, Ketzerbach 63, 35037 Marburg (Germany); Sommer, Frank O., E-mail: sommerf@med.uni-marburg.de [Institute for Medical Microbiology and Hospital Hygiene, Marburg University, Hans Meerwein Str 2, 35032 Marburg (Germany); Lohoff, Michael D., E-mail: lohoff@med.uni-marburg.de [Institute for Medical Microbiology and Hospital Hygiene, Marburg University, Hans Meerwein Str 2, 35032 Marburg (Germany); Bakowsky, Udo, E-mail: ubakowsky@aol.com [Department of Pharmaceutical Technology and Biopharmaceutics, Marburg University, Ketzerbach 63, 35037 Marburg (Germany)

2016-01-01

Graft-associated infections entirely determine the short-term patency of polyethylene terephthalate PET cardiovascular graft. We attempted to enzymatically inhibit the initial bacterial adhesion to PET grafts using lysozyme. Lysozyme was covalently immobilized onto woven and knitted forms of crimped PET grafts by the end-point method. Our figures of merit revealed lysozyme immobilization yield of 15.7 μg/cm{sup 2}, as determined by the Bradford assay. The activity of immobilized lysozyme on woven and knitted PET manifested 58.4% and 55.87% using Micrococcus lysodeikticus cells, respectively. Noteworthy, the adhesion of vein catheter-isolated Staphylococcus epidermidis decreased by 6- to 8-folds and of Staphylococcus aureus by 11- to 12-folds, while the Gram-negative Escherichia coli showed only a decrease by 3- to 4-folds. The anti-adhesion efficiency was specific for bacterial cells and no significant effect was observed on adhesion and growth of L929 cells. In conclusion, immobilization of lysozyme onto PET grafts can inhibit the graft-associated infection. - Highlights: • Lysozyme was covalently immobilized on crimped polyethylene terephthalate (PET). • The activity of immobilized lysozyme was meaningfully reduced. • The maintained activity significantly declined the adhesion of Gram-positive stains. • The enzymatic anti-adhesion efficiency reported lesser extent against Gram-negative. • The anti-bacterial activity displayed no significant effect on cells compatibility.

Laser ablation of lysozyme with UV, visible and infrared femto- and nanosecond pulses

DEFF Research Database (Denmark)

Schou, Jørgen; Canulescu, Stela; Matei, Andreea

Lysozyme is an interesting molecule for laser ablation of organic materials, because the ablation has been comprehensively studied, it is a medium heavy molecule with a mass of 14305 Da, which can be detected by standard techniques, and because it is used as a bactericidal protein in the food...... industry. Lysozyme molecules do not absorb energy for wavelengths above 310 nm, but nevertheless there is a strong mass loss by ablation for laser irradiation in the visible regime. The total ablation yield of lysozyme at 355 nm and at 2 J/cm2 is about 155 µg/pulse, possibly one of the highest ablation...... the ablation process for different wavelengths and time duration. Measurements for 6-7-ns laser ablation were carried out at DTU on Risø Campus, while measurements with pulses of 300 fs were carried out at the University of Naples in a similar setup. For all wavelengths except at nanosecond laser pulses at 355...

Positron annihilation studies in lysozyme and catalase

International Nuclear Information System (INIS)

Rohilla, Y.; Singh, K.P.; Roy Choudhury, S.; Jain, P.C.

1992-01-01

Positron annihilation studies have been carried out in two enzymes, lysozyme and catalase. Temperature dependence of the positron lifetimes in these two enzymes has been investigated. The results explained in terms of the free volume model and fluctuations between different conformational micro states of enzyme structures provide a new insight into the mechanism of bio-activity of these enzymes. (author). 15 refs., 4 figs

Transport properties of MnTe films with cracks produced in thermal cycling process

Energy Technology Data Exchange (ETDEWEB)

Yang, Liang; Wang, Zhenhua; Zhang, Zhidong [Institute of Metal Research, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shenyang National Laboratory for Materials Science, Shenyang (China)

2017-10-15

As a promising material in antiferromagnetic spintronics, MnTe films manifested complex characteristics according to previous reports. In this work, we investigate in details the temperature dependence of resistivity of MnTe films grown on SiO{sub 2}/Si substrate and focus on the divaricating of cooling and warming resistivity-temperature (R-T) curves. It is found that such a divaricating in resistivity is associated with cracks produced in thermal cycles. By comparing the crystalline character and the morphology before and after the cycles, we verify the appearance of cracks and the release of stress in the films. Based on the temperature dependence of thermal-expansion coefficient of Si and MnTe, the origin of the cracks is the mismatched thermal-expansion coefficient (α). The humps, which only appear in the R-T curve of the first cooling process, are attributed to the produced cracks and/or the unreleased stress. (orig.)

Structural basis of protein oxidation resistance: a lysozyme study.

Directory of Open Access Journals (Sweden)

Marion Girod

Full Text Available Accumulation of oxidative damage in proteins correlates with aging since it can cause irreversible and progressive degeneration of almost all cellular functions. Apparently, native protein structures have evolved intrinsic resistance to oxidation since perfectly folded proteins are, by large most robust. Here we explore the structural basis of protein resistance to radiation-induced oxidation using chicken egg white lysozyme in the native and misfolded form. We study the differential resistance to oxidative damage of six different parts of native and misfolded lysozyme by a targeted tandem/mass spectrometry approach of its tryptic fragments. The decay of the amount of each lysozyme fragment with increasing radiation dose is found to be a two steps process, characterized by a double exponential evolution of their amounts: the first one can be largely attributed to oxidation of specific amino acids, while the second one corresponds to further degradation of the protein. By correlating these results to the structural parameters computed from molecular dynamics (MD simulations, we find the protein parts with increased root-mean-square deviation (RMSD to be more susceptible to modifications. In addition, involvement of amino acid side-chains in hydrogen bonds has a protective effect against oxidation Increased exposure to solvent of individual amino acid side chains correlates with high susceptibility to oxidative and other modifications like side chain fragmentation. Generally, while none of the structural parameters alone can account for the fate of peptides during radiation, together they provide an insight into the relationship between protein structure and susceptibility to oxidation.

Effects of prolonged running in the heat and cool environments on selected physiological parameters and salivary lysozyme responses

Directory of Open Access Journals (Sweden)

Nur S. Ibrahim

2017-12-01

Conclusion: This study found similar lysozyme responses between both hot and cool trials. Thus, room/ambient temperature did not affect lysozyme responses among recreational athletes. Nevertheless, the selected physiological parameters were significantly affected by room temperature.

Dynamics of hydration in hen egg white lysozyme.

Science.gov (United States)

Sterpone, F; Ceccarelli, M; Marchi, M

2001-08-10

We investigate the hydration dynamics of a small globular protein, hen egg-white lysozyme. Extensive simulations (two trajectories of 9 ns each) were carried out to identify the time-scales and mechanism of water attachment to this protein. The location of the surface and integral water molecules in lysozyme was also investigated. Three peculiar temporal scales of the hydration dynamics can be discerned: two among these, with sub-nanosecond mean residence time, tau(w), are characteristic of surface hydration water; the slower time-scale (tau(w) approximately 2/3 ns) is associated with buried water molecules in hydrophilic pores and in superficial clefts. The computed tau(w) values in the two independent runs fall in a similar range and are consistent with each other, thus adding extra weight to our result. The tau(w) of surface water obtained from the two independent trajectories is 20 and 24 ps. In both simulations only three water molecules are bound to lysozyme for the entire length of the trajectories, in agreement with nuclear magnetic relaxation dispersion estimates. Locations other than those identified in the protein crystal are found to be possible for these long-residing water molecules. The dynamics of the hydration water molecules observed in our simulations implies that each water molecule visits a multitude of residues during the lifetime of its bound with the protein. The number of residues seen by a single water molecule increases with the time-scale of its residence time and, on average, is equal to one only for the water molecules with shorter residence time. Thus, tau(w) values obtained from inelastic neutron scattering and based on jump-diffusion models are likely not to account for the contribution of water molecules with longer residence time. Copyright 2001 Academic Press.

Lysozyme as an alternative to antibiotics improves growth performance and small intestinal morphology in nursery pigs.

Science.gov (United States)

Oliver, W T; Wells, J E

2013-07-01

Lysozyme is a 1,4-β-N-acetylmuramidase that has antimicrobial properties. The objective of this experiment was to determine if lysozyme in nursery diets improved growth performance and gastrointestinal health of pigs weaned from the sow at 24 d of age. Two replicates of 96 pigs (192 total; 96 males, 96 females) were weaned from the sow at 24 d of age, blocked by BW and gender, and then assigned to 1 of 24 pens (4 pigs/pen). Each block was randomly assigned 1 of 3 dietary treatments for 28 d: control (two 14-d phases), control + antibiotics (carbadox/copper sulfate), or control + lysozyme (100 mg/kg diet). Pigs were weighed and blood sampled on d 0, 14, and 28 of treatment. Blood was analyzed for plasma urea nitrogen (PUN) and IgA. At 28 d, pigs were killed, and samples of jejunum and ileum were collected and fixed for intestinal morphology measurements. An additional jejunum sample was taken from the 12 pigs with the median BW per treatment to determine transepithelial electrical resistance (TER). Pigs consuming antibiotics or lysozyme grew at a faster rate than control pigs (0.433 ± 0.009 and 0.421 ± 0.008 vs. 0.398 ± 0.008 kg/d, respectively; P 0.48), but G:F was improved in pigs consuming antibiotics or lysozyme (0.756 ± 0.014, 0.750 ± 0.021, and 0.695 ± 0.019 kg/kg; P 0.48). Dietary treatment did not affect TER (P > 0.37), but gilts had lower TER compared with barrows (P 0.53). However, jejunum villi height was increased and crypt depth was decreased in pigs consuming antibiotics or lysozyme (P pigs weaned from the sow at 24 d of age.

Corneal cell adhesion to contact lens hydrogel materials enhanced via tear film protein deposition.

Directory of Open Access Journals (Sweden)

Claire M Elkins

Full Text Available Tear film protein deposition on contact lens hydrogels has been well characterized from the perspective of bacterial adhesion and viability. However, the effect of protein deposition on lens interactions with the corneal epithelium remains largely unexplored. The current study employs a live cell rheometer to quantify human corneal epithelial cell adhesion to soft contact lenses fouled with the tear film protein lysozyme. PureVision balafilcon A and AirOptix lotrafilcon B lenses were soaked for five days in either phosphate buffered saline (PBS, borate buffered saline (BBS, or Sensitive Eyes Plus Saline Solution (Sensitive Eyes, either pure or in the presence of lysozyme. Treated contact lenses were then contacted to a live monolayer of corneal epithelial cells for two hours, after which the contact lens was sheared laterally. The apparent cell monolayer relaxation modulus was then used to quantify the extent of cell adhesion to the contact lens surface. For both lens types, lysozyme increased corneal cell adhesion to the contact lens, with the apparent cell monolayer relaxation modulus increasing up to an order of magnitude in the presence of protein. The magnitude of this increase depended on the identity of the soaking solution: lenses soaked in borate-buffered solutions (BBS, Sensitive Eyes exhibited a much greater increase in cell attachment upon protein addition than those soaked in PBS. Significantly, all measurements were conducted while subjecting the cells to moderate surface pressures and shear rates, similar to those experienced by corneal cells in vivo.

Surface-enhanced Raman spectroscopy (SERS) using Ag nanoparticle films produced by pulsed laser deposition

Energy Technology Data Exchange (ETDEWEB)

Smyth, C.A., E-mail: smythc2@tcd.ie [School of Physics, Trinity College Dublin, Dublin 2 (Ireland); Mirza, I.; Lunney, J.G.; McCabe, E.M. [School of Physics, Trinity College Dublin, Dublin 2 (Ireland)

2013-01-01

Highlights: Black-Right-Pointing-Pointer Pulsed laser deposition (PLD) produces silver nanoparticle films. Black-Right-Pointing-Pointer These films can be used for surface-enhanced Raman spectroscopy (SERS). Black-Right-Pointing-Pointer Commercial film shows good SERS reproducibility but poor signal intensity. Black-Right-Pointing-Pointer PLD shows a good SERS response coupled with good reproducibility. - Abstract: Thin silver nanoparticle films, of thickness 7 nm, were deposited onto glass microslides using pulsed laser deposition (PLD). The films were then characterised using UV-vis spectroscopy and scanning transmission electron microscopy before Rhodamine 6G was deposited onto them for investigation using surface-enhanced Raman spectroscopy (SERS). The sensitivity obtained using SERS was compared to that obtained using a colloidal silver suspension and also to a commercial SERS substrate. The reproducibility of the films is also examined using statistical analysis.

Combined pulmonary involvement in hereditary lysozyme amyloidosis with associated pulmonary sarcoidosis: a case report.

Science.gov (United States)

McCarthy, Cormac; Deegan, Alexander P; Garvey, John F; McDonnell, Timothy J

2013-12-17

Sarcoidosis is a multisystem inflammatory disorder of unknown cause which can affect any organ system. Autosomal dominant lysozyme amyloidosis is a very rare form of hereditary amyloidosis. The Arg64 variant is extraordinarily rare with each family showing a particular pattern of organ involvement, however while Sicca syndrome, gastrointestinal involvement and renal failure are common, lymph node involvement is very rare. In this case report we describe the first reported case of sarcoidosis in association with hereditary lysozyme amyloidosis.

Gold nanoparticles and films produced by a laser ablation/gas deposition (LAGD) method

International Nuclear Information System (INIS)

Kawakami, Yuji; Seto, Takafumi; Yoshida, Toshinobu; Ozawa, Eiichi

2002-01-01

Gold nanoparticles have great potential for various nanoelectronic applications such as single electron transistors, an infrared absorption sensor and so on. It is very important to understand and control the size distribution of the particles for such a variety of applications. In this paper, we report the size distribution of gold nanoparticles and the relationship between the nanoparticle-films and the electrical property produced by a laser ablation method. Gold nanoparticle-films were prepared by a technique, which sprays nanoparticles on the substrate through a nozzle. We call it a gas deposition method. The nanoparticles were generated by the nanosecond pulsed Nd:YAG laser ablation of a gold substrate under a low-pressure inert gas atmosphere. The ambient pressure was changed to control the average size and their distribution. The particles produced in the generation chamber were transported by a helium carrier gas to the deposition chamber and deposited on a substrate to form the films composed of gold nanoparticles. The electrical resistivity of the generated gold nanoparticle-films on the glass substrates was measured using a four-probe method. The size distribution of the nanoparticles was examined using transmission electron microscopy (TEM) and a low-pressure differential mobility analyzer (LP-DMA). The relationship between the particle size and the electrical properties of each film made by the different synthesis conditions were analyzed. The electrical resistivity changed from the order of 10 -5 to 10 -1 Ω cm depending on the ambient pressure and the size distribution

Type II textured molybdenum disulphide films produced by direct vapour transport and rf-magnetron sputtering

International Nuclear Information System (INIS)

Bohlken, S.F.; Lemon, K.D.; Jakovidis, G.; Taheri, E.H.

1999-01-01

Full text: Molybdenum disulphide (MoS 2 ) is one of the few naturally occurring Layered Transition Metal Dichalcogenides and is the primary source for elemental molybdenum. It displays exceptional lubrication performance in both vacuum and atmospheric conditions over a wide temperature range. An important emerging application of MoS 2 and related materials is photovoltaics. Films of MoS 2 exhibit several morphologies described by the orientation of platelets with respect to the substrate. Films with platelets perpendicular or parallel to the substrate are referred to by their morphology, which is type-I or type-II respectively. Production of exclusive type-II films is highly desirable in applications involving lubrication and photovoltaics. For example, type-II morphology reduces friction and minority carrier recombination centres, thus improving tribological and photovoltaic performance. We have successfully produced type-II films using both direct vapour transport and rf-magnetron sputtering Continuous polycrystalline films (∼ 10 μm thick) grown in our laboratory using vapour transport have typical areas 1000 mm 2 . A novel ejecta filtration technique was applied to rf-magnetron sputtering. Films produced using this approach retain exclusive type-II morphology at thicknesses where type-I would normally be observed (∼ 200nm)

Lysozyme's lectin-like characteristics facilitates its immune defense function

KAUST Repository

Zhang, Ruiyan

2017-06-06

Interactions between human lysozyme (HL) and the lipopolysaccharide (LPS) of Klebsiella pneumoniae O1, a causative agent of lung infection, were identified by surface plasmon resonance. To characterize the molecular mechanism of this interaction, HL binding to synthetic disaccharides and tetrasaccharides representing one and two repeating units, respectively, of the O-chain of this LPS were studied. pH-dependent structural rearrangements of HL after interaction with the disaccharide were observed through nuclear magnetic resonance. The crystal structure of the HL-tetrasaccharide complex revealed carbohydrate chain packing into the A, B, C, and D binding sites of HL, which primarily occurred through residue-specific, direct or water-mediated hydrogen bonds and hydrophobic contacts. Overall, these results support a crucial role of the Glu35/Asp53/Trp63/Asp102 residues in HL binding to the tetrasaccharide. These observations suggest an unknown glycan-guided mechanism that underlies recognition of the bacterial cell wall by lysozyme and may complement the HL immune defense function.

Lysozyme's lectin-like characteristics facilitates its immune defense function

KAUST Repository

Zhang, Ruiyan; Wu, Lisha; Eckert, Thomas; Burg-Roderfeld, Monika; Rojas-Macias, Miguel A.; Lü tteke, Thomas; Krylov, Vadim B.; Argunov, Dmitry A.; Datta, Aritreyee; Markart, Philipp; Guenther, Andreas; Norden, Bengt; Schauer, Roland; Bhunia, Anirban; Enani, Mushira Abdelaziz; Billeter, Martin; Scheidig, Axel J.; Nifantiev, Nikolay E.; Siebert, Hans-Christian

2017-01-01

Interactions between human lysozyme (HL) and the lipopolysaccharide (LPS) of Klebsiella pneumoniae O1, a causative agent of lung infection, were identified by surface plasmon resonance. To characterize the molecular mechanism of this interaction, HL binding to synthetic disaccharides and tetrasaccharides representing one and two repeating units, respectively, of the O-chain of this LPS were studied. pH-dependent structural rearrangements of HL after interaction with the disaccharide were observed through nuclear magnetic resonance. The crystal structure of the HL-tetrasaccharide complex revealed carbohydrate chain packing into the A, B, C, and D binding sites of HL, which primarily occurred through residue-specific, direct or water-mediated hydrogen bonds and hydrophobic contacts. Overall, these results support a crucial role of the Glu35/Asp53/Trp63/Asp102 residues in HL binding to the tetrasaccharide. These observations suggest an unknown glycan-guided mechanism that underlies recognition of the bacterial cell wall by lysozyme and may complement the HL immune defense function.

Complex coacervate core micelles with a lysozyme-modified corona

NARCIS (Netherlands)

Danial, Maarten; Klok, Harm-Anton; Norde, Willem; Stuart, Martien A. Cohen

2007-01-01

This paper describes the preparation, characterization, and enzymatic activity of complex coacervate core micelles (C3Ms) composed of poly(acrylic acid) (PAA) and poly(N-methyl-2-vinyl pyridinium iodide)-b-poly(ethylene oxide) (PQ2VP-PEO) to which the antibacterial enzyme lysozyme is end-attached.

Expression and Activity of Lysozyme in Apis Mellifera Carnica Brood Infested with Varroa Destructor

OpenAIRE

Zaobidna Ewa A.; Żółtowska Krystyna; Łopieńska-Biernat Elżbieta

2017-01-01

Varroa destructor is a parasitic mite that attacks the honey bee, and previous studies have suggested that parasitosis caused by this mite is accompanied by immunosuppresion in the host. In this study, the effect of mite infestation on the expression of the lysozyme-1 (lys-1) gene and lysozyme activity in Apis mellifera carnica was determined. The experiment was carried out on the five developmental stages of honey bee workers and drones. Developmental and gender-related differences in gene e...

Modeling growth of Alicyclobacillus acidoterrestris DSM 3922 type strain vegetative cells in the apple juice with nisin and lysozyme

Directory of Open Access Journals (Sweden)

Celenk Molva

2017-05-01

Full Text Available In the present study, the effect of storage temperature on A. acidoterrestris DSM 3922 cells (105 CFU/mL was examined during growth in reconstituted apple juice (pH 3.8, °Brix 11.3 containing nisin (0–100 IU/mL and lysozyme (0–100 mg/L. The growth curves were obtained at three temperatures of 27, 35 and 43 °C using absorbance data (OD600 nm. Based on the results, the minimal inhibitory concentrations (MICs of nisin were found as 10 IU/mL at all tested temperatures. On the other hand, increasing the temperature decreased the amount of lysozyme for growth inhibition. The MICs of lysozyme were found as 10, 2.5 and 1.25 mg/L at 27, 35 and 43 °C, respectively. At selected non-inhibitory doses, nisin (1.25–5 IU/mL and lysozyme (0.3–2.5 mg/L prolonged the lag time compared to the controls at the corresponding temperatures. In addition, there was a strong linear relationship between the lag time and lysozyme concentrations at 27 and 35 °C (R2 > 0.98. The results of this study demonstrated that both nisin and lysozyme could be used to inhibit the growth of A. acidoterrestris cells in the apple juice. The results also indicated that the growth parameters were variable depending on the storage temperature and the type of the antimicrobial agent used in the apple juice.

The state of immunological reactivity and nonspecific protection factor (lysozyme in children with reactive arthritis

Directory of Open Access Journals (Sweden)

Vladimir Savvo

2016-03-01

Full Text Available Aim. An improvement of diagnostics and prognostication of ReA clinical course in children on the base of studying the immunological reactivity and non-specific protection factor (lysozyme.Materials and methods. Examination of children took place in the municipal children’s cardiorheumatologic department of MHPI “Kharkov municipal children’s clinical hospital № 24" and municipal children’s polyclinic of the Kharkov city (№ 1, 2, 7, 12, 13, 14, 23.40 children with ReA underwent immunological examinations, detection of sIgA in the saliva and lysozyme in the blood serum in acute period and in 9–12 months after the beginning of disease. 19 children (47,5 % – 2–6 years old, 21 children (52,5 %. – 7–14 years old. Boys – 22 (55,0 %, girls – 18 (45,0 %. The mean age of children in group was 7,2±0,32. The control group included 32 healthy children. The mean age of children in group was 7,4±0,54.The ReA diagnosis was set according to the order of Ukrainian MHP of 19.07.2005 № 362 “Protocol of diagnostics and treatment of disease of musculoskeletal system and connective tissue in children ICD-D М00-М25 arthropathies”.Immunological examinations included the study of indices of cellular, humoral, monocytic-phagocytic links of immunity, content of cytokines (IL-1β, IL-6, detection of sIgA and index of nonspecific protection factor (lysozyme.Assessment of results of researches was carried out using STATISTICА program for Windows (version 10.0, Microsoft Excel 2012, MATLAB 2015a.Results. In the ReA acute period in children was observed depression of T-system on the background of activation of immunity B-system as the reliable decrease of СD8, СD25 and increase of СD21. There was revealed an increase of IL-6, increase of phagocytic number, spontaneous NBT-test and spontaneous neutrophils activity index.The sIgA level reliably exceeded the standard. At determination of lysozyme the blood serum of patients with ReA in acute

[Partially unfolded state of lysozyme with a developed secondary structure in dimethylsulfoxide].

Science.gov (United States)

Timchenko, A A; Kirkitadze, M D; Prokhorov, D A; Potekhin, S A; Serdiuk, I N

1996-06-01

The conformation of a chicken egg lysozyme molecule (dimensions, stoichiometry of its associates, and the degree of helicity) in DMSO was studied by small-angle neutron scattering, dynamic light scattering, and optical rotatory dispersion in the visible region of the spectrum. At high DMSO concentrations (70%), the protein was shown to exist as a dimer. The monomer molecules in the dimer adopt a partially unfolded conformation, with dimensions substantially greater than those in the native state and a high content of secondary structure (the degree of helicity is close to that of native lysozyme). This approach provides a unique possibility to assess the compactness of molecules in associates, which may be very useful in studying protein self-organization.

Electromagnetic Fields Effects on the Secondary Structure of Lysozyme and Bioprotective Effectiveness of Trehalose

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Emanuele Calabrò

2012-01-01

Full Text Available FTIR spectroscopy was used to investigate the effects of extremely low frequency (50 Hz electromagnetic field and of microwaves at 900 MHz on the secondary structure of a typical protein, the lysozyme, evaluating the bioprotective effectiveness of trehalose. Lysozyme in D2O solution (60 mg/ml was exposed to 50 Hz frequency electromagnetic field at 180 μT. The FTIR spectra indicated an increase of CH2 group at 1921 and 1853 cm−1 after 3 h of exposure. Such effect was not observed after the addition of trehalose (150 mg/mL at the same exposure conditions. Lysozyme dissolved in D2O at the concentration of 100 mg/mL was exposed up to 4 h to 900 MHz mobile phone microwaves at 25 mA/m. A significant increase in intensity of the amide I vibration band in the secondary structure of the protein was observed after 4 h exposure to microwaves. This effect was inhibited by the presence of trehalose at the concentration of 150 mg/mL. Fourier self-deconvolution spectral analysis of lysozyme in D2O solution after exposure to microwaves revealed an increase in intensity of the conformational components of amide I mode, particularly of β-sheet and turn that can be attributed to disorder and unfolding processes of the protein.

Fibrous polymer grafted magnetic chitosan beads with strong poly(cation-exchange) groups for single step purification of lysozyme.

Science.gov (United States)

Bayramoglu, Gulay; Tekinay, Turgay; Ozalp, V Cengiz; Arica, M Yakup

2015-05-15

Lysozyme is an important polypetide used in medical and food applications. We report a novel magnetic strong cation exchange beads for efficient purification of lysozyme from chicken egg white. Magnetic chitosan (MCHT) beads were synthesized via phase inversion method, and then grafted with poly(glycidyl methacrylate) (p(GMA)) via the surface-initiated atom transfer radical polymerization (SI-ATRP). Epoxy groups of the grafted polymer, were modified into strong cation-exchange groups (i.e., sulfonate groups) in the presence of sodium sulfite. The MCTH and MCTH-g-p(GMA)-SO3H beads were characterized by ATR-FTIR, SEM, and VSM. The sulphonate groups content of the modified MCTH-g-p(GMA)-4 beads was found to be 0.53mmolg(-1) of beads by the potentiometric titration method. The MCTH-g-p(GMA)-SO3H beads were first used as an ion-exchange support for adsorption of lysozyme from aqueous solution. The influence of different experimental parameters such as pH, contact time, and temperature on the adsorption process was evaluated. The maximum adsorption capacity was found to be 208.7mgg(-1) beads. Adsorption of lysozyme on the MCTH-g-p(GMA)-SO3H beads fitted to Langmuir isotherm model and followed the pseudo second-order kinetic. More than 93% of the adsorbed lysozyme was desorbed using Na2CO3 solution (pH 11.0). The purity of the lysozyme was checked by HPLC and SDS gel electrophoresis. In addition, the MCTH-g-p(GMA)-SO3H beads prepared in this work showed promising potential for separation of various anionic molecules. Copyright © 2015 Elsevier B.V. All rights reserved.

Multiple I-Type Lysozymes in the Hydrothermal Vent Mussel Bathymodiolus azoricus and Their Role in Symbiotic Plasticity.

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Camille Detree

Full Text Available The aim of this study was first to identify lysozymes paralogs in the deep sea mussel Bathymodiolus azoricus then to measure their relative expression or activity in different tissue or conditions. B. azoricus is a bivalve that lives close to hydrothermal chimney in the Mid-Atlantic Ridge (MAR. They harbour in specialized gill cells two types of endosymbiont (gram-bacteria: sulphide oxidizing bacteria (SOX and methanotrophic bacteria (MOX. This association is thought to be ruled by specific mechanism or actors of regulation to deal with the presence of symbiont but these mechanisms are still poorly understood. Here, we focused on the implication of lysozyme, a bactericidal enzyme, in this endosymbiosis. The relative expression of Ba-lysozymes paralogs and the global anti-microbial activity, were measured in natural population (Lucky Strike--1700 m, Mid-Atlantic Ridge, and in in situ experimental conditions. B. azoricus individuals were moved away from the hydrothermal fluid to induce a loss of symbiont. Then after 6 days some mussels were brought back to the mussel bed to induce a re-acquisition of symbiotic bacteria. Results show the presence of 6 paralogs in B. azoricus. In absence of symbionts, 3 paralogs are up-regulated while others are not differentially expressed. Moreover the global activity of lysozyme is increasing with the loss of symbiont. All together these results suggest that lysozyme may play a crucial role in symbiont regulation.

Isolation and partial purification of lysozyme from saliva of Bali Cattle ...

African Journals Online (AJOL)

SND

2012-01-26

Jan 26, 2012 ... 2Faculty of Mathematics and Life Sciences, Mataram University, Jl. Majapahit No. ... The method adopted for isolation of the lysozyme was from Su and ... Vis spectrophotometer (Shimadzu UV-160, Japan) using a BSA.

Label-Free Aptasensor for Lysozyme Detection Using Electrochemical Impedance Spectroscopy

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Dionisia Ortiz-Aguayo

2018-01-01

Full Text Available This research develops a label-free aptamer biosensor (aptasensor based on graphite-epoxy composite electrodes (GECs for the detection of lysozyme protein using Electrochemical Impedance Spectroscopy (EIS technique. The chosen immobilization technique was based on covalent bonding using carbodiimide chemistry; for this purpose, carboxylic moieties were first generated on the graphite by electrochemical grafting. The detection was performed using [Fe(CN6]3−/[Fe(CN6]4− as redox probe. After recording the frequency response, values were fitted to its electric model using the principle of equivalent circuits. The aptasensor showed a linear response up to 5 µM for lysozyme and a limit of detection of 1.67 µM. The sensitivity of the established method was 0.090 µM−1 in relative charge transfer resistance values. The interference response by main proteins, such as bovine serum albumin and cytochrome c, has been also characterized. To finally verify the performance of the developed aptasensor, it was applied to wine analysis.

Label-Free Aptasensor for Lysozyme Detection Using Electrochemical Impedance Spectroscopy.

Science.gov (United States)

Ortiz-Aguayo, Dionisia; Del Valle, Manel

2018-01-26

This research develops a label-free aptamer biosensor (aptasensor) based on graphite-epoxy composite electrodes (GECs) for the detection of lysozyme protein using Electrochemical Impedance Spectroscopy (EIS) technique. The chosen immobilization technique was based on covalent bonding using carbodiimide chemistry; for this purpose, carboxylic moieties were first generated on the graphite by electrochemical grafting. The detection was performed using [Fe(CN)₆] 3- /[Fe(CN)₆] 4- as redox probe. After recording the frequency response, values were fitted to its electric model using the principle of equivalent circuits. The aptasensor showed a linear response up to 5 µM for lysozyme and a limit of detection of 1.67 µM. The sensitivity of the established method was 0.090 µM -1 in relative charge transfer resistance values. The interference response by main proteins, such as bovine serum albumin and cytochrome c, has been also characterized. To finally verify the performance of the developed aptasensor, it was applied to wine analysis.

Complexation of lysozyme with adsorbed PtBS-b-SCPI block polyelectrolyte micelles on silver surface.

Science.gov (United States)

Papagiannopoulos, Aristeidis; Christoulaki, Anastasia; Spiliopoulos, Nikolaos; Vradis, Alexandros; Toprakcioglu, Chris; Pispas, Stergios

2015-01-20

We present a study of the interaction of the positively charged model protein lysozyme with the negatively charged amphiphilic diblock polyelectrolyte micelles of poly(tert-butylstyrene-b-sodium (sulfamate/carboxylate)isoprene) (PtBS-b-SCPI) on the silver/water interface. The adsorption kinetics are monitored by surface plasmon resonance, and the surface morphology is probed by atomic force microscopy. The micellar adsorption is described by stretched-exponential kinetics, and the micellar layer morphology shows that the micelles do not lose their integrity upon adsorption. The complexation of lysozyme with the adsorbed micellar layers depends on the micelles arrangement and density in the underlying layer, and lysozyme follows the local morphology of the underlying roughness. When the micellar adsorbed amount is small, the layers show low capacity in protein complexation and low resistance in loading. When the micellar adsorbed amount is high, the situation is reversed. The adsorbed layers both with or without added protein are found to be irreversibly adsorbed on the Ag surface.

Effect of guar gum conjugation on functional, antioxidant and antimicrobial activity of egg white lysozyme.

Science.gov (United States)

Hamdani, Afshan Mumtaz; Wani, Idrees Ahmed; Bhat, Naseer Ahmad; Siddiqi, Raushid Ahmad

2018-02-01

This study was undertaken to analyze the effect of conjugation of egg-white lysozyme with guar gum. Lysozyme is an antimicrobial polypeptide that can be used for food preservation. Its antibacterial activity is limited to gram positive bacteria. Conjugation with polysaccharides like guar gum may broaden its activity against gram negatives. Conjugate was developed through Maillard reaction. Assays carried out included sugar estimation, SDS-PAGE, GPC, color, FT-IR, DSC, thermal stability, solubility, emulsifying, foaming and antioxidant activity. In addition, antimicrobial activity of the conjugate was determined against two gram positive (Staphyllococcus aureus and Enterococcus) and two gram negative pathogens (E. coli and Salmonella). Results showed higher functional properties of lysozyme-guar gum conjugate. The antioxidant properties increased from 2.02-35.80% (Inhibition of DPPH) and 1.65-4.93AAE/g (reducing power) upon guar gum conjugation. Conjugate significantly inhibited gram negative bacteria and the antibacterial activity also increased significantly against gram positive pathogens. Copyright © 2017 Elsevier Ltd. All rights reserved.

Tunable pinning effects produced by non-uniform antidot arrays in YBCO thin films

Energy Technology Data Exchange (ETDEWEB)

George, J.; Jones, A.; Al-Qurainy, M. [Institute for Superconducting and Electronic Materials, University of Wollongong, NSW (Australia); Fedoseev, S.A. [Institute for Superconducting and Electronic Materials, University of Wollongong, NSW (Australia); Centre for Medical Radiation Physics, University of Wollongong, NSW (Australia); Rosenfeld, A. [Centre for Medical Radiation Physics, University of Wollongong, NSW (Australia); Pan, A.V. [Institute for Superconducting and Electronic Materials, University of Wollongong, NSW (Australia); National Research Nuclear University MEPhI (Moscow Engineering Physics Institute), Moscow (Russian Federation)

2017-04-15

Uniform, graded and spaced arrays of 3 μm triangular antidots in pulsed laser deposited YBa{sub 2}Cu{sub 3}O{sub 7} (YBCO) superconducting thin films are compared by examining the improvements in the critical current density J{sub c} they produced. The comparison is made to establish the role of their lithographically defined (non-)uniformity and the effectiveness to control and/or enhance the critical current density. It is found that almost all types of non-uniform arrays, including graded ones enhance J{sub c} over the broad applied magnetic field and temperature range due to the modified critical state. Whereas uniform arrays of antidots either reduce or produce no effect on J{sub c} compared to the original (as-deposited) thin films. (copyright 2017 by WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

White shrimp (Litopenaeus vannamei) recombinant lysozyme has antibacterial activity against Gram negative bacteria: Vibrio alginolyticus, Vibrio parahemolyticus and Vibrio cholerae.

Science.gov (United States)

de-la-Re-Vega, Enrique; García-Galaz, Alfonso; Díaz-Cinco, Martha E; Sotelo-Mundo, Rogerio R

2006-03-01

C-type lysozyme has been described as an antibacterial component of the shrimp innate defence system. We determined quantitatively the antibacterial activity of white shrimp (Litopenaeus vannamei) recombinant lysozyme against three Gram negative bacteria: Vibrio alginolyticus, Vibrio parahemolyticus and Vibrio cholerae, using a turbidimetric assay with live bacteria and differential bacterial viable count after interaction with the protein. In conclusion, the antibacterial activity of recombinant shrimp lysozyme against Vibrio sp. is at least equal to the values against the Gram positive M. luteus and more active against the shrimp pathogens V. alginolyticus and V. parahemolyticus.

IBA of ZrO2:Yb/Si thin films produced by the spray pyrolysis method

International Nuclear Information System (INIS)

Andrade, E.; Ramirez, E.B.; Alonso, J.C.; Rocha, M.F.

2008-01-01

A spray pyrolysis method was used to produce thin films of ZrO 2 doped with different Yb concentrations on Si(1 0 0). The films of these ionic semiconductors have potential applications as solid electrolytes in modern ceramic fuel cells of second generation. The determination of the atomic composition of the films is very important because it strongly affects the chemical and thermal stability, as well as electrical properties of the films. A combination of two Ion Beam Analysis (IBA) methods was applied to obtain the atomic composition of the films. A nuclear reaction analysis (NRA) method using a low energy deuterium beam was applied to measure the oxygen content of the films. Heavy ion Rutherford backscattering (HI-RBS) method using a 12 C 3+ beam was applied to measure the Yb and Zr atomic profiles of the samples. X-ray diffraction (XRD) and ellipsometry were also employed to determine structural properties and refractive index of the films, respectively. The IBA, XRD and the ellipsometry supply a wide range of information about the film layers, which can be used for qualification as well as for feedback to the films production

Expression of T4 Lysozyme Gene (gene e) in Streptococcus ...

African Journals Online (AJOL)

pL2 plasmid isolated from E. coli was introduced into S. salivarius subsp. thermophilus and Lactococcus lactis cells by electro-transformation. The lysozyme enzymes expressing by these bacteria were found to be active on Micrococcus luteus cells and thereby preventing their growth on assay plates. Thermostability of ...

Small-angle neutron scattering study of differences in phase behavior of silica nanoparticles in the presence of lysozyme and bovine serum albumin proteins

Science.gov (United States)

Yadav, Indresh; Kumar, Sugam; Aswal, V. K.; Kohlbrecher, J.

2014-03-01

The differences in phase behavior of anionic silica nanoparticles (88 Å) in the presence of two globular proteins [cationic lysozyme (molecular weight (MW) 14.7 kD) and anionic bovine serum albumin (BSA) (MW 66.4 kD)] have been studied by small-angle neutron scattering. The measurements were carried out on a fixed concentration (1 wt %) of Ludox silica nanoparticles with varying concentrations of proteins (0-5 wt %) at pH = 7. It is found that, despite having different natures (opposite charges), both proteins can render to the same kind of aggregation of silica nanoparticles. However, the concentration regions over which the aggregation is observed are widely different for the two proteins. Lysozyme with very small amounts (e.g., 0.01 wt %) leads to the aggregation of silica nanoparticles. On the other hand, silica nanoparticles coexist with BSA as independent entities at low protein concentrations and turn to aggregates at high protein concentrations (>1 wt %). In the case of lysozyme, the charge neutralization by the protein on the nanoparticles gives rise to the protein-mediated aggregation of the nanoparticles. The nanoparticle aggregates coexist with unaggregated nanoparticles at low protein concentrations, whereas, they coexist with a free protein at higher protein concentrations. For BSA, the nonadsorbing nature of the protein produces the depletion force that causes the aggregation of the nanoparticles at higher protein concentrations. The evolution of the interaction is modeled by the two Yukawa potential, taking account of both attractive and repulsive terms of the interaction in these systems. The nanoparticle aggregation is found to be governed by the short-range attraction for lysozyme and the long-range attraction for BSA. The aggregates are characterized by the diffusion limited aggregate type of mass fractal morphology.

Electron loss from multiply protonated lysozyme ions in high energy collisions with molecular oxygen

DEFF Research Database (Denmark)

Hvelplund, P; Nielsen, SB; Sørensen, M

2001-01-01

We report on the electron loss from multiply protonated lysozyme ions Lys-Hn(n)+ (n = 7 - 17) and the concomitant formation of Lys-Hn(n+1)+. in high-energy collisions with molecular oxygen (laboratory kinetic energy = 50 x n keV). The cross section for electron loss increases with the charge state...... of the precursor from n = 7 to n = 11 and then remains constant when n increases further. The absolute size of the cross section ranges from 100 to 200 A2. The electron loss is modeled as an electron transfer process between lysozyme cations and molecular oxygen....

Method of producing thin cellulose nitrate film

International Nuclear Information System (INIS)

Lupica, S.B.

1975-01-01

An improved method for forming a thin nitrocellulose film of reproducible thickness is described. The film is a cellulose nitrate film, 10 to 20 microns in thickness, cast from a solution of cellulose nitrate in tetrahydrofuran, said solution containing from 7 to 15 percent, by weight, of dioctyl phthalate, said cellulose nitrate having a nitrogen content of from 10 to 13 percent

Purification and Characterization of Recombinant Human Lysozyme from Eggs of Transgenic Chickens.

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Hanyu Wu

Full Text Available Transgenic chickens as bioreactors have several advantages, such as the simple establishment procedure, correct glycosylation profile of expressed proteins, etc. Lysozyme is widely used in food industry, livestock farming, and medical field as a replacement of antibiotics because of its antibacterial and complement system-modulating activity. In this study, we used RT-PCR, Western blot, and immunofluorescence to detect the expression of recombinant human lysozyme (rhLY in the transgenic chicken. We demonstrated that the transgene of rhLY was genetically stable across different generations. We next optimized the purification procedure of rhLY from the transgenic eggs by utilizing two steps of cation-exchange chromatography and one gel-filtration chromatography. About 6 mg rhLY with the purity exceeding 90% was obtained from ten eggs, and the purification efficiency was about 75%. The purified rhLY had similar physicochemical and biological properties in molecular mass and antibacterial activity compared to the commercial human lysozyme. Additionally, both of them exhibited thermal stability at 60°C and tolerated an extensive pH range of 2 to 11. In conclusion, our study proved that the transgenic chickens we have previously generated were genetically stable and suitable for the production of active rhLY. We also provided a pipeline for purifying the recombinant proteins from transgenic eggs, which could be useful for other studies.

Behavior of lysozyme adsorbed onto biological liquid crystal lipid monolayer at the air/water interface

Science.gov (United States)

Lu, Xiaolong; Shi, Ruixin; Hao, Changchun; Chen, Huan; Zhang, Lei; Li, Junhua; Xu, Guoqing; Sun, Runguang

2016-09-01

The interaction between proteins and lipids is one of the basic problems of modern biochemistry and biophysics. The purpose of this study is to compare the penetration degree of lysozyme into 1,2-diapalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dipalmitoyl-sn-glycero-3-phosphoethano-lamine (DPPE) by analyzing the data of surface pressure-area (π-A) isotherms and surface pressure-time (π-T) curves. Lysozyme can penetrate into both DPPC and DPPE monolayers because of the increase of surface pressure at an initial pressure of 15 mN/m. However, the changes of DPPE are larger than DPPC, indicating stronger interaction of lysozyme with DPPE than DPPC. The reason may be due to the different head groups and phase state of DPPC and DPPE monolayers at the surface pressure of 15 mN/m. Atomic force microscopy reveals that lysozyme was absorbed by DPPC and DPPE monolayers, which leads to self-aggregation and self-assembly, forming irregular multimers and conical multimeric. Through analysis, we think that the process of polymer formation is similar to the aggregation mechanism of amyloid fibers. Project supported by the National Natural Science Foundation of China (Grant Nos. 21402114 and 11544009), the Natural Science Basic Research Plan in Shaanxi Province of China (Grant No. 2016JM2010), the Fundamental Research Funds for the Central Universities of China (Grant No. GK201603026), and the National University Science and Technology Innovation Project of China (Grant No. 201610718013).

Studies on isolation and partial purification of lysozyme from egg ...

African Journals Online (AJOL)

STORAGESEVER

2009-01-05

Jan 5, 2009 ... nine species of lovebirds, and many different colour var- ieties. Their scientific name Agapornis comes from the. Greek Agapa meaning, “Love” and ornis meaning “bird”. This paper describes the isolation, partial purification and determination of molecular weight of lysozyme of Agapor- nis sp. egg white.

Molecular Insight into Human Lysozyme and Its Ability to Form Amyloid Fibrils in High Concentrations of Sodium Dodecyl Sulfate: A View from Molecular Dynamics Simulations.

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Majid Jafari

Full Text Available Changes in the tertiary structure of proteins and the resultant fibrillary aggregation could result in fatal heredity diseases, such as lysozyme systemic amyloidosis. Human lysozyme is a globular protein with antimicrobial properties with tendencies to fibrillate and hence is known as a fibril-forming protein. Therefore, its behavior under different ambient conditions is of great importance. In this study, we conducted two 500000 ps molecular dynamics (MD simulations of human lysozyme in sodium dodecyl sulfate (SDS at two ambient temperatures. To achieve comparative results, we also performed two 500000 ps human lysozyme MD simulations in pure water as controls. The aim of this study was to provide further molecular insight into all interactions in the lysozyme-SDS complexes and to provide a perspective on the ability of human lysozyme to form amyloid fibrils in the presence of SDS surfactant molecules. SDS, which is an anionic detergent, contains a hydrophobic tail with 12 carbon atoms and a negatively charged head group. The SDS surfactant is known to be a stabilizer for helical structures above the critical micelle concentration (CMC [1]. During the 500000 ps MD simulations, the helical structures were maintained by the SDS surfactant above its CMC at 300 K, while at 370 K, human lysozyme lost most of its helices and gained β-sheets. Therefore, we suggest that future studies investigate the β-amyloid formation of human lysozyme at SDS concentrations above the CMC and at high temperatures.

Electrical and optical properties of thin indium tin oxide films produced by pulsed laser ablation in oxygen or rare gas atmospheres

DEFF Research Database (Denmark)

Thestrup, B.; Schou, Jørgen; Nordskov, A.

1999-01-01

Films of indium tin oxide (ITO) have been produced in different background gases by pulsed laser deposition (PLD). The Films deposited in rare gas atmospheres on room temperature substrates were metallic, electrically conductive, but had poor transmission of visible light. For substrate temperatu......Films of indium tin oxide (ITO) have been produced in different background gases by pulsed laser deposition (PLD). The Films deposited in rare gas atmospheres on room temperature substrates were metallic, electrically conductive, but had poor transmission of visible light. For substrate...

Prophenoloxidase system, lysozyme and protease inhibitor distribution in the common cuttlefish Sepia officinalis.

Science.gov (United States)

Le Pabic, Charles; Safi, Georges; Serpentini, Antoine; Lebel, Jean-Marc; Robin, Jean-Paul; Koueta, Noussithé

2014-01-01

The immune system of cephalopods remains poorly understood. The aim of this study was to determine the specific activity of immune enzymes in epithelial barriers, circulatory and digestive systems of the common cuttlefish Sepia officinalis. Three enzyme groups with putative functions in immunity were investigated: phenoloxidases (POs), lysozymes and protease inhibitors (PIs). Consistent with a role in immunity, highest PO activities were found in the integument as well as the respiratory and circulatory organs under zymogenic (proPO) and active form. Surprisingly, high PO activities were also found in the digestive gland and its appendages. Similarly, high lysozyme activities were detected in the integument and circulatory organs, but also in the posterior salivary glands, highlighting the implication of this antibacterial enzyme group in most tissues exposed to the environment but also within the circulatory system. Albeit highest in digestive organs, the ubiquitous detection of PI activity in assayed compartments suggests immune function(s) in a wide range of tissues. Our study reports proPO/PO, lysozyme and PI distributions in S. officinalis body compartments for the first time, and thus provides the fundamental basis for a better understanding of the humoral immune system in cephalopods as well as invertebrates. Copyright © 2014 Elsevier Inc. All rights reserved.

The extracytoplasmic function sigma factor SigV plays a key role in the original model of lysozyme resistance and virulence of Enterococcus faecalis.

Directory of Open Access Journals (Sweden)

André Le Jeune

Full Text Available BACKGROUND: Enterococcus faecalis is one of the leading agents of nosocomial infections. To cause diseases, pathogens or opportunistic bacteria have to adapt and survive to the defense systems encountered in the host. One of the most important compounds of the host innate defense response against invading microorganisms is lysozyme. It is found in a wide variety of body fluids, as well as in cells of the innate immune system. Lysozyme could act either as a muramidase and/or as a cationic antimicrobial peptide. Like Staphylococcus aureus, E. faecalis is one of the few bacteria that are completely lysozyme resistant. RESULTS: This study revealed that oatA (O-acetyl transferase and dlt (D-Alanylation of lipoteicoic acids genes contribute only partly to the lysozyme resistance of E. faecalis and that a specific transcriptional regulator, the extracytoplasmic function SigV sigma factor plays a key role in this event. Indeed, the sigV single mutant is as sensitive as the oatA/dltA double mutant, and the sigV/oatA/dltA triple mutant displays the highest level of lysozyme sensitivity suggesting synergistic effects of these genes. In S. aureus, mutation of both oatA and dlt genes abolishes completely the lysozyme resistance, whereas this is not the case in E. faecalis. Interestingly SigV does not control neither oatA nor dlt genes. Moreover, the sigV mutants clearly showed a reduced capacity to colonize host tissues, as they are significantly less recovered than the parental JH2-2 strain from organs of mice subjected to intravenous or urinary tract infections. CONCLUSIONS: This work led to the discovery of an original model of lysozyme resistance mechanism which is obviously more complex than those described for other Gram positive pathogens. Moreover, our data provide evidences for a direct link between lysozyme resistance and virulence of E. faecalis.

Crystallization and preliminary x-ray structure analysis of the egg-white lysozyme from a Taiwanese Soft-Shelled Turtle (Tri onyx Sinensis Wiegmann)

International Nuclear Information System (INIS)

Siritapetawee, Jaruwan; Thammasirirak, Sompong; Yuvaniyama, Jirudon

2005-10-01

Lysozyme has been purified from the egg-white of a Taiwanese soft-shelled turtle. This soft-shelled turtle ' s egg-white lysozyme migrated on 12.5% SDS-PAGE at about 14.8 kDa. The lysozyme has been crystallized using the sitting drop vapor diffusion technique and 30% (w/v) polyethylene glycol 8000 in 0.1 M sodium cacodylate, p H 6.5 containing 0.2 M ammonium sulfate as a precipitant. One of the crystals diffracted X rays beyond 2 angstrom unit resolution and belonged to the orthorhombic, space group P212121, with unit cell dimensions of a = 37.8 angstrom unit, b = 55.6 angstrom unit, and c 72.2 angstrom unit and one molecule of the enzyme per asymmetric unit. The data were collected to 1.9 angstrom resolution with an R merge of 4.6%, suitable for high resolution structure analysis. The single-crystal X-ray structure of lysozyme has been initially phased with the Molecular Replacement technique using pheasant egg-white lysozyme (PDB ID 1GHL) as a search template. Model rebuilding and refinement are in progress

High-pressure protein crystallography of hen egg-white lysozyme

Energy Technology Data Exchange (ETDEWEB)

Yamada, Hiroyuki; Nagae, Takayuki [Nagoya University, Chikusa, Nagoya, Aichi 464-8603 (Japan); Watanabe, Nobuhisa, E-mail: nobuhisa@nagoya-u.jp [Nagoya University, Chikusa, Nagoya, Aichi 464-8603 (Japan); Nagoya University, Chikusa, Nagoya, Aichi 464-8603 (Japan)

2015-04-01

The crystal structure of hen egg-white lysozyme (HEWL) was analyzed under pressures of up to 950 MPa. The high pressure modified the conformation of the molecule and induced a novel phase transition in the tetragonal crystal of HEWL. Crystal structures of hen egg-white lysozyme (HEWL) determined under pressures ranging from ambient pressure to 950 MPa are presented. From 0.1 to 710 MPa, the molecular and internal cavity volumes are monotonically compressed. However, from 710 to 890 MPa the internal cavity volume remains almost constant. Moreover, as the pressure increases to 950 MPa, the tetragonal crystal of HEWL undergoes a phase transition from P4{sub 3}2{sub 1}2 to P4{sub 3}. Under high pressure, the crystal structure of the enzyme undergoes several local and global changes accompanied by changes in hydration structure. For example, water molecules penetrate into an internal cavity neighbouring the active site and induce an alternate conformation of one of the catalytic residues, Glu35. These phenomena have not been detected by conventional X-ray crystal structure analysis and might play an important role in the catalytic activity of HEWL.

10 GHz surface impedance measurements of (Y9Er)BaCuO films produced by MOCVD, laser ablation, and sputtering

International Nuclear Information System (INIS)

Luine, J.; Daly, K.; Hu, R.; Kain, A.; Lee, A.; Manasevit, H.; Pettiette-Hall, C.; Simon, R.; St John, D.; Wagner, M.

1991-01-01

This paper reports on a parallel-plate resonator technique previously used to measure microwave surface resistance R s (T) extended to also measure absolute penetration depth λ(T). Measurements of both quantities near 10 GHz from 4.2 K to Tc are reported for ErBaCuO thin films produced by metal-organic chemical vapor deposition (MOCVD) and YBaCuO think films produced by laser ablation and single-target off-axis sputtering. All the films were made at TRW. Each production method gives rise to films whose surface resistance is below 1 milliohm at temperatures below 40K. The low temperature penetration depths range from 250 nm for the laser ablation and sputtered films to 800 nm for the MOCVD films. The penetration depths in all cases increase with temperature according to the Gorter-Casimir temperature dependence

Evaluation of lysozyme, complement C3, and total protein in different developmental stages of Caspian kutum (Rutilus frisii kutum K.

Directory of Open Access Journals (Sweden)

Abdollahi Razieh

2016-03-01

Full Text Available In this study, non–specific immune parameters in fertilized eggs, eyed embryos, larvae 10, 25, 50, 60, and 70 days post hatch (DPH, and female broodstock of Caspian kutum, Rutilus frisii kutum (Kamensky, were evaluated. The lysozyme activity, complement C3, and total protein levels were measured with the turbidimetric, immunoturbidimetric, and Bradford methods, respectively. The results showed that lysozyme levels decreased from levels noted in the fertilized eggs until the larvae were 10 days old. Subsequently, significant increases in lysozyme levels were observed until 70 DPH. An increasing trend of complement component C3 was noted from the levels in fertilized eggs to 10 DPH, following which it decreased significantly. Total protein levels differed significantly in early developmental stages of Caspian kutum. The higher values of complement component C3 than of lysozyme in the early life stages could be indicative of the former’s more fundamental role.

Resonance scattering spectra of micrococcus lysodeikticus and its application to assay of lysozyme activity.

Science.gov (United States)

Jiang, Zhi-Liang; Huang, Guo-Xia

2007-02-01

Several methods, including turbidimetric and colorimetric methods, have been reported for the detection of lysozyme activity. However, there is no report about the resonance scattering spectral (RSS) assay, which is based on the catalytic effect of lysozyme on the hydrolysis of micrococcus lysodeikticus (ML) and its resonance scattering effect. ML has 5 resonance scattering peaks at 360 400, 420, 470, and 520 nm with the strongest one at 470 nm. The concentration of ML in the range of 2.0x10(6)-9.3x10(8) cells/ml is proportional to the RS intensity at 470 nm (I(470 nm)). A new catalytic RSS method has been proposed for 0.24-40.0 U/ml (or 0.012-2.0 mug/ml) lysozyme activity, with a detection limit (3sigma) of 0.014 U/ml (or 0.0007 microg/ml). Saliva samples were assayed by this method, and it is in agreement with the results of turbidimetric method. The slope, intercept and the correlation coefficient of the regression analysis of the 2 assays were 0.9665, -87.50, and 0.9973, respectively. The assay has high sensitivity and simplicity.

Lysozyme as an alternative to antibiotics improves growth performance and tumor necrosis factor-a levels during an indirect immune challenge

Science.gov (United States)

Lysozyme is a 1,4-ß-N-acetylmuramidase that has antimicrobial properties. The objective of this study was to determine the effect of lysozyme and antibiotics on growth performance and immune response during an indirect disease challenge. Two replicates of 720 pigs each were weaned from the sow at ...

Electrical properties of multilayer (DLC-TiC) films produced by pulsed laser deposition

Science.gov (United States)

Alawajji, Raad A.; Kannarpady, Ganesh K.; Nima, Zeid A.; Kelly, Nigel; Watanabe, Fumiya; Biris, Alexandru S.

2018-04-01

In this work, pulsed laser deposition was used to produce a multilayer diamond like carbon (ML (DLC-TiC)) thin film. The ML (DLC-TiC) films were deposited on Si (100) and glass substrates at various substrate temperatures in the range of 20-450 °C. Raman spectroscopy, x-ray photoelectron spectroscopy (XPS), and atomic force microscopy were utilized to characterize the prepared films. Raman analysis revealed that as the substrate temperature increased, the G-peak position shifted to a higher raman shift and the full width at half maximum of the G and D bands decreased. XPS analysis indicated a decrease in sp3/sp2 ratio and an increase in Ti-C bond intensity when the substrate temperature was increased. Additionally, the surface roughness of ML (DLC-TiC) filmswas affected by the type and temperature of the substrate. The electrical measurement results indicated that the electrical resistivity of the ML (DLC-TiC) film deposited on Si and glass substrates showed the same behavior-the resistivity decreased when substrate temperature increased. Furthermore, the ML (DLC-TiC) films deposited on silicon showed lower electrical resistivity, dropping from 8.39E-4 Ω-cm to 5.00E-4 Ω-cm, and, similarly, the films on the glass substrate displayed a drop in electrical resistivity from 1.8E-2 Ω-cm to 1.2E-3 Ω-cm. These enhanced electrical properties indicate that the ML (DLC-TiC) films have widespread potential as transducers for biosensors in biological research; electrochemical electrodes, because these films can be chemically modified; biocompatible coatings for medicals tools; and more.

Production, crystallization and X-ray characterization of chemically glycosylated hen egg-white lysozyme

International Nuclear Information System (INIS)

López-Jaramillo, F. J.; Pérez-Banderas, F.; Hernández-Mateo, F.; Santoyo-González, F.

2005-01-01

The feasibility of glycosylation post-purification has been demonstrated by introducing glucose into the model protein lysozyme via a novel reaction that is compatible with biological samples. The crystallization of glycoproteins is one of the challenges to be confronted by the crystallographic community in the frame of what is known as glycobiology. The state of the art for the crystallization of glycoproteins is not promising and removal of the carbohydrate chains is generally suggested since they are flexible and a source of heterogeneity. In this paper, the feasibility of introducing glucose into the model protein hen egg-white lysozyme via a post-purification glycosylation reaction that may turn any protein into a model glycoprotein whose carbohydrate fraction can be manipulated is demonstrated

Spectroscopic determination of lysozyme conformational changes in the presence of trehalose and guanidine.

Science.gov (United States)

Barreca, Davide; Laganà, Giuseppina; Ficarra, Silvana; Gattuso, Giuseppe; Magazù, Salvatore; La Torre, Roberto; Tellone, Ester; Bellocco, Ersilia

2013-06-01

The bioprotective action of the disaccharide trehalose has been studied against the well-known denaturating agent, guanidine hydrochloride. The results indicated a direct influence of trehalose on both enzymatic activity and conformational changes of lysozyme, as shown by the decrease of the inactivation rate constant of about 1.48-fold and the loss of α-helix structure of lysozyme. In addition, ESI-MS hydrogen-deuterium (H/D) exchange experiments allowed us to correlate the structural and dynamic features of the protein in the presence of the two additives, highlighting as trehalose remarkably influenced this exchange by decreasing local protein environment changes and solvent accessibility to the amide peptide backbone, as further evidenced by circular dichroism and (1)H NMR measurements.

Lysozyme-magnesium aluminum silicate microparticles: Molecular interaction, bioactivity and release studies

DEFF Research Database (Denmark)

Kanjanakawinkul, Watchara; Medlicott, Natalie J.; Rades, Thomas

2015-01-01

The objectives of this study were to investigate the adsorption behavior of lysozyme (LSZ) onto magnesium aluminum silicate (MAS) at various pHs and to characterize the LSZ–MAS microparticles obtained from the molecular interaction between LSZ and MAS. The results showed that LSZ could be bound...

Clearing of suspensions of Micrococcus lysodeikticus catalysed by lysozymes from hen, goose, and turkey egg whites, human milk, and phage T4. Assessment of potential as signal generators for homogeneous enzyme immunoassays for urinary steroids.

Science.gov (United States)

Cooke, Delwyn G; Blackwell, Leonard F

2007-01-01

Lysozymes (3.2.1.17) from goose (Anser anser) egg white, turkey (Melagris gallopavo) egg white, phage T4 and human milk were compared with hen egg white lysozyme in their ability to clear a suspension of Micrococcus lysodeikticus. All of the lysozymes, except hen egg white lysozyme, catalysed the clearing of the Micrococcus lysodeikticus suspension in a biphasic fashion. Compared to hen egg white lysozyme, the total absorbance or transmission change over 5 and 20 minutes was less in all cases, except for human lysozyme. Human lysozyme was, therefore, a potential alternative, more rapid signal generator for the measurement of urinary estrone glucuronide excretion rates because of its structural similarity to hen egg white lysozyme. The apparent K(M) values for hen egg white lysozyme increased with the enzyme concentration.

Chronopotentiometric sensing of specific interactions between lysozyme and the DNA aptamer

Czech Academy of Sciences Publication Activity Database

Ostatná, Veronika; Vargová, Veronika; Kekedy-Nagy, L.; Černocká, Hana; Ferapontova, E.E.

2017-01-01

Roč. 114, APR2017 (2017), s. 42-47 ISSN 1567-5394 R&D Projects: GA ČR GA13-00956S Institutional support: RVO:68081707 Keywords : self-assembled monolayers * protein interactions * lysozyme Subject RIV: CE - Biochemistry OBOR OECD: Biochemistry and molecular biology Impact factor: 3.346, year: 2016

Magnetic cation exchange isolation of lysozyme from native hen egg white

Czech Academy of Sciences Publication Activity Database

Šafařík, Ivo; Maděrová, Zdeňka; Tokar, O.; Šafaříková, Miroslava

2007-01-01

Roč. 45, - (2007), s. 355-359 ISSN 1330-9862 R&D Projects: GA MŠk(CZ) 1P05OC053 Institutional research plan: CEZ:AV0Z60870520 Keywords : magnetic separation * lysozyme * hen egg white Subject RIV: EI - Biotechnology ; Bionics Impact factor: 0.906, year: 2007

Lysozyme complexes with thermo- and pH-responsive PNIPAM-b-PAA block copolymer

Energy Technology Data Exchange (ETDEWEB)

Pippa, Natassa [National and Kapodistrian University of Athens, Department of Pharmaceutical Technology, Faculty of Pharmacy (Greece); Meristoudi, Anastasia; Pispas, Stergios, E-mail: pispas@eie.gr [National Hellenic Research Foundation, Theoretical and Physical Chemistry Institute (Greece); Demetzos, Costas [National and Kapodistrian University of Athens, Department of Pharmaceutical Technology, Faculty of Pharmacy (Greece)

2017-02-15

Lysozyme is an enzyme responsible for the damage of bacterial cell walls and is abundant in a number of secretions such as tears and human milk. In the present study, we investigated the structure, the physicochemical characteristics, and the temperature-responsiveness of lysozyme complexes with poly(N-isopropylacrylamide)-b-poly(acrylic acid) block polyelectrolyte in aqueous media. A gamut of light-scattering techniques and fluorescence spectroscopy were used in order to examine the complexation process, as well as the structure, solution behavior, and temperature response of the nanosized complexes. The concentration of copolymer polyelectrolyte was kept constant. The values of the scattering intensity, I{sub 90}, which is proportional to the mass of the species in solution, increased gradually as a function of C{sub LYS,} providing proof of the occurring complexation, while the size of the nanostructures decreased. The structure of the complexes became more open as the C{sub LYS} increased. The increase of the salinity did not affect the structural characteristics of the supramolecular nanoparticulate aggregates. On the other hand, the physicochemical and structural characteristics of the complexes changed upon increasing temperature, and the changes depended on the initial ratio block polyelectrolyte/lysozyme. The knowledge on developing block polyelectrolyte/protein complexes through electrostatic interactions, obtained from this investigation, may be applied to the design of nutraceuticals.

Purification of Egg White Lysozyme from Indonesian Kampung Chicken and Ducks

Directory of Open Access Journals (Sweden)

Z. Wulandari

2015-04-01

Full Text Available Egg white lysozyme (EWL has considerably a wide functional protein exhibiting antibacterial activity mainly against Gram-positive bacteria. The EWL is widely applied in food industry and is considerably safe. Despite its high potency, EWL of Indonesian poultry has never been studied and exploited. This study was aimed to purify EWL from two Indonesian poultry: kampung chicken and Cihateup duck, and compared to egg of commercial laying hens. The eggs in this study were obtained from field laboratory of Faculty of Animal Science, Bogor Agricultural University (IPB and classified in AA quality based on the interior quality. First attempt to purify the EWL was performed by using ethanol precipitation yielding purified EWL which was still contaminated by other proteins, hence designated as partially purified EWL. Final concentrations of partially purified EWL of kampung chicken, commercial laying hens, and Cihateup duck were about 5800, 5400, and 5500 μg/mL, respectively. To confirm whether the use of ethanol in the purification affecting EWL antibacterial activities, the activities were examined against Staphylococcus aureus. It demonstrated that the partially purified EWL exhibited ability to inhibit S. aureus at 6 and 26 h suggesting that the method was feasible as it did not interfere EWL antibacterial activities. Yet, based on SDS-Page, purity was the issue in ethanol precipitation method. Further attempt using ion exchange chromatography at pH 10 successfully purified lysozyme as indicated by a single band corresponding to lysozyme size (~14 kD free from bands of other proteins. Altogether, a single step of ion exchange chromatography is sufficient and promising to isolate EWL from Indonesian poultry for various industrial purposes.

Complex formation in mixtures of lysozyme-stabilized emulsions and human saliva

NARCIS (Netherlands)

Silletti, E.; Vingerhoeds, M.H.; Norde, W.; Aken, van G.A.

2007-01-01

In this paper, we studied the interaction between human unstimulated saliva and lysozyme-stabilized oil-in-water emulsions (10 wt/wt% oil phase, 10 mM NaCl, pH 6.7), to reveal the driving force for flocculation of these emulsions. Confocal scanning laser microscopy (CSLM) showed formation of

Active biodegradable films produced with blends of rice flour and poly(butylene adipate co-terephthalate): effect of potassium sorbate on film characteristics.

Science.gov (United States)

Sousa, G M; Soares Júnior, M S; Yamashita, F

2013-08-01

The objective of work was to produce and characterize biodegradable films from rice flour, poly(butylene adipate co-terephthalate) (PBAT), glycerol and potassium sorbate, for application as active packaging for fresh lasagna pasta. The films were evaluated with respect to their optical, water vapor barrier, mechanical and microstructural properties. The mechanical properties and microstructure were evaluated after use as packaging material for fresh pasta for 45 days at 7°C. The blends of rice flour, PBAT, glycerol and potassium sorbate showed good processability and allowed for the pilot scale production of films by blow extrusion process. The addition of 1 to 5% potassium sorbate as plasticizer agent of films in place of glycerol did not alter the film mechanical properties and a sorbate concentration greater or equal than 3% reduced the opacity, although increasing the water vapor permeability. The films could be used as active packaging for fresh food pasta, since they remained integral and easy to handle after application. The rice flour was shown to be an excellent material for the formulation of biodegradable films, since it is a low-cost raw material from a renewable source. The addition of potassium sorbate did not affect the extrusion process, and could be used in the production of packaging for use with foods. Copyright © 2013 Elsevier B.V. All rights reserved.

Improvement of wine aromatic quality using mixtures of lysozyme and dimethyl dicarbonate, with low SO2 concentration.

Science.gov (United States)

Nieto-Rojo, Rodrigo; Luquin, Asuncion; Ancín-Azpilicueta, Carmen

2015-01-01

The use of sulphur dioxide (SO2) in the treatment of foodstuffs presents some problems as it could lead to pseudo-allergies in some people. The aim of this research work was to study the addition of different preservative mixtures and their influence on the concentration of volatile compounds and sensorial quality in wine. To do so, vinifications were carried out using Garnacha must to which lysozyme, dimethyl dicarbonate (DMDC) and mixtures of these with SO2 were added at different doses (25 and 50 mg l(-1)). The results were compared with a control sample to which only SO2 had been added (50 mg l(-1)). In general, mixtures of SO2 with lysozyme and DMDC favoured the formation of volatile compounds in the wines. Wines obtained from the mixtures of lysozyme and DMDC with 25 mg l(-1) of SO2 had better sensorial quality than the wines obtained with 50 mg l(-1) as the only preservative used.

Effects of mannose, fructose, and fucose on the structure, stability, and hydration of lysozyme in aqueous solution

DEFF Research Database (Denmark)

Rahim, Abdoul; Peters, Günther H.J.; Jalkanen, Karl J.

2013-01-01

The bio-protective properties of monosaccharaides, namely mannose, fructose and fucose, on the stability and dynamical properties of the NMR determined hen egg-white lysozyme structure have been investigated by means of molecular dynamics simulations at room temperature in aqueous solution and in...... of the solvent and sugar distributions around lysozyme was used to investigate the interfacial solvent and sugar structure near the protein surface.......The bio-protective properties of monosaccharaides, namely mannose, fructose and fucose, on the stability and dynamical properties of the NMR determined hen egg-white lysozyme structure have been investigated by means of molecular dynamics simulations at room temperature in aqueous solution and in 7...... and 13 wt % concentrations of the three sugars. Results are discussed in the framework of the bio-protective phenomena. The three sugars show similar bio-protective behaviours at room temperature (300 K) in the concentration range studied as shown by the small RMSDs of the resulting MD structures from...

Leukocytes respiratory burst and lysozyme level in pacu (Piaractus mesopotamicus Holmberg, 1887

Directory of Open Access Journals (Sweden)

JS. Abreu

Full Text Available Innate immune responses are useful to determine the health status of fish and to evaluate the effect of immunomodulatory substances in fish farming. Leukocytes respiratory burst was measured in pacu (Piaractus mesopotamicus using chemiluminescence assay and nitroblue tetrazolium (NBT reduction assay. The nitroblue tetrazolium reduction seemed more adequate than chemiluminescence assay for leukocytes oxidative burst determination, since it was difficult to isolate the blood leucocytes for chemiluminescence assay. Plasma and serum lysozyme were measured using a turbidimetric assay. The heating of serum and plasma samples (56 ºC for 30 minutes for complement system inactivation darkened the plasma samples and interfered in the results. The lysozyme activity in serum was higher than in plasma, suggesting that serum samples are more appropriate for the analysis. This study established protocols that can be useful tools in the study of immune mechanisms of the tropical fish pacu.

Relationship between salivary immunoglobulin a, lactoferrin and lysozyme flow rates and lifestyle factors in Japanese children: a cross-sectional study.

Science.gov (United States)

Ide, Momo; Saruta, Juri; To, Masahiro; Yamamoto, Yuko; Sugimoto, Masahiro; Fuchida, Shinya; Yokoyama, Mina; Kimoto, Shigenari; Tsukinoki, Keiichi

2016-10-01

The antimicrobial substances in saliva contribute to the maintenance of both oral health and overall health of the body. Therefore, the associations among immunoglobulin A (IgA), lactoferrin and lysozyme flow rates in the saliva of children, and their relationships with the physical attributes and lifestyle factors of children, were examined. Saliva was collected from 90 children who visited the Kanagawa Dental University Hospital Pediatric Dentistry, and questionnaires were completed by guardians. IgA, lactoferrin and lysozyme concentrations were measured in the saliva samples using enzyme-linked immunosorbent assays (ELISAs). The IgA flow rate in saliva increased as age, height and weight increased. A correlation was found between lactoferrin and lysozyme flow rates. When the antimicrobial substance flow rates in the saliva were divided into two groups of 22 children each based on the highest and lowest quartiles, children with either a low or high IgA flow rate also had a high or low lactoferrin flow rate, respectively. The same pattern was observed for lactoferrin and lysozyme flow rates. There is a high probability that the IgA flow rate in the saliva of children reflects and corresponds to the developmental status of immune function as the child ages and increases in height and weight. The flow rates of lactoferrin and lysozyme were correlated in children. In addition, regarding lifestyle factors, the duration of sleep and lactoferrin flow rate were also related.

Specific delivery of captopril to the kidney with the prodrug captopril-lysozyme

NARCIS (Netherlands)

Kok, R.J; Moolenaar, Frits; de Zeeuw, D; Meijer, D.K F

Low-molecular-weight proteins (LMWPs) accumulate in the proximal tubular cells of the kidney, which makes these proteins interesting tools for renal drug targeting. We studied this approach using the LMWP lysozyme as a carrier for the angiotensin-converting enzyme inhibitor captopril. Captopril was

Effect of histatin-5 and lysozyme on the ability of Streptococcus mutans to form biofilms in in vitro conditions.

Science.gov (United States)

Krzyściak, Wirginia; Jurczak, Anna; Piątkowski, Jakub; Kościelniak, Dorota; Gregorczyk-Maga, Iwona; Kołodziej, Iwona; Papież, Monika A; Olczak-Kowalczyk, Dorota

2015-09-20

The mechanisms of adhesion to solid surfaces enable S. mutans to colonize oral cavities and form biofilms, which play an important role in caries development. Additional properties enabling the survival of S. mutans in the oral cavity include its ability to survive in acidic environments and specific interactions with other microorganisms inhabiting this ecosystem. The aim of this study was to determine the antibacterial activity of saliva histatin-5 (peptide) and lysozyme (protein) against S. mutans and L. rhamnosus, as representatives of physiological flora. The study involved strains of physiological (L. rhamnosus) and cariogenic (S. mutans) flora isolated from one patient with diagnosed early caries of the deciduous teeth. It was proved that the presence of probiotic L. rhamnosus bacteria in the environment had a negative impact on the ability of S. mutans to produce biofilm. Moreover, the antibacterial activity of histatin-5 was confirmed, and it inhibited S. mutans growth at concentrations of 27.2 μg/ml and 54.4 μg/ml, both individually and in a mixture with lysozyme (in a total concentration of 54.4 μg/ml). The data obtained constitute a promising result due to their potential future application in the prevention and early diagnosis of caries.

Effect of histatin-5 and lysozyme on the ability of Streptococcus mutans to form biofilms in in vitro conditions

Directory of Open Access Journals (Sweden)

Wirginia Krzyściak

2015-09-01

Full Text Available The mechanisms of adhesion to solid surfaces enable S. mutans to colonize oral cavities and form biofilms, which play an important role in caries development. Additional properties enabling the survival of S. mutans in the oral cavity include its ability to survive in acidic environments and specific interactions with other microorganisms inhabiting this ecosystem.The aim of this study was to determine the antibacterial activity of saliva histatin-5 (peptide and lysozyme (protein against S. mutans and L. rhamnosus, as representatives of physiological flora.The study involved strains of physiological (L. rhamnosus and cariogenic (S. mutans flora isolated from one patient with diagnosed early caries of the deciduous teeth.It was proved that the presence of probiotic L. rhamnosus bacteria in the environment had a negative impact on the ability of S. mutans to produce biofilm. Moreover, the antibacterial activity of histatin-5 was confirmed, and it inhibited S. mutans growth at concentrations of 27.2 μg/ml and 54.4 μg/ml, both individually and in a mixture with lysozyme (in a total concentration of 54.4 μg/ml.The data obtained constitute a promising result due to their potential future application in the prevention and early diagnosis of caries.

Structure dependent resistivity and dielectric characteristics of tantalum oxynitride thin films produced by magnetron sputtering

Energy Technology Data Exchange (ETDEWEB)

Cristea, D., E-mail: daniel.cristea@unitbv.ro [Department of Materials Science, Transilvania University, 500036 Brasov (Romania); Crisan, A. [Department of Materials Science, Transilvania University, 500036 Brasov (Romania); Cretu, N. [Electrical Engineering and Applied Physics Department, Transilvania University, 500036 Brasov (Romania); Borges, J. [Centro de Física, Universidade do Minho, Campus de Gualtar, 4710 - 057 Braga (Portugal); Instituto Pedro Nunes, Laboratório de Ensaios, Desgaste e Materiais, Rua Pedro Nunes, 3030-199 Coimbra (Portugal); SEG-CEMUC, Mechanical Engineering Department, University of Coimbra, 3030-788 Coimbra (Portugal); Lopes, C.; Cunha, L. [Centro de Física, Universidade do Minho, Campus de Gualtar, 4710 - 057 Braga (Portugal); Ion, V.; Dinescu, M. [National Institute for Lasers, Plasma and Radiation Physics, Lasers Department, “Photonic Processing of Advanced Materials” Group, PO Box MG-16, RO 77125 Magurele-Bucharest (Romania); Barradas, N.P. [Centro de Ciências e Tecnologias Nucleares, Instituto Superior Técnico, Universidade de Lisboa, E.N. 10 ao km 139,7, 2695-066 Bobadela LRS (Portugal); Alves, E. [Instituto de Plasmas e Fusão Nuclear, Instituto Superior Técnico, Universidade de Lisboa, E.N. 10 ao km 139,7, 2695-066 Bobadela LRS (Portugal); Apreutesei, M. [MATEIS Laboratory-INSA de Lyon, 21 Avenue Jean Capelle, 69621 Villeurbanne cedex (France); Université de Lyon, Institut des Nanotechnologies de Lyon INL-UMR5270, CNRS, Ecole Centrale de Lyon, Ecully F-69134 (France); Munteanu, D. [Department of Materials Science, Transilvania University, 500036 Brasov (Romania)

2015-11-01

Highlights: • Tantalum oxynitride thin films have been deposited by magnetron sputtering, in various configurations. • The rising of the reactive gases mixture flow has the consequence of a gradual increase in the non-metallic content in the films, which results in a 10 orders of magnitude resistivity domain. • The higher resistivity films exhibit dielectric constants up to 41 and quality factors up to 70. - Abstract: The main purpose of this work is to present and to interpret the change of electrical properties of Ta{sub x}N{sub y}O{sub z} thin films, produced by DC reactive magnetron sputtering. Some parameters were varied during deposition: the flow of the reactive gases mixture (N{sub 2} and O{sub 2}, with a constant concentration ratio of 17:3); the substrate voltage bias (grounded, −50 V or −100 V) and the substrate (glass, (1 0 0) Si or high speed steel). The obtained films exhibit significant differences. The variation of the deposition parameters induces variations of the composition, microstructure and morphology. These differences cause variation of the electrical resistivity essentially correlated with the composition and structural changes. The gradual decrease of the Ta concentration in the films induces amorphization and causes a raise of the resistivity. The dielectric characteristics of some of the high resistance Ta{sub x}N{sub y}O{sub z} films were obtained in the samples with a capacitor-like design (deposited onto high speed steel, with gold pads deposited on the dielectric Ta{sub x}N{sub y}O{sub z} films). Some of these films exhibited dielectric constant values higher than those reported for other tantalum based dielectric films.

Polymorphous silicon thin films produced in dusty plasmas: application to solar cells

International Nuclear Information System (INIS)

Roca i Cabarrocas, Pere; Chaabane, N; Kharchenko, A V; Tchakarov, S

2004-01-01

We summarize our current understanding of the optimization of PIN solar cells produced by plasma enhanced chemical vapour deposition from silane-hydrogen mixtures. To increase the deposition rate, the discharge is operated under plasma conditions close to powder formation, where silicon nanocrystals contribute to the deposition of so-called polymorphous silicon thin films. We show that the increase in deposition rate can be achieved via an accurate control of the plasma parameters. However, this also results in a highly defective interface in the solar cells due to the bombardment of the P-layer by positively charged nanocrystals during the deposition of the I-layer. We show that decreasing the ion energy by increasing the total pressure or by using silane-helium mixtures allows us to increase both the deposition rate and the solar cells efficiency, as required for cost effective thin film photovoltaics

Use of europium ions for SAD phasing of lysozyme at the Cu Kα wavelength

International Nuclear Information System (INIS)

Vijayakumar, Balakrishnan; Velmurugan, Devadasan

2012-01-01

Europium(III) ions bound to the surface of hen egg-white lysozyme were found to exhibit good anomalous signal facilitating SAD phasing using laboratory-source data and automated model building. The europium ion-binding sites were observed up to the 15σ level. Europium is shown to be a good anomalous scatterer in SAD phasing for solving the structure of biological macromolecules. The large value of the anomalous contribution of europium, f′′ = 11.17 e − , at the Cu Kα wavelength is an advantage in de novo phasing and automated model building. Tetragonal crystals of hen egg-white lysozyme (HEWL) incorporating europium(III) chloride (50 mM) were obtained which diffracted to a resolution of 2.3 Å at a wavelength of 1.54 Å (Cu Kα). The master data set (360° frames) was split and analyzed for anomalous signal-to-noise ratio, multiplicity, completeness, SAD phasing and automated building. The structure solution and model building of the split data sets were carried out using phenix.autosol and phenix.autobuild. The contributions of the Eu ions to SAD phasing using in-house data collection are discussed. This study revealed successful lysozyme phasing by SAD using laboratory-source data involving Eu ions, which are mainly coordinated by the side chains of Asn46, Asp52 and Asp101 together with some water molecules

Complexation of lysozyme with sodium caseinate and micellar casein in aqueous buffered solutions

NARCIS (Netherlands)

Antonov, Y.A.; Moldenaers, P.; Cardinaels, R.M.

We present an extended structural and morphological study of the complexation of lysozyme (Lys) with sodium caseinate (SC) and micellar casein (MC) by means of turbidity measurements, phase analysis, dynamic, static and electrophoretic light scattering, bright-field and confocal laser scanning

[ACID-BASE MODULATION OF LYSOZYME ACTIVITY IN MEDIUM FOR CULTIVATION OF ENTEROBACTERIA].

Science.gov (United States)

Andryuschenko, S V; Perunova, N B

2015-01-01

Determination of modulating effect of acid-base state of medium for cultivation of enterobacteria on activity of C-type lysozyme. Escherichia coli BL21 (DE3) strain for protein expression, Escherichia coli K12 MG1655 model strain, Escherichia coli No. 242 strain, isolated from intestine biotope; 2 Klebsiella pneumoniae strains, one of those contained plasmid homologue of periplasmatic lysozyme inhibitor gene pliC; 1 typical Salmonella enterica ATCC 14028 strain and a Micrococcus luteus ATCC 15307 strain as a control--served as material for the study. The bacteria were cultivated for 24 hours in 2 ml of liquid medium LB at 37 degrees C, 250 rpm. Determination of antilysozyme activity (ALA) was carried out by a photonepehlometrical method according to O.V. Bukharin et al. (1999) with alterations. All the studied microorganisms, including Micrococcus luteus, at the specified conditions 24 hours after cultivation were established to change the pH of the liquid nutrient medium LB from the initial value of 6.6 ± 0.1 to 8.2 ± 0.2 units. ALA determination in the cultivation medium without buffer correction was accompanied by a decline of lysozyme activity at an order of magnitude. The effect was absent during ALA measurement by a standard technique. The local shift of acid-base state of biotope under the conditions of buffer system insufficiency results in a reversible alteration of antimicrobial activity of muramidase, that among other non-specific factors of the environment determines the background of interactions on the level of associative symbiosis. This aspect should be taken into consideration during development of models, that are close to real conditions of microsymbiocenotical interactions.

Use of {gamma}-irradiation to produce films from whey, casein and soya proteins: structure and functionals characteristics

Energy Technology Data Exchange (ETDEWEB)

Lacroix, M. E-mail: monique.lacroix@inrs-iaf.uquebec.ca; Le, T.C.; Ouattara, B.; Yu, H.; Letendre, M.; Sabato, S.F.; Mateescu, M.A.; Patterson, G

2002-03-01

{gamma}-irradiation and thermal treatments have been used to produce sterilized cross-linked films. Formulations containing variable concentrations of calcium caseinate and whey proteins (whey protein isolate (WPI) and commercial whey protein concentrate) or mixture of soya protein isolate (SPI) with WPI was investigated on the physico-chemical properties of these films. Results showed that the mechanical properties of cross-linked films improved significantly the puncture strength for all types of films. Size-exclusion chromatography showed for no cross-linked proteins, a molecular mass of around 40 kDa. The soluble fractions of the cross-linked proteins molecular distributions were between 600 and 3800 kDa. {gamma}-irradiation seems to modify to a certain extent the conformation of proteins which will adopt structures more ordered and more stable, as suggested by X-ray diffraction analysis. Microstructure observations showed that the mechanical characteristics of these films are closely related to their microscopic structure. Water vapor permeability of films based on SPI was also significantly decreased when irradiated. Microbial resistance was also evaluated for cross-linked films. Results showed that the level of biodegradation of cross-linked films was 36% after 60 d of fermentation in the presence of Pseudomonas aeruginosa.

Use of γ-irradiation to produce films from whey, casein and soya proteins: structure and functionals characteristics

International Nuclear Information System (INIS)

Lacroix, M.; Le, T.C.; Ouattara, B.; Yu, H.; Letendre, M.; Sabato, S.F.; Mateescu, M.A.; Patterson, G.

2002-01-01

γ-irradiation and thermal treatments have been used to produce sterilized cross-linked films. Formulations containing variable concentrations of calcium caseinate and whey proteins (whey protein isolate (WPI) and commercial whey protein concentrate) or mixture of soya protein isolate (SPI) with WPI was investigated on the physico-chemical properties of these films. Results showed that the mechanical properties of cross-linked films improved significantly the puncture strength for all types of films. Size-exclusion chromatography showed for no cross-linked proteins, a molecular mass of around 40 kDa. The soluble fractions of the cross-linked proteins molecular distributions were between 600 and 3800 kDa. γ-irradiation seems to modify to a certain extent the conformation of proteins which will adopt structures more ordered and more stable, as suggested by X-ray diffraction analysis. Microstructure observations showed that the mechanical characteristics of these films are closely related to their microscopic structure. Water vapor permeability of films based on SPI was also significantly decreased when irradiated. Microbial resistance was also evaluated for cross-linked films. Results showed that the level of biodegradation of cross-linked films was 36% after 60 d of fermentation in the presence of Pseudomonas aeruginosa

Pluronic-lysozyme conjugates as anti-adhesive and antibacterial bifunctional polymers for surface coating

NARCIS (Netherlands)

Muszanska, A.K.; Busscher, H.J.; Herrmann, A.; Mei, van der H.C.; Norde, W.

2011-01-01

This paper describes the preparation and characterization of polymer protein conjugates composed of a synthetic triblock copolymer with a central polypropylene oxide (PPO) block and two terminal polyethylene oxide (PEO) segments, Pluronic F-127, and the antibacterial enzyme lysozyme attached to the

Newly identified invertebrate-type lysozyme (Splys-i) in mud crab (Scylla paramamosain) exhibiting muramidase-deficient antimicrobial activity.

Science.gov (United States)

Zhou, Jian; Zhao, Shu; Fang, Wen-Hong; Zhou, Jun-Fang; Zhang, Jing-Xiao; Ma, Hongyu; Lan, Jiang-Feng; Li, Xin-Cang

2017-09-01

Lysozymes are widely distributed immune effectors exerting muramidase activity against the peptidoglycan of the bacterial cell wall to trigger cell lysis. However, some invertebrate-type (i-type) lysozymes deficient of muramidase activity still exhibit antimicrobial activity. To date, the mechanism underlying the antimicrobial effect of muramidase-deficient i-type lysozymes remains unclear. Accordingly, this study characterized a novel i-type lysozyme, Splys-i, in the mud crab Scylla paramamosain. Splys-i shared the highest identity with the Litopenaeus vannamei i-type lysozyme (Lvlys-i2, 54% identity) at the amino acid level. Alignment analysis and 3D structure comparison show that Splys-i may be a muramidase-deficient i-type lysozyme because it lacks the two conserved catalytic residues (Glu and Asp) that are necessary for muramidase activity. Splys-i is mainly distributed in the intestine, stomach, gills, hepatopancreas, and hemocytes, and it is upregulated by Vibrio harveyi or Staphylococcus aureus challenge. Recombinant Splys-i protein (rSplys-i) can inhibit the growth of Gram-negative bacteria (V. harveyi, Vibrio alginolyticus, Vibrio parahemolyticus, and Escherichia coli), Gram-positive bacteria (S. aureus, Bacillus subtilis, and Bacillus megaterium), and the fungus Candida albicans to varying degrees. In this study, two binding assays and a bacterial agglutination assay were conducted to elucidate the potential antimicrobial mechanisms of Splys-i. Results demonstrated that rSplys-i could bind to all nine aforementioned microorganisms. It also exhibited a strong binding activity to lipopolysaccharide from E. coli and lipoteichoic acid and peptidoglycan (PGN) from S. aureus but a weak binding activity to PGN from B. subtilis and β-glucan from fungi. Moreover, rSplys-i could agglutinate these nine types of microorganisms in the presence of Ca 2+ at different protein concentrations. These results suggest that the binding activity and its triggered

Influence of the ionic liquid 1-butyl-3-methylimidazolium bromide on amyloid fibrillogenesis in lysozyme: Evidence from photophysical and imaging studies.

Science.gov (United States)

Basu, Anirban; Bhattacharya, Subhash Chandra; Kumar, Gopinatha Suresh

2018-02-01

Many proteins can abnormally fold to form pathological amyloid deposits/aggregates that are responsible for various degenerative disorders called amyloidosis. Here we have examined the anti-amyloidogenic potency of an ionic liquid, 1-butyl-3-methylimidazolium bromide, using lysozyme as a model system. Thioflavin T fluorescence assay demonstrated that the ionic liquid suppressed the formation of lysozyme fibrils significantly. This observation was further confirmed by the Congo red assay. Fluorescence microscopy, intrinsic fluorescence studies, nile red fluorescence assay, ANS binding assay and circular dichroism studies also testified diminishing of the fibrillogenesis in the presence of ionic liquid. Formation of amyloid fibrils was also characterized by α to β conformational transition. From far-UV circular dichroism studies it was observed that the β-sheet content of the lysozyme samples decreased in the presence of the ionic liquid which in turn implied that fibrillogenesis was supressed by the ionic liquid. Atomic force microscopy imaging unequivocally established that the ionic liquid attenuated fibrillogenesis in lysozyme. These results may be useful for the development of more effective therapeutics for amyloidosis. Copyright © 2017 Elsevier B.V. All rights reserved.

Structural basis for the appearance of a molten globule state in chimeric molecules derived from lysozyme and alpha-lactalbumin.

Science.gov (United States)

Joniau, M; Haezebrouck, P; Noyelle, K; Van Dael, H

2001-07-01

The problem as to why alpha-lactalbumin, in the absence of Ca(2+), forms a molten globule intermediate, in contrast to its structural homologue lysozyme, has been addressed by the construction of chimeras of human lysozyme in which either the Ca(2+)-binding loop or a part of helix C of bovine alpha-lactalbumin were transplanted. Previously, we have shown that the introduction of both structural elements together in the lysozyme matrix causes the apo form of the resulting chimera to display molten globule behavior during the course of thermal denaturation. In this article, we demonstrate that this molten globule character is not correlated with the Ca(2+)-binding loop. Also, the Del 101 mutant in which Arg101 was deleted to simulate the alpha-lactalbumin conformation of the connecting loop between helix C and helix D, does not show a stable equilibrium intermediate. Rather, the molten globule character of the chimeras has to be related with a specific part of helix C. More particularly, attention is drawn to the four hydrophobic side-chains I93, V96, I99, and L100, the lysozyme counterparts of which are constituted of less bulky valines and alanine. Our observations are discussed in terms of decreased stability of the native form and increased stability of the intermediate molten globule. Copyright 2001 Wiley-Liss, Inc.

Targeting the human lysozyme gene on bovine αs1- casein gene ...

African Journals Online (AJOL)

Targeting an exogenous gene into a favorable gene locus and for expression under endogenous regulators is an ideal method in mammary gland bioreactor research. For this purpose, a gene targeting vector was constructed to targeting the human lysozyme gene on bovine αs1-casein gene locus. In this case, the ...

Targeting the human lysozyme gene on bovine αs1- casein gene ...

African Journals Online (AJOL)

ajl yemi

2011-11-28

Nov 28, 2011 ... Targeting an exogenous gene into a favorable gene locus and for expression under endogenous regulators is ... case, the expression of human lysozyme could be regulated by the endogenous cis-element of αs1- casein gene in .... Mouse mammary epithelial C127 cells (Cell Bank, Chinese. Academy of ...

Label-Free Aptasensor for Lysozyme Detection Using Electrochemical Impedance Spectroscopy

OpenAIRE

Dionisia Ortiz-Aguayo; Manel del Valle

2018-01-01

This research develops a label-free aptamer biosensor (aptasensor) based on graphite-epoxy composite electrodes (GECs) for the detection of lysozyme protein using Electrochemical Impedance Spectroscopy (EIS) technique. The chosen immobilization technique was based on covalent bonding using carbodiimide chemistry; for this purpose, carboxylic moieties were first generated on the graphite by electrochemical grafting. The detection was performed using [Fe(CN)6]3−/[Fe(CN)6]4− as redox probe. Afte...

Producing Films, Producing Meanings: Some Theoretical Reflections

Directory of Open Access Journals (Sweden)

Giorgio Grignaffini

2015-01-01

Full Text Available L'articolo si pone l'obiettivo di stimolare una maggiore attenzione da parte della semiotica del cinema alle pratiche produttive e ai materiali preparatori del film, finora poco studiati rispetto alle dinamiche che i testi stessi attivano sulla ricezione. Tale proposta di allargamento del campo di ricerca, viene approfondita nelle sue premesse teoriche in due momenti. Dapprima viene contestualizzata rispetto alla pratica dell'analisi semiotica "tradizionale"; poi, utilizzando gli strumenti della sociosemiotica elaborata da Eric Landowski, in particolare sfruttando il concetto di "aggiustamento" sviluppato nel quadro della riflessione sulle modalità di interazione, vengono evidenziati campi di applicazione (ad es. il lavoro del regista con l’attore e possibili vantaggi euristici.

Characterizations of arsenic-doped zinc oxide films produced by atmospheric metal-organic chemical vapor deposition

Energy Technology Data Exchange (ETDEWEB)

Weng, Li-Wei, E-mail: onlyway54@hotmail.com [Department of Electronic Engineering, College of Electrical Engineering and Computer Science, Chung Yuan Christian University, Chung-Li 32023, Taiwan (China); Uen, Wu-Yih, E-mail: uenwuyih@ms37.hinet.net [Department of Electronic Engineering, College of Electrical Engineering and Computer Science, Chung Yuan Christian University, Chung-Li 32023, Taiwan (China); Lan, Shan-Ming; Liao, Sen-Mao [Department of Electronic Engineering, College of Electrical Engineering and Computer Science, Chung Yuan Christian University, Chung-Li 32023, Taiwan (China); Yang, Tsun-Neng; Wu, Chih-Hung; Hong, Hwe-Fen; Ma, Wei-Yang [Institute of Nuclear Energy Research, P.O. Box 3-11, Lungtan 32500, Taiwan (China); Shen, Chin-Chang [Chemical Engineering Division, Institute of Nuclear Energy Research, Longtan Township, Taoyuan 32546, Taiwan (China)

2013-07-15

p-type ZnO films were prepared by atmospheric metal-organic chemical vapor deposition technique using arsine (AsH{sub 3}) as the doping source. The electrical and optical properties of arsenic-doped ZnO (ZnO:As) films fabricated at 450–600 °C with various AsH{sub 3} flow rates ranging from 8 to 21.34 μmol/min were analyzed and compared. Hall measurements indicate that stable p-type ZnO films with hole concentrations varying from 7.2 × 10{sup 15} to 5.8 × 10{sup 18} cm{sup −3} could be obtained. Besides, low temperature (17 K) photoluminescence spectra of all ZnO:As films also demonstrate the dominance of the line related to the neutral acceptor-bound exciton. Moreover, the elemental identity and chemical bonding information for ZnO:As films were examined by X-ray photoelectron spectroscopy. Based on the results obtained, the effects of doping conditions on the mechanism responsible for the p-type conduction were studied. Conclusively, a simple technique to fabricate good-quality p-type ZnO films has been recognized in this work. Depositing the film at 550 °C with an AsH{sub 3} flow rate of 13.72 μmol/min is appropriate for producing hole concentrations on the order of 10{sup 17} cm{sup −3} for it. Ultimately, by increasing the AsH{sub 3} flow rate to 21.34 μmol/min for doping and depositing the film at 600 °C, ZnO:As films with a hole concentration over 5 × 10{sup 18} cm{sup −3} together with a mobility of 1.93 cm{sup 2}V{sup −1} s{sup −1} and a resistivity of 0.494 ohm-cm can be achieved.

Engineering Pichia pastoris for Efficient Production of a Novel Bifunctional Strongylocentrotus purpuratus Invertebrate-Type Lysozyme.

Science.gov (United States)

Huang, Peng; Shi, Jinlei; Sun, Qingwen; Dong, Xianping; Zhang, Ning

2018-04-13

Lysozymes are known as ubiquitously distributed immune effectors with hydrolytic activity against peptidoglycan, the major bacterial cell wall polymer, to trigger cell lysis. In the present study, the full-length cDNA sequence of a novel sea urchin Strongylocentrotus purpuratus invertebrate-type lysozyme (sp-iLys) was synthesized according to the codon usage bias of Pichia pastoris and was cloned into a constitutive expression plasmid pPIC9K. The resulting plasmid, pPIC9K-sp-iLys, was integrated into the genome of P. pastoris strain GS115. The bioactive recombinant sp-iLys was successfully secreted into the culture broth by positive transformants. The highest lytic activity of 960 U/mL of culture supernatant was reached in fed-batch fermentation. Using chitin affinity chromatography and gel-filtration chromatography, recombinant sp-iLys was produced with a yield of 94.5 mg/L and purity of > 99%. Recombinant sp-iLys reached its peak lytic activity of 8560 U/mg at pH 6.0 and 30 °C and showed antimicrobial activities against Gram-negative bacteria (Vibrio vulnificus, Vibrio parahemolyticus, and Aeromonas hydrophila) and Gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis). In addition, recombinant sp-iLys displayed isopeptidase activity which reached the peak at pH 7.5 and 37 °C with the presence of 0.05 M Na + . In conclusion, this report describes the heterologous expression of recombinant sp-iLys in P. pastoris on a preparative-scale, which possesses lytic activity and isopeptidase activity. This suggests that sp-iLys might play an important role in the innate immunity of S. purpuratus.

Lysozyme and bilirubin bind to ACE and regulate its conformation and shedding

Science.gov (United States)

Danilov, Sergei M.; Lünsdorf, Heinrich; Akinbi, Henry T.; Nesterovitch, Andrew B.; Epshtein, Yuliya; Letsiou, Eleftheria; Kryukova, Olga V.; Piegeler, Tobias; Golukhova, Elena Z.; Schwartz, David E.; Dull, Randal O.; Minshall, Richard D.; Kost, Olga A.; Garcia, Joe G. N.

2016-01-01

Angiotensin I-converting enzyme (ACE) hydrolyzes numerous peptides and is a critical participant in blood pressure regulation and vascular remodeling. Elevated tissue ACE levels are associated with increased risk for cardiovascular and respiratory disorders. Blood ACE concentrations are determined by proteolytic cleavage of ACE from the endothelial cell surface, a process that remains incompletely understood. In this study, we identified a novel ACE gene mutation (Arg532Trp substitution in the N domain of somatic ACE) that increases blood ACE activity 7-fold and interrogated the mechanism by which this mutation significantly increases blood ACE levels. We hypothesized that this ACE mutation disrupts the binding site for blood components which may stabilize ACE conformation and diminish ACE shedding. We identified the ACE-binding protein in the blood as lysozyme and also a Low Molecular Weight (LMW) ACE effector, bilirubin, which act in concert to regulate ACE conformation and thereby influence ACE shedding. These results provide mechanistic insight into the elevated blood level of ACE observed in patients on ACE inhibitor therapy and elevated blood lysozyme and ACE levels in sarcoidosis patients. PMID:27734897

A comparative study for adsorption of lysozyme from aqueous samples onto Fe3O4 magnetic nanoparticles using different ionic liquids as modifier.

Science.gov (United States)

Kamran, Sedigheh; Absalan, Ghodratollah; Asadi, Mozaffar

2015-12-01

In this paper, nanoparticles of Fe3O4 as well as their modified forms with different ionic liquids (IL-Fe3O4) were prepared and used for adsorption of lysozyme. The mean size and the surface morphology of the nanoparticles were characterized by TEM, XRD and FTIR techniques. Adsorption studies of lysozyme were performed under different experimental conditions in batch system on different modified magnetic nanoparticles such as, lysozyme concentration, pH of the solution, and contact time. Experimental results were obtained under the optimum operational conditions of pH 9.0 and a contact time of 10 min when initial protein concentrations of 0.05-2.0 mg mL(-1) were used. The isotherm evaluations revealed that the Langmuir model attained better fits to the equilibrium data than the Freundlich model. The maximum obtained adsorption capacities were 370.4, 400.0 500.0 and 526.3 mg of lysozyme for adsorption onto Fe3O4 and modified magnetic nanoparticles by [C4MIM][Br], [C6MIM][Br] and [C8MIM][Br] per gram of adsorbent, respectively. The Langmuir adsorption constants were 0.004, 0.019, 0.024 and 0.012 L mg(-1) for adsorptions of lysozyme onto Fe3O4 and modified magnetic nanoparticles by [C4MIM][Br], [C6MIM][Br] and [C8MIM][Br], respectively. The adsorption capacity of lysozyme was found to be dependent on its chemical structure, pH of the solution, temperature and type of ionic liquid as modifier. The applicability of two kinetic models including pseudo-first order and pseudo-second order model was estimated. Furthermore, the thermodynamic parameters were calculated. Protein could desorb from IL-Fe3O4 nanoparticles by using NaCl solution at pH 9.5 and was reused.

Role of annealing temperature on microstructural and electro-optical properties of ITO films produced by sputtering

Science.gov (United States)

Senol, Abdulkadir; Gulen, Mahir; Yildirim, Gurcan; Ozturk, Ozgur; Varilci, Ahmet; Terzioglu, Cabir; Belenli, Ibrahim

2013-03-01

In this study, we investigate the effect of annealing temperature on electrical, optical and microstructural properties of indium tin oxide (ITO) films deposited onto Soda lime glass substrates by conventional direct current (DC) magnetron reactive sputtering technique at 100 watt using an ITO ceramic target (In2O3:SnO2, 90:10 wt. %) in argon atmosphere at room temperature. The films obtained are exposed to the calcination process at different temperature up to 700 ° C. Resistivity, Hall Effect, X-ray diffractometer (XRD), ultra violet-visible spectrometer (UV-vis) and atomic force microscopy (AFM) measurements are performed to characterize the samples. Moreover, phase purity, surface morphology, optical and photocatalytic properties of the films are compared with each other. Furthermore, mobility, carrier density and conductivity characteristics of the samples prepared are carried out as function of temperature in the range of 80-300 K at the magnetic field of 0.550 T. The results obtained show that all the properties depend strongly on the annealing temperature and in fact the film annealed at 400 ° C obtains the better optical properties due to the high refractive index while the film produced at 100 °C exhibits much better photoactivity than the other films as a result of the large optical energy band gap.

Oxidation of nanostructured Ti films produced by low energy cluster beam deposition: An X-ray Photoelectron Spectroscopy characterization

International Nuclear Information System (INIS)

Simone, Monica de; Snidero, Elena; Coreno, Marcello; Bongiorno, Gero; Giorgetti, Luca; Amati, Matteo; Cepek, Cinzia

2012-01-01

We used in-situ X-ray Photoelectron Spectroscopy (XPS) to study the oxidation process of a cluster-assembled metallic titanium film exposed to molecular oxygen at room temperature. The nanostructured film has been grown on a Si(111) substrate, in ultra high vacuum conditions, by coupling a supersonic cluster beam deposition system with an XPS experimental chamber. Our results show that upon in-situ oxygen exposure Ti 3+ is the first oxidation state observed, followed by Ti 4+ , whereas Ti 2+ is practically absent during the whole process. Our results compare well with the existing literature on Ti films produced using other techniques.

Enzymatic degradation of polycaprolactone–gelatin blend

International Nuclear Information System (INIS)

Banerjee, Aditi; Chatterjee, Kaushik; Madras, Giridhar

2015-01-01

Blends of polycaprolactone (PCL), a synthetic polymer and gelatin, natural polymer offer a optimal combination of strength, water wettability and cytocompatibility for use as a resorbable biomaterial. The enzymatic degradation of PCL, gelatin and PCL–gelatin blended films was studied in the presence of lipase (Novozym 435, immobilized) and lysozyme. Novozym 435 degraded the PCL films whereas lysozyme degraded the gelatin. Though Novozym 435 and lysozyme individually could degrade PCL–gelatin blended films, the combination of these enzymes showed the highest degradation of these blended films. Moreover, the enzymatic degradation was much faster when fresh enzymes were added at regular intervals. The changes in physico-chemical properties of polymer films due to degradation were studied by scanning electron microscopy, Fourier transform infrared spectroscopy and differential scanning calorimetry. These results have important implications for designing resorbable biomedical implants. (paper)

Tetragonal Lysozyme Interactions Studied by Site Directed Mutagenesis

Science.gov (United States)

Crawford, Lisa; Karr, Laurel J.; Nadarajah, Arunan; Pusey, Marc

1999-01-01

A number of recent experimental and theoretical studies have indicated that tetragonal lysozyme crystal growth proceeds by the addition of aggregates, formed by reversible self association of the solute molecules in the bulk solution. Periodic bond chain and atomic force microscopy studies have indicated that the probable growth unit is at minimum a 43 tetramer, and most likely an octamer composed of two complete turns about the 43 axis. If these results are correct, then there are intermolecular interactions which are only formed in the solution and others only formed at the joining of the growth unit to the crystal surface. We have set out to study these interactions, and the correctness of this hypothesis, using site directed mutagenesis of specific amino acid residues involved in the different bonds. We had initially expressed wild type lysozyme in S. cervasiae with yields of approximately 5 mg/L, which were eventually raised to approximately 40 mg/L. We are now moving the expression to the Pichia system, with anticipated yields of 300 to (3)500 mg/L, comparable to what can be obtained from egg whites. An additional advantage of using recombinant protein is the greater genetic homogeneity of the material obtained and the absence of any other contaminating egg proteins. The first mutation experiments are TYR 23 (Registered) PHE or ALA and ASN 113 (Registered) ALA or ASP. Both TYR 23 and ASN 113 form part of the postulated dimerization intermolecular binding site which lead to the formation of the 43 helix. Tyrosine also participates in an intermolecular hydrogen bond with ARG 114. The results of these and subsequent experiments will be discussed.

Crystalline and amorphous carbon nitride films produced by high-energy shock plasma deposition

International Nuclear Information System (INIS)

Bursilll, L.A.; Peng, Julin; Gurarie, V.N.; Orlov, A.V.; Prawer, S.

1995-01-01

High-energy shock plasma deposition techniques are used to produce carbon-nitride films containing both crystalline and amorphous components. The structures are examined by high-resolution transmission electron microscopy, parallel-electron-energy loss spectroscopy and electron diffraction. The crystalline phase appears to be face-centered cubic with unit cell parameter approx. a=0.63nm and it may be stabilized by calcium and oxygen at about 1-2 at % levels. The carbon atoms appear to have both trigonal and tetrahedral bonding for the crystalline phase. There is PEELS evidence that a significant fraction of the nitrogen atoms have sp 2 trigonal bonds in the crystalline phase. The amorphous carbon-nitride film component varies from essentially graphite, containing virtually no nitrogen, to amorphous carbon-nitride containing up to 10 at % N, where the fraction of sp 3 bonds is significant. 15 refs., 5 figs

Investigation of factors affecting the stability of lysozyme spray dried from ethanol-water solutions

DEFF Research Database (Denmark)

Ji, Shuying; Thulstrup, Peter Waaben; Mu, Huiling

2017-01-01

-ethanol mixtures. The effect of formulation additives (trehalose, Tween 20 and phosphate-buffered saline) and processing conditions (inlet temperature and storage time of lysozyme in the feed solution before the spray drying process) on the protein bioactivity was investigated. The results showed...

The Effect of Ethylene Glycol, Glycine Betaine, and Urea on Lysozyme Thermal Stability

Science.gov (United States)

Schwinefus, Jeffrey J.; Leslie, Elizabeth J.; Nordstrom, Anna R.

2010-01-01

The four-week student project described in this article is an extension of protein thermal denaturation experiments to include effects of added cosolutes ethylene glycol, glycine betaine, and urea on the unfolding of lysozyme. The transition temperatures and van't Hoff enthalpies for unfolding are evaluated for six concentrations of each cosolute,…

Susceptibility of bifidobacteria to lysozyme as a possible selection criterion for probiotic bifidobacterial strains

Czech Academy of Sciences Publication Activity Database

Rada, V.; Šplíchal, Igor; Rocková, S.; Grmanová, M.; Vlková, E.

2010-01-01

Roč. 32, č. 3 (2010), s. 451-455 ISSN 0141-5492 R&D Projects: GA ČR GA523/07/0572 Institutional research plan: CEZ:AV0Z50200510 Keywords : Bifidobacteria * human milk * Lysozyme Subject RIV: EE - Microbiology, Virology Impact factor: 1.768, year: 2010

Cross-linked lysozyme crystal templated synthesis of Au nanoparticles as high-performance recyclable catalysts

International Nuclear Information System (INIS)

Liang Miao; Liu Xia; Qi Wei; Su Rongxin; Huang Renliang; Yu Yanjun; He Zhimin; Wang Libing

2013-01-01

Bio-nanomaterials fabricated using a bioinspired templating technique represent a novel class of composite materials with diverse applications in biomedical, electronic devices, drug delivery, and catalysis. In this study, Au nanoparticles (NPs) are synthesized within the solvent channels of cross-linked lysozyme crystals (CLLCs) in situ without the introduction of extra chemical reagents or physical treatments. The as-prepared AuNPs-in-protein crystal hybrid materials are characterized by light microscopy, transmission electron microscopy, x-ray diffraction, and Fourier-transform infrared spectroscopy analyses. Small AuNPs with narrow size distribution reveal the restriction effects of the porous structure in the lysozyme crystals. These composite materials are proven to be active heterogeneous catalysts for the reduction of 4-nitrophenol to 4-aminophenol. These catalysts can be easily recovered and reused at least 20 times because of the physical stability and macro-dimension of CLLCs. This work is the first to use CLLCs as a solid biotemplate for the preparation of recyclable high-performance catalysts. (paper)

Fourier transform infrared studies in solid egg white lysozyme

International Nuclear Information System (INIS)

Rivzi, T.Z.

1994-12-01

Fourier Transform Infrared (FTIR) Spectroscopy is the most recent addition to the arsenal of bioanalytical techniques capable of providing information about the secondary structure of proteins in a variety of environments. FTIR spectra have been obtained in solid egg white lysozyme. The spectra display the usual amide I, II and III bands. Secondary structural information obtained from the spectra after applying resolution enhancement techniques to the amide I band has been found consistent with the x-ray crystallographic data of the protein and also to the spectroscopic data of the protein in aqueous solution. (author). 17 refs, 6 figs, 2 tabs

Antimicrobial Performance of Two Different Packaging Materials on the Microbiological Quality of Fresh Salmon

Directory of Open Access Journals (Sweden)

Manuela Rollini

2016-01-01

Full Text Available In the present research the antimicrobial activity of two active packaging materials on the spoilage microbiota of fresh salmon fillets was tested. A PET-coated film (PET: Polyethylene Terephthalate containing lysozyme and lactoferrin was tested in parallel with a carvacrol-coextruded multilayer film. Salmon fillet samples were stored up to four days at 0 and 5 °C, comparatively. The carvacrol multilayer film was found effective in preventing mesophiles and psychrotrophs at shorter storage time and at lower temperature (4.0 compared to 5.0 log CFU/g in the control sample—CFU: Colony Forming Units. Lysozyme/lactoferrin-coated PET was instead efficient in decreasing H2S-producing bacteria at longer storage time and higher temperature (2.7 instead of 4.7 log CFU/g in the control sample. Even if is not intended as a way to “clean” a contaminated food product, an active package solution can indeed contribute to reducing the microbial population in food items, thus lowering the risk of food-related diseases.

Educational Films: Writing, Directing, and Producing for Classroom, Television, and Industry.

Science.gov (United States)

Herman, Lewis

Intended for beginning and nonprofessional film makers interested in making the educational film, this book explains the technical aspects of film making, the roles of the specialists responsible for it, the types of film treatments (expository, narrative, and dramatic), and the various types of educational films. Some of the technical aspects…

Size Exclusion Chromatography Studies of the Initial Self-Association Steps of Chicken Egg White Lysozyme Nucleation

Science.gov (United States)

Ewing, Felecia; Donovan, David; Pusey, Marc

2000-01-01

Nucleation is one of the least understood aspects of crystallogenesis. In the case of macromolecule nucleation, this understanding is further hampered by uncertainty over what precisely is being discussed. We define the process of solute self-association (aggregation, oligomerization, interaction, clustering, etc.) whereby n-mers (n > or = 2) having a crystallographic or nascent crystallographic arrangement leading to the critical nucleus reversibly form in the solution, to be part of the nucleation process. This reversible self-association process is a fundamental part of the nucleation process, and occurs as a function of the solute concentration. In the case of chicken egg white lysozyme, a considerable body of experimental evidence leads us to the conclusion that it also forms the crystal growth units. Size exclusion chromatography is a simple and direct method for determining the equilibrium constants for the self-association process. A Pharmacia FPLC system was used to provide accurate solution flow rates. The column, injection valve, and sample loop were all mounted within a temperature-controlled chamber. Chromatographically re-purified lysozyme was first dialyzed against the column equilibration buffer, with injection onto the column after several hours pre-incubation at the running temperature. Preliminary experiments, were carried out using a Toyopearl HW-50F column (1 x 50cm), equilibrated with 0.1 M sodium acetate, 5% sodium chloride, pH 4.6, at 15C. Protein concentrations from 0.1 to 4 mg/ml were employed (C(sub sat) = 1.2 mg/ml). The data from several different protein preparations consistently shows a progressively decreasing elution volume with increasing protein concentration, indicating that reversible self-association is occurring. The dotted line indicates the monomeric lysozyme elution volume. However, lysozyme interacts with the column matrix in these experiments, which complicates data analysis.Accordingly, we are testing silica-based HPLC

Relationship between β-relaxation and structural stability of lysozyme: Microscopic insight on thermostabilization mechanism by trehalose from Raman spectroscopy experiments

Energy Technology Data Exchange (ETDEWEB)

Hédoux, Alain, E-mail: alain.hedoux@univ-lille1.fr; Paccou, Laurent; Guinet, Yannick [Université Lille Nord de France, F-59000 Lille France, USTL UMET UMR 8207 F-59655 Villeneuve d’Ascq (France)

2014-06-14

Raman investigations were carried out in the low-frequency and amide I regions on lysozyme aqueous solutions in absence and presence of trehalose. Raman spectroscopy gives the unique opportunity to analyze the protein and solvent dynamics in the low-frequency range while monitoring the unfolding process by capturing the spectrum of the amide I band. From the analysis of the quasielastic intensity, a dynamic change is firstly observed in a highly hydrated protein, around 70 °C, and interpreted in relation with the denaturation mechanism of the protein. The use of heavy water and partly deuterated trehalose gives clear information on protein–trehalose interactions in the native state of lysozyme (at room temperature) and during the thermal denaturation process of lysozyme. At room temperature, it was found that trehalose is preferentially excluded from the protein surface, and has a main effect on the tetrahedral local order of water molecules corresponding to a stiffening of the H-bond network in the solvent. The consequence is a significant reduction of the amplitude of fast relaxational motions, inducing a less marked dynamic transition shifted toward the high temperatures. Upon heating, interaction between trehalose and lysozyme is detected during the solvent penetration within the protein, i.e., while the native globular state softens into a molten globule (MG) state. Addition of trehalose reduces the protein flexibility in the MG state, improving the structural stability of the protein, and inhibiting the protein aggregation.

Establishing Antibacterial Multilayer Films on the Surface of Direct Metal Laser Sintered Titanium Primed with Phase-Transited Lysozyme

Science.gov (United States)

Guan, Binbin; Wang, Haorong; Xu, Ruiqing; Zheng, Guoying; Yang, Jie; Liu, Zihao; Cao, Man; Wu, Mingyao; Song, Jinhua; Li, Neng; Li, Ting; Cai, Qing; Yang, Xiaoping; Li, Yanqiu; Zhang, Xu

2016-11-01

Direct metal laser sintering is a technology that allows the fabrication of titanium (Ti) implants with a functional gradation of porosity and surface roughness according to three-dimensional (3D) computer data. The surface roughness of direct metal laser sintered titanium (DMLS-Ti) implants may provide abundant binding sites for bacteria. Bacterial colonization and subsequent biofilm formation can cause unsatisfactory cell adhesion and implant-related infections. To prevent such infections, a novel phase-transited lysozyme (PTL) was utilized as an initial functional layer to simply and effectively prime DMLS-Ti surfaces for subsequent coating with antibacterial multilayers. The purpose of the present study was to establish a surface with dual biological functionality. The minocycline-loaded polyelectrolyte multilayers of hyaluronic acid (HA) and chitosan (CS) formed via a layer-by-layer (LbL) self-assembly technique on PTL-functionalized DMLS-Ti were designed to inhibit pathogenic microbial infections while allowing the DMLS-Ti itself and the modified coatings to retain acceptable biocompatibility. The experimental results indicate that the DMLS-Ti and the hydrogel treated surfaces can inhibit early bacterial adhesion while completely preserving osteoblast functions. This design is expected to gain considerable interest in the medical field and to have good potential for applications in multifunctional DMLS-Ti implants.

Establishing Antibacterial Multilayer Films on the Surface of Direct Metal Laser Sintered Titanium Primed with Phase-Transited Lysozyme.

Science.gov (United States)

Guan, Binbin; Wang, Haorong; Xu, Ruiqing; Zheng, Guoying; Yang, Jie; Liu, Zihao; Cao, Man; Wu, Mingyao; Song, Jinhua; Li, Neng; Li, Ting; Cai, Qing; Yang, Xiaoping; Li, Yanqiu; Zhang, Xu

2016-11-08

Direct metal laser sintering is a technology that allows the fabrication of titanium (Ti) implants with a functional gradation of porosity and surface roughness according to three-dimensional (3D) computer data. The surface roughness of direct metal laser sintered titanium (DMLS-Ti) implants may provide abundant binding sites for bacteria. Bacterial colonization and subsequent biofilm formation can cause unsatisfactory cell adhesion and implant-related infections. To prevent such infections, a novel phase-transited lysozyme (PTL) was utilized as an initial functional layer to simply and effectively prime DMLS-Ti surfaces for subsequent coating with antibacterial multilayers. The purpose of the present study was to establish a surface with dual biological functionality. The minocycline-loaded polyelectrolyte multilayers of hyaluronic acid (HA) and chitosan (CS) formed via a layer-by-layer (LbL) self-assembly technique on PTL-functionalized DMLS-Ti were designed to inhibit pathogenic microbial infections while allowing the DMLS-Ti itself and the modified coatings to retain acceptable biocompatibility. The experimental results indicate that the DMLS-Ti and the hydrogel treated surfaces can inhibit early bacterial adhesion while completely preserving osteoblast functions. This design is expected to gain considerable interest in the medical field and to have good potential for applications in multifunctional DMLS-Ti implants.

Proteins at fluid interfaces: adsorption layers and thin liquid films.

Science.gov (United States)

Yampolskaya, Galina; Platikanov, Dimo

2006-12-21

A review in which many original published results of the authors as well as many other papers are discussed. The structure and some properties of the globular proteins are shortly presented, special accent being put on the alpha-chymotrypsin (alpha-ChT), lysozyme (LZ), human serum albumin (HSA), and bovine serum albumin (BSA) which have been used in the experiments with thin liquid films. The behaviour of protein adsorption layers (PAL) is extensively discussed. The dynamics of PAL formation, including the kinetics of adsorption as well as the time evolution of the surface tension of protein aqueous solutions, are considered. A considerable place is devoted to the surface tension and adsorption isotherms of the globular protein solutions, the simulation of PAL by interacting hard spheres, the experimental surface tension isotherms of the above mentioned proteins, and the interfacial tension isotherms for the protein aqueous solution/oil interface. The rheological properties of PAL at fluid interfaces are shortly reviewed. After a brief information about the experimental methods for investigation of protein thin liquid (foam or emulsion) films, the properties of the protein black foam films are extensively discussed: the conditions for their formation, the influence of the electrolytes and pH on the film type and stability, the thermodynamic properties of the black foam films, the contact angles film/bulk and their dynamic hysteresis. The next center of attention concerns some properties of the protein emulsion films: the conditions for formation of emulsion black films, the formation and development of a dimpling in microscopic, circular films. The protein-phospholipid mixed foam films are also briefly considered.

Effect of intracrystalline water on micro-Vickers hardness in tetragonal hen egg-white lysozyme single crystals

International Nuclear Information System (INIS)

Koizumi, H; Kawamoto, H; Tachibana, M; Kojima, K

2008-01-01

Mechanical properties of high quality tetragonal hen egg-white lysozyme single crystals which are one type of protein crystal were investigated by the indentation method. The indentation marks were clearly observed on the crystal surface and no elastic recovery of them occurred. The value of the micro-Vickers hardness in the wet condition was estimated to be about 20 MPa at room temperature. The hardness greatly depended on the amount of intracrystalline water (mobile water) contained in the crystals. The hardness increased with increasing evaporation time to air at room temperature. It reached the maximum at about 260 MPa, which is 13 times as much as that in the wet condition. The origin of such a change in hardness was explained in terms of the dislocation mechanisms in lysozyme single crystals

Properties of plasmonic arrays produced by pulsed-laser nanostructuring of thin Au films

Directory of Open Access Journals (Sweden)

Katarzyna Grochowska

2014-11-01

Full Text Available A brief description of research advances in the area of short-pulse-laser nanostructuring of thin Au films is followed by examples of experimental data and a discussion of our results on the characterization of structural and optical properties of gold nanostructures. These consist of partially spherical or spheroidal nanoparticles (NPs which have a size distribution (80 ± 42 nm and self-organization characterized by a short-distance order (length scale ≈140 nm. For the NP shapes produced, an observably broader tuning range (of about 150 nm of the surface plasmon resonance (SPR band is obtained by renewal thin film deposition and laser annealing of the NP array. Despite the broadened SPR bands, which indicate damping confirmed by short dephasing times not exceeding 4 fs, the self-organized Au NP structures reveal quite a strong enhancement of the optical signal. This was consistent with the near-field modeling and micro-Raman measurements as well as a test of the electrochemical sensing capability.

THE PRIMARY STRUCTURE OF HEVAMINE, AN ENZYME WITH LYSOZYME CHITINASE ACTIVITY FROM HEVEA-BRASILIENSIS LATEX

NARCIS (Netherlands)

JEKEL, PA; HARTMANN, JBH; BEINTEMA, JJ

1991-01-01

The primary structure of hevamine, an enzyme with lysozyme/chitinase activity from Hevea brasiliensis latex, has been determined predominantly with conventional non-automatic methods. The positions of three disulfide bridges have been determined. The sequence has about 60% identity with that of a

Two-Step Cycle for Producing Multiple Anodic Aluminum Oxide (AAO) Films with Increasing Long-Range Order.

Science.gov (United States)

Choudhary, Eric; Szalai, Veronika

2016-01-01

Nanoporous anodic aluminum oxide (AAO) membranes are being used for an increasing number of applications. However, the original two-step anodization method in which the first anodization is sacrificial to pre-pattern the second is still widely used to produce them. This method provides relatively low throughput and material utilization as half of the films are discarded. An alternative scheme that relies on alternating anodization and cathodic delamination is demonstrated that allows for the fabrication of several AAO films with only one sacrificial layer thus greatly improving total aluminum to alumina yield. The thickness for which the cathodic delamination performs best to yield full, unbroken AAO sheets is around 85 μm. Additionally, an image analysis method is used to quantify the degree of long-range ordering of the unit cells in the AAO films which was found to increase with each successive iteration of the fabrication cycle.

Preparation and antifouling property of polyurethane film modified by chondroitin sulfate

Science.gov (United States)

Yuan, Huihui; Xue, Jing; Qian, Bin; Chen, Huaying; Zhu, Yonggang; Lan, Minbo

2017-02-01

An antifouling polyurethane film modified by chondroitin sulfate (PU-CS) was prepared by chemical grafting with N-Boc-1,3-propanediamine as a spacer. The different mass fraction of N-Boc-1,3-propanediamine was investigated to obtain PU-CS films with different CS grafting density. The surface properties of PU-CS films were comprehensively characterized. Proteins adsorption and glycosaminoglycans adhesion on films were evaluated. Moreover, inorganic salt deposition on film with highest CS grafting density (3.70 μg/cm2) was briefly investigated. The results showed that the increase of CS grafting density improved not only the hydrophilicity but the antifouling performance of films. The best antifouling film reduced the adsorption of fibrinogen (BFG), human serum albumin (HSA) and lysozyme (LYS) by 81.4%, 95.0% and 76.5%, respectively, and the adhesion of chondroitin (CS), heparin (HP) and hyaluronic acid (HA) by 70.6%, 87.4% and 81.3%, respectively. In addition, the co-adsorption of proteins and glycosaminoglycans reduced up to 86.9% and 75.5%, respectively. Changes in inorganic salt deposition after co-adsorption of proteins and glycosaminoglycans on PU-CS(3) suggested that the proteins promoted the inorganic salt deposition, while glycosaminoglycans inhibited the crystal growth. The negatively charged polysaccharides might promote the generation of smaller crystals which could be conducive to provide theoretical and practical guide to develop novel urinary stents with significant anti-encrustation properties.

A calorimetric study of the interactions in the aqueous solutions of lysozyme in the presence of denaturing cosolvents

Energy Technology Data Exchange (ETDEWEB)

Castronuovo, Giuseppina, E-mail: giuseppina.castronuovo@unina.it [Department of Chemistry, University Federico II of Naples, Complesso Universitario Monte S. Angelo, via Cintia, 80126 Naples (Italy); Niccoli, Marcella [Department of Chemistry, University Federico II of Naples, Complesso Universitario Monte S. Angelo, via Cintia, 80126 Naples (Italy)

2012-09-10

Highlights: Black-Right-Pointing-Pointer A thermodynamic method is reported to monitor the chemical denaturation of lysozyme. Black-Right-Pointing-Pointer The enthalpic interaction coefficients are very useful parameters to gain information about the mechanism through which two hydrated molecules interact in solution. Black-Right-Pointing-Pointer Hypotheses are proposed about the mechanism underlying the denaturation of lysozyme induced by high concentrations of urea or ethanol. - Abstract: A thermodynamic method is reported to monitor the chemical denaturation of lysozyme. Heats of dilution of the protein in concentrated aqueous solutions of urea or ethanol have been determined at 298.15 K by flow microcalorimetry. The pairwise enthalpic interaction coefficients of the protein in the different solvent media are derived. These parameters allow to gain information about the influence of the cosolvents on the interactions acting between two interacting hydrated molecules of lysozyme, hence on the denaturation process. At increasing urea concentration, up to about 6 mol kg{sup -1}, the values of the interaction coefficients are large and negative and remain almost unaltered. The invariance of the coefficients underlines that, even in highly concentrated urea, the hydration shell of the protein is such to maintain essentially unaltered the native conformation. At higher urea concentrations, a sudden change in the sign of the coefficients monitors the variation in the interactions between two hydrated denatured protein molecules. The same trend is found when ethanol is the cosolvent. At increasing concentration of the cosolvent, coefficients are, at first, almost invariant. After that, denaturation occurs, detected as a jump toward much more negative values. The results obtained are rationalized on the basis of those previously found for small model molecules in concentrated solutions of urea or ethanol. The thermodynamic framework allows useful comments to be made on

A calorimetric study of the interactions in the aqueous solutions of lysozyme in the presence of denaturing cosolvents

International Nuclear Information System (INIS)

Castronuovo, Giuseppina; Niccoli, Marcella

2012-01-01

Highlights: ► A thermodynamic method is reported to monitor the chemical denaturation of lysozyme. ► The enthalpic interaction coefficients are very useful parameters to gain information about the mechanism through which two hydrated molecules interact in solution. ► Hypotheses are proposed about the mechanism underlying the denaturation of lysozyme induced by high concentrations of urea or ethanol. - Abstract: A thermodynamic method is reported to monitor the chemical denaturation of lysozyme. Heats of dilution of the protein in concentrated aqueous solutions of urea or ethanol have been determined at 298.15 K by flow microcalorimetry. The pairwise enthalpic interaction coefficients of the protein in the different solvent media are derived. These parameters allow to gain information about the influence of the cosolvents on the interactions acting between two interacting hydrated molecules of lysozyme, hence on the denaturation process. At increasing urea concentration, up to about 6 mol kg −1 , the values of the interaction coefficients are large and negative and remain almost unaltered. The invariance of the coefficients underlines that, even in highly concentrated urea, the hydration shell of the protein is such to maintain essentially unaltered the native conformation. At higher urea concentrations, a sudden change in the sign of the coefficients monitors the variation in the interactions between two hydrated denatured protein molecules. The same trend is found when ethanol is the cosolvent. At increasing concentration of the cosolvent, coefficients are, at first, almost invariant. After that, denaturation occurs, detected as a jump toward much more negative values. The results obtained are rationalized on the basis of those previously found for small model molecules in concentrated solutions of urea or ethanol. The thermodynamic framework allows useful comments to be made on the possible mode of action of the two cosolvents on the stability of proteins

Reduced Position Effect in Mature Transgenic Plants Conferred by the Chicken Lysozyme Matrix-Associated Region

NARCIS (Netherlands)

Mlynárová, Ľudmila; Loonen, Annelies; Heldens, Jos; Jansen, Ritsert C.; Keizer, Paul; Stiekema, Willem J.; Nap, Jan-Peter

1994-01-01

Matrix-associated regions may be useful for studying the role of chromatin architecture in transgene activity of transformed plants. The chicken lysozyme A element was shown to have specific affinity for tobacco nuclear matrices, and its influence on the variability of transgene expression in

Optical performance of thin films produced by the pulsed laser deposition of SiAlON and Er targets

Energy Technology Data Exchange (ETDEWEB)

Camps, I., E-mail: camps@io.cfmac.csic.es [Laser Processing Group, Instituto de Óptica, CSIC, C/Serrano 121, 28006 Madrid (Spain); Ramírez, J.M. [MIND-IN2UB, Departament d’Electrònica, Universitat de Barcelona, c/Martí i Franqués 1, 08028 Barcelona (Spain); Mariscal, A.; Serna, R. [Laser Processing Group, Instituto de Óptica, CSIC, C/Serrano 121, 28006 Madrid (Spain); Garrido, B. [MIND-IN2UB, Departament d’Electrònica, Universitat de Barcelona, c/Martí i Franqués 1, 08028 Barcelona (Spain); Perálvarez, M.; Carreras, J. [IREC, Fundació Privada Institut de Recerca en Energia de Catalunya (Spain); Barradas, N.P.; Alves, L.C. [C" 2TN, Instituto Superior Técnico, Universidade de Lisboa, E.N. 10, 2695-066 Bobadela (Portugal); Alves, E. [IPFN, Instituto Superior Técnico, Universidade de Lisboa, E.N. 10, 2695-066 Bobadela (Portugal)

2015-05-01

Highlights: • PLD production of Er-doped thin films from a low cost commercial SiAlON target. • The role of the ablation fluence on the composition, optical properties as well as on the light emission performance at 1.5 μm. • The optimized performance is obtained for the samples deposited at the higher used ablation energy density. Further improvement was achieved through annealing. - Abstract: We report the preparation and optical performance of thin films produced by pulsed laser deposition in vacuum at room temperature, by focusing an ArF excimer laser onto two separate targets: a commercial ceramic SiAlON and a metallic Er target. As a result of the alternate deposition Er:SiAlON films were formed. The as grown films exhibited an Er-related emission peaking at 1532 nm. The role of the PLD energy density during deposition on the final matrix film was investigated, in order to achieve an optimized matrix composition with enhanced optical properties, and its effect on the light emission performance.

Thin films for precision optics

International Nuclear Information System (INIS)

Araujo, J.F.; Maurici, N.; Castro, J.C. de

1983-01-01

The technology of producing dielectric and/or metallic thin films for high precision optical components is discussed. Computer programs were developed in order to calculate and register, graphically, reflectance and transmittance spectra of multi-layer films. The technology of vacuum evaporation of several materials was implemented in our thin-films laboratory; various films for optics were then developed. The possibility of first calculate film characteristics and then produce the film is of great advantage since it reduces the time required to produce a new type of film and also reduces the cost of the project. (C.L.B.) [pt

Generation of bi-transgenic pigs overexpressing human lactoferrin and lysozyme in milk.

Science.gov (United States)

Cui, Dan; Li, Jia; Zhang, Linlin; Liu, Shen; Wen, Xiao; Li, Qiuyan; Zhao, Yaofeng; Hu, Xiaoxiang; Zhang, Ran; Li, Ning

2015-04-01

Intensive swine production industry uses antibiotics to treat diseases and improve pig growth. This can not only cause antibiotic resistance, but can also pollute the environment or eventually affect human public health. To date, human lactoferrin (hLF) and human lysozyme (hLZ) have been known as non-adaptive but interactive antimicrobial members and could act in concert against bacteria, which contribute to host defense. Therefore, their expression in pigs might be an alternative strategy for replacing antibiotics in the pig production industry. In our study, we produced hLF and hLZ bi-transgenic pigs and assessed the milk's antibacterial ability. Integration of both transgenes was confirmed by PCR and southern blot. Both the hLF and hLZ were expressed in the mammary gland of bi-transgenic pigs, as detected by western blotting. The expression amounts were 6.5 g/L for hLF and 1.1 mg/L for hLZ using ELISA. Interestingly, pig milk containing hLF and hLZ had synergistic antimicrobial activity. Our results suggest an alternative approach for avoiding the use of antibiotics in the pig industry, which would be of great benefit to the commercial swine production.

Multilayered Films Produced by Layer-by-Layer Assembly of Chitosan and Alginate as a Potential Platform for the Formation of Human Adipose-Derived Stem Cell aggregates

Directory of Open Access Journals (Sweden)

Javad Hatami

2017-09-01

Full Text Available The construction of multilayered films with tunable properties could offer new routes to produce biomaterials as a platform for 3D cell cultivation. In this study, multilayered films produced with five bilayers of chitosan and alginate (CHT/ALG were built using water-soluble modified mesyl and tosyl–CHT via layer-by-layer (LbL self-assembly. NMR results demonstrated the presences of mesyl (2.83 ppm and tosyl groups (2.39, 7.37 and 7.70 ppm in the chemical structure of modified chitosans. The buildup of multilayered films was monitored by quartz-crystal-microbalance (QCM-D and film thickness was estimated using the Voigt-based viscoelastic model. QCM-D results demonstrated that CHT/ALG films constructed using mesyl or tosyl modifications (mCHT/ALG were significantly thinner in comparison to the CHT/ALG films constructed with unmodified chitosan (p < 0.05. Adhesion analysis demonstrated that human adipose stem cells (hASCs did not adhere to the mCHT/ALG multilayered films and formed aggregates with sizes between ca. 100–200 µm. In vitro studies on cell metabolic activity and live/dead staining suggested that mCHT/ALG multilayered films are nontoxic toward hACSs. Multilayered films produced via LbL assembly of ALG and off-the-shelf, water-soluble modified chitosans could be used as a scaffold for the 3D aggregates formation of hASCs in vitro.

[Do lactoferrin, lysozyme and the lactoperoxidase-thiocyanate-hydrogen peroxide-system cause negative microbiological results in mastitis secretions?].

Science.gov (United States)

Schmedt Auf Der Günne, H; Tenhagen, B A; Kutzer, P; Forderung, D; Heuwieser, W

2002-07-01

Lactoferrin, lysozyme and the lactoperoxidase-thiocyanate-peroxide-system are naturally occurring antimicrobial components of milk. The objective of this study was to examine, whether these components were responsible for negative results, when mastitis milk is cultured microbiologically. Quarter milk samples from 75 cows with clinical mastitis on a dairy farm in Brandenburg were submitted for microbiological culture and analysed for the content and the activities of the three components. Animals from all stages of lactation with clinical mastitis were included in the study. Animals were examined clinically and milk samples were collected prior to first treatment. Secretions from quarters with clinical mastitis were compared to those of neighbouring quarters without clinical mastitis. Secretions with positive cultural results were compared to those with negative results. The concentrations or activities of the three factors were significantly higher in the diseased quarters than in the quarters without clinical signs of mastitis. The concentration of lysozyme increased with severity of the clinical signs (local swelling and changes in secretion). The concentration of lactoferrin was significantly higher in quarters with slight alterations of glandular tissue than in quarters with medium or severe alterations (P mastitis with positive and negative culture results. The concentration of lysozyme was even higher in culturally positive samples than in negative samples (P culture of milk samples from quarters with clinical mastitis.

Crystallization and evaluation of hen egg-white lysozyme crystals for protein pH titration in the crystalline state

International Nuclear Information System (INIS)

Iwai, Wakari; Yagi, Daichi; Ishikawa, Takuya; Ohnishi, Yuki; Tanaka, Ichiro; Niimura, Nobuo

2008-01-01

Hen egg-white lysozyme was crystallized over a wide pH range (2.5–8.0) and the quality of the crystals was characterized. Crystallization phase diagrams at pH 2.5, 6.0 and 7.5 were determined To observe the ionized status of the amino acid residues in proteins at different pH (protein pH titration in the crystalline state) by neutron diffraction, hen egg-white lysozyme was crystallized over a wide pH range (2.5–8.0). Crystallization phase diagrams at pH 2.5, 6.0 and 7.5 were determined. At pH < 4.5 the border between the metastable region and the nucleation region shifted to the left (lower precipitant concentration) in the phase diagram, and at pH > 4.5 the border shifted to the right (higher precipitant concentration). The qualities of these crystals were characterized using the Wilson plot method. The qualities of all crystals at different pH were more or less equivalent (B-factor values within 25–40). It is expected that neutron diffraction analysis of these crystals of different pH provides equivalent data in quality for discussions of protein pH titration in the crystalline state of hen egg-white lysozyme

Polymorphism of Lysozyme Condensates.

Science.gov (United States)

Safari, Mohammad S; Byington, Michael C; Conrad, Jacinta C; Vekilov, Peter G

2017-10-05

Protein condensates play essential roles in physiological processes and pathological conditions. Recently discovered mesoscopic protein-rich clusters may act as crucial precursors for the nucleation of ordered protein solids, such as crystals, sickle hemoglobin polymers, and amyloid fibrils. These clusters challenge settled paradigms of protein condensation as the constituent protein molecules present features characteristic of both partially misfolded and native proteins. Here we employ the antimicrobial enzyme lysozyme and examine the similarities between mesoscopic clusters, amyloid structures, and disordered aggregates consisting of chemically modified protein. We show that the mesoscopic clusters are distinct from the other two classes of aggregates. Whereas cluster formation and amyloid oligomerization are both reversible, aggregation triggered by reduction of the intramolecular S-S bonds is permanent. In contrast to the amyloid structures, protein molecules in the clusters retain their enzymatic activity. Furthermore, an essential feature of the mesoscopic clusters is their constant radius of less than 50 nm. The amyloid and disordered aggregates are significantly larger and rapidly grow. These findings demonstrate that the clusters are a product of limited protein structural flexibility. In view of the role of the clusters in the nucleation of ordered protein solids, our results suggest that fine-tuning the degree of protein conformational stability is a powerful tool to control and direct the pathways of protein condensation.

Towards producing novel fish gelatin films by combination treatments of ultraviolet radiation and sugars (ribose and lactose) as cross-linking agents.

Science.gov (United States)

Bhat, Rajeev; Karim, A A

2014-07-01

Developing novel fish gelatin films with better mechanical properties than mammalian gelatin is a challenging but promising endeavor. Studies were undertaken to produce fish gelatin films by combining treatments with different sugars (ribose and lactose) followed 'by' 'and' ultraviolet (UV) radiation, as possible cross-linking agents. Increase in tensile strength and percent elongation at break was recorded, which was more significant in films without sugars that were exposed to UV radiation. Films with added ribose showed decreased solubility after UV treatment and exhibited higher swelling percentage than films with added lactose, which readily dissolved in water. FTIR spectra of all the films showed identical patterns, which indicated no major changes to have occurred in the functional groups as a result of interaction between gelatin, sugars and UV irradiation. The results of this study could be explored for commercial use, depending on industrial needs for either production of edible films or for food packaging purposes.

Functionalization of multiwalled carbon nanotubes by microwave irradiation for lysozyme attachment: comparison of covalent and adsorption methods by kinetics of thermal inactivation

Science.gov (United States)

Puentes-Camacho, Daniel; Velázquez, Enrique F.; Rodríguez-Félix, Dora E.; Castillo-Ortega, Mónica; Sotelo-Mundo, Rogerio R.; del Castillo-Castro, Teresa

2017-12-01

Proteins suffer changes in their tertiary structure when they are immobilized, and enzymatic activity is affected due to the low biocompatibility of some supporting materials. In this work immobilization of lysozyme on carbon nanotubes previously functionalized by microwave irradiation was studied. The effectiveness of the microwave-assisted acid treatment of carbon nanotubes was evaluated by XPS, TEM, Raman and FTIR spectroscopy. The carboxylic modification of nanotube surfaces by this fast, simple and feasible method allowed the physical adsorption and covalent linking of active lysozyme onto the carbonaceous material. Thermal inactivation kinetics, thermodynamic parameters and storage stability were studied for adsorbed and covalent enzyme complexes. A major stability was found for lysozyme immobilized by the covalent method, the activation energy for inactivation of the enzyme was higher for the covalent method and it was stable after 50 d of storage at 4 °C. The current study highlights the effect of protein immobilization method on the biotechnological potential of nanostructured biocatalysts.

DIRECTOR/PRODUCER ROBERT ZEMECKIS DURING FILMING OF 'CONTACT' AT LC39 PRESS SITE

Science.gov (United States)

1997-01-01

Robert Zemeckis, director/producer, and other Warner Bros. crew members oversee the filming of scenes for the movie 'Contact' at Kennedy Space Center's Launch Complex 39 Press Site on January 30. The screenplay for 'Contact' is based on the best-selling novel by the late astronomer Carl Sagan. The cast includes Jodie Foster, Matthew McConaughey, John Hurt, James Woods, Tom Skerritt, David Morse, William Fichtner, Rob Lowe and Angela Bassett. Described by Warner Bros. as a science fiction drama, 'Contact' will depict humankind's first encounter with evidence of extraterrestrial life.

Effect of the incorporation of antimicrobial/antioxidant proteins on the properties of potato starch films.

Science.gov (United States)

Moreno, Olga; Atarés, Lorena; Chiralt, Amparo

2015-11-20

Glycerol plasticized potato starch films containing bioactive proteins (lactoferrin (LF) and/or lysozyme (LZ), at 0.1 and 0.2 ratio with respect to starch) were obtained by casting method and characterized as to their microstructural, thermal and physical (water content, mechanical, water and oxygen barrier, optical) properties. The bioactive properties, named antioxidant and antimicrobial, of the proteins and the films were also characterized. The incorporation of proteins affected the structural and physical properties of potato starch films, while modifying their thermal behavior and increasing the glass transition temperature. Both proteins showed a certain degree of compatibility with starch chains through the bond formations (increase in Tg), while a part is separated and migrates to the film surface. Their incorporation, especially that of lactoferrin, greatly increased the film's brittleness, regardless of the films water content, although they enhance the water vapor and oxygen barrier properties, whatever the age of the film. Protein also reduced the film's transparency and gloss, while lactoferrin induced color changes. The thermal degradation of blend films and isolated proteins occurred at temperatures of over 250°C, which means that blend starch films can be thermoprocessed, according to their thermoplastic properties and following the usual practices of the plastics industries. A synergistic antimicrobial action against Escherichia coli and coliforms was observed when both LZ and LF were simultaneously applied. Both of these exhibited antioxidant capacity. Copyright © 2015. Published by Elsevier Ltd.

Slow aggregation of lysozyme in alkaline pH monitored in real time employing the fluorescence anisotropy of covalently labelled dansyl probe.

Science.gov (United States)

Homchaudhuri, Lopamudra; Kumar, Satish; Swaminathan, Rajaram

2006-04-03

The onset of hen egg white lysozyme aggregation on exposure to alkaline pH of 12.2 and subsequent slow growth of soluble lysozyme aggregates (at 298 K) was directly monitored by steady-state and time-resolved fluorescence anisotropy of covalently attached dansyl probe over a period of 24 h. The rotational correlation time accounting for tumbling of lysozyme in solution (40 microM) increased from approximately 3.6 ns (in pH 7) to approximately 40ns on exposure to pH 12.2 over a period of 6 h and remained stable thereafter. The growth of aggregates was strongly concentration dependent, irreversible after 60 min and inhibited by the presence of 0.9 M l-arginine in the medium. The day old aggregates were resistant to denaturation by 6 M guanidine.HCl. Our results reveal slow segmental motion of the dansyl probe in day old aggregates in the absence of L-arginine (0.9 M), but a much faster motion in its presence, when growth of aggregates is halted.

Salt-specific effects in lysozyme solutions

Directory of Open Access Journals (Sweden)

T. Janc

2016-03-01

Full Text Available The effects of additions of low-molecular-mass salts on the properties of aqueous lysozyme solutions are examined by using the cloud-point temperature, T_{cloud}, measurements. Mixtures of protein, buffer, and simple salt in water are studied at pH=6.8 (phosphate buffer and pH=4.6 (acetate buffer. We show that an addition of buffer in the amount above I_{buffer} = 0.6 mol dm^{-3} does not affect the T_{cloud} values. However, by replacing a certain amount of the buffer electrolyte by another salt, keeping the total ionic strength constant, we can significantly change the cloud-point temperature. All the salts de-stabilize the solution and the magnitude of the effect depends on the nature of the salt. Experimental results are analyzed within the framework of the one-component model, which treats the protein-protein interaction as highly directional and of short-range. We use this approach to predict the second virial coefficients, and liquid-liquid phase diagrams under conditions, where T_{cloud} is determined experimentally.

Attempt to produce both thick and thinned flowing superfluid films

International Nuclear Information System (INIS)

Kwoh, D.S.W.; Goodstein, D.L.

1977-01-01

As discussed in the preceding paper by Graham, a controversy has arisen over conflicting reports of whether a superfluid film becomes thinned when it is set into motion. We have performed an experiment designed to reproduce as nearly as possible two previous measurements giving opposite results. Our experiment is also designed to test directly a theory proposed by Goodstein and Saffman which would have reconciled the apparently contradictory observations. We are unable to reproduce the thick-film result, finding kinetic thinning in all cases, even where the Goodstein--Saffman theory would lead us to expect a thick film. We conclude, in agreement with Graham, that the film is always thinned when it flows, and that the theory is therefore unnecessary

Adsorption of human tear lipocalin to human meibomian lipid films.

Science.gov (United States)

Millar, Thomas J; Mudgil, Poonam; Butovich, Igor A; Palaniappan, Chendur K

2009-01-01

Tear lipocalin (Tlc) is a major lipid binding protein in tears and is thought to have an important role in stabilizing the Meibomian lipid layer by transferring lipids to it from the aqueous layer or ocular surface, or by adsorbing to it directly. These possible roles have been investigated in vitro using human Tlc. Tlc was purified from human tears by size exclusion chromatography followed by ion exchange chromatography. Three additional samples of the Tlc were prepared by lipidation, delipidation, and relipidation. The lipids extracted from the purified Tlc were analyzed by HPLC-MS followed by fragmentation. Adsorption of these different forms of Tlc to a human Meibomian lipid film spread on the surface of an artificial tear buffer in a Langmuir trough were observed by recording changes in the pressure with time (Pi-T profile) and monitoring the appearance of the film microscopically. These results were compared with similar experiments using a bovine Meibomian lipid film. The results indicated that Tlc binds slowly to a human Meibomian lipid film compared with lysozyme or lactoferrin, even at 37 degrees C. The adsorption of Tlc to a human Meibomian lipid film was very different from its adsorption to a bovine Meibomian lipid film, indicating the nature of the lipids in the film is critical to the adsorption process. Similarly, the different forms of Tlc had quite distinct adsorption patterns, as indicated both by changes in Pi-T profiles and the microscopic appearance of the films. It was concluded that human Tlc was capable of adsorbing to and penetrating into a Meibomian lipid layer, but this process is very complex and depends on both the types of lipids bound to Tlc and the lipid complement comprising the Meibomian lipid film.

Adsorption of Human Tear Lipocalin to Human Meibomian Lipid Films

Science.gov (United States)

Millar, Thomas J.; Mudgil, Poonam; Butovich, Igor A.; Palaniappan, Chendur K.

2009-01-01

Purpose Tear lipocalin (Tlc) is a major lipid binding protein in tears and is thought to have an important role in stabilizing the Meibomian lipid layer by transferring lipids to it from the aqueous layer or ocular surface, or by adsorbing to it directly. These possible roles have been investigated in vitro using human Tlc. Methods Tlc was purified from human tears by size exclusion chromatography followed by ion exchange chromatography. Three additional samples of the Tlc were prepared by lipidation, delipidation, and relipidation. The lipids extracted from the purified Tlc were analyzed by HPLC-MS followed by fragmentation. Adsorption of these different forms of Tlc to a human Meibomian lipid film spread on the surface of an artificial tear buffer in a Langmuir trough were observed by recording changes in the pressure with time (∏-T profile) and monitoring the appearance of the film microscopically. These results were compared with similar experiments using a bovine Meibomian lipid film. Results The results indicated that Tlc binds slowly to a human Meibomian lipid film compared with lysozyme or lactoferrin, even at 37°C. The adsorption of Tlc to a human Meibomian lipid film was very different from its adsorption to a bovine Meibomian lipid film, indicating the nature of the lipids in the film is critical to the adsorption process. Similarly, the different forms of Tlc had quite distinct adsorption patterns, as indicated both by changes in ∏-T profiles and the microscopic appearance of the films. Conclusions It was concluded that human Tlc was capable of adsorbing to and penetrating into a Meibomian lipid layer, but this process is very complex and depends on both the types of lipids bound to Tlc and the lipid complement comprising the Meibomian lipid film. PMID:18757516

Niobium films produced by magnetron sputtering using an Ar-He mixture as discharge gas

CERN Document Server

Schucan, G M; Calatroni, Sergio

1995-01-01

Superconducting RF accelerating cavities have been produced at CERN by sputter-coating, with a thin niobium layer, cavities made of copper. In the present work, the discharge behaviour and niobium film properties have been investigated when part of the argon sputtering gas is replaced with helium. Helium is chosen because of its low mass, which reduces the energy lost by the niobium atoms colliding with the sputter gas atoms. The higher niobium atom energy should lead to higher adatom mobility on the substrate and, hence, to a larger grain size, a feature which is highly desirable to reduce the cavity surface resistance. It has been found that helium addition effectively helps to maintain the discharge at considerably lower argon pressures, via metastable-neutral ionisation and high secondary electron yield. However, a large amount of helium is trapped in the film, amount which is proportional to the helium partial pressure during the discharge, resulting in a reduction of both Residual Resistivity Ratio and ...

Layer-by-layer films assembled from natural polymers for sustained release of neurotrophin

International Nuclear Information System (INIS)

Zhang, Zhiling; Li, Qianqi; Han, Lin; Zhong, Yinghui

2015-01-01

Cortical neural prostheses (CNPs) hold great promise for paralyzed patients by recording neural signals from the brain and translating them into movement commands. However, these electrodes normally fail to record neural signals weeks to months after implantation due to inflammation and neuronal loss around the implanted neural electrodes. Sustained local delivery of neurotrophins from biocompatible coatings on CNPs can potentially promote neuron survival and attract the nearby neurons to migrate toward the electrodes to increase neuron density at the electrode/brain interface, which is important for maintaining the recording quality and long-term performance of the implanted CNPs. However, sustained release of neurotrophins from biocompatible ultrathin coatings is very difficult to achieve. In this study, we investigated the potential of several biocompatible natural polyanions including heparin, dextran sulfate, and gelatin to form layer-by-layer (LbL) assembly with positively charged neurotrophin nerve growth factor (NGF) and its model protein lysozyme, and whether sustained release of NGF and lysozyme can be achieved from the nanoscale thin LbL coatings. We found that gelatin, which is less negatively charged than heparin and dextran sulfate, showed the highest efficacy in loading proteins into the LbL films because other interactions in addition to electrostatic interactions were involved in LbL assembly. Sustained release of NGF and lysozymes for approximately 2 weeks was achieved from the gelatin-based LbL coatings. Released NGF maintained the bioactivity to stimulate neurite outgrowth from PC12 cells. Gelatin is generally recognized as safe by the FDA. Thus, the biocompatible LbL coating developed in this study is highly promising to be used for implanted CNPs to improve their long-term performance in human patients. (paper)

EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA); Scientific Opinion related to a notification from the Oenological Products and Practices International Association (OENOPPIA) on lysozyme from hen‟s egg to be used in the manufacture of wine as an anti-microbial stabilizer

DEFF Research Database (Denmark)

Tetens, Inge

Following a request from the European Commission, the Panel on Dietetic Products, Nutrition and Allergies (NDA) was asked to deliver a scientific opinion related to a notification from the Oenological Products and Practices International Association (OENOPPIA) on lysozyme from hen‟s egg used in t...... reactions in susceptible individuals under the conditions of use proposed by the applicant. © European Food Safety Authority, 2011...... in the manufacture of wine as an anti-microbial stabilizer/additive pursuant to Article 6, paragraph 11 of Directive 2000/13/EC – for permanent exemption from labelling. Allergic sensitisation against lysozyme is common among egg allergic individuals. In winemaking, lysozyme is used for the control of lactic acid...... individuals have been demonstrated in wines treated with lysozyme, and a number of clinical reports (including one double-blind placebo-controlled food challenge with lysozyme) described clinical allergic reactions to lysozyme. The Panel concludes that wines treated with lysozyme may trigger adverse allergic...

Re-refinement of 4xan: hen egg-white lysozyme with carboplatin in sodium bromide solution

NARCIS (Netherlands)

Tanley, Simon W. M.; Schreurs, Antoine M. M.; Kroon - Batenburg, Louise; Helliwell, John R.

2016-01-01

A re-refinement of 4xan, hen egg white lysozyme (HEWL) with carboplatin crystallised in NaBr solution, has been made (Tanley et al 2016). This follows our Response article (Tanley et al 2015) to the Critique article of Shabalin et al 2015, suggesting the need for corrections to some solute molecule

IAEA film library

International Nuclear Information System (INIS)

1959-01-01

Most of the scientific and technical films shown during the Second Geneva Conference for the Peaceful Uses of Atomic Energy were donated to IAEA by the producing countries at the end of the Conference. They will form the basic stock for the Agency's loan service intended to provide atomic energy institutions in Member States with film material. A detailed catalogue of the films, classified according to subject and giving conditions of loan or purchase, is now being prepared. In addition to this, information on all films produced in Member Countries dealing with the peaceful uses cf atomic energy is being assembled. The documentary information contained in the films in IAEA's possession relates to the following subjects: national programmes; nuclear physics; accelerators; plasma and fusion; reactors (power, research, material testing and experimental); prospecting and mining; ore dressing; metallurgy; production of fuel elements; treatment of irradiated fuel elements; protection against radiation; detection and counting; uses of radiation in medicine, biochemistry, agriculture and industry; industrial application of nuclear explosions. Most of the commentaries are in the language of the producing country. A few films are available in a choice of two languages. The films donated to the Agency total 82, two of which have been produced in Canada, 13 in France, one in India, one in Romania, one in Spain, 14 in the United Kingdom, one in the Union of South Africa, 47 in the United States of America and two in the USSR: they are mostly illustrations of papers presented at the Second Geneva Conference. In arranging for the circulation of scientific and technical films IAEA wishes to help meet some of the training and information needs of Member States. It is hoped that all organizations producing films on the peaceful uses of atomic energy will entrust copies to the IAEA with a view to their widest possible circulation. In the meantime, the Agency's films have been given

IAEA film library

Energy Technology Data Exchange (ETDEWEB)

NONE

1959-01-15

Most of the scientific and technical films shown during the Second Geneva Conference for the Peaceful Uses of Atomic Energy were donated to IAEA by the producing countries at the end of the Conference. They will form the basic stock for the Agency's loan service intended to provide atomic energy institutions in Member States with film material. A detailed catalogue of the films, classified according to subject and giving conditions of loan or purchase, is now being prepared. In addition to this, information on all films produced in Member Countries dealing with the peaceful uses cf atomic energy is being assembled. The documentary information contained in the films in IAEA's possession relates to the following subjects: national programmes; nuclear physics; accelerators; plasma and fusion; reactors (power, research, material testing and experimental); prospecting and mining; ore dressing; metallurgy; production of fuel elements; treatment of irradiated fuel elements; protection against radiation; detection and counting; uses of radiation in medicine, biochemistry, agriculture and industry; industrial application of nuclear explosions. Most of the commentaries are in the language of the producing country. A few films are available in a choice of two languages. The films donated to the Agency total 82, two of which have been produced in Canada, 13 in France, one in India, one in Romania, one in Spain, 14 in the United Kingdom, one in the Union of South Africa, 47 in the United States of America and two in the USSR: they are mostly illustrations of papers presented at the Second Geneva Conference. In arranging for the circulation of scientific and technical films IAEA wishes to help meet some of the training and information needs of Member States. It is hoped that all organizations producing films on the peaceful uses of atomic energy will entrust copies to the IAEA with a view to their widest possible circulation. In the meantime, the Agency's films have been given

Natural stimulus responsive scaffolds/cells for bone tissue engineering: influence of lysozyme upon scaffold degradation and osteogenic differentiation of cultured marrow stromal cells induced by CaP coatings.

Science.gov (United States)

Martins, Ana M; Pham, Quynh P; Malafaya, Patrícia B; Raphael, Robert M; Kasper, F Kurtis; Reis, Rui L; Mikos, Antonios G

2009-08-01

This work proposes the use of nonporous, smart, and stimulus responsive chitosan-based scaffolds for bone tissue engineering applications. The overall vision is to use biodegradable scaffolds based on chitosan and starch that present properties that will be regulated by bone regeneration, with the capability of gradual in situ pore formation. Biomimetic calcium phosphate (CaP) coatings were used as a strategy to incorporate lysozyme at the surface of chitosan-based materials with the main objective of controlling and tailoring their degradation profile as a function of immersion time. To confirm the concept, degradation tests with a lysozyme concentration similar to that incorporated into CaP chitosan-based scaffolds were used to study the degradation of the scaffolds and the formation of pores as a function of immersion time. Degradation studies with lysozyme (1.5 g/L) showed the formation of pores, indicating an increase of porosity ( approximately 5-55% up to 21 days) resulting in porous three-dimensional structures with interconnected pores. Additional studies investigated the influence of a CaP biomimetic coating on osteogenic differentiation of rat marrow stromal cells (MSCs) and showed enhanced differentiation of rat MSCs seeded on the CaP-coated chitosan-based scaffolds with lysozyme incorporated. At all culture times, CaP-coated chitosan-based scaffolds with incorporated lysozyme demonstrated greater osteogenic differentiation of MSCs, bone matrix production, and mineralization as demonstrated by calcium deposition measurements, compared with controls (uncoated scaffolds). The ability of these CaP-coated chitosan-based scaffolds with incorporated lysozyme to create an interconnected pore network in situ coupled with the demonstrated positive effect of these scaffolds upon osteogenic differentiation of MSCs and mineralized matrix production illustrates the strong potential of these scaffolds for application in bone tissue engineering strategies.

Thin HTSC films produced by a polymer metal precursor technique

Science.gov (United States)

Lampe, L. v.; Zygalsky, F.; Hinrichsen, G.

In precursors the metal ions are combined with acid groups of polymethacrylic acid (PMAA), polyacrylic acid (PAA) or novolac. Compared to thermal degradation temperature of pure polymers those of precursors are low. Precursors films were patterned by UV lithography. Diffractometric investigations showed that the c-axis oriented epitaxial films of YBa 2Cu 3O x and Bi 2Sr 2CaCu 2O x originated from amorphous metal oxide films, which were received after thermal degradation of the precursor. Transition temperatures and current densities were determined by electric resistivity measurements.

Optical and structural properties of protein/gold hybrid bio-nanofilms prepared by layer-by-layer method.

Science.gov (United States)

Pál, Edit; Hornok, Viktória; Sebok, Dániel; Majzik, Andrea; Dékány, Imre

2010-08-01

Lysozyme/gold thin layers were prepared by layer-by-layer (LbL) self-assembly method. The build-up of the films was followed by UV-vis-absorbance spectra, quartz crystal microbalance (QCM) and surface plasmon resonance (SPR) techniques. The structural property of films was examined by X-ray diffraction (XRD) measurements, while their morphology was studied by scanning electron microscopy (SEM) and atomic force microscopy (AFM). It was found that gold nanoparticles (NPs) had cubic crystalline structure, the primary particles form aggregates in the thin layer due to the presence of lysozyme molecules. The UV-vis measurements prove change in particle size while the colour of the film changes from wine-red to blue. The layer thickness of films was determined using the above methods and the loose, porous structure of the films explains the difference in the results. The vapour adsorption property of hybrid layers was also studied by QCM using different saturated vapours and ammonia gas. The lysozyme/Au films were most sensitive for ammonia gas among the tested gases/vapours due to the strongest interaction between the functional groups of the protein. Copyright 2010 Elsevier B.V. All rights reserved.

The Timing of cotE Expression Affects Bacillus subtilis Spore Coat Morphology but Not Lysozyme Resistance▿

Science.gov (United States)

Costa, Teresa; Serrano, Mónica; Steil, Leif; Völker, Uwe; Moran, Charles P.; Henriques, Adriano O.

2007-01-01

The synthesis of structural components and morphogenetic factors required for the assembly of the Bacillus subtilis spore coat is governed by a mother cell-specific transcriptional cascade. The first two temporal classes of gene expression, which involve RNA polymerase sigma σE factor and the ancillary regulators GerR and SpoIIID, are deployed prior to engulfment of the prespore by the mother cell. The two last classes rely on σK, whose activation follows engulfment completion, and GerE. The cotE gene codes for a morphogenetic protein essential for the assembly of the outer coat layer and spore resistance to lysozyme. cotE is expressed first from a σE-dependent promoter and, in a second stage, from a promoter that additionally requires SpoIIID and that remains active under σK control. CotE localizes prior to engulfment completion close to the surface of the developing spore, but formation of the outer coat is a late, σK-controlled event. We have transplanted cotE to progressively later classes of mother cell gene expression. This created an early class of mutants in which cotE is expressed prior to engulfment completion and a late class in which expression of cotE follows the complete engulfment of the prespore. Mutants of the early class assemble a nearly normal outer coat structure, whereas mutants of the late class do not. Hence, the early expression of CotE is essential for outer coat assembly. Surprisingly, however, all mutants were fully resistant to lysozyme. The results suggest that CotE has genetically separable functions in spore resistance to lysozyme and spore outer coat assembly. PMID:17172339

26 CFR 1.992-4 - Coordination with personal holding company provisions in case of certain produced film rents.

Science.gov (United States)

2010-04-01

... 26 Internal Revenue 10 2010-04-01 2010-04-01 false Coordination with personal holding company provisions in case of certain produced film rents. 1.992-4 Section 1.992-4 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Domestic International...

Rapid localized crystallization of lysozyme by laser trapping.

Science.gov (United States)

Yuyama, Ken-Ichi; Chang, Kai-Di; Tu, Jing-Ru; Masuhara, Hiroshi; Sugiyama, Teruki

2018-02-28

Confining protein crystallization to a millimetre size was achieved within 0.5 h after stopping 1 h intense trapping laser irradiation, which shows excellent performance in spatial and temporal controllability compared to spontaneous nucleation. A continuous-wave near-infrared laser beam is tightly focused into a glass/solution interfacial layer of a supersaturated buffer solution of hen egg-white lysozyme (HEWL). The crystallization is not observed during laser trapping, but initiated by stopping the laser irradiation. The generated crystals are localized densely in a circular area with a diameter of a few millimetres around the focal spot and show specific directions of the optical axes of the HEWL crystals. To interpret this unique crystallization, we propose a mechanism that nucleation and the subsequent growth take place in a highly concentrated domain consisting of HEWL liquid-like clusters after turning off laser trapping.

The interaction of equine lysozyme:oleic acid complexes with lipid membranes suggests a cargo off-loading mechanism

DEFF Research Database (Denmark)

Nielsen, Søren Bang; Wilhelm, Kristina; Vad, Brian

2010-01-01

The normal function of equine lysozyme (EL) is the hydrolysis of peptidoglycan residues of bacterial cell walls. EL is closely related to alpha-lactalbumins with respect to sequence and structure and further possesses the calcium binding site of alpha-lactalbumins. Recently, EL multimeric complexes...

Reinforcement of LENRA film by in-situ generated silica produced by sol gel process

International Nuclear Information System (INIS)

Mahathir Mohamed; Eda Yuhana Ariffin; Dahlan Mohd; Ibrahim Abdullah

2008-08-01

Liquid epoxidised natural rubber acrylate (LENRA) film was reinforced with silica-siloxane structures formed in-situ via sol gel process. Combination of these two components produces organic-inorganic composites. Tetraethylorthosilicate (TEOS) was used as precursor material for silica generation. Sol gel reaction was carried out at different concentrations of TEOS i.e. between 10 and 50 phr. Instrumental analysis was carried out by dynamic mechanical analysis (DMA), thermogravimetry analysis (TGA) and FTIR. It was found that miscibility between organic and inorganic components improved with the presence of silanol groups (Si-OH) and polar solvent i.e. THF, via hydrogen bonding formation between siloxane and LENRA. In this work, the effects of TEOS composition on mechanical properties and interaction that occurs between fillers and matrix have also been studied. It was observed that increasing the concentration of TEOS improved the scratch and stress properties of the film. Morphology study by the scanning electron microscopy (SEM) showed in-situ generated silica particles were homogenous and well dispersed at low concentrations of TEOS. (Author)

The Educational Film Industry

Science.gov (United States)

Tortora, Vincent R.; Schillaci, Peter

1975-01-01

Increased dialog is needed among educational film producers, distributors, and consumers in order to be sure that what is being produced meets educators' needs and also to help solve the financial problems of the film industry. (LS)

The Disruptive Effect of Lysozyme on the Bacterial Cell Wall Explored by an "In-Silico" Structural Outlook

Science.gov (United States)

Primo, Emiliano D.; Otero, Lisandro H.; Ruiz, Francisco; Klinke, Sebastián; Giordano, Walter

2018-01-01

The bacterial cell wall, a structural unit of peptidoglycan polymer comprised of glycan strands consisting of a repeating disaccharide motif [N-acetylglucosamine (NAG) and N-acetylmuramylpentapeptide (NAM pentapeptide)], encases bacteria and provides structural integrity and protection. Lysozymes are enzymes that break down the bacterial cell wall…

Correlates between Models of Virulence for Mycobacterium tuberculosis among Isolates of the Central Asian Lineage: a Case for Lysozyme Resistance Testing?

Science.gov (United States)

Casali, Nicola; Clark, Simon O.; Hooper, Richard; Williams, Ann; Velji, Preya; Gonzalo, Ximena

2015-01-01

Virulence factors (VFs) contribute to the emergence of new human Mycobacterium tuberculosis strains, are lineage dependent, and are relevant to the development of M. tuberculosis drugs/vaccines. VFs were sought within M. tuberculosis lineage 3, which has the Central Asian (CAS) spoligotype. Three isolates were selected from clusters previously identified as dominant in London, United Kingdom. Strain-associated virulence was studied in guinea pig, monocyte-derived macrophage, and lysozyme resistance assays. Whole-genome sequencing, single nucleotide polymorphism (SNP) analysis, and a literature review contributed to the identification of SNPs of interest. The animal model revealed borderline differences in strain-associated pathogenicity. Ex vivo, isolate C72 exhibited statistically significant differences in intracellular growth relative to C6 and C14. SNP candidates inducing lower fitness levels included 123 unique nonsynonymous SNPs, including three located in genes (lysX, caeA, and ponA2) previously identified as VFs in the laboratory-adapted reference strain H37Rv and shown to confer lysozyme resistance. C72 growth was most affected by lysozyme in vitro. A BLAST search revealed that all three SNPs of interest (C35F, P76Q, and P780R) also occurred in Tiruvallur, India, and in Uganda. Unlike C72, however, no single isolate identified through BLAST carried all three SNPs simultaneously. CAS isolates representative of three medium-sized human clusters demonstrated differential outcomes in models commonly used to estimate strain-associated virulence, supporting the idea that virulence varies within, not just across, M. tuberculosis lineages. Three VF SNPs of interest were identified in two additional locations worldwide, which suggested independent selection and supported a role for these SNPs in virulence. The relevance of lysozyme resistance to strain virulence remains to be established. PMID:25776753

Antimicrobial polycaprolactone/polyethylene glycol embedded lysozyme coatings of Ti implants for osteoblast functional properties in tissue engineering

Science.gov (United States)

Visan, A.; Cristescu, R.; Stefan, N.; Miroiu, M.; Nita, C.; Socol, M.; Florica, C.; Rasoga, O.; Zgura, I.; Sima, L. E.; Chiritoiu, M.; Chifiriuc, M. C.; Holban, A. M.; Mihailescu, I. N.; Socol, G.

2017-09-01

In this study, coatings based on lysozyme embedded into a matrix of polyethylene glycol (PEG) and polycaprolactone (PCL) were fabricated by two different methods (Matrix Assisted Pulsed Laser Evaporation - MAPLE and Dip Coating) for obtaining antimicrobial coatings envisaged for long term medical applications. Coatings with different PEG:PCL compositions (3:1; 1:1; 1:3) were synthesized in order to evaluate the antimicrobial activity of lysozyme embedded into the polymeric matrix. The main surface features, such as roughness and wettability, with impact on the microbial adhesion as well as on the eukaryote cell function were measured. The obtained composite coatings exhibited a significant antibacterial activity against Escherichia coli, Bacillus subtilis, Enterococcus faecalis and Staphylococcus aureus strains. As well, specific blended coatings showed appropriate viability, good spreading and normal cell morphology of SaOs2 human osteoblasts and mesenchymal stem cells (MSCs). These investigations highlight the suitability of biodegradable composites as implant coatings for decreasing the risk of bacterial contamination associated with prosthetic procedures.

Method for producing textured substrates for thin-film photovoltaic cells

Science.gov (United States)

Lauf, Robert J.

1994-01-01

The invention pertains to the production of ceramic substrates used in the manufacture of thin-film photovoltaic cells used for directly converting solar energy to electrical energy. Elongated ribbon-like sheets of substrate precursor containing a mixture of ceramic particulates, a binder, and a plasticizer are formed and then while green provided with a mechanically textured surface region used for supporting the thin film semiconductor of the photovoltaic cell when the sheets of the substrate precursor are subsequently cut into substrate-sized shapes and then sintered. The textured surface pattern on the substrate provides enhanced light trapping and collection for substantially increasing the solar energy conversion efficiency of thin-film photovoltaic cells.

Amino acids and glycine ethyl ester as new crystallization reagents for lysozyme

International Nuclear Information System (INIS)

Ito, Len; Shiraki, Kentaro; Yamaguchi, Hiroshi

2010-01-01

During the past two decades, amino acids and amino-acid derivatives have been applied in various fields of protein chemistry. The potential use of amino acids and their derivatives as new precipitating agents is described. Several amino acids and their derivatives are prominent additives in the field of protein chemistry. This study reports the use of charged amino acids and glycine ethyl ester as precipitants in protein crystallization, using hen egg-white lysozyme (HEWL) as a model. A discussion of the crystallization of HEWL using these reagents as precipitating agents is given

Assessment of antimicrobial activity of c-type lysozyme from Indian shrimp Fenneropenaeus indicus

Directory of Open Access Journals (Sweden)

Viswanathan Karthik

2014-10-01

Full Text Available Objective: To assess the multitudinal antimicrobial effects of recombinant lysozyme from Fenneropenaeus indicus (rFi-Lyz in comparison with commercially available recombinant hen egg white lysozyme (rHEWL. Methods: Antimicrobial activity of the recombinant rFi-Lyz using several Gram positive, Gram negative bacteria and fungi in comparison with rHEWL has been evaluated. rFi-Lyz was expressed and purified using Ni2+ affinity chromatography. The effect of rFi-Lyz in the growth of yeast Candida krusei, plant molds Rhizoctonia solani and Fusarium solani was assessed by well diffusion assay in petri plates with potato dextrose agar. Results: rFi-Lyz exhibited high inhibitory activity on Gram positive bacteria such as Staphylococcus aureus and Bacillus subtilis. Among various Gram negative bacteria tested Klebsiella pneumoniae exhibited the highest inhibition followed by Pseudomonas aeruginosa and Shigella dysenteriae. rFi-Lyz also exhibited significant inhibition on two marine pathogens Aeromonas veronii and Vibrio alginolyticus. Among the various fungal strains tested, rFi-Lyz inhibited the growth of budding yeast Candida krusei significantly. Further the growth of two other plants fungus Rhizoctonia solani and Fusarium oxysporum were retarded by rFi-Lyz in the plate inhibition assay. Conclusions: rFi-Lyz exhibits a broad spectrum of antimicrobial activity like a natural antibiotic on various pathogenic bacteria and fungal strains.

Crystallization and evaluation of hen egg-white lysozyme crystals for protein pH titration in the crystalline state.

Science.gov (United States)

Iwai, Wakari; Yagi, Daichi; Ishikawa, Takuya; Ohnishi, Yuki; Tanaka, Ichiro; Niimura, Nobuo

2008-05-01

To observe the ionized status of the amino acid residues in proteins at different pH (protein pH titration in the crystalline state) by neutron diffraction, hen egg-white lysozyme was crystallized over a wide pH range (2.5-8.0). Crystallization phase diagrams at pH 2.5, 6.0 and 7.5 were determined. At pH diagram, and at pH > 4.5 the border shifted to the right (higher precipitant concentration). The qualities of these crystals were characterized using the Wilson plot method. The qualities of all crystals at different pH were more or less equivalent (B-factor values within 25-40). It is expected that neutron diffraction analysis of these crystals of different pH provides equivalent data in quality for discussions of protein pH titration in the crystalline state of hen egg-white lysozyme.

Influence of deposition parameters on the optical and structural properties of TiO/sub 2/ films produced by reactive dc plasmatron sputtering

Energy Technology Data Exchange (ETDEWEB)

Schiller, S; Beister, G; Sieber, W; Schirmer, G; Hacker, E

1981-09-11

We investigated the variations in the structure and optical properties of TiO/sub 2/ films produced by reactive d.c. plasmatron sputtering with the most important deposition parameters. Over a wide range, the phase composition (ratio of rutile to anatase) and the grain size of the TiO/sub 2/ films can be influenced in a controlled manner by variations in the substrate temperature and the oxygen partial pressure. Because of their high refractive index and low light-scattering losses, plasmatron-sputtered TiO/sub 2/ films are of great interest in the field of optical interference coatings, e.g. for dielectric multilayer stacks.

Uniformly irradiated polymer film

International Nuclear Information System (INIS)

Fowler, S.L.

1979-01-01

Irradiated film having substantial uniformity in the radiation dosage profile is produced by irradiating the film within a trough having lateral deflection blocks disposed adjacent the film edges for deflecting electrons toward the surface of the trough bottom for further deflecting the electrons toward the film edge

Local pH at the surface of hen egg white lysozyme

Science.gov (United States)

Otosu, Takuhiro; Kobayashi, Kaito; Yamaguchi, Shoichi

2018-02-01

The microenvironment at the surface of hen-egg-white lysozyme (HEWL) was examined by analyzing the change in pKa of fluorescein isothiocyanate (FITC) upon binding to the N-terminus of HEWL. The result showed that the local pH at the HEWL surface is higher than the bulk pH. Furthermore, the data showed that the difference between the local and bulk pH becomes larger with decreasing pH, suggesting HEWL repels more protons at lower pH. Because the local pH affects the protonation states of functional amino-acids at the protein surface, the results provide the fundamental insight into the microenvironment at the protein surface.

Methods for using atomic layer deposition to produce a film for solid state electrolytes and protective electrode coatings for lithium batteries

Science.gov (United States)

Elam, Jeffrey W.; Meng, Xiangbo

2018-03-13

A method for using atomic layer deposition to produce a film configured for use in an anode, cathode, or solid state electrolyte of a lithium-ion battery or a lithium-sulfur battery. The method includes repeating a cycle for a predetermined number of times in an inert atmosphere. The cycle includes exposing a substrate to a first precursor, purging the substrate with inert gas, exposing the substrate to a second precursor, and purging the substrate with inert gas. The film is a metal sulfide.

Growth and properties of the CuInS2 thin films produced by glancing angle deposition

International Nuclear Information System (INIS)

Akkari, F. Chaffar; Kanzari, M.; Rezig, B.

2008-01-01

We use the glancing angle deposition technique (GLAD) to grow CuInS 2 thin films by a vacuum thermal method onto glass substrates. During deposition, the substrate temperature was maintained at 200 deg. C. Due to shadowing effect the oblique angle deposition technique can produce nanorods tilted toward the incident deposition flux. The evaporated atoms arrive at the growing interface at a fixed angle θ measured from the substrate normal. The substrate is rotated with rotational speed ω fixed at 0.033 rev s -1 . We show that the use of this growth technique leads to an improvement in the optical properties of the films. Indeed high absorption coefficients (10 5 -3.10 5 cm -1 ) in the visible range and near-IR spectral range are reached. In the case of the absence of the substrate rotation, scanning electron microscopy pictures show that the structure of the resulting film consists of nanocolumns that are progressively inclined towards the evaporation source as the incident angle was increased. If a rapid azimuthal rotation accompanies the substrate tilt, the resulting nanostructure is composed of an array of pillars normal to the substrate. The surface morphology show an improvement without presence of secondary phases for higher incident angles (θ > 60 deg.)

Electrical initiation of an energetic nanolaminate film

Science.gov (United States)

Tringe, Joseph W.; Gash, Alexander E.; Barbee, Jr., Troy W.

2010-03-30

A heating apparatus comprising an energetic nanolaminate film that produces heat when initiated, a power source that provides an electric current, and a control that initiates the energetic nanolaminate film by directing the electric current to the energetic nanolaminate film and joule heating the energetic nanolaminate film to an initiation temperature. Also a method of heating comprising providing an energetic nanolaminate film that produces heat when initiated, and initiating the energetic nanolaminate film by directing an electric current to the energetic nanolaminate film and joule heating the energetic nanolaminate film to an initiation temperature.

Lysozyme from the gut of the soft tick Ornithodoros moubata: the sequence, phylogeny and post-feeding regulation

Czech Academy of Sciences Publication Activity Database

Grunclová, L.; Fouquier, H.; Hypša, Václav; Kopáček, Petr

2003-01-01

Roč. 27, č. 8 (2003), s. 651-660 ISSN 0145-305X R&D Projects: GA ČR GA206/00/0266 Institutional research plan: CEZ:AV0Z6022909; CEZ:MSM 123100003 Keywords : Ornithodoros moubata * lysozyme * innate immunity Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.390, year: 2003

Accuracy and efficiency of published film dosimetry techniques using a flat-bed scanner and EBT3 film.

Science.gov (United States)

Spelleken, E; Crowe, S B; Sutherland, B; Challens, C; Kairn, T

2018-03-01

Gafchromic EBT3 film is widely used for patient specific quality assurance of complex treatment plans. Film dosimetry techniques commonly involve the use of transmission scanning to produce TIFF files, which are analysed using a non-linear calibration relationship between the dose and red channel net optical density (netOD). Numerous film calibration techniques featured in the literature have not been independently verified or evaluated. A range of previously published film dosimetry techniques were re-evaluated, to identify whether these methods produce better results than the commonly-used non-linear, netOD method. EBT3 film was irradiated at calibration doses between 0 and 4000 cGy and 25 pieces of film were irradiated at 200 cGy to evaluate uniformity. The film was scanned using two different scanners: The Epson Perfection V800 and the Epson Expression 10000XL. Calibration curves, uncertainty in the fit of the curve, overall uncertainty and uniformity were calculated following the methods described by the different calibration techniques. It was found that protocols based on a conventional film dosimetry technique produced results that were accurate and uniform to within 1%, while some of the unconventional techniques produced much higher uncertainties (> 25% for some techniques). Some of the uncommon methods produced reliable results when irradiated to the standard treatment doses (film analysis technique which uses transmission scanning, red colour channel analysis, netOD and a non-linear calibration curve for measuring doses up to 4000 cGy when using EBT3 film.

New sub-family of lysozyme-like proteins shows no catalytic activity: crystallographic and biochemical study of STM3605 protein from Salmonella Typhimurium

Energy Technology Data Exchange (ETDEWEB)

Michalska, Karolina; Brown, Roslyn N.; Li, Hui; Jedrzejczak, Robert; Niemann, George; Heffron, Fred; Cort, John R.; Adkins, Joshua N.; Babnigg, Gyorgy; Joachimiak, Andrzej

2013-03-01

Phage viruses that infect prokaryotes integrate their genome into the host chromosome; thus, microbial genomes typically contain genetic remnants of both recent and ancient phage infections. Often phage genes occur in clusters of atypical G+C content that reflect integration of the foreign DNA. However, some phage genes occur in isolation without other phage gene neighbors, probably resulting from horizontal gene transfer. In these cases, the phage gene product is unlikely to function as a component of a mature phage particle, and instead may have been co-opted by the host for its own benefit. The product of one such gene from Salmonella enterica serovar Typhimurium, STM3605, encodes a protein with modest sequence similarity to phage-like lysozyme (N-acetylmuramidase) but appears to lack essential catalytic residues that are strictly conserved in all lysozymes. Close homologs in other bacteria share this characteristic. The structure of the STM3605 protein was characterized by X-ray crystallography, and functional assays showed that it is a stable, folded protein whose structure closely resembles lysozyme. However, this protein is unlikely to hydrolyze peptidoglycan. Instead, STM3605 is presumed to have evolved an alternative function because it shows some lytic activity and partitions to micelles.

Effect of mobile phone use on salivary concentrations of protein, amylase, lipase, immunoglobulin A, lysozyme, lactoferrin, peroxidase and C-reactive protein of the parotid gland.

Science.gov (United States)

Hashemipour, M S; Yarbakht, M; Gholamhosseinian, A; Famori, H

2014-05-01

The possibility of side effects associated with the electromagnetic waves emitted from mobile phones is a controversial issue. The present study aimed to evaluate the effect of mobile phone use on parotid gland salivary concentrations of protein, amylase, lipase, immunoglobulin A, lysozyme, lactoferrin, peroxidase and C-reactive protein. Stimulated salivary samples were collected simultaneously from both parotid glands of 86 healthy volunteers. Salivary flow rate and salivary concentrations of proteins, amylase, lipase, lysozyme, lactoferrin, peroxidase, C-reactive protein and immunoglobulin A, were measured. Data were analysed using t-tests and one-way analyses of variance. Salivary flow rate and parotid gland salivary concentrations of protein were significantly higher on the right side compared to the left in those that predominantly held mobile phones on the right side. In addition, there was a decrease in concentrations of amylase, lipase, lysozyme, lactoferrin and peroxidase. The side of dominant mobile phone use was associated with differences in salivary flow rate and parotid gland salivary concentrations, in right-dominant users. Although mobile phone use influenced salivary composition, the relationship was not significant.

Effects of the aromatase inhibitor Letrozole on serum immunoglobulin and lysozyme levels in immunized rainbow trout (Oncorhynchus mykiss Walbaum females

Directory of Open Access Journals (Sweden)

Paria Akbary

2013-12-01

Full Text Available Letrozole is a synthetic aromatase inhibitor and interfere in the committed step in the synthesis of endogenous estrogens from androgens. Also estrogens regulate the immune system in teleost. Changes of 17- β- esrtradiol (E2, serum immunoglobulin and lysozyme levels were measured using a method based on the ability of lysozyme to lyse the bacterium Micrococcus lysodeikticus, enzyme-linked immunosorbent assay (ELISA and ELISA respectively. Twelve broodstocks were injected weekly with 2.5 mg kg-1 letrozole (an endocrine disrupter component two months before spawning season and vaccinated intraperitoneally (i.p with a bacterin (inactivated L. garviae one month before spawning. Twelve broodstocks for vaccination and twelve female rainbow trout as control group were also immiunised (i.p with the bacterin and injected (i.p with PBS, respectively. In the group received 2.5 mg AI kg-1 per week, serum E2 levels were significantly lower than that of other groups. Total immunoglobulin level and lysozyme activity were significantly higher in the parents received 2.5 mg kg-1 per week and were immunized with 10-9 cells ml-1 Lactococcus garvieae  compared to the group which immunized with L. garvieae and the control (non- immunized. The present study, suggests that aromatase inhibitors such as letrozole may be a potential tool to regulate the synthesis of E2, is involved in the hormone- immune system interaction in rainbow trout.

Protein adsorption resistance of PVP-modified polyurethane film prepared by surface-initiated atom transfer radical polymerization

International Nuclear Information System (INIS)

Yuan, Huihui; Qian, Bin; Zhang, Wei; Lan, Minbo

2016-01-01

Highlights: • Antifouling PVP brushes were successfully grafted on PU films by SI-ATRP. • The effect of polymerization time on surface property and topography was studied. • Hydrophilicity and protein fouling resistance of PVP–PU films were greatly promoted. • Competitive adsorption of three proteins on PVP–PU films was evaluated. - Abstract: An anti-fouling surface of polyurethane (PU) film grafted with Poly(N-vinylpyrrolidone) (PVP) was prepared through surface-initiated atom transfer radical polymerization (SI-ATRP). And the polymerization time was investigated to obtain PU films with PVP brushes of different lengths. The surface properties and protein adsorption of modified PU films were evaluated. The results showed that the hydrophilicity of PU–PVP films were improved with the increase of polymerization time, which was not positive correlation with the surface roughness due to the brush structure. Additionally, the protein resistance performance was promoted when prolonging the polymerization time. The best antifouling PU–PVP (6.0 h) film reduced the adsoption level of bovine serum albumin (BSA), lysozyme (LYS), and brovin serum fibrinogen (BFG) by 93.4%, 68.3%, 85.6%, respectively, compared to the unmodified PU film. The competitive adsorption of three proteins indicated that LYS preferentially adsorbed on the modified PU film, while BFG had the lowest adsorption selectivity. And the amount of BFG on PU–PVP (6.0 h) film reduced greatly to 0.08 μg/cm"2, which was almost one-tenth of its adsorption from the single-protein system. Presented results suggested that both hydrophilicity and surface roughness might be the important factors in all cases of protein adsorption, and the competitive or selective adsorption might be related to the size of the proteins, especially on the non-charged films.

Protein adsorption resistance of PVP-modified polyurethane film prepared by surface-initiated atom transfer radical polymerization

Energy Technology Data Exchange (ETDEWEB)

Yuan, Huihui; Qian, Bin; Zhang, Wei [Shanghai Key Laboratory of Functional Materials Chemistry and Research Center of Analysis and Test, East China University of Science and Technology, Shanghai 200237 (China); Lan, Minbo, E-mail: minbolan@ecust.edu.cn [Shanghai Key Laboratory of Functional Materials Chemistry and Research Center of Analysis and Test, East China University of Science and Technology, Shanghai 200237 (China); State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237 (China)

2016-02-15

Highlights: • Antifouling PVP brushes were successfully grafted on PU films by SI-ATRP. • The effect of polymerization time on surface property and topography was studied. • Hydrophilicity and protein fouling resistance of PVP–PU films were greatly promoted. • Competitive adsorption of three proteins on PVP–PU films was evaluated. - Abstract: An anti-fouling surface of polyurethane (PU) film grafted with Poly(N-vinylpyrrolidone) (PVP) was prepared through surface-initiated atom transfer radical polymerization (SI-ATRP). And the polymerization time was investigated to obtain PU films with PVP brushes of different lengths. The surface properties and protein adsorption of modified PU films were evaluated. The results showed that the hydrophilicity of PU–PVP films were improved with the increase of polymerization time, which was not positive correlation with the surface roughness due to the brush structure. Additionally, the protein resistance performance was promoted when prolonging the polymerization time. The best antifouling PU–PVP (6.0 h) film reduced the adsoption level of bovine serum albumin (BSA), lysozyme (LYS), and brovin serum fibrinogen (BFG) by 93.4%, 68.3%, 85.6%, respectively, compared to the unmodified PU film. The competitive adsorption of three proteins indicated that LYS preferentially adsorbed on the modified PU film, while BFG had the lowest adsorption selectivity. And the amount of BFG on PU–PVP (6.0 h) film reduced greatly to 0.08 μg/cm{sup 2}, which was almost one-tenth of its adsorption from the single-protein system. Presented results suggested that both hydrophilicity and surface roughness might be the important factors in all cases of protein adsorption, and the competitive or selective adsorption might be related to the size of the proteins, especially on the non-charged films.

As Film Goes Byte: The Change From Analog to Digital Film Perception

OpenAIRE

Loertscher, Miriam Laura; Weibel, David; Spiegel, Simon; Flueckiger, Barbara; Mennel, Pierre; Mast, Fred; Iseli, Christian

2016-01-01

The digital revolution changed film production in many ways. Until the end of the 20th century, most film professionals and critics preferred celluloid film. However, no previous empirical study compared complete narrative films recorded with analog and digital cinematography. Three short narrative films were produced with an analog and a digital camera attached to a 3D rig in order to control all optical parameters. In postproduction, a third version of a digital film was created to mimic th...

As Film Goes Byte: The Change From Analog to Digital Film Perception.

OpenAIRE

Loertscher M. L. Weibel D. Spiegel S. Flueckiger B. Mennel P. Mast F. W. & Iseli C.

2016-01-01

The digital revolution changed film production in many ways. Until the end of the 20th century most film professionals and critics preferred celluloid film. However no previous empirical study compared complete narrative films recorded with analog and digital cinematography. Three short narrative films were produced with an analog and a digital camera attached to a 3D rig in order to control all optical parameters. In postproduction a third version of a digital film was created to mimic the a...

Interaction between lysozyme and humic acid in layer-by-layer assemblies: Effects of pH and ionic strength

NARCIS (Netherlands)

Tan, W.F.; Norde, W.; Koopal, L.K.

2014-01-01

The interaction between protein and soluble organic matter is studied through layer-by-layer assembly of lysozyme (LSZ) and purified Aldrich humic acid (PAHA) at a solid surface (2-D) and in solution (3-D). By bringing a silica surface in alternating contact with solutions of LSZ and PAHA a

Interaction between lysozyme and humic acid in layer-by-layer assemblies : Effects of pH and ionic strength

NARCIS (Netherlands)

Tan, Wenfeng; Norde, Willem; Koopal, Luuk K.

2014-01-01

The interaction between protein and soluble organic matter is studied through layer-by-layer assembly of lysozyme (LSZ) and purified Aldrich humic acid (PAHA) at a solid surface (2-D) and in solution (3-D). By bringing a silica surface in alternating contact with solutions of LSZ and PAHA a

Film processing

International Nuclear Information System (INIS)

Abdul Nassir Ibrahim; Azali Muhammad; Ab. Razak Hamzah; Abd. Aziz Mohamed; Mohamad Pauzi Ismail

2008-01-01

The processing was made not only to show what are in the film but also to produce radiograph with high quality where the information gathered really presented level of the quality of the object inspected. Besides that, good procedure will make the film with good quality can keep the film in long time for reference. Here, more detailed on how the dark room functioned and its design. So, the good procedure while processed the film will be discussed detailed in this chapter from entering the dark room to exit from there.

Amorphous Terfenol-D films using nanosecond pulsed laser deposition

International Nuclear Information System (INIS)

Ma, James; O'Brien, Daniel T.; Kovar, Desiderio

2009-01-01

Thin films of Terfenol-D were produced by nanosecond pulsed laser deposition (PLD) at two fluences. Electron dispersive spectroscopy conducted using scanning electron and transmission electron microscopes showed that the film compositions were similar to that of the PLD target. Contrary to previous assertions that suggested that nanosecond PLD results in crystalline films, X-ray diffraction and transmission electron microscopy analysis showed that the films produced at both fluences were amorphous. Splatters present on the film had similar compositions to the overall film and were also amorphous. Magnetic measurements showed that the films had high saturation magnetization and magnetostriction, similar to high quality films produced using other physical vapor deposition methods.

In situ study of the growth and degradation processes in tetragonal lysozyme crystals on a silicon substrate by high-resolution X-ray diffractometry

Science.gov (United States)

Kovalchuk, M. V.; Prosekov, P. A.; Marchenkova, M. A.; Blagov, A. E.; D'yakova, Yu. A.; Tereshchenko, E. Yu.; Pisarevskii, Yu. V.; Kondratev, O. A.

2014-09-01

The results of an in situ study of the growth of tetragonal lysozyme crystals by high-resolution X-ray diffractometry are considered. The crystals are grown by the sitting-drop method on crystalline silicon substrates of different types: both on smooth substrates and substrates with artificial surface-relief structures using graphoepitaxy. The crystals are grown in a special hermetically closed crystallization cell, which enables one to obtain images with an optical microscope and perform in situ X-ray diffraction studies in the course of crystal growth. Measurements for lysozyme crystals were carried out in different stages of the crystallization process, including crystal nucleation and growth, developed crystals, the degradation of the crystal structure, and complete destruction.

Lysozyme-loaded lipid-polymer hybrid nanoparticles: preparation, characterization and colloidal stability evaluation.

Science.gov (United States)

Devrim, Burcu; Kara, Aslı; Vural, İmran; Bozkır, Asuman

2016-11-01

Lipid-polymer hybrid nanoparticles (LPNPs) are polymeric nanoparticles enveloped by lipid layers, which have emerged as a potent therapeutic nanocarrier alternative to liposomes and polymeric nanoparticles. The aim of this work was to develop, characterize and evaluate LPNPs to deliver a model protein, lysozyme. Lysozyme-loaded LPNPs were prepared by using the modified w/o/w double-emulsion-solvent-evaporation method. Poly-ɛ-caprolactone (PCL) was used as polymeric core material and tripalmitin:lechitin mixture was used to form a lipid shell around the LPNPs. LPNPs were evaluated for particle size distribution, zeta potential, morphology, encapsulation efficiency, in vitro drug release, stability and cytotoxicity. The DLS measurement results showed that the particle size of LPNPs ranged from 58.04 ± 1.95 nm to 2009.00 ± 0.52 nm. The AFM and TEM images of LPNPs demonstrate that LPNPs are spherical in shape. The protein-loading capacity of LPNPs ranged from 5.81% to 60.32%, depending on the formulation parameters. LPNPs displayed a biphasic drug release pattern with a burst release within 1 h, followed by sustained release afterward. Colloidal stability results of LPNPs in different media showed that particle size and zeta potential values of particles did not change significantly in all media except of FBS 100% for 120 h. Finally, the results of a cellular uptake study showed that LPNPs were significantly taken up by 83.3% in L929 cells. We concluded that the LPNPs prepared with PCL as polymeric core material and tripalmitin:lechitin mixture as lipid shell should be a promising choice for protein delivery.

Metal ions-binding T4 lysozyme as an intramolecular protein purification tag compatible with X-ray crystallography

Czech Academy of Sciences Publication Activity Database

Bouřa, Evžen; Bäumlová, Adriana; Chalupská, Dominika; Dubánková, Anna; Klíma, Martin

2017-01-01

Roč. 26, č. 6 (2017), s. 1116-1123 ISSN 0961-8368 R&D Projects: GA ČR(CZ) GJ17-07058Y; GA MŠk LO1302 Institutional support: RVO:61388963 Keywords : phage T4 * lysozyme * endolysin * histidine tag * protein purification * crystal structure Subject RIV: CE - Biochemistry OBOR OECD: Biochemistry and molecular biology Impact factor: 2.523, year: 2016

A comparative study on microstructure and tribological properties of Si3N4 and TiN thin films produced by IBED method

International Nuclear Information System (INIS)

Zhuang Daming; Liu Jiajun; Zhu Baoliang; Li Wenzhi; Zhang Xushou; Yang Shengrong

1995-01-01

In this paper, the tribological properties of Si 3 N 4 and TiN thin films produced by ion beam enhanced deposition (IBED) method were compared on an SRV friction and wear testing machine. In order to understand the reasons of their excellent properties the microstructure, microhardness and bonding strength with the substrate were analysed by SEM, X-ray diffraction, Knoop hardness test and scratching test methods separately. The results show that the TiN(1) films exhibits the best tribological properties, which are closely related with its higher hardness and bonding strength. (author)

Short-time dynamics of lysozyme solutions with competing short-range attraction and long-range repulsion: Experiment and theory

Science.gov (United States)

Riest, Jonas; Nägele, Gerhard; Liu, Yun; Wagner, Norman J.; Godfrin, P. Douglas

2018-02-01

Recently, atypical static features of microstructural ordering in low-salinity lysozyme protein solutions have been extensively explored experimentally and explained theoretically based on a short-range attractive plus long-range repulsive (SALR) interaction potential. However, the protein dynamics and the relationship to the atypical SALR structure remain to be demonstrated. Here, the applicability of semi-analytic theoretical methods predicting diffusion properties and viscosity in isotropic particle suspensions to low-salinity lysozyme protein solutions is tested. Using the interaction potential parameters previously obtained from static structure factor measurements, our results of Monte Carlo simulations representing seven experimental lysoyzme samples indicate that they exist either in dispersed fluid or random percolated states. The self-consistent Zerah-Hansen scheme is used to describe the static structure factor, S(q), which is the input to our calculation schemes for the short-time hydrodynamic function, H(q), and the zero-frequency viscosity η. The schemes account for hydrodynamic interactions included on an approximate level. Theoretical predictions for H(q) as a function of the wavenumber q quantitatively agree with experimental results at small protein concentrations obtained using neutron spin echo measurements. At higher concentrations, qualitative agreement is preserved although the calculated hydrodynamic functions are overestimated. We attribute the differences for higher concentrations and lower temperatures to translational-rotational diffusion coupling induced by the shape and interaction anisotropy of particles and clusters, patchiness of the lysozyme particle surfaces, and the intra-cluster dynamics, features not included in our simple globular particle model. The theoretical results for the solution viscosity, η, are in qualitative agreement with our experimental data even at higher concentrations. We demonstrate that semi

Patterned transparent zinc oxide films produced by sol-gel embossing

Energy Technology Data Exchange (ETDEWEB)

Rao, J.; Koh, L.H.K.; Crean, G.M. [Tyndall National Institute, Lee Maltings, Prospect Row, Cork (Ireland); Department of Microelectronic Engineering, University College Cork, Cork (Ireland); O' Brien, S. [Tyndall National Institute, Lee Maltings, Prospect Row, Cork (Ireland); Winfield, R.J.

2008-08-15

A low cost zinc oxide embossing technique is reported as a method of fabricating structures relevant to a variety of applications. A zinc based sol-gel material was prepared from zinc acetate[Zn(C{sub 2}H{sub 3}O{sub 2}){sub 2}], monoethanolamine[H{sub 2}NC{sub 2}H{sub 4}OH] and isopropanol. The sol-gel was cast into a polydimethylsiloxane (PDMS) mould a track design, placed in contact with the substrate and dried under vacuum at 70 C for 3 hours. The formed track pattern was further densified to provide a stable conductor film that retained the embossed shape. An optimum Zn sol-gel content of 0.6 M was identified. The embossed films had a transparency of greater than 83% in the visible region. The optical bandgap energy was evaluated to be 3.306 eV. The influence of ZnO sol-gel film synthesis and embossing parameters on the microstructure, morphology and optical transparency of fabricated structures is described. (copyright 2008 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

Fundamentals of laser-assisted fabrication of inorganic and organic films

DEFF Research Database (Denmark)

Schou, Jørgen

2008-01-01

The standard method for producing films by laser-assisted methods, Pulsed Laser Deposition (PLD) will be reviewed. The films considered are usually inorganic films, but also films of organic materials have been produced. Also the deposition of organic films by MAPLE (Matrix Assisted Pulsed Laser...

Densification of zirconia films by coevaporation with silica

International Nuclear Information System (INIS)

Feldman, A.; Farabaugh, E.N.

1985-04-01

Optical films of zirconia have been receiving considerable attention because of their potential use as the high-index layer in multilayer optical coatings for the ultraviolet portion of the spectrum. Several problems are associated with electron-beam deposited zirconia films, including index instability and index inhomogeneity. The index instability is caused by the adsorption and the desorption of water in the porous columnar structure of the zirconia films. Index inhomogeneity is due to the inhomogeneous structure in the films. Recent work has shown that the first several tens of nanometers of a film possess a cubic structure, whereas the outmost layers possess a monoclinic structure. One approach for producing bulk-like zirzonia films that is receiving considerable attention at present is ion-assisted electron-beam deposition. This is because the method has successfully produced zirconia films having bulk-like densities and refractive indices that show insignificant sensitivity to water adsorption. In this paper a similar effect is demonstrated when mixed zirconia:silica films are produced by coevaporation from independent electron-beam sources, and, in particular, it is shown that the admixture of a small amount of silica with the zirconia produces a film possessing a higher refractive index than a pure zirconia film

Josephson tunnel junctions in niobium films

International Nuclear Information System (INIS)

Wiik, Tapio.

1976-12-01

A method of fabricating stable Josephson tunnel junctions with reproducible characteristics is described. The junctions have a sandwich structure consisting of a vacuum evaporated niobium film, a niobium oxide layer produced by the glow discharge method and a lead film deposited by vacuum evaporation. Difficulties in producing thin-film Josephson junctions are discussed. Experimental results suggest that the lower critical field of the niobium film is the most essential parameter when evaluating the quality of these junctions. The dependence of the lower critical field on the film thickness and on the Ginzburg-Landau parameter of the film is studied analytically. Comparison with the properties of the evaporated films and with the previous calculations for bulk specimens shows that the presented model is applicable for most of the prepared samples. (author)

Film Sensor Device Fabricated by a Piezoelectric Poly(L-lactic acid) Film

Science.gov (United States)

Ando, Masamichi; Kawamura, Hideki; Kageyama, Keisuke; Tajitsu, Yoshiro

2012-09-01

Synthetic piezoelectric polymer films produced from petroleum feedstock have long been used as thin-film sensors and actuators. However, the fossil fuel requirements for synthetic polymer production and carbon dioxide emission from its combustion have raised concern about the environmental impact of its continued use. Eco-friendly biomass polymers, such as poly(L-lactic acid) (PLLA), are made from plant-based (vegetable starch) plastics and, thus, have a much smaller carbon footprint. Additionally, PLLA does not exhibit pyroelectricity or unnecessary poling. This suggests the usefulness of PLLA films for the human-machine interface (HMI). As an example of a new HMI, we have produced a TV remote control using a PLLA film. The intuitive operation provided by this PLLA device suggests that it is useful for the elderly or handicapped.

Specific effect of the linear charge density of the acid polysaccharide on thermal aggregation/ disaggregation processes in complex carrageenan/lysozyme systems

NARCIS (Netherlands)

Antonov, Y.; Zhuravleva, I.; Cardinaels, R.M.; Moldenaers, P.

2017-01-01

We study thermal aggregation and disaggregation processes in complex carrageenan/lysozyme systems with a different linear charge density of the sulphated polysaccharide. To this end, we determine the temperature dependency of the turbidity and the intensity size distribution functions in complex

Transparent and Printable Regenerated Kenaf Cellulose/PVA Film

Directory of Open Access Journals (Sweden)

Hatika Kaco

2014-02-01

Full Text Available Cellulose was extracted from kenaf core powder by a series of bleaching processes and subsequently dissolved using an alkaline LiOH/urea solvent at low temperatures. The produced cellulose solution was mixed with polyvinyl alcohol (PVA with different ratios of cellulose/PVA and coagulated to produce regenerated transparent films. The films were then air dried to produce transparent film. The effects of PVA content on tensile index, transparency, pore size, and printability of the films were studied. A slight reduction of 7% on the tensile index of the film was observed when the content of PVA increased to 10%. Nevertheless, the addition of 10% of PVA increased the porosity of the regenerated cellulose/PVA film, while the transparency of the film increased by 10%. The films were color-printed using a laser printer and can be recycled, in which the printed ink can be removed easily from the films with higher amount of PVA content. In addition, the films can be reprinted repeatedly several times.

Importance of the CMAP Correction to the CHARMM22 Protein Force Field: Dynamics of Hen Lysozyme

OpenAIRE

Buck, Matthias; Bouguet-Bonnet, Sabine; Pastor, Richard W.; MacKerell, Alexander D.

2005-01-01

The recently developed CMAP correction to the CHARMM22 force field (C22) is evaluated from 25 ns molecular dynamics simulations on hen lysozyme. Substantial deviations from experimental backbone root mean-square fluctuations and N-H NMR order parameters obtained in the C22 trajectories (especially in the loops) are eliminated by the CMAP correction. Thus, the C22/CMAP force field yields improved dynamical and structural properties of proteins in molecular dynamics simulations.

Silk film biomaterials for ocular surface repair

Science.gov (United States)

Lawrence, Brian David

Current biomaterial approaches for repairing the cornea's ocular surface upon injury are partially effective due to inherent material limitations. As a result there is a need to expand the biomaterial options available for use in the eye, which in turn will help to expand new clinical innovations and technology development. The studies illustrated here are a collection of work to further characterize silk film biomaterials for use on the ocular surface. Silk films were produced from regenerated fibroin protein solution derived from the Bombyx mori silkworm cocoon. Methods of silk film processing and production were developed to produce consistent biomaterials for in vitro and in vivo evaluation. A wide range of experiments was undertaken that spanned from in vitro silk film material characterization to in vivo evaluation. It was found that a variety of silk film properties could be controlled through a water-annealing process. Silk films were then generated that could be use in vitro to produce stratified corneal epithelial cell sheets comparable to tissue grown on the clinical standard substrate of amniotic membrane. This understanding was translated to produce a silk film design that enhanced corneal healing in vivo on a rabbit injury model. Further work produced silk films with varying surface topographies that were used as a simplified analog to the corneal basement membrane surface in vitro. These studies demonstrated that silk film surface topography is capable of directing corneal epithelial cell attachment, growth, and migration response. Most notably epithelial tissue development was controllably directed by the presence of the silk surface topography through increasing cell sheet migration efficiency at the individual cellular level. Taken together, the presented findings represent a comprehensive characterization of silk film biomaterials for use in ocular surface reconstruction, and indicate their utility as a potential material choice in the

Human Lysozyme Synergistically Enhances Bactericidal Dynamics and Lowers the Resistant Mutant Prevention Concentration for Metronidazole to Helicobacter pylori by Increasing Cell Permeability

Directory of Open Access Journals (Sweden)

Xiaolin Zhang

2016-10-01

Full Text Available Metronidazole (MNZ is an effective agent that has been employed to eradicate Helicobacter pylori (H. pylori. The emergence of broad MNZ resistance in H. pylori has affected the efficacy of this therapeutic agent. The concentration of MNZ, especially the mutant prevention concentration (MPC, plays an important role in selecting or enriching resistant mutants and regulating therapeutic effects. A strategy to reduce the MPC that can not only effectively treat H. pylori but also prevent resistance mutations is needed. H. pylori is highly resistant to lysozyme. Lysozyme possesses a hydrolytic bacterial cell wall peptidoglycan and a cationic dependent mode. These effects can increase the permeability of bacterial cells and promote antibiotic absorption into bacterial cells. In this study, human lysozyme (hLYS was used to probe its effects on the integrity of the H. pylori outer and inner membranes using as fluorescent probe hydrophobic 1-N-phenyl-naphthylamine (NPN and the release of aspartate aminotransferase. Further studies using a propidium iodide staining method assessed whether hLYS could increase cell permeability and promote cell absorption. Finally, we determined the effects of hLYS on the bactericidal dynamics and MPC of MNZ in H. pylori. Our findings indicate that hLYS could dramatically increase cell permeability, reduce the MPC of MNZ for H. pylori, and enhance its bactericidal dynamic activity, demonstrating that hLYS could reduce the probability of MNZ inducing resistance mutations.

Boundary singularities produced by the motion of soap films.

Science.gov (United States)

Goldstein, Raymond E; McTavish, James; Moffatt, H Keith; Pesci, Adriana I

2014-06-10

Recent work has shown that a Möbius strip soap film rendered unstable by deforming its frame changes topology to that of a disk through a "neck-pinching" boundary singularity. This behavior is unlike that of the catenoid, which transitions to two disks through a bulk singularity. It is not yet understood whether the type of singularity is generally a consequence of the surface topology, nor how this dependence could arise from an equation of motion for the surface. To address these questions we investigate experimentally, computationally, and theoretically the route to singularities of soap films with different topologies, including a family of punctured Klein bottles. We show that the location of singularities (bulk or boundary) may depend on the path of the boundary deformation. In the unstable regime the driving force for soap-film motion is the mean curvature. Thus, the narrowest part of the neck, associated with the shortest nontrivial closed geodesic of the surface, has the highest curvature and is the fastest moving. Just before onset of the instability there exists on the stable surface the shortest closed geodesic, which is the initial condition for evolution of the neck's geodesics, all of which have the same topological relationship to the frame. We make the plausible conjectures that if the initial geodesic is linked to the boundary, then the singularity will occur at the boundary, whereas if the two are unlinked initially, then the singularity will occur in the bulk. Numerical study of mean curvature flows and experiments support these conjectures.

Crystalline and amorphous phases in carbon nitride films produced by intense high-pressure plasma

International Nuclear Information System (INIS)

Gurarie, V.N.; Orlov, A.V.; Bursill, L.A.; JuLin, P.; Nugent, K.W.; Chon, J.W.; Prawer, S.

1997-01-01

Carbon-nitride films are prepared using a high-intensity pulsed plasma deposition technique. A wide range of nitrogen pressure and discharge intensity are used to investigate their effect on the morphology, nitrogen content, structure, bonding, phase composition and mechanical characteristics of the CN films deposited. Increasing the nitrogen pressure from 0.1 atm to 10 atm results in an increase of nitrogen incorporation into CN films to maximum of 45 at %. Under the high-energy density deposition conditions which involve ablation of the quartz substrate the CN films are found to incorporate in excess of 60 at %N. Raman spectra of these films contain sharp peaks characteristic of a distinct crystalline CN phase. TEM diffraction patterns for the films deposited below 1 atm unambiguously show the presence of micron-sized crystals displaying a cubic symmetry. (authors)

Pseudomonas aeruginosa outer membrane vesicles triggered by human mucosal fluid and lysozyme can prime host tissue surfaces for bacterial adhesion

Directory of Open Access Journals (Sweden)

Matteo Maria Emiliano Metruccio

2016-06-01

Full Text Available Pseudomonas aeruginosa is a leading cause of human morbidity and mortality that often targets epithelial surfaces. Host immunocompromise, or the presence of indwelling medical devices, including contact lenses, can predispose to infection. While medical devices are known to accumulate bacterial biofilms, it is not well understood why resistant epithelial surfaces become susceptible to P. aeruginosa. Many bacteria, including P. aeruginosa, release Outer Membrane Vesicles (OMVs in response to stress that can fuse with host cells to alter their function. Here, we tested the hypothesis that mucosal fluid can trigger OMV release to compromise an epithelial barrier. This was tested using tear fluid and corneal epithelial cells in vitro and in vivo. After 1 h both human tear fluid, and the tear component lysozyme, greatly enhanced OMV release from P. aeruginosa strain PAO1 compared to PBS controls (~100 fold. TEM and SDS-PAGE showed tear fluid and lysozyme-induced OMVs were similar in size and protein composition, but differed from biofilm-harvested OMVs, the latter smaller with fewer proteins. Lysozyme-induced OMVs were cytotoxic to human corneal epithelial cells in vitro and murine corneal epithelium in vivo. OMV exposure in vivo enhanced Ly6G/C expression at the corneal surface, suggesting myeloid cell recruitment, and primed the cornea for bacterial adhesion (~4-fold, P < 0.01. Sonication disrupted OMVs retained cytotoxic activity, but did not promote adhesion, suggesting the latter required OMV-mediated events beyond cell killing. These data suggest that mucosal fluid induced P. aeruginosa OMVs could contribute to loss of epithelial barrier function during medical device-related infections.

Discovery in Film, Book Two.

Science.gov (United States)

Gordon, Malcolm W.

Approximately 80 16 millimeter (16mm) short films are reviewed in this introduction and guide which attempts to be comprehensive in touching the major areas and styles of 16mm films now being produced. An attempt is made to describe as carefully as possible the style and content of each film and suggest ways in which the films might be used. Films…

Quantitative characterization of silicon- and aluminium oxynitride films produced by reactive dc-magnetron sputtering

International Nuclear Information System (INIS)

Dreer, S.

2000-05-01

The deposition of aluminum and silicon oxynitride films by reactive dc-magnetron sputtering was systematically planned by design of experiments, carried out and evaluated with the application of specialized statistics software. The influence of the deposition parameters on the resulting films was evaluated by multiple regression analysis. With the obtained data a model of the deposition process for the quantitative prediction of the deposition parameters necessary to obtain films with desired composition was built. This is also of technological importance, since the physical properties of the films strongly depend on their composition. Furthermore, the long term repeatability of the deposition process was implemented into the model. A precise and economic way for quantitative bulk analysis of silicon/aluminum, oxygen and nitrogen based on EPMA was presented and the use of data gained by the latter method is discussed for the calculation of relative sensitivity factors for SIMS and hf-SNMS. Advantages and disadvantages of SIMS, hf-SNMS, hf-GD-OES, and sputter AES were compared. The combination FT-IR/EPMA/SIMS at present offers the best possibility for a quantitative bulk and in depth distribution analysis of such films in the range of 20 to 1000 nm thickness. The films were also characterized by XRD and PAA. The refractive index and the growth rate of the films were determined by spectroscopic ellipsometry. With indentation by a nano hardness tester the hardness and the Young's modulus of the films were obtained. The results of these measurements were evaluated by statistical software. The dependencies of the physical properties on the deposition parameters and on the film thickness were evaluated and quantified. Furthermore, the dependencies of the physical properties on the film composition represented by the oxygen content were evaluated. (author)

Radiochromic film and polarization effects

International Nuclear Information System (INIS)

Yu, P.K.N.; Cheung, T.; Butson, M.J.; Cancer Services, Wollongong, NSW; Inwood, D.

2004-01-01

Full text: A new high sensitivity radiochromic film has been tested for its polarization properties. Gafchromic HS film has been shown to produce a relatively small (less than 3%) variation in measured optical density measured at 660nm wavelength when the light source is fully linear polarized and the film is rotated through 360 deg angle. Similar variations are seen when the detector is linearly polarized. If both light source and detector is linearly polarised variations in measured optical density can reach 15% when the film is rotated through 360 deg angle. This seems to be due to a phase shift in polarised light caused by the radiochromic film resulting in the polarised light source becoming out of phase with the polarised detector. Gafchromic HS radiochromic film produces a minimal polarization response with varying angle of rotation however we recommend that a polarization test be performed on a densitometry system to establish the extent of its polarization properties before accuracy dosimetry is performed with radiochromic HS film. Copyright (2004) Australasian College of Physical Scientists and Engineers in Medicine

Biaxial stretching of film principles and applications

CERN Document Server

Demeuse, M T

2011-01-01

Biaxial (having two axes) stretching of film is used for a range of applications and is the primary manufacturing process by which products are produced for the food packaging industry. Biaxial stretching of film: principles and applications provides an overview of the manufacturing processes and range of applications for biaxially stretched films. Part one reviews the fundamental principles of biaxial stretching. After an introductory chapter which defines terms, chapters discuss equipment design and requirements, laboratory evaluations, biaxial film structures and typical industrial processes for the biaxial orientation of films. Additional topics include post production processing of biaxially stretched films, the stress-strain behaviour of poly(ethylene terephthalate) and academic investigations of biaxially stretched films. Part two investigates the applications of biaxial films including fresh cut produce, snack packaging and product labelling. A final chapter investigates potential future trends for bi...

Hybrid perovskite solar cells: In situ investigation of solution-processed PbI2 reveals metastable precursors and a pathway to producing porous thin films

KAUST Repository

Barrit, Dounya

2017-04-17

The successful and widely used two-step process of producing the hybrid organic-inorganic perovskite CH3NH3PbI3, consists of converting a solution deposited PbI2 film by reacting it with CH3NH3I. Here, we investigate the solidification of PbI2 films from a DMF solution by performing in situ grazing incidence wide angle X-ray scattering (GIWAXS) measurements. The measurements reveal an elaborate sol–gel process involving three PbI2⋅DMF solvate complexes—including disordered and ordered ones—prior to PbI2 formation. The ordered solvates appear to be metastable as they transform into the PbI2 phase in air within minutes without annealing. Morphological analysis of air-dried and annealed films reveals that the air-dried PbI2 is substantially more porous when the coating process produces one of the intermediate solvates, making this more suitable for subsequent conversion into the perovskite phase. The observation of metastable solvates on the pathway to PbI2 formation open up new opportunities for influencing the two-step conversion of metal halides into efficient light harvesting or emitting perovskite semiconductors.

Message films in Africa: A look into the past

Directory of Open Access Journals (Sweden)

Rachael Diang′a

2016-12-01

Full Text Available Message Film-making has characterised much of films produced in post-independent Kenya. The country produced very few films in the 1980s, when indigenous film-making actually began to take root. Liberalisation of the economy, embracement of the digital technology and democratization in the 1990s paved way for a more stable film culture in the decade. A more promising growth of the film industry has been largely witnessed since the turn of the 21st century. Through this period, I note a strong tendency to produce films that are loaded with social messages deemed urgent and important to the target audience. In making these films, the film-makers hope to make a positive impact in the lives of the target audiences. These films tend to valorise the message, sometimes, neglecting the basic filmic codes, a practice that renders the films less entertaining. This endangers the growth of the industry since local films find very stiff competition from foreign films that are common on Kenyan screens. This study therefore investigates possible roots of message film-making in Africa that directly influence the tendencies in Kenya by making references to other African countries’ film experiences. My assumption is that Kenya’s cultural experiences are shared by other African countries.

Diffusion in and around alginate and chitosan films with embedded sub-millimeter voids

Energy Technology Data Exchange (ETDEWEB)

Patra, Subhajit; Bal, Dharmendra Kumar; Ganguly, Somenath, E-mail: snganguly@che.iitkgp.ernet.in

2016-02-01

Hydrogel scaffolds from biopolymers have potential use in the controlled release of drugs, and as 3-D structure for the formation of tissue matrix. This article describes the solute release behavior of alginate and chitosan films with embedded voids of sub-millimeter dimensions. Nitrogen gas was bubbled in a fluidic arrangement to generate bubbles, prior to the crosslinking. The crosslinked gel was dried in a vacuum oven, and subsequently, soaked in Vitamin B-12 solution. The dimensions of the voids immediately after the cross-linking of gel, and also after complete drying were obtained using a digital microscope and scanning electron microscope respectively. The porosity of the gel was measured gravimetrically. The release of Vitamin B-12 in PBS buffer on a shaker was studied. The release experiments were repeated at an elevated temperature of 37 °C in the presence of lysozyme. The diffusion coefficient within the gel layer and the mass transfer coefficient at the interface with the bulk-liquid were estimated using a mathematical model. For comparison, the experiment was repeated with a film that does not have any embedded void. The enhancement in diffusion coefficient due to the presence of voids is discussed in this article. - Highlights: • Formation of sub-millimeter voids in biopolymer films using fluidic arrangement • The retention of self-assembled bubbles in films after crosslinking, and drying • The enhancement observed in release of model drug with introduction of voids • The diffusion coefficients in and around biopolymer films from model regression • Use of classical model in explaining release profiles from dual porosity media.

Two duplicated chicken-type lysozyme genes in disc abalone Haliotis discus discus: molecular aspects in relevance to structure, genomic organization, mRNA expression and bacteriolytic function.

Science.gov (United States)

Umasuthan, Navaneethaiyer; Bathige, S D N K; Kasthuri, Saranya Revathy; Wan, Qiang; Whang, Ilson; Lee, Jehee

2013-08-01

Lysozymes are crucial antibacterial proteins that are associated with catalytic cleavage of peptidoglycan and subsequent bacteriolysis. The present study describes the identification of two lysozyme genes from disc abalone Haliotis discus discus and their characterization at sequence-, genomic-, transcriptional- and functional-levels. Two cDNAs and BAC clones bearing lysozyme genes were isolated from abalone transcriptome and BAC genomic libraries, respectively and sequences were determined. Corresponding deduced amino acid sequences harbored a chicken-type lysozyme (LysC) family profile and exhibited conserved characteristics of LysC family members including active residues (Glu and Asp) and GS(S/T)DYGIFQINS motif suggested that they are LysC counterparts in disc abalone and designated as abLysC1 and abLysC2. While abLysC1 represented the homolog recently reported in Ezo abalone [1], abLysC2 shared significant identity with LysC homologs. Unlike other vertebrate LysCs, coding sequence of abLysCs were distributed within five exons interrupted by four introns. Both abLysCs revealed a broader mRNA distribution with highest levels in mantle (abLysC1) and hepatopancreas (abLysC2) suggesting their likely main role in defense and digestion, respectively. Investigation of temporal transcriptional profiles post-LPS and -pathogen challenges revealed induced-responses of abLysCs in gills and hemocytes. The in vitro muramidase activity of purified recombinant (r) abLysCs proteins was evaluated, and findings indicated that they are active in acidic pH range (3.5-6.5) and over a broad temperature range (20-60 °C) and influenced by ionic strength. When the antibacterial spectra of (r)abLysCs were examined, they displayed differential activities against both Gram positive and Gram negative strains providing evidence for their involvement in bacteriolytic function in abalone physiology. Copyright © 2013 Elsevier Ltd. All rights reserved.

The influence of lysozyme on mannitol polymorphism in freeze-dried and spray-dried formulations depends on the selection of the drying process

DEFF Research Database (Denmark)

Grohganz, Holger; Lee, Yan-Ying; Rantanen, Jukka

2013-01-01

Freeze-drying and spray-drying are often applied drying techniques for biopharmaceutical formulations. The formation of different solid forms upon drying is often dependent on the complex interplay between excipient selection and process parameters. The purpose of this study was to investigate...... the influence of the chosen drying method on the solid state form. Mannitol-lysozyme solutions of 20mg/mL, with the amount of lysozyme varying between 2.5% and 50% (w/w) of total solid content, were freeze-dried and spray-dried, respectively. The resulting solid state of mannitol was analysed by near......-dried formulations an increase in protein concentration resulted in a shift from ß-mannitol to a-mannitol. An increase in final drying temperature of the freeze-drying process towards the temperature of the spray-drying process did not lead to significant changes. It can thus be concluded that it is the drying...

Tribological properties of nitrogen-containing amorphous carbon film produced by dc plasma chemical vapor deposition

International Nuclear Information System (INIS)

Zhang Wei; Wazumi, Koichiro; Tanaka, Akihiro; Koga, Yoshinori

2003-01-01

The nitrogen-contained amorphous carbon (a-C:N) films were deposited in a dc plasma chemical vapor deposition system with different substrate bias voltages. The structural, mechanical, and tribological properties of the a-C:N films were investigated. The influence of the bias voltage on the tribological behaviors of the a-C:N films was evaluated under various environments (dry air, O 2 , N 2 , and vacuum) using a ball-on-disk friction tester. It showed that the sp 3 C and hydrogen concentration of the a-C:N films decreases with increasing the bias voltage. However, the nitrogen concentration increases with increasing the bias voltage. As a result, the hardness and internal stress decrease and the critical load for fracturing increases as the substrate bias increases. For the tribological properties of the a-C:N films, the friction coefficient of the films slightly decreases in the environments of N 2 , O 2 , or dry air, but increases slightly in the vacuum environment by increasing the bias voltage. It indicates that the incorporated nitrogen in the a-C:N films would decrease the friction coefficient of the films in N 2 or O 2 environments, but slightly increases the friction coefficient of the films in a vacuum. The excellent wear resistance of the a-C:N films, in the level of 10 -9 -10 -8 mm 3 /Nm, can be observed in N 2 , vacuum, and dry air environments. In addition, the effect of the bias voltage on the wear rate of the a-C:N films becomes less obvious by nitrogen incorporation. So, we suggest the incorporated nitrogen, which bonded to carbon and restrained the increase of the fraction of sp 2 C-C, would restrain the wear of the a-C:N films in different environments, especially in dry air

Physical–chemical and biological behavior of an amorphous calcium phosphate thin film produced by RF-magnetron sputtering

International Nuclear Information System (INIS)

Santos, Euler A. dos; Moldovan, Simona; Mateescu, Mihaela; Faerber, Jacques; Acosta, Manuel; Pelletier, Hervé; Anselme, Karine; Werckmann, Jacques

2012-01-01

This work evaluates the thermal reactivity and the biological reactivity of an amorphous calcium phosphate thin film produced by radio frequency (RF) magnetron sputtering onto titanium substrates. The analyses showed that the sputtering conditions used in this work led to the deposition of an amorphous calcium phosphate. The thermal treatment of this amorphous coating in the presence of H 2 O and CO 2 promoted the formation of a carbonated HA crystalline coating with the entrance of CO 3 2− ions into the hydroxyl HA lattice. When immersed in culture medium, the amorphous and carbonated coatings exhibited a remarkable instability. The presence of proteins increased the dissolution process, which was confirmed by scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS) analyses. Moreover, the carbonated HA coating induced precipitation independently of the presence of proteins under dynamic conditions. Despite this surface instability, this reactive calcium phosphate significantly improved the cellular behavior. The cell proliferation was higher on the Ticp than on the calcium phosphate coatings, but the two coatings increased cellular spreading and stress fiber formation. In this sense, the presence of reactive calcium phosphate coatings can stimulate cellular behavior. - Highlights: ► Functionalization of Ti with reactive CaP thin film by RF-magnetron sputtering. ► De-hydroxylation facilitating the insertion of CO 3 2− into the HA lattice. ► High surface reactivity in the presence of culture medium. ► Cell behavior improved by the presence of reactive films.

Imaging transport phenomena during lysozyme protein crystal growth by the hanging drop technique

Science.gov (United States)

Sethia Gupta, Anamika; Gupta, Rajive; Panigrahi, P. K.; Muralidhar, K.

2013-06-01

The present study reports the transport process that occurs during the growth of lysozyme protein crystals by the hanging drop technique. A rainbow schlieren technique has been employed for imaging changes in salt concentration. A one dimensional color filter is used to record the deflection of the light beam. An optical microscope and an X-ray crystallography unit are used to characterize the size, tetragonal shape and Bravais lattice constants of the grown crystals. A parametric study on the effect of drop composition, drop size, reservoir height and number of drops on the crystal size and quality is reported. Changes in refractive index are not large enough to create a meaningful schlieren image in the air gap between the drop and the reservoir. However, condensation of fresh water over the reservoir solution creates large changes in the concentration of NaCl, giving rise to clear color patterns in the schlieren images. These have been analyzed to obtain salt concentration profiles near the free surface of the reservoir solution as a function of time. The diffusion of fresh water into the reservoir solution at the early stages of crystal growth followed by the mass flux of salt from the bulk solution towards the free surface has been recorded. The overall crystal growth process can be classified into two regimes, as demarcated by the changes in slope of salt concentration within the reservoir. The salt concentration in the reservoir equilibrates at long times when the crystallization process is complete. Thus, transport processes in the reservoir emerge as the route to monitor protein crystal growth in the hanging drop configuration. Results show that crystal growth rate is faster for a higher lysozyme concentration, smaller drops, and larger reservoir heights.

Correlation between calculated molecular descriptors of excipient amino acids and experimentally observed thermal stability of lysozyme

DEFF Research Database (Denmark)

Meng-Lund, Helena; Friis, Natascha; van de Weert, Marco

2017-01-01

for lysozyme in combination with 13 different amino acids using high throughput fluorescence spectroscopy and kinetic static light scattering measurements. On the theoretical side, around 200 2D and 3D molecular descriptors were calculated based on the amino acids' chemical structure. Multivariate data...... prominent stabilizing factor for both responses, whereas hydrophilic surface properties and high molecular mass density mostly had a positive influence on the unfolding temperature. A high partition coefficient (logP(o/w)) was identified as the most prominent destabilizing factor for both responses...

Method for producing nanowire-polymer composite electrodes

Energy Technology Data Exchange (ETDEWEB)

Pei, Qibing; Yu, Zhibin

2017-11-21

A method for producing flexible, nanoparticle-polymer composite electrodes is described. Conductive nanoparticles, preferably metal nanowires or nanotubes, are deposited on a smooth surface of a platform to produce a porous conductive layer. A second application of conductive nanoparticles or a mixture of nanoparticles can also be deposited to form a porous conductive layer. The conductive layer is then coated with at least one coating of monomers that is polymerized to form a conductive layer-polymer composite film. Optionally, a protective coating can be applied to the top of the composite film. In one embodiment, the monomer coating includes light transducing particles to reduce the total internal reflection of light through the composite film or pigments that absorb light at one wavelength and re-emit light at a longer wavelength. The resulting composite film has an active side that is smooth with surface height variations of 100 nm or less.

The combat film : the death and rebirth of an American film tradition

OpenAIRE

Greve, Henrik Havdal

2009-01-01

Abstract This thesis analyzes how four popular combat films produced by Hollywood portray and interprets important events in American history. The films exemplify how American cinema has varied in its portrayal of the American army in relation to both the perspectives of the film makers and widely held attitudes toward war and American foreign policy. In regard to social attitudes, militarism has been an instrument to create support for war. The term describes a process where a societ...

Stability and activity of lysozyme in stoichiometric and non-stoichiometric protic ionic liquid (PIL)-water systems

Science.gov (United States)

Wijaya, Emmy C.; Separovic, Frances; Drummond, Calum J.; Greaves, Tamar L.

2018-05-01

There has been a substantial increase in enzyme applications within the biochemical and pharmaceutical industries, for example, as industrial biocatalysts. However, enzymes have narrow marginal stability which makes them prone to become inactive and/or denature with a slight change in the solvent environment. Typically industrial applications require harsher solvent environments than enzyme native environments, and hence there is a need to understand solvent-protein interactions in order to develop strategies to maintain, or enhance, the enzymatic activity under industrially relevant solvent conditions. Previously we have shown that protic ionic liquids (PILs) with water can have a stabilising effect on lysozyme, with a large variation dependent on which PIL ions are present, and the water concentration [E. C. Wijaya et al., Phys. Chem. Chem. Phys. 18(37), 25926-25936 (2016)]. Here we extend on this work using non-stoichiometric aqueous PIL solvents to investigate, and isolate, the role of pH and ionicity on enzymes. We have used the PILs ethylammonium nitrate (EAN) and ethanolammonium formate (EOAF) since our previous work has identified these as good solvents for lysozyme. Solvent libraries were made from these two PILs with an additional precursor acid or base to modify the acidity/basicity of the neutral stoichiometric PIL, and with water added, to have solutions with 4-17 mol. % of the PIL ions in water. Molar ratios of base:acid were varied between 1:1.05 and 2:1 for EAN and 1:1.25 and 2:1 for EOAF, which enabled from highly basic to highly acidic solutions to be obtained. This was to modify the acidity/basicity of the neutral stoichiometric PILs, without the addition of buffers. The structure and stability of hen egg white lysozyme (HEWL) were explored under these solvent conditions using synchrotron small angle X-ray scattering (SAXS), Fourier transform infrared (FTIR), and activity assays. The radius of gyration and Kratky plots obtained from the SAXS data

Chemical modification of lysine residues in lysozyme may dramatically influence its amyloid fibrillation.

Science.gov (United States)

Morshedi, Dina; Ebrahim-Habibi, Azadeh; Moosavi-Movahedi, Ali Akbar; Nemat-Gorgani, Mohsen

2010-04-01

Studies on the aggregation of mutant proteins have provided new insights into the genetics of amyloid diseases and the role of the net charge of the protein on the rate, extent, and type of aggregate formation. In the present work, hen egg white lysozyme (HEWL) was employed as the model protein. Acetylation and (separately) citraconylation were employed to neutralize the charge on lysine residues. Acetylation of the lysine residues promoted amyloid formation, resulting in more pronounced fibrils and a dramatic decline in the nucleation time. In contrast, citraconylation produced the opposite effect. In both cases, native secondary and tertiary structures appeared to be retained. Studies on the effect of pH on aggregation suggested greater possibilities for amorphous aggregate formation rather than fibrillation at pH values closer to neutrality, in which the protein is known to take up a conformation more similar to its native form. This is in accord with reports in the literature suggesting that formation of amorphous aggregates is more favored under relatively more native conditions. pH 5 provided a critical environment in which a mixture of amorphous and fibrillar structures were observed. Use of Tango and Aggrescan software which describe aggregation tendencies of different parts of a protein structure suggested critical importance of some of the lysine residues in the aggregation process. Results are discussed in terms of the importance of the net charge in control of protein-protein interactions leading to aggregate formation and possible specific roles of lysine residues 96 and 97. Copyright 2009 Elsevier B.V. All rights reserved.

Uniform and Conformal Carbon Nanofilms Produced Based on Molecular Layer Deposition

Directory of Open Access Journals (Sweden)

Peng Yang

2013-12-01

Full Text Available Continuous and uniform carbon nanofilms (CNFs are prepared by pyrolysis of polyimide films which are produced by molecular layer deposition (MLD. The film thickness can be easily controlled at nanometer scale by altering the cycle numbers. During the annealing process at 600 °C, the polyimide film is subject to shrinkage of 70% in thickness. The obtained CNFs do not exhibit a well-graphitized structure due to the low calcination temperature. No clear pore structures are observed in the produced films. CNFs grown on a glass substrate with a thickness of about 1.4 nm shows almost 98% optical transmittance in the visible spectrum range. Au nanoparticles coated with CNFs are produced by this method. Carbon nanotubes with uniform wall thickness are obtained using anodic aluminum oxide as a template by depositing polyimide films into its pores. Our results demonstrate that this method is very effective to coat conformal and uniform CNFs on various substrates, such as nanoparticles and porous templates, to produce functional composite nanomaterials.

Film Handbook. Communication Manual. 2nd Edition.

Science.gov (United States)

Beuthner, Reginald, Comp.; And Others

Reference material is provided in this book for editors, producers, camera and sound technicians, others working in film, and film students. Each chapter is a self-contained unit about specific aspects of film-making, designed to accompany film courses being offered by the University of the West Indies, the Jamaica Broadcasting Corporation, and…

Molecular packing, hydrogen bonding, and fast dynamics in lysozyme/trehalose/glycerol and trehalose/glycerol glasses at low hydration

OpenAIRE

Lerbret, Adrien; Affouard, Frédéric

2017-01-01

Water and glycerol are well-known to facilitate the structural relaxation of amorphous protein matrices. However, several studies evidenced that they may also limit fast ($\\sim$ pico-nanosecond, ps-ns) and small-amplitude ($\\sim$ \\AA ) motions of proteins, which govern their stability in freeze-dried sugar mixtures. To determine how they interact with proteins and sugars in glassy matrices and, thereby, modulate their fast dynamics, we performed molecular dynamics (MD) simulations of lysozyme...

Quantitative film radiography

International Nuclear Information System (INIS)

Devine, G.; Dobie, D.; Fugina, J.; Hernandez, J.; Logan, C.; Mohr, P.; Moss, R.; Schumacher, B.; Updike, E.; Weirup, D.

1991-01-01

We have developed a system of quantitative radiography in order to produce quantitative images displaying homogeneity of parts. The materials that we characterize are synthetic composites and may contain important subtle density variations not discernible by examining a raw film x-radiograph. In order to quantitatively interpret film radiographs, it is necessary to digitize, interpret, and display the images. Our integrated system of quantitative radiography displays accurate, high-resolution pseudo-color images in units of density. We characterize approximately 10,000 parts per year in hundreds of different configurations and compositions with this system. This report discusses: the method; film processor monitoring and control; verifying film and processor performance; and correction of scatter effects

Ceramic Composite Thin Films

Science.gov (United States)

Ruoff, Rodney S. (Inventor); Stankovich, Sasha (Inventor); Dikin, Dmitriy A. (Inventor); Nguyen, SonBinh T. (Inventor)

2013-01-01

A ceramic composite thin film or layer includes individual graphene oxide and/or electrically conductive graphene sheets dispersed in a ceramic (e.g. silica) matrix. The thin film or layer can be electrically conductive film or layer depending the amount of graphene sheets present. The composite films or layers are transparent, chemically inert and compatible with both glass and hydrophilic SiOx/silicon substrates. The composite film or layer can be produced by making a suspension of graphene oxide sheet fragments, introducing a silica-precursor or silica to the suspension to form a sol, depositing the sol on a substrate as thin film or layer, at least partially reducing the graphene oxide sheets to conductive graphene sheets, and thermally consolidating the thin film or layer to form a silica matrix in which the graphene oxide and/or graphene sheets are dispersed.

Oral transmucosal delivery of domperidone from immediate release films produced via hot-melt extrusion technology.

Science.gov (United States)

Palem, Chinna Reddy; Kumar Battu, Sunil; Maddineni, Sindhuri; Gannu, Ramesh; Repka, Michael A; Yamsani, Madhusudan Rao

2013-02-01

The objective of the study was to prepare and characterize the domperidone (DOM) hot-melt extruded (HME) buccal films by both in vitro and in vivo techniques. The HME film formulations contained PEO N10 and/or its combination with HPMC E5 LV or Eudragit RL100 as polymeric carriers, and PEG3350 as a plasticizer. The blends were co-processed at a screw speed of 50 rpm with the barrel temperatures ranging from 120-160°C utilizing a bench top co-rotating twin-screw hot-melt extruder using a transverse-slit die. The HME films were evaluated for drug content, drug excipient interaction, in vitro drug release, mechanical properties, in vivo residence time, in vitro bioadhesion, swelling and erosion, ex vivo permeation from HME films and the selected optimal formulation was subjected for bioavailability studies in healthy human volunteers. The extruded films demonstrated no drug excipient interaction and excellent content uniformity. The selected HME film formulation (DOM2) exhibited a tensile strength (0.72 Kg/mm(2)), elongation at break (28.4% mm(2)), in vivo residence time (120 min), peak detachment force (1.55 N), work of adhesion (1.49 mJ), swelling index (210.2%), erosion (10.5%) and in vitro drug release of 84.8% in 2 h. Bioavailability from the optimized HME buccal films was 1.5 times higher than the oral dosage form and the results showed statistically significant (p buccal-adhesive films with improved bioavailability characteristics.

Polyurethane coating with thin polymer films produced by plasma polymerization of diglyme

International Nuclear Information System (INIS)

Ribeiro, M A; Ramos, A S; Manfredini, M I; Alves, H A; Ramos, E C T; Honda, R Y; Kostov, K G; Lucena, E F; Mota, R P; Algatti, M A; Kayama, M E

2009-01-01

Aqueous-based polyurethane dispersions have been widely utilized as lubricants in textile, shoes, automotive, biomaterial and many other industries because they are less aggressive to surrounding environment. In this work thin films with different thickness were deposited on biocompatible polyurethane by plasma polymerization process using diethylene glycol dimethyl ether (Diglyme) as monomer. Molecular structure of the films was analyzed by Fourier Transform Infrared spectroscopy. The spectra exhibited absorption bands of O-H (3500-3200cm -1 ), C-H (3000-2900cm -1 ), C=O (1730-1650cm -1 ), C-O and C-O-C bonds at 1200-1600cm -1 . The samples wettability was evaluated by measurements of contact angle using different liquids such as water, glycerol, poly-ethane and CMC. The polyurethane surface showed hydrophilic behavior after diglyme plasma-deposition with contact angle dropping from 85 deg. to 22 deg. Scanning Electron Microscopy revealed that diglyme films covered uniformly the polyurethane surfaces ensuring to it a biocompatible characteristic.

Polyurethane coating with thin polymer films produced by plasma polymerization of diglyme

Science.gov (United States)

Ribeiro, M. A.; Ramos, A. S.; Manfredini, M. I.; Alves, H. A.; Y Honda, R.; Kostov, K. G.; Lucena, E. F.; Ramos, E. C. T.; Mota, R. P.; Algatti, M. A.; Kayama, M. E.

2009-05-01

Aqueous-based polyurethane dispersions have been widely utilized as lubricants in textile, shoes, automotive, biomaterial and many other industries because they are less aggressive to surrounding environment. In this work thin films with different thickness were deposited on biocompatible polyurethane by plasma polymerization process using diethylene glycol dimethyl ether (Diglyme) as monomer. Molecular structure of the films was analyzed by Fourier Transform Infrared spectroscopy. The spectra exhibited absorption bands of O-H (3500-3200cm-1), C-H (3000-2900cm-1), C=O (1730-1650cm-1), C-O and C-O-C bonds at 1200-1600cm-1. The samples wettability was evaluated by measurements of contact angle using different liquids such as water, glycerol, poly-ethane and CMC. The polyurethane surface showed hydrophilic behavior after diglyme plasma-deposition with contact angle dropping from 85° to 22°. Scanning Electron Microscopy revealed that diglyme films covered uniformly the polyurethane surfaces ensuring to it a biocompatible characteristic.

Polyurethane coating with thin polymer films produced by plasma polymerization of diglyme

Energy Technology Data Exchange (ETDEWEB)

Ribeiro, M A; Ramos, A S; Manfredini, M I; Alves, H A; Ramos, E C T [UNIVAP, Sao Jose dos Campos, SP (Brazil); Honda, R Y; Kostov, K G; Lucena, E F; Mota, R P; Algatti, M A; Kayama, M E, E-mail: rmota@feg.unesp.b [FEG-DFQ-UNESP, Av. Ariberto Pereira da Cunha 333, 12516-410 - Guaratingueta, SP (Brazil)

2009-05-01

Aqueous-based polyurethane dispersions have been widely utilized as lubricants in textile, shoes, automotive, biomaterial and many other industries because they are less aggressive to surrounding environment. In this work thin films with different thickness were deposited on biocompatible polyurethane by plasma polymerization process using diethylene glycol dimethyl ether (Diglyme) as monomer. Molecular structure of the films was analyzed by Fourier Transform Infrared spectroscopy. The spectra exhibited absorption bands of O-H (3500-3200cm{sup -1}), C-H (3000-2900cm{sup -1}), C=O (1730-1650cm{sup -1}), C-O and C-O-C bonds at 1200-1600cm{sup -1}. The samples wettability was evaluated by measurements of contact angle using different liquids such as water, glycerol, poly-ethane and CMC. The polyurethane surface showed hydrophilic behavior after diglyme plasma-deposition with contact angle dropping from 85 deg. to 22 deg. Scanning Electron Microscopy revealed that diglyme films covered uniformly the polyurethane surfaces ensuring to it a biocompatible characteristic.

Direct-current substrate bias effects on amorphous silicon sputter-deposited films for thin film transistor fabrication

International Nuclear Information System (INIS)

Jun, Seung-Ik; Rack, Philip D.; McKnight, Timothy E.; Melechko, Anatoli V.; Simpson, Michael L.

2005-01-01

The effect that direct current (dc) substrate bias has on radio frequency-sputter-deposited amorphous silicon (a-Si) films has been investigated. The substrate bias produces a denser a-Si film with fewer defects compared to unbiased films. The reduced number of defects results in a higher resistivity because defect-mediated conduction paths are reduced. Thin film transistors (TFTs) that were completely sputter deposited were fabricated and characterized. The TFT with the biased a-Si film showed lower leakage (off-state) current, higher on/off current ratio, and higher transconductance (field effect mobility) than the TFT with the unbiased a-Si film

Lactoferrin, myeloperoxidase, lysozyme and eosinophil cationic protein in exudate in delayed type hypersensitivity

DEFF Research Database (Denmark)

Lerche, A; Bisgaard, H; Christensen, J D

1988-01-01

allergic patients with nickel challenge in the chamber medium showed a time-dependent increase of mononuclear cells, eosinophils and basophils and a concomitant decrease of polymorphonuclear granulocytes, characteristic of a combined specific and unspecific inflammation. The morphology of the exudate...... in contact allergic patients exposed to nickel showed a dominance of polymorphonuclear granulocytes throughout the study period, while mononuclear cells, eosinophils and basophils were detected at a much lower quantity and with a considerable delay. Further, we studied the kinetics of the leucocyte granule...... proteins: lactoferrin, myeloperoxidase, lysozyme and eosinophil cationic protein in exudate fluid in a parallel test. A significant higher flux was found for all during the second day of allergen exposure compared to contact allergic patients without allergen challenge as well as normal volunteers...

A Study on Environmental Education Films.

Science.gov (United States)

Gerba, Diana

The degree of communication between the film industry and educators and its effect on the future directions of environmental education films are the focus of this report. Separate surveys were mailed to 100 film industry producers and distributors and 150 elementary and secondary educators in Maine, Kansas, Pennsylvania, California, and Alabama.…

[Alterations in tears aqueous layer during cytostatics treatment].

Science.gov (United States)

Wojciechowska, Katarzyna; Wieckowska-Szakiel, Marzena; Rózalska, Barbara; Jurowski, Piotr

2013-01-01

The aim of the study was to evaluate tears secretion, pH and lysozyme activity in tears aqueous layer during chemotherapy in lung, breast and bowel cancer. 36 patients were enrolled to the study. Depending on the type of cancer and type of chemotherapy patients were divided into three groups. Group I (12 patients) diagnosed with non-small-cell lung cancer treated with PE schema (cisplatin, etoposide), Group II (12 patients) with breast cancer treated with FAC schema (fluorouracil, doxorubicin, cyclophosphamide), Group III (12 patients) with bowel cancer treated with FU/LV schema (fluorouracil, leucovorin). In all the patients: Schirmer's I test, pH measurements and lysozyme test were performed. Patients were examined before chemotherapy, after 2nd, 4th, 6th cycle. In group I and II lowering of tears secretion (p tears secretion (p tears film in all groups (p tears aqueous layer causing alterations of tears secretions. PH alterations depending on type of chemotherapy was observed. Lowering of lysozyme activity in tears was observed. All the deteriorations aggravate with duration of chemotherapy. Alterations of tears film parameters during chemotherapy may influence upon eye surface homeostasis and infectious complication. tears aqueous layer, Schirmer's test, lysozyme activity, tears pH.

X-ray exposures given with various combinations of film and screen

International Nuclear Information System (INIS)

Schlenker, R.A.; Kirsh, I.E.

A comparison of entrance exposure doses of patients for various film and screen combinations and xeroradiographs showed that moderate speed screen and film gave exposures 4 to 7 times greater than fast film and screen and that xeroradiographs produced exposures 20 to 40 times greater than fast film and screen. Xeroradiographs produced table top incident exposures which exceeded Illinois Department of Health limits, while conventional film-screen combinations did not. (U.S.)

Zirconia thin films from aqueous precursors: Processing, microstructural development, and epitaxial growth

International Nuclear Information System (INIS)

Miller, K.T.

1991-01-01

Thin films of ZrO 2 (Y 2 O 3 ) were prepared from aqueous salt precursors by spin coating. Films were pyrolyzed to produce porous polycrystalline thin films of 5-10 nm grain size. Subsequent microstructural development depends greatly upon the nature of the substrate. Upon randomly oriented sapphire, the films initially sintered to full density; further heat treatment and grain growth causes these films to break into interconnected islands and finally isolated particles. Thermodynamic calculations predict that breakup is energetically favorable when the grain-size film-thickness ratio exceeds a critical value. Upon basal-plane-oriented sapphire, grain growth and breakup prefer the (100) oriented grains, presumably because this orientation is a special interface of low energy. The isolated, oriented grains produced by film breakup act as seeds for the growth of newly deposited material. Upon (100) cubic zirconia, true epitaxial films develop. Epitaxial growth was observed for lattice mismatches up to 1.59%. Growth proceeds from a fine epitaxial layer which is produced during the initial stages of heat treatment, consuming the porous polycrystalline material and producing a dense epitaxial thin film whose misfit is accommodated by a combination of film strain and misfit dislocations

Supercritical fluid molecular spray thin films and fine powders

Science.gov (United States)

Smith, Richard D.

1988-01-01

Solid films are deposited, or fine powders formed, by dissolving a solid material into a supercritical fluid solution at an elevated pressure and then rapidly expanding the solution through a short orifice into a region of relatively low pressure. This produces a molecular spray which is directed against a substrate to deposit a solid thin film thereon, or discharged into a collection chamber to collect a fine powder. The solvent is vaporized and pumped away. Solution pressure is varied to determine, together with flow rate, the rate of deposition and to control in part whether a film or powder is produced and the granularity of each. Solution temperature is varied in relation to formation of a two-phase system during expansion to control porosity of the film or powder. A wide variety of film textures and powder shapes are produced of both organic and inorganic compounds. Films are produced with regular textural feature dimensions of 1.0-2.0 .mu.m down to a range of 0.01 to 0.1 .mu.m. Powders are formed in very narrow size distributions, with average sizes in the range of 0.02 to 5 .mu.m.

Elemental redistribution in coloured films on SUS304 stainless steel produced by current pulse method

International Nuclear Information System (INIS)

Lin, C.J.; Duh, J.G.

1996-01-01

Coloured films are deposited on an SUS304 substrate by the square wave current pulse method in a 2.5 M CrO 3 + 5 M H 2 SO 4 solution at 75 C. The absolute ratio of anodic to cathodic charge density and the corresponding final value of cathodic potential provide an easy and appropriate approach to the control of colour in the current pulse colouring method. This is proved in this study for a frequency region lower than 1 Hz. The corresponding cathodic potential increases with increasing dwell time in the negative applied current region and is related to the colour tone of the deposited film. The total time required to obtain the same thickness of coloured film is shorter at lower frequency than at higher frequency, which takes more time to charge the double layer. The thickness of the coloured films is determined by both Auger electron spectroscopy (AES) and secondary ion mass spectrometry (SIMS) depth profiling and is of submicron order. The film thickness is proportional to the introduced total charge per unit area in the current pulse colouring process. CrO + and FeO + ions are observed in the coloured films and their concentrations increase as the colouring time is increased. The Fe concentration in the coloured films is lower than that in the substrate and decreases with the colouring time. It is the iron species that first becomes involved in the anodic reaction and the spinel oxide structure of iron and chromium is present in the coloured films. (orig.)

Impurities in chromium deposits produced by electroplating and physical vapor deposition

Energy Technology Data Exchange (ETDEWEB)

Dini, J.W.

1994-05-01

Impurity contents in electrodeposited (hexavalent and trivalent) chromium deposits and physically vapor deposited (thermal evaporation, electron beam evaporation and rf-sputtering) were compared. Oxygen is the key impurity obtained in electrodeposited films but it can be minimized in hexavalent plating solutions by operating at high temperature, e. g., 85 C. Electrodeposits produced in trivalent chromium plating solutions and physically vapor deposited films have much higher oxygen contents than electrodeposits produced in hexavalent chromium solutions operated at temperatures around 85 C. Depending on the target material used for physically vapor deposited films, these films can also have high amounts of other impurities.

PVA/Polysaccharides Blended Films: Mechanical Properties

Directory of Open Access Journals (Sweden)

Fábio E. F. Silva

2013-01-01

Full Text Available Blends of polyvinyl alcohol (PVA and angico gum (AG and/or cashew gum (CG were used to produce films by casting method. Morphological and mechanical properties of these films were studied and compared to the properties of a commercial collagen membrane of bovine origin (MBO. The films presented thickness varying from 70 to 140 μm (PVA/AG and 140 to 200 μm (PVA/CG. Macroscopic analysis showed that a PVA/CG film was very similar to MBO regarding the color and transparency. The higher values of tensile strength (TS and elastic modulus (EM were observed in the film. On the other hand, PVA/CG and PVA/CG-AG presented the highest value of percentage of elongation (E%. Pearson’s Correlation Analysis revealed a positive correlation between TS and EM and a negative correlation between E% and EM. The PVA/CG film presented mechanical properties very similar to MBO, with the advantage of a higher E% (11.96 than MBO (2.94. The properties of the PVA blended films depended on the polysaccharide added in the blend, as well as the acid used as a catalyst. However, all produced films presented interesting mechanical characteristics which enables several biotechnological applications.

The effect of protein-coated contact lenses on the adhesion and viability of gram negative bacteria.

Science.gov (United States)

Williams, Timothy J; Schneider, Rene P; Willcox, Mark D P

2003-10-01

Gram negative bacterial adhesion to contact lenses can cause adverse responses. During contact lens wear, components of the tear film adsorb to the contact lens. This study aimed to investigate the effect of this conditioning film on the viability of bacteria. Bacteria adhered to contact lenses which were either unworn, worn for daily-, extended- or overnight-wear or coated with lactoferrin or lysozyme. Numbers of viable and total cells were estimated. The number of viable attached cells was found to be significantly lower than the total number of cells on worn (50% for strain Paer1 on daily-wear lenses) or lactoferrin-coated lenses (56% for strain Paer1). Lysozyme-coated lenses no statistically significant effect on adhesion. The conditioning film gained through wear may not inhibit bacterial adhesion, but may act adversely upon those bacteria that succeed in attaching.

[Enthalpy stabilization of chicken egg lysozyme in aqueous dimethylsulfoxide solutions].

Science.gov (United States)

Kovrigin, E L; Kirkitadze, M D; Potekhin, S A

1996-01-01

Scanning microcalorimetry data have been used to plot the dependences of the denaturation enthalpy of hen egg lysozyme on dimethylsulfoxide concentration at fixed temperatures. It has been shown that at dimethylsulfoxide concentrations below 40% (v/v) the enthalpy does not depend on pH of the medium. An increase of dimethylsulfoxide concentrations in this range leads to a linear growth of enthalpy. The rate of enthalpy growth decreases with the temperature increase. The denaturation enthalpy begins to considerably depend on pH at dimethylsulfoxide concentrations more than 40%. Spectroscopy data indicate that conformational changes occur in the protein in this range of concentrations already at room temperature, whereas according to scanning microcalorimetry, they take place at much higher temperatures. This difference is probably due to a decrease of the real temperature of protein melting below room temperature and a very inhibited character of the denaturational transition. This results in a decrease of calorimetric enthalpy at acidic pH owing to incomplete protein renaturation upon calorimeter cooling to the starting point.

Improved Dielectric Films For Capacitors

Science.gov (United States)

Yen, Shiao-Ping S.; Lewis, Carol R.; Cygan, Peter J.; Jow, T. Richard

1994-01-01

Dielectric films made from blends of some commercially available high-dielectric-constant cyanoresins with each other and with cellulose triacetate (CTA) have both high dielectric constants and high breakdown strengths. Dielectric constants as high as 16.2. Films used to produce high-energy-density capacitors.

The Effect of Thermal Annealing on the Optical Properties of a-SiC:H Films Produced by DC Sputtering Methods: I

Directory of Open Access Journals (Sweden)

Lusitra Munisa

2003-04-01

Full Text Available Silicon Target Case. The effects of thermal annealing treatment on the optical properties of amorphous silicon carbon films deposited by silicon target in an argon, methane and hydrogen gas mixture have been studied using ultra violet-visible (uv-vis spectroscopy. Both n and α, and consequently the real and imaginary parts of the dielectric constant, show a considerable variation with subsequent annealing up to annealing temperature 500 °C, with the most rapid changes occurring for temperature 300 °C. The films tend denser as the annealing temperature increased up to 500°C. The optical gap improved slightly upon annealing, where as the disorder of the amorphous network reduced. The annealing treatment produces reorganization of the amorphous network since thermal annealing results in dissociation of hydrogenated bonds (Si-H and C-H.

Scalable organic solvent free supercritical fluid spray drying process for producing dry protein formulations.

Science.gov (United States)

Nuchuchua, O; Every, H A; Hofland, G W; Jiskoot, W

2014-11-01

In this study, we evaluated the influence of supercritical carbon dioxide (scCO2) spray drying conditions, in the absence of organic solvent, on the ability to produce dry protein/trehalose formulations at 1:10 and 1:4 (w/w) ratios. When using a 4L drying vessel, we found that decreasing the solution flow rate and solution volume, or increasing the scCO2 flow rate resulted in a significant reduction in the residual water content in dried products (Karl Fischer titration). The best conditions were then used to evaluate the ability to scale the scCO2 spray drying process from 4L to 10L chamber. The ratio of scCO2 and solution flow rate was kept constant. The products on both scales exhibited similar residual moisture contents, particle morphologies (SEM), and glass transition temperatures (DSC). After reconstitution, the lysozyme activity (enzymatic assay) and structure (circular dichroism, HP-SEC) were fully preserved, but the sub-visible particle content was slightly increased (flow imaging microscopy, nanoparticle tracking analysis). Furthermore, the drying condition was applicable to other proteins resulting in products of similar quality as the lysozyme formulations. In conclusion, we established scCO2 spray drying processing conditions for protein formulations without an organic solvent that holds promise for the industrial production of dry protein formulations. Copyright © 2014 Elsevier B.V. All rights reserved.

Effective interactions in lysozyme aqueous solutions: a small-angle neutron scattering and computer simulation study.

Science.gov (United States)

Abramo, M C; Caccamo, C; Costa, D; Pellicane, G; Ruberto, R; Wanderlingh, U

2012-01-21

We report protein-protein structure factors of aqueous lysozyme solutions at different pH and ionic strengths, as determined by small-angle neutron scattering experiments. The observed upturn of the structure factor at small wavevectors, as the pH increases, marks a crossover between two different regimes, one dominated by repulsive forces, and another one where attractive interactions become prominent, with the ensuing development of enhanced density fluctuations. In order to rationalize such experimental outcome from a microscopic viewpoint, we have carried out extensive simulations of different coarse-grained models. We have first studied a model in which macromolecules are described as soft spheres interacting through an attractive r(-6) potential, plus embedded pH-dependent discrete charges; we show that the uprise undergone by the structure factor is qualitatively predicted. We have then studied a Derjaguin-Landau-Verwey-Overbeek (DLVO) model, in which only central interactions are advocated; we demonstrate that this model leads to a protein-rich/protein-poor coexistence curve that agrees quite well with the experimental counterpart; experimental correlations are instead reproduced only at low pH and ionic strengths. We have finally investigated a third, "mixed" model in which the central attractive term of the DLVO potential is imported within the distributed-charge approach; it turns out that the different balance of interactions, with a much shorter-range attractive contribution, leads in this latter case to an improved agreement with the experimental crossover. We discuss the relationship between experimental correlations, phase coexistence, and features of effective interactions, as well as possible paths toward a quantitative prediction of structural properties of real lysozyme solutions. © 2012 American Institute of Physics

Irradiation gamma on chitosan films

Energy Technology Data Exchange (ETDEWEB)

Mello, Luana Miranda Lopes de; Souza, Adriana Regia Marques de; Arthur, Valter, E-mail: lumilopes@hotmail.com, E-mail: drilavras@yahoo.com.br, E-mail: arthur@cena.usp.br [Universidade Federal do Tocantins (UFT), Palmas,TO (Brazil). Departmento de Ciencia e Tecnologia de Alimentos; Universidade Federal de Goias (UFGO), Goiania (Brazil). Departmento de Ciencia e Tecnologia de Alimentos; Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil)

2017-11-01

Films are preformed structures, independent, that are used to wrap food after processing, increasing their shelf life and enhancing its bright and attractive appearance. They are prepared from biological materials as an alternative to the plastic synthetic containers to improve the quality of the environment. Chitosan is a biodegradable polymer composed of β-(1-4) linked D-glucosamine (deacetylated unit) and N-acetyl-D- glucosamine (acetylated unit). It is produced commercially by deacetylation of chitin, which is a structural component of the exoskeleton of crustaceans. She is able to form films and edible and/or biodegradable coatings. With the objective to evaluate the effect of different doses of gamma radiation (0, 5, 10 and 15 kGy) and chitosan concentrations (1 and 2%) in film properties, it was evaluated its optical, mechanical and morphological properties. The films were produced by casting. Irradiation did not affect the thickness of the films, but influenced its colors, increasing the tone of the film for a stronger yellowish color. This fact can be attributed to the increased concentration of C = O bonds of chitosan due to the breakdown of the chain reaction and the Maillard reaction. Irradiated films showed smoother surface and less rough, due to the degradation of the chitosan molecule and poor mechanical properties, not showing good flexibility and stretching. (author)

Irradiation gamma on chitosan films

International Nuclear Information System (INIS)

Mello, Luana Miranda Lopes de; Souza, Adriana Regia Marques de; Arthur, Valter

2017-01-01

Films are preformed structures, independent, that are used to wrap food after processing, increasing their shelf life and enhancing its bright and attractive appearance. They are prepared from biological materials as an alternative to the plastic synthetic containers to improve the quality of the environment. Chitosan is a biodegradable polymer composed of β-(1-4) linked D-glucosamine (deacetylated unit) and N-acetyl-D- glucosamine (acetylated unit). It is produced commercially by deacetylation of chitin, which is a structural component of the exoskeleton of crustaceans. She is able to form films and edible and/or biodegradable coatings. With the objective to evaluate the effect of different doses of gamma radiation (0, 5, 10 and 15 kGy) and chitosan concentrations (1 and 2%) in film properties, it was evaluated its optical, mechanical and morphological properties. The films were produced by casting. Irradiation did not affect the thickness of the films, but influenced its colors, increasing the tone of the film for a stronger yellowish color. This fact can be attributed to the increased concentration of C = O bonds of chitosan due to the breakdown of the chain reaction and the Maillard reaction. Irradiated films showed smoother surface and less rough, due to the degradation of the chitosan molecule and poor mechanical properties, not showing good flexibility and stretching. (author)

Microstructure and mechanical properties of ion-beam-produced Fe-Ti-(N), Fe-Ti-(C), and Fe-Ti-(C,N) surface films

Science.gov (United States)

Hirvonen, J.-P.; Nastasi, M.; Zocco, T. G.; Jervis, T. R.

1990-06-01

Ion-mixed films of Fe53 Ti47 were produced by ion irradiating a Fe-Ti multilayer structure on AISI 304 stainless steel. The ion-mixed films were subsequently implanted with nitrogen, carbon, or both carbon and nitrogen. The microstructure following nitrogen implantation consisted of a bcc solid solution of iron and titanium and finely dispersed TiN precipitates. In the cases of carbon or carbon and nitrogen implantation, a two-phase structure consisting of an amorphous matrix with TiC or Ti(C,N) precipitates was found. All these films initially possessed improved tribological properties as revealed by lowered friction and increased wear resistance. However, after an extended test of 1000 wear cycles, a reduced friction was only observed for the carbon or carbon and nitrogen implanted samples. The wear track on the dual implanted surface was extremely smooth, while the surface of the nitrogen-implanted sample was partly worn through, causing the friction to increase to the level of the untreated sample. The improved tribological properties of the implanted films are attributed to an increase in surface hardness. However, the surface hardness is unable to explain differences between different implantations. In the case of the dual carbon and nitrogen implantation, improvements appear to be in part the result from an increased capability to accommodate plastic deformation. These conclusions are supported by transmission electron microscope studies of the wear tracks as well as by nanoindentation measurements.

Fluorinated ionic liquids for protein drug delivery systems: Investigating their impact on the structure and function of lysozyme.

Science.gov (United States)

Alves, Márcia; Vieira, Nicole S M; Rebelo, Luís Paulo N; Araújo, João M M; Pereiro, Ana B; Archer, Margarida

2017-06-30

Since the approval of recombinant human insulin by FDA in 1982, more than 200 proteins are currently available for pharmaceutical use to treat a wide range of diseases. However, innovation is still required to develop effective approaches for drug delivery. Our aim is to investigate the potential use of fluorinated ionic liquids (FILs) as drug delivery systems (DDS) for therapeutic proteins. Some initial parameters need to be assessed before further studies can proceed. This work evaluates the impact of FILs on the stability, function, structure and aggregation state of lysozyme. Different techniques were used for this purpose, which included differential scanning fluorimetry (DSF), spectrophotometric assays, circular dichroism (CD), dynamic light scattering (DLS), and scanning and transmission electron microscopy (SEM/TEM). Ionic liquids composed of cholinium-, imidazolium- or pyridinium- derivatives were combined with different anions and analysed at different concentrations in aqueous solutions (below and above the critical aggregation concentration, CAC). The results herein presented show that the addition of ionic liquids had no significant effect on the stability and hydrolytic activity of lysozyme. Moreover, a distinct behaviour was observed in DLS experiments for non-surfactant and surfactant ionic liquids, with the latter encapsulating the protein at concentrations above the CAC. These results encourage us to further study ionic liquids as promising tools for DDS of protein drugs. Copyright © 2017 Elsevier B.V. All rights reserved.

Associations between microsatellite polymorphism within the macrophage expressed lysozyme (mLys gene and milk Indices Properties in Polish Black-and-White Cows

Directory of Open Access Journals (Sweden)

Małgorzata Walczak-Wójciak

2005-01-01

Full Text Available The experiment was performed in the years 1998–2000 on two farms located near to Vistula Lowlands. The experimental materials were comprised of 52 daughters of bull Paran, among which 21 possessed the mLys-mic 7 allele and 31 – the mLys-mic 3 allele. The serum and whey bacteriolytic activity of the lysozyme, concentrations of selected mineral elements, technological properties of milk at successive the first lactation stages (30, 100, 150 and 200 days, somatic cell count and bacterial count were investigated in this study. The results obtained indicate that Lys-mic polymorphism has low suitability as a marker for milk production capacity in cows. The lysozyme gene, treated as a mastitis resistance factor, showed no effects on somatic cell count and bacterial count. Further investigations, conducted not only during the first lactation, but also during the next lactation in a herd threatened by a variety of pathogenic factors, would be carried to validate this Lys-mic gene polymorphism on mastitis resistance.

Purification and Characterization of a Novel Anti-Campylobacter Bacteriocin Produced by Lactobacillus curvatus DN317.

Science.gov (United States)

Zommiti, Mohamed; Almohammed, Hamdan; Ferchichi, Mounir

2016-12-01

The lactic acid bacteria (LAB) microbiota of Saudi chicken ceca was determined. From 60 samples, 204 isolates of lactic acid bacteria were obtained. Three isolates produced antimicrobial activities against Campylobacter jejuni, Listeria monocytogenes, and Bacillus subtilis. The isolate DN317, which had the highest activity against Campylobacter jejuni ATCC 33560, was identified as Lactobacillus curvatus (GenBank accession numbers: KX353849 and KX353850). Full inhibitory activity was observed after a 2-h incubation with the supernatant at pH values between 4 and 8. Only 16% of the activity was conserved after a treatment at 121 °C for 15 min. The use of proteinase K, pepsin, chymotrypsin, trypsin, papain, and lysozyme drastically reduced the antimicrobial activity. However, lipase, catalase, and lysozyme had no effect on this activity. The active peptide produced by Lactobacillus curvatus DN317 was purified by precipitation with an 80% saturated ammonium sulfate solution, and two steps of reversed phase HPLC on a C18 column. The molecular weight of this peptide was 4448 Da as determined by MALDI-ToF. N-terminal sequence analysis using Edman degradation revealed 47 amino acid residues (UniProt Knowledgebase accession number C0HK82) revealing homology with the amino acid sequences of sakacin P and curvaticin L442. The antimicrobial activity of the bacteriocin, namely curvaticin DN317, was found to be bacteriostatic against Campylobacter jejuni ATCC 33560. The use of microbial antagonism by LAB is one of the best ways to control microorganisms safely in foods. This result constitutes a reasonable advance in the antimicrobial field because of its potential applications in food technology.

Special equipment for etching nitrocellulose film

International Nuclear Information System (INIS)

Domanus, J.C.

1983-08-01

Nitrocellulose film and converter screens used for neutron radiography are described. Difficulties in visualization of radiographs on those films are mentioned. Because there is no equipment for etching nitrocellulose film available on the market Risoe has designed and produced such equipment at an estimated cost of Dkr. 15,000. Design criteria for this equipment are given and its performance described

The active site of hen egg-white lysozyme: flexibility and chemical bonding

Energy Technology Data Exchange (ETDEWEB)

Held, Jeanette, E-mail: jeanette.netzel@uni-bayreuth.de; Smaalen, Sander van [University of Bayreuth, D-95440 Bayreuth (Germany)

2014-04-01

Chemical bonding at the active site of lysozyme is analyzed on the basis of a multipole model employing transferable multipole parameters from a database. Large B factors at low temperatures reflect frozen-in disorder, but therefore prevent a meaningful free refinement of multipole parameters. Chemical bonding at the active site of hen egg-white lysozyme (HEWL) is analyzed on the basis of Bader’s quantum theory of atoms in molecules [QTAIM; Bader (1994 ▶), Atoms in Molecules: A Quantum Theory. Oxford University Press] applied to electron-density maps derived from a multipole model. The observation is made that the atomic displacement parameters (ADPs) of HEWL at a temperature of 100 K are larger than ADPs in crystals of small biological molecules at 298 K. This feature shows that the ADPs in the cold crystals of HEWL reflect frozen-in disorder rather than thermal vibrations of the atoms. Directly generalizing the results of multipole studies on small-molecule crystals, the important consequence for electron-density analysis of protein crystals is that multipole parameters cannot be independently varied in a meaningful way in structure refinements. Instead, a multipole model for HEWL has been developed by refinement of atomic coordinates and ADPs against the X-ray diffraction data of Wang and coworkers [Wang et al. (2007), Acta Cryst. D63, 1254–1268], while multipole parameters were fixed to the values for transferable multipole parameters from the ELMAM2 database [Domagala et al. (2012), Acta Cryst. A68, 337–351] . Static and dynamic electron densities based on this multipole model are presented. Analysis of their topological properties according to the QTAIM shows that the covalent bonds possess similar properties to the covalent bonds of small molecules. Hydrogen bonds of intermediate strength are identified for the Glu35 and Asp52 residues, which are considered to be essential parts of the active site of HEWL. Furthermore, a series of weak C�

The active site of hen egg-white lysozyme: flexibility and chemical bonding

International Nuclear Information System (INIS)

Held, Jeanette; Smaalen, Sander van

2014-01-01

Chemical bonding at the active site of lysozyme is analyzed on the basis of a multipole model employing transferable multipole parameters from a database. Large B factors at low temperatures reflect frozen-in disorder, but therefore prevent a meaningful free refinement of multipole parameters. Chemical bonding at the active site of hen egg-white lysozyme (HEWL) is analyzed on the basis of Bader’s quantum theory of atoms in molecules [QTAIM; Bader (1994 ▶), Atoms in Molecules: A Quantum Theory. Oxford University Press] applied to electron-density maps derived from a multipole model. The observation is made that the atomic displacement parameters (ADPs) of HEWL at a temperature of 100 K are larger than ADPs in crystals of small biological molecules at 298 K. This feature shows that the ADPs in the cold crystals of HEWL reflect frozen-in disorder rather than thermal vibrations of the atoms. Directly generalizing the results of multipole studies on small-molecule crystals, the important consequence for electron-density analysis of protein crystals is that multipole parameters cannot be independently varied in a meaningful way in structure refinements. Instead, a multipole model for HEWL has been developed by refinement of atomic coordinates and ADPs against the X-ray diffraction data of Wang and coworkers [Wang et al. (2007), Acta Cryst. D63, 1254–1268], while multipole parameters were fixed to the values for transferable multipole parameters from the ELMAM2 database [Domagala et al. (2012), Acta Cryst. A68, 337–351] . Static and dynamic electron densities based on this multipole model are presented. Analysis of their topological properties according to the QTAIM shows that the covalent bonds possess similar properties to the covalent bonds of small molecules. Hydrogen bonds of intermediate strength are identified for the Glu35 and Asp52 residues, which are considered to be essential parts of the active site of HEWL. Furthermore, a series of weak C�

Shift in aggregation, ROS generation, antioxidative defense, lysozyme and acetylcholinesterase activities in the cells of an Indian freshwater sponge exposed to washing soda (sodium carbonate).

Science.gov (United States)

Mukherjee, Soumalya; Ray, Mitali; Ray, Sajal

2016-09-01

Washing soda, chemically identified as anhydrous sodium carbonate, is a popular cleaning agent among the rural and urban populations of India which often contaminates the freshwater ponds and lakes, the natural habitat of sponge Eunapius carteri. Present investigation deals with estimation of cellular aggregation, generation of ROS and activities of antioxidant enzymes, lysozyme and acetylcholinesterase in the cells of E. carteri under the environmentally realistic concentrations of washing soda. Prolonged treatment of washing soda inhibited the degree of cellular aggregation. Experimental exposure of 8 and 16mg/l of sodium carbonate for 48h elevated the physiological level of reactive oxygen species (ROS) generation in the agranulocytes, semigranulocytes and granulocytes of E. carteri, whereas, treatment of 192h inhibited the ROS generation in three cellular morphotypes. Activities of superoxide dismutase, catalase and glutathione-S-transferase were recorded to be inhibited under prolonged exposure of washing soda. Washing soda mediated inhibition of ROS generation and depletion in the activities of antioxidant enzymes were indicative to an undesirable shift in cytotoxic status and antioxidative defense in E. carteri. Inhibition in the activity of lysozyme under the treatment of sodium carbonate was suggestive to a severe impairment of the innate immunological efficiency of E. carteri distributed in the washing soda contaminated habitat. Washing soda mediated inhibition in the activity of acetylcholinesterase indicated its neurotoxicity in E. carteri. Washing soda, a reported environmental contaminant, affected adversely the immunophysiological status of E. carteri with reference to cellular aggregation, oxidative stress, antioxidative defense, lysozyme and acetylcholinesterase activity. Copyright © 2016 Elsevier Inc. All rights reserved.

Effect of enzymatic degradation of chitosan in polyhydroxybutyrate/chitosan/calcium phosphate composites on in vitro osteoblast response.

Science.gov (United States)

Giretova, Maria; Medvecky, Lubomir; Stulajterova, Radoslava; Sopcak, Tibor; Briancin, Jaroslav; Tatarkova, Monika

2016-12-01

Polyhydroxybutyrate/chitosan/calcium phosphate composites are interesting biomaterials for utilization in regenerative medicine and they may by applied in reconstruction of deeper subchondral defects. Insufficient informations were found in recent papers about the influence of lysozyme degradation of chitosan in calcium phosphate/chitosan based composites on in vitro cytotoxicity and proliferation activity of osteoblasts. The effect of enzymatic chitosan degradation on osteoblasts proliferation was studied on composite films in which the porosity of origin 3D scaffolds was eliminated and the surface texture was modified. The significantly enhanced proliferation activity with faster population growth of osteoblasts were found on enzymatically degraded biopolymer composite films with α-tricalcium phosphate and nanohydroxyapatite. No cytotoxicity of composite films prepared from lysozyme degraded scaffolds containing a large fraction of low molecular weight chitosans (LMWC), was revealed after 10 days of cultivation. Contrary to above in the higher cytotoxicity origin untreated nanohydroxyapatite films and porous composite scaffolds. The results showed that the synergistic effect of surface distribution, morphology of nanohydroxyapatite particles, microtopography and the presence of LMWC due to chitosan degradation in composite films were responsible for compensation of the cytotoxicity of nanohydroxyapatite composite films or porous composite scaffolds.

Development, characterization and potential applications of edible film from seaweed (Kappaphycus alvarezii)

Science.gov (United States)

Moey, Siah Watt; Abdullah, Aminah; Ahmad, Ishak

2014-09-01

A new patent pending process is proposed in this study to produce edible film directly from seaweed (Kappaphycus alvarezii). Seaweed together with other ingredients had been used to produce the film through casting technique. Physical and mechanical tests were performed on the edible film to examine the thickness, colour, transparency, solubility, tensile strength, elongation at break, water permeability rate, oxygen permeability rate and surface morphology. The produced film was transparent, stretchable, sealable and have basic properties for applications in food, pharmaceutical, cosmetic, toiletries and also agricultural industries. Edible film was successfully developed directly from dry seaweed instead of using alginate and carrageenan. The edible film processing method developed in this research was easier and cheaper compared with the method by using alginate and carrageenan.

Edible Film from Polyblend of Ginger Starch, Chitosan, and Sorbitol as Plasticizer

Science.gov (United States)

Sariningsih, N.; Putra, Y. P.; Pamungkas, W. P.; Kusumaningsih, T.

2018-03-01

Polyblend ginger starch/chitosan based edible film has been succesfully prepared and characterized. The purpose of this research was to produce edible film from polyblend of ginger starch, chitosan, and sorbitol as plasticizer. The resulted edible film were characterized by using FTIR, TGA and UTM. Edible film of ginger starch had OH vibration (3430 cm-1). Besides, edible film had elongation up to 15.63%. The thermal degradation of this material reached 208°C indicating high termal stability. The water uptake of the edible film was 42.85%. It concluded that edible film produce in this research has potential as a packaging.

Exercise increases lactoferrin, but decreases lysozyme in salivary granulocytes.

Science.gov (United States)

Gillum, Trevor; Kuennen, Matthew; McKenna, Zachary; Castillo, Micaela; Jordan-Patterson, Alex; Bohnert, Caitlin

2017-05-01

Intracellular lactoferrin (Lac) and lysozyme (Lys) content play an important role in regulating inflammation and promoting host protection. While exercise has demonstrated an increase in Lac and Lys concentration in exocrine solutions, little is known regarding intracellular concentration changes in response to exercise. To quantify intracellular Lac and Lys concentration before and after exercise in salivary CD45 + CD15 + cells. 11 males (20.3 ± 0.8 years, 57.2 ± 7.6 mL/kg/min V̇O 2pk , 11.1 ± 3.9% body fat) ran for 45 min at 75% of VO 2pk . 12 mL of stimulated saliva were collected pre and immediately post exercise. Saliva was filtered through a 30-µm filter before analysis of leukocytes (CD45 + ) and granulocytes (CD45 + CD15 + ) using flow cytometry. Median fluorescent intensity (MFI) of Lac increased from pre (64,268 ± 46,036 MFI) to post (117,134 ± 88,115 MFI) exercise (p exercise (pre: 16,933 ± 8249; post: 11,616 ± 6875) (p exercise. Conversely, the exercise-associated decrease of intracellular Lys content could be the cause of increased Lys in exocrine solutions.

Inactivation of gram-negative bacteria in milk and banana juice by hen egg white and lambda lysozyme under high hydrostatic pressure.

Science.gov (United States)

Nakimbugwe, Dorothy; Masschalck, Barbara; Anim, Grace; Michiels, Chris W

2006-10-15

The effect of hen egg white lysozyme (HEWL) and bacteriophage lambda lysozyme (LaL) in combination with high pressure (HP) treatment on the inactivation of four gram-negative bacteria (Escherichia coli O157:H7, Shigella flexneri, Yersinia enterocolitica and Salmonella typhimurium), was studied in skim milk (pH 6.8; a(w) 0.997) and in banana juice (pH 3.8; a(w) 0.971). In the absence of lysozymes, S. flexneri was more sensitive to HP in milk than in banana juice, while the opposite was observed for the other three bacteria. In combination with HP treatment, LaL was more effective than HEWL on all bacteria in both milk and banana juice. Depending on the bacteria, inactivation levels in banana juice were increased from 0.4-2.7 log units by HP treatment alone to 3.6-6.5 log units in the presence of 224 U/ml LaL. Bacterial inactivation in milk was also enhanced by LaL but only by 0.5-2.1 log units. Under the experimental conditions used, LaL was more effective in banana juice than in milk, while the effectiveness of HEWL under the same conditions was not significantly affected by the food matrix. This effect could be ascribed to the low pH of the banana juice since LaL was also more effective on E. coli in buffer at pH 3.8 than at pH 6.8. Since neither LaL nor HEWL are enzymatically active at pH 3.8, we analysed bacterial lysis after HP treatment in the presence of these enzymes, and found that inactivation proceeds through a non-lytic mechanism at pH 3.8 and a lytic mechanism at pH 6.8. Based on these results, LaL may offer interesting perspectives for use as an extra hurdle in high pressure food preservation.

Shanghai and Globalization through the Lens of Film Noir: Lou Ye’s 2000 Film, Suzhou River

Directory of Open Access Journals (Sweden)

Hongwei Lu

2010-10-01

Full Text Available In the 1990s, the film industry in China decentralized with the bankruptcy of the state-owned studio system. Privatized independent film companies took over where the government had left off and a more independent film culture emerged. Although obstacles such as political censorship, financial pressures, and Hollywood infiltration were still in the way for Chinese filmmakers, privatization of the film industry was under way. As a result of this process, new film productions of controversial subject matter came into being. In 1998 one of China’s first independent film production companies—Dream Factory—was founded. Dream Factory’s first production, in association with Berlin-based German producer Philippe Bober, was the 'Suzhou River, 'directed by its founder Lou Ye'.'1 The 2000 film, though winning prizes at international film festivals such as the Rotterdam Film Festival and the Paris Film Festival, has been banned by the Chinese government since its production.

Molecular characterization, transcriptional profiling, and antibacterial potential of G-type lysozyme from seahorse (Hippocampus abdominalis).

Science.gov (United States)

Ko, Jiyeon; Wan, Qiang; Bathige, S D N K; Lee, Jehee

2016-11-01

Lysozymes are a family of enzymes that catalyze the hydrolysis of bacterial cell wall, acting as antimicrobial effectors of the innate immune system. In the present study, an ortholog of goose-type lysozyme (ShLysG) from the big-belly seahorse (Hippocampus abdominalis) was identified and characterized structurally and functionally. The full-length cDNA sequence (1213 bp) of ShLysG is comprised of an open reading frame made up of 552 bp, encoding a polypeptide of 184 amino acid (aa) with a predicted molecular mass of 20 kDa. In silico analysis of ShLysG revealed the absence of signal peptide and the presence of a characteristic bacterial soluble lytic transglycosylase (SLT) domain bearing three catalytic residues (Glu 71 , Asp 84 , and Asp 95 ) and seven N-acetyl-d-glucosamine binding sites (Glu 71 , Asp 95 , Tyr 98 , His 99 , Ile 117 , Tyr 145 , and Asn 146 ). Homology analysis demonstrated that the aa sequence of ShLysG shared 60.7-67.4% identity and 72.6-79.3% similarity with the orthologs of other teleosts. Phylogenetic analysis of ShLysG indicated a closest relationship with the ortholog from Gadus morhua. In healthy seahorse, ShLysG mRNA showed a constitutive expression in all the tissues examined, with the highest expression in kidney and the least expression in liver. The ShLysG mRNA levels were also shown significant elevation upon the bacterial and pathogen-associated molecular pattern (PAMPs) challenges. Furthermore, lytic activities of ShLysG recombinant protein were detected against several Gram-negative and Gram-positive bacterial species. Taken together, these results suggest that ShLysG might possess a potential immune defensive role against invading microbial pathogens in seahorse. Copyright © 2016 Elsevier Ltd. All rights reserved.

Simulated Thin-Film Growth and Imaging

Science.gov (United States)

Schillaci, Michael

2001-06-01

Thin-films have become the cornerstone of the electronics, telecommunications, and broadband markets. A list of potential products includes: computer boards and chips, satellites, cell phones, fuel cells, superconductors, flat panel displays, optical waveguides, building and automotive windows, food and beverage plastic containers, metal foils, pipe plating, vision ware, manufacturing equipment and turbine engines. For all of these reasons a basic understanding of the physical processes involved in both growing and imaging thin-films can provide a wonderful research project for advanced undergraduate and first-year graduate students. After producing rudimentary two- and three-dimensional thin-film models incorporating ballsitic deposition and nearest neighbor Coulomb-type interactions, the QM tunneling equations are used to produce simulated scanning tunneling microscope (SSTM) images of the films. A discussion of computational platforms, languages, and software packages that may be used to accomplish similar results is also given.

Ultrashort pulse laser deposition of thin films

Science.gov (United States)

Perry, Michael D.; Banks, Paul S.; Stuart, Brent C.

2002-01-01

Short pulse PLD is a viable technique of producing high quality films with properties very close to that of crystalline diamond. The plasma generated using femtosecond lasers is composed of single atom ions with no clusters producing films with high Sp.sup.3 /Sp.sup.2 ratios. Using a high average power femtosecond laser system, the present invention dramatically increases deposition rates to up to 25 .mu.m/hr (which exceeds many CVD processes) while growing particulate-free films. In the present invention, deposition rates is a function of laser wavelength, laser fluence, laser spot size, and target/substrate separation. The relevant laser parameters are shown to ensure particulate-free growth, and characterizations of the films grown are made using several diagnostic techniques including electron energy loss spectroscopy (EELS) and Raman spectroscopy.

The spatial thickness distribution of metal films produced by large area pulsed laser deposition

DEFF Research Database (Denmark)

Pryds, Nini; Schou, Jørgen; Linderoth, Søren

2007-01-01

Thin films of metals have been deposited in the large-area Pulsed Laser Deposition (PLD) Facility at Riso National Laboratory. Thin films of Ag and Ni were deposited with laser pulses from an excimer laser at 248 nm with a rectangular beam spot at a fluence of 10 J/cm(2) on glass substrates of 127...

Filme autosuportado de polianilina desdopada para aplicações anticorrosivas Characterization of self-standing films of undoped polyanilina

Directory of Open Access Journals (Sweden)

Rodrigo S. Silva

2012-01-01

Full Text Available Os polímeros intrinsecamente condutores (PIC, como a polianilina (PAni, têm sido estudados com alternativa em filmes protetores de metais oxidáveis contra a corrosão. Dada a dificuldade de fusão ou dissolução da PAni para a produção de filmes, investigou-se a possibilidade de obter filmes pela mistura de PAni no estado oxidado e desdopado, conhecida como base esmeraldina, com plastificante não dopante 4-cloro-3-metilfenol (CMF e solvente conveniente. Filmes produzidos desta forma foram caracterizados por espectroscopia FTIR e RAMAN, por TGA e ensaio de voltametria cíclica. A caracterização mostrou filmes termicamente estáveis até a temperatura de 200 ºC com indícios de interação da PAni com o CMF, com a PAni se mantendo no estado oxidado no filme produzido, condição necessária para futura aplicação como filme anticorrosivo de proteção anódica.The intrinsically conductive polymers (PCI, such as polyaniline (PAni, have been studied as an alternative in the development of protective films of oxidizable metals against corrosion. Given the difficulty of mixing and dissolving PAni for the production of films, here we investigated the possibility of obtaining a mixture of PAni films in the oxidized, undoped state, referred to as emeraldine base, with the non-doping plasticizer 4-chloro-3-methylphenol (CMF and a convenient solvent. Films produced in this way were characterized by FTIR and Raman spectroscopy, TGA and cyclic voltammetry. The characterization showed films thermally stable up to 200 ºC with evidence of interaction between PAni and CMF, with PAni remaining in the oxidized state in the film produced, which a necessary condition for application as anodic protection in anticorrosion films.

Efficacy and Safety of Oral Spray Containing Lysozyme and Cetylpyridinium: Subjective Determination of Patients with Tonsillopharyngitis.

Science.gov (United States)

Catic, Tarik; Mehic, Meliha; Binakaj, Zahida; Sahman, Bilsena; Cordalija, Vedina; Kerla, Amra; Martinovic, Igor; Eskic, Hajra

2016-12-01

Tonsillopharyngitis (sore throat) is a common disease mainly related to the seasonal common cold. To relieve unpleasant symptoms and discomfort of acute tonsillopharyngitis associated with common cold, patients usually take some non-prescription drugs. The primary aim of this study was to assess subjective determinations of the efficacy and the safety/tolerability of an oral spray comprising a combination of lysozyme chloride and cetylpyridinium chloride in those patients. The study involved 1727 patients with tonsillopharyngitis associated with common cold and treated with the studied drug, in the period from December 2014 through March 2015. In total, 95% of patients rated the studied drug to be well, very well and excellently effective. In 32% of patients, the symptoms were relieved 10 minutes after the application of the spray. Significant correlations were found between the two subjective assessments of the drug efficiency with the total of 74.11% (95% CI: 73.41, 77.47%) of patients who said that the feeling of pain in the throat completely disappeared after the drug administration, evaluated the impact/effect of the drug was very good or good (Pearson Chi Square=391.401, p<0.001). The effectiveness was significantly better in patients with up to two episodes of common cold a year (Pearson Chi Square=6.101; p=0.014). The studied drug was rated to be well, very well and excellently tolerated by 97% of patients. According to patients' subjective assessment, the combination of lysozyme chloride and cetylpyridinium chloride in a formulation of spray can quickly, efficiently and safely resolve the symptoms of acute tonsillopharyngitis associated with common cold.

Our Favorite Film Shocks

DEFF Research Database (Denmark)

Willerslev, Rane; Suhr, Christian

2014-01-01

The modern medium of film has long been hailed for its capacity for producing shocks of an entertaining, thought-provoking, or even politically emancipative nature. But what is a shock, how and when does it occur, how long does it last, and are there particular techniques for producing cinematic...

Application of composition modulated thin films

International Nuclear Information System (INIS)

Hilliard, J.E.

1979-01-01

Film produced by evaporating two components through a rotating pinwheel shutter which cuts off the vapor first from one source and then the other are evaluated. These films have a modulated composition rather than a layered structure. Mechanical properties were determined using a bulge tester

Effect of self-bias voltage on the wettability, chemical functionality and nanomechanical properties of hexamethyldisiloxane films

Energy Technology Data Exchange (ETDEWEB)

Albuquerque, M.D.F. [Program of Metallurgical and Materials Engineering, COPPE, Federal University of Rio de Janeiro (UFRJ), P.O. Box 68505, 21941-972 Rio de Janeiro, RJ (Brazil); Santos, E. [Program of Metallurgical and Materials Engineering, COPPE, Federal University of Rio de Janeiro (UFRJ), P.O. Box 68505, 21941-972 Rio de Janeiro, RJ (Brazil); Faculty of Civil Engineering, University Center of Volta Redonda (UniFOA), Volta Redonda, RJ (Brazil); Perdone, R.R.T. [Program of Metallurgical and Materials Engineering, COPPE, Federal University of Rio de Janeiro (UFRJ), P.O. Box 68505, 21941-972 Rio de Janeiro, RJ (Brazil); Simao, R.A., E-mail: renata@metalmat.ufrj.br [Program of Metallurgical and Materials Engineering, COPPE, Federal University of Rio de Janeiro (UFRJ), P.O. Box 68505, 21941-972 Rio de Janeiro, RJ (Brazil)

2014-08-01

Copper and silicon substrates were coated by chemical vapor deposition using hexamethyldisiloxane (HMDSO) as the precursor gas. Substrates were placed both at the anode and cathode of a glow discharge reactor, and films were deposited using different self-bias voltages. This study focuses on comparing the differences between the hydrophilicity, polymeric character, chemical structure and nanomechanical properties of HMDSO films produced at the cathode and anode of the reactor at different self-bias voltages. Fourier transform infrared spectroscopy and Raman confocal spectroscopy indicated a significant increase in the content of organic groups when films were deposited at the anode. Analyzing the nanomechanical properties of the cathode and anode films indicated that the penetration depth was higher for samples prepared at the cathode (lower hardness) compared with the samples produced at the anode. The measured contact angles indicated that all samples became hydrophobic with water contact angles close to 100°; however, a different lyophobic character was observed when diiodomethane was used. Films produced at the anode with diiodomethane exhibited higher contact angles than did films produced at the cathode. - Highlights: • Hexamethyldisiloxane (HMDSO) films deposited by CVD on Si and Cu substrates • HMDSO films produced at the anode have greater content of organic SiO{sub 4} groups. • Films produced at the anode are harder than those deposited at the cathode. • HMDSO films produced at the cathode exhibited higher elastic recovery. • All films are hydrophobic (θ close to 100°)

Effect of self-bias voltage on the wettability, chemical functionality and nanomechanical properties of hexamethyldisiloxane films

International Nuclear Information System (INIS)

Albuquerque, M.D.F.; Santos, E.; Perdone, R.R.T.; Simao, R.A.

2014-01-01

Copper and silicon substrates were coated by chemical vapor deposition using hexamethyldisiloxane (HMDSO) as the precursor gas. Substrates were placed both at the anode and cathode of a glow discharge reactor, and films were deposited using different self-bias voltages. This study focuses on comparing the differences between the hydrophilicity, polymeric character, chemical structure and nanomechanical properties of HMDSO films produced at the cathode and anode of the reactor at different self-bias voltages. Fourier transform infrared spectroscopy and Raman confocal spectroscopy indicated a significant increase in the content of organic groups when films were deposited at the anode. Analyzing the nanomechanical properties of the cathode and anode films indicated that the penetration depth was higher for samples prepared at the cathode (lower hardness) compared with the samples produced at the anode. The measured contact angles indicated that all samples became hydrophobic with water contact angles close to 100°; however, a different lyophobic character was observed when diiodomethane was used. Films produced at the anode with diiodomethane exhibited higher contact angles than did films produced at the cathode. - Highlights: • Hexamethyldisiloxane (HMDSO) films deposited by CVD on Si and Cu substrates • HMDSO films produced at the anode have greater content of organic SiO 4 groups. • Films produced at the anode are harder than those deposited at the cathode. • HMDSO films produced at the cathode exhibited higher elastic recovery. • All films are hydrophobic (θ close to 100°)

A Study on the Origins of Cultural Films in Korea: A Focus on Films by the Japanese Government-General of Korea

Directory of Open Access Journals (Sweden)

Ryeosil Kim

2014-08-01

Full Text Available The term "cultural film" was coined in Korea under the Japanese rule, and it had been used as a general term for nonfiction films since the Korean liberation until the 1980s. However, unlike the term "documentary film," cultural film contains implications of propaganda film, controlled and produced as part of cultural and public relations policies of the government, thus it had been seldom studied for a long time. The term “cultural film” did not mean a film genre in the beginning. Therefore it is impossible to conceptualize the term based on common formats or contents of the films which have been called “cultural films.” Cultural films have been generally recognized as nonfiction films, and therefore documentary films. Yet in the production and distribution sites, fiction films and even animations were considered cultural films. In order to delve into the nature of cultural film, we have to reexamine the process in which the proper noun “Cultural Film” became a common noun. This paper attempts to explore the historical context in which the term “cultural film” was coined and used in the colonial Korea, when cultural films began to be produced. I examine the overall time period in which: 1 the concept of nonfiction film was first introduced in the Motion Picture Censorship Regulation, 2 the term “cultural film” became legally stipulated at a time when national policies were being established as Korea experienced the Manchurian Incident and the Sino-Japanese War, and 3 “cultural films” were screened mandatorily as “national films” that the nation had to foster through the Japan Film Law and Chosôn Film Act.

Improvement of physical properties of IGZO thin films prepared by excimer laser annealing of sol–gel derived precursor films

International Nuclear Information System (INIS)

Tsay, Chien-Yie; Huang, Tzu-Teng

2013-01-01

Indium gallium zinc oxide (IGZO) transparent semiconductor thin films were prepared by KrF excimer laser annealing of sol–gel derived precursor films. Each as-coated film was dried at 150 °C in air and then annealed using excimer laser irradiation. The influence of laser irradiation energy density on surface conditions, optical transmittances, and electrical properties of laser annealed IGZO thin films were investigated, and the physical properties of the excimer laser annealed (ELA) and the thermally annealed (TA) thin films were compared. Experimental results showed that two kinds of surface morphology resulted from excimer laser annealing. Irradiation with a lower energy density (≤250 mJ cm −2 ) produced wavy and irregular surfaces, while irradiation with a higher energy density (≥350 mJ cm −2 ) produced flat and dense surfaces consisting of uniform nano-sized amorphous particles. The explanation for the differences in surface features and film quality is that using laser irradiation energy to form IGZO thin films improves the film density and removes organic constituents. The dried IGZO sol–gel films irradiated with a laser energy density of 350 mJ/cm 2 had the best physical properties of all the ELA IGZO thin films. The mean resistivity of the ELA 350 thin films (4.48 × 10 3 Ω cm) was lower than that of TA thin films (1.39 × 10 4 Ω cm), and the average optical transmittance in the visible range (90.2%) of the ELA 350 thin films was slightly higher than that of TA thin films (89.7%). - Highlights: • IGZO semiconductor films were prepared by laser annealing of sol–gel derived films. • Surface roughness and resistivity of ELA samples were affected by energy density. • The ELA 350 IGZO film exhibited the best properties among all of ELA IGZO films. • Transmittance and resistivity of ELA 350 films are greater than those of TA films

Improvement of physical properties of IGZO thin films prepared by excimer laser annealing of sol–gel derived precursor films

Energy Technology Data Exchange (ETDEWEB)

Tsay, Chien-Yie, E-mail: cytsay@fcu.edu.tw; Huang, Tzu-Teng

2013-06-15

Indium gallium zinc oxide (IGZO) transparent semiconductor thin films were prepared by KrF excimer laser annealing of sol–gel derived precursor films. Each as-coated film was dried at 150 °C in air and then annealed using excimer laser irradiation. The influence of laser irradiation energy density on surface conditions, optical transmittances, and electrical properties of laser annealed IGZO thin films were investigated, and the physical properties of the excimer laser annealed (ELA) and the thermally annealed (TA) thin films were compared. Experimental results showed that two kinds of surface morphology resulted from excimer laser annealing. Irradiation with a lower energy density (≤250 mJ cm{sup −2}) produced wavy and irregular surfaces, while irradiation with a higher energy density (≥350 mJ cm{sup −2}) produced flat and dense surfaces consisting of uniform nano-sized amorphous particles. The explanation for the differences in surface features and film quality is that using laser irradiation energy to form IGZO thin films improves the film density and removes organic constituents. The dried IGZO sol–gel films irradiated with a laser energy density of 350 mJ/cm{sup 2} had the best physical properties of all the ELA IGZO thin films. The mean resistivity of the ELA 350 thin films (4.48 × 10{sup 3} Ω cm) was lower than that of TA thin films (1.39 × 10{sup 4} Ω cm), and the average optical transmittance in the visible range (90.2%) of the ELA 350 thin films was slightly higher than that of TA thin films (89.7%). - Highlights: • IGZO semiconductor films were prepared by laser annealing of sol–gel derived films. • Surface roughness and resistivity of ELA samples were affected by energy density. • The ELA 350 IGZO film exhibited the best properties among all of ELA IGZO films. • Transmittance and resistivity of ELA 350 films are greater than those of TA films.

Testing Solutions for Adult Film Performers.

Science.gov (United States)

Bergman, Zachary R

2014-01-01

The majority of the nation's adult films are produced in California, and within California, most production occurs in Los Angeles. In order to regulate that content, the County of Los Angeles passed the Safer Sex in the Adult Film Industry Act (Measure B) by way of referendum in November 2012. Measure B requires that adult film producers wishing to film in Los Angeles County obtain permits from the Los Angeles County Department of Public Health, and it also mandates that adult film performers use condoms while filming and "engaging in anal or vaginal sexual intercourse." Nevertheless, between August 2013 and January 2014, several adult film performers in California tested positive for HIV, and the threat of infection remains. Although Measure B is not the best way forward for Los Angeles County, elements of the ordinance should be incorporated into future legislative efforts. Given the economic ramifications of industry flight due to more localized regulations, this Note concludes that California should pass statewide comprehensive reform. Any such new legislation must treat "independent contractors," the classification generally used for adult film performs, as if they were regular employees. Legislation should also couple mandatory testing mechanisms with provisions granting performers the right to choose whether they use condoms. Finally, legislation must include mechanisms that ensure performers' preferences are not improperly tainted by outside forces and pressures. While there will always be risks associated with the production of adult content, if undertaken, these reforms could significantly mitigate those hazards.

Properties and practical application of thin CeOx films

Directory of Open Access Journals (Sweden)

Maksimchuk N. V.

2010-10-01

Full Text Available The properties of CeOx films produced by various methods have been investigated. According to the comparative analisys “metallic mirror oxidation” method allows to produce films with significantly better characteristics than the «explosive evaporation» method. Though the latter method yields higher photosensitivity of CeOx films and structures on their base. In the process the optimal value of the substrate temperature was determined. Obtained data expand the CeOx application potential in microelectronic sensor sphere.

Theoretical study of phase behaviour of DLVO model for lysozyme and γ-crystalline aqueous electrolyte solutions

Directory of Open Access Journals (Sweden)

R. Melnyk

2015-03-01

Full Text Available Mean spherical approximation (MSA, second-order Barker-Henderson (BH perturbation theory and thermodynamic perturbation theory (TPT for associating fluids in combination with BH perturbation theory are applied to the study of the structural properties and phase behaviour of the Derjaguin-Landau-Verwey-Overbeek (DLVO model of lysozyme and γ-cristalline aqueous electrolyte solutions. Predictions of the MSA for the structure factors are in good agreement with the corresponding computer simulation predictions. The agreement between theoretical results for the liquid-gas phase diagram and the corresponding results of the experiment and computer simulation is less satisfactory, with predictions of the combined BH-TPT approach being the most accurate.

New radiochroic film densitometry system

International Nuclear Information System (INIS)

Tran, T.; Freeman, N.; Johnston, P.

2004-01-01

Full text: The advantages of radiochromic film in radiation dosimetry are well known. They include dosimetry with high spatial resolution, response less dependent on incident beam energy than common radiotherapy films (such as the Kodak XV films), tissue equivalence and the ability to be handled and developed in room light. This study entails the design and testing of a new radiochromic densitometry system. The system consists of a single light emitting diode (LED), opaque 'diffuser' and digital camera. Customised software was developed to analyse images obtained from the digital camera. Standard characteristics of a commercially available super bright red LED (peak wavelength 625nm) was analysed in order to determine the voltage, current and intensity settings. Various methods in diffusing the single LED light source were investigated and it was determined that an opaque transmission 'diffuser' was the best alternative. While the intensity of the LED was kept constant, the digital camera exposure times were varied in order to determine a setting which would produce the best image exposure. The system was designed and built and preliminary tests were carried using the standard radiochromic film GafChromic MD-55-2. LED current vs. voltage curves were characteristically exponential for positive voltage. Studies into LED intensity versus camera exposure produced an unexpected result. At high exposures the camera saturates and if even higher exposures are used the LED intensity apparently decreases. This was thought to be due to the pixels in the charge couple device (CCD) saturating and eventually electronically 'bleeding' into adjacent pixels. Using the opaque transmission 'diffuser' enabled successful use of the single LED light producing an area of homogenous light intensity in which images of radiochromic films can be obtained. Preliminary results from radiochromic film characteristic studies show no unusual results. The single LED, diffuser and digital camera

PVA/Polysaccharides Blended Films: Mechanical Properties

OpenAIRE

Silva, Fábio E. F.; Di-Medeiros, Maria Carolina B.; Batista, Karla A.; Fernandes, Kátia F.

2013-01-01

Blends of polyvinyl alcohol (PVA) and angico gum (AG) and/or cashew gum (CG) were used to produce films by casting method. Morphological and mechanical properties of these films were studied and compared to the properties of a commercial collagen membrane of bovine origin (MBO). The films presented thickness varying from 70 to 140 μm (PVA/AG) and 140 to 200 μm (PVA/CG). Macroscopic analysis showed that a PVA/CG film was very similar to MBO regarding the color and transparency. The higher valu...

Comparative carbon-13 nuclear magnetic resonance study at 67. 9 MHz on lysozyme (human and egg-white) and. cap alpha. -lactalbumin (human and bovine) in their native and denatured state

Energy Technology Data Exchange (ETDEWEB)

van Binst, G; Biesemans, M [Brussels Univ. (Belgium). Faculte des Sciences

1975-01-01

A first detailed comparison of the /sup 13/C spectra at high field of two lysozymes (human and egg-white) and two ..cap alpha..-lactalbumins (human and bovine milk) is presented. Assignments were made on most of the resonance peaks in the aliphatic and aromatic regions of the denatured proteins. The relative peak intensities clearly demonstrate the differences in the amino acid composition of the related proteins. The broadening and the complexity of the spectra of the native proteins reflect the non equivalence of the chemical groups in the folded conformation. The usefulness of /sup 13/C nmr spectroscopy in the study of the interaction between small molecules and proteins was tested on N-acteyl-glucosamine in the presence of lysozyme.

227 Rituals in Nollywood Films: A Discourse in Ideology Asiegbu ...

African Journals Online (AJOL)

USER

Introduction. Nollywood movies refer to the video films produced in Nigeria. The ... Consequently, the Nigerian video film industry is a highly placed commercial ..... video film industry: Technological fascination and the domestication game. In.

Radiographic film: surface dose extrapolation techniques

International Nuclear Information System (INIS)

Cheung, T.; Yu, P.K.N.; Butson, M.J.; Cancer Services, Wollongong, NSW; Currie, M.

2004-01-01

Full text: Assessment of surface dose delivered from radiotherapy x-ray beams for optimal results should be performed both inside and outside the prescribed treatment fields An extrapolation technique can be used with radiographic film to perform surface dose assessment for open field high energy x-ray beams. This can produce an accurate 2 dimensional map of surface dose if required. Results have shown that surface % dose can be estimated within ±3% of parallel plate ionisation chamber results with radiographic film using a series of film layers to produce an extrapolated result. Extrapolated percentage dose assessment for 10cm, 20cmand 30cm square fields was estimated to be 15% ± 2%, 29% ± 3% and 38% ± 3% at the central axis and relatively uniform across the treatment field. Corresponding parallel plate ionisation chamber measurement are 16%, 27% and 37% respectively. Surface doses are also measured outside the treatment field which are mainly due to scattered electron contamination. To achieve this result, film calibration curves must be irradiated to similar x-ray field sizes as the experimental film to minimize quantitative variations in film optical density caused by varying x-ray spectrum with field size. Copyright (2004) Australasian College of Physical Scientists and Engineers in Medicine

Production, Characterization And Tribological Properties Of Molybdenum Doped Diamond-like Carbon Films

OpenAIRE

Alp, Emre

2012-01-01

Thin films whose thickness is typically less than several microns are produced by the deposition of individual atoms on any substrate. Historically, thin films have been used for about half a century in producing instrument hard coatings, optical coatings, thin-film batteries, electronic devices, photovoltaic devices, memory devices and decorative parts. Thin film technology is still being developed by a technological advancement since it is a key factor in the twenty-first century developmen...

Levitation of a drop over a film flow

Science.gov (United States)

Sreenivas, K. R.; de, P. K.; Arakeri, Jaywant H.

1999-02-01

A vertical jet of water impinging on a horizontal surface produces a radial film flow followed by a circular hydraulic jump. We report a phenomenon where fairly large (1 ml) drops of liquid levitate just upstream of the jump on a thin air layer between the drop and the film flow. We explain the phenomenon using lubrication theory. Bearing action both in the air film and the water film seems to be necessary to support large drops. Horizontal support is given to the drop by the hydraulic jump. A variety of drop shapes is observed depending on the volume of the drop and liquid properties. We show that interaction of the forces due to gravity, surface tension, viscosity and inertia produces these various shapes.

Adherent zirconia films by reactive ion implantation

International Nuclear Information System (INIS)

Bunker, S.N.; Armini, A.J.

1993-01-01

Conventional methods of forming ceramic coatings on metal substrates, such as CVD or plasma spray, typically retain a sharp interface and may have adhesion problems. In order to produce a completely mixed interface for better adhesion, a method using reactive ion implantation was used which can grow a thick stoichiometric film of an oxide ceramic starting from inside the substrate. Zirconium oxide ceramic films have been produced by this technique using a high-energy zirconium ion beam in an oxygen gas ambient. Compositional data are shown based on Auger electron spectroscopy of the film. Tribological properties of the layer were determined from wear and friction measurements using a pin-on-disk test apparatus. The adhesion was measured both by a scratch technique as well as by thermal shock. Results show an extremely adherent ZrO 2 film with good tribological properties

Determination of gel content and SEM morphology for sago-PVA blends film

International Nuclear Information System (INIS)

Sarada Idris; Zulkafli Ghazali; Kamarudin Hashim

2006-01-01

Blends of polyvinyl alcohol and sago starch have been prepared to evaluate the potential of producing biodegradable products. Glycerol was introduced in the blends to improve the flexibility of the films as plasticizer in order more flexible film. These blends have been subjected to electron beam irradiation to evaluate and characterized radiation effect on the blends. Subsequently films were produced from this blend. The gel content of un-irradiated and irradiated films as evidence of cross linking was measured and discussed. This paper also discuss the films morphology from Scanning Electron Microscopy(SEM) observation. (Author)

Nigerian popular films: copyright and originality | Effiong | Lwati: A ...

African Journals Online (AJOL)

This essay attempts to examine the effect of copyright activities on Nigerian popular films. To an extent, it is to show how these activities have affected the distribution of films produced by Nigerians and the rising wave of returning to the cinema halls. Movie producers have expressed disappointment in the loss of their ...

Selected Films for Young Adults, 1984.

Science.gov (United States)

Top of the News, 1984

1984-01-01

This 22-item filmography of 16mm films recommended for use in programs planned for young adults was compiled by the Selected Films for Young Adults Committee, Young Adult Services Division, American Library Association. Producers, directors, distributors, length, price, and brief annotations are provided. Addresses for 12 distributors are…

T- and B-cell response analysis following calf immunisation with experimental Mycoplasma bovis vaccine containing saponin and lysozyme dimer

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Dudek Katarzyna

2017-12-01

Full Text Available Introduction:Mycoplasma bovis is a well-known cause of various disorders in cattle, such as pneumonia, arthritis, mastitis kerato-conjunctivitis, pharyngitis, laryngitis, otitis media, meningitis, and reproductive disorders. There are no commercial vaccines against M. bovis in Europe, therefore, experimental ones are still under investigation. The aim of this study was to evaluate the effect of experimental M. bovis vaccine, containing the Polish field M. bovis strain as well as saponin and lysozyme dimer adjuvants, on the T- and B-cell response in calves.

Surface Modification of Aerosol-Assisted CVD Produced TiO2 Thin Film for Dye Sensitised Solar Cell

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SuPei Lim

2014-01-01

Full Text Available We report a simple and convenient method for the preparation of Ag/TiO2 thin films supported on indium tin oxide, which was achieved by sonochemical deposition of Ag+ on aerosol-assisted chemical vapour deposited TiO2 thin films. Posttreatment was performed on the film by immersion in HCl. The as-prepared composite film was characterised by X-ray diffraction, ultraviolet-visible absorption spectroscopy, Raman spectroscopy, and field emission scanning electron microscopy. The photoelectrochemical measurements and J-V characterisation showed approximately fivefold increase in photocurrent density generation and approximately sevenfold enhancement in dye sensitiser solar cell (DSSC conversion efficiency, which was achieved after modification of the TiO2 film with HCl posttreatment and Ag particle deposition. The improved photocurrent density of 933.30 μA/cm2, as well as DSSC power conversion efficiency of 3.63% with high stability, is an indication that the as-synthesised thin film is a potential candidate for solar energy conversion applications.