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Sample records for lysozyme activity serum

  1. A novel anti-inflammatory activity of lysozyme: modulation of serum complement activation

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    M. O. Ogundele

    1998-01-01

    Full Text Available Lysozyme is an ubiquitous enzyme found in most biological secretions and leukocytes. This study was aimed at investigating its interaction with other inflammatory mediators on mucosa surfaces, particularly the complement system. Lysozyme has been shown in our present study, to inhibit the haemolytic activity of serum complement in a dose-dependent fashion, when tested within the levels present in normal and inflamed breast-milk samples, and other mucosal secretions. This represents a new antiinflammatory action of lysozym e in relation to the serum complement, and the exact mode of the interaction need further studies.

  2. Variations of serum and mucus lysozyme activity and total protein content in the male and female Caspian kutum (Rutilus frisii kutum, Kamensky 1901) during reproductive period.

    Science.gov (United States)

    Ghafoori, Zomorod; Heidari, Behrooz; Farzadfar, Fariba; Aghamaali, Mahmoudreza

    2014-03-01

    Serum and mucus lysozyme were measured in male and female Caspian kutum (Rutilus frisii kutum) under seasonal temperature, gonadal growth and reproductive migration. Significant difference with almost similar trend in serum and mucus lysozyme of the female Caspian kutum in sampling time and ovarian growth was observed. However, while there was no significant difference in serum lysozyme of the male specimen in sampling time and testicular growth, significant variations was observed in mucus lysozyme. In addition, there was significant difference in mucus total protein both for male and female specimens. The effectiveness ratio of factors on lysozyme variations followed in descending order by seasonal temperature (main factor), reproductive activity and migration with negligible effect and the lysozyme level was not significantly different in male and female Caspian kutum.

  3. Studies of bactericidal activity to Escherichia coli of porcine serum and colostral immunoglobulins and the role of lysozyme with secretory IgA

    Science.gov (United States)

    Hill, I. R.; Porter, P.

    1974-01-01

    Gel filtration and immune inhibition techniques were used to study bactericidal activities of IgG, IgM and IgA against smooth strains of Escherichia coli 0141 and 08 in sow serum and colostrum and post-colostral piglet serum. Bactericidal activity in sow sera was primarily associated with IgM and a low molecular weight IgG component, 7S IgG activity was less frequently observed. In colostral whey fractions and post-colostral piglet sera, in the absence of lysozyme, bactericidal antibody activity was associated with IgM and 7S IgG. In post-colostral serum bactericidal antibody was also attributable to a low molecular weight form of IgG. IgA in serum from the sow and neonate showed no bactericidal activity, even in the presence of lysozyme, whereas in colostrum secretory 11S IgA had bactericidal activity, but only in the presence of complement and lysozyme. PMID:4212358

  4. Measurement of Content and Activity of Lysozyme in Serum of Piglets%仔猪血清中溶菌酶含量和活力的测定

    Institute of Scientific and Technical Information of China (English)

    王海霞; 迟瑞宾

    2011-01-01

    溶菌酶参与机体内多种免疫反应,测定它在血清中的含量可作为了解机体免疫功能的一个重要指标.试验用36头22日龄仔猪血清为试验样本,利用比浊法和ELISA两种方法测定血清中的溶菌酶.结果表明:ELISA测定法对同一样本3次测定值间的变异系数和对36个仔猪血清样本溶菌酶测定值变异系数均极显著(P<0.01)小于比浊法测定结果的变异系数,表明ELISA方法比比浊法的精确度高.两种方法测定结果间的Pearson相关系数为0.33,未达到显著差异水平(P>0.05).比浊法测定的是溶菌酶的活性,ELISA测定的是血清中溶菌酶的净含量,两种溶菌酶测定方法具有不同的测定原理,导致二者测定结果之间的相关系数很低.ELISA受血清的颜色影响程度低,易于实现批量检测,但是需要酶标仪、洗板机等较昂贵设备.在实际研究中,研究者应该根据自己的试验目的、样本量以及价格等因素综合考虑溶菌酶的最佳测定方法.%Lysozyme participates in many immune responses, and its content in serum is an important indicator of immune function of the animal. In the study,the lysozyme in serum of 36 piglets(20 day after birth) was measured with two methods, nephelometry and enzyme-linked immunosorbent assay (ELISA). The results showed that both the coefficients of variance (C. V. ) among the three tests of the same samples of ELISA and that of the testing results of 36 samples were significantly lower than those of nephelometry, indicating the higher testing precision of ELISA than nephelometry. The coefficient of correlation between the results of two methods was 0. 33, which was not significantly correlated (P>0. 05). Nephelometry tests the enzyme activity of lysozyme, and ELISA tests the actual net content in serum. Hence, the two methods have different testing principles, which lead to their low coefficient of correlation. Additionally, ELISA was less affected by the color of

  5. Immunoassay of serum muramidase (lysozyme) in ocular diseases.

    Science.gov (United States)

    Sen, D K; Sarin, G S

    1987-01-01

    Serum levels of muramidase activity were measured in 162 patients with different ocular diseases and 84 healthy subjects by electro-immuno-diffusion technique. We demonstrated for the first time that electro-immuno-diffusion technique could be successfully applied for the estimation of serum muramidase concentrations. Serum muramidase was found to be high in significant number of cases with granulomatous uveitis, tuberculous keratitis, central serous retinopathy and Eales' disease. Tuberculosis was presumed to be the cause in them by the process of exlusion. Patients with high serum muramidase activity were subjected to anti-tubercular treatment with a marked clinical improvement. It is suggested that high serum muramidase could be an useful parameter in deciding the line of treatment in patients with ocular diseases of uncertain etiology. Serum muramidase concentrations showed return to normal levels with the clinical improvement of the diseases with treatment. It increased again with the re-appearance of the activity of the diseases.

  6. Recombinant goose-type lysozyme in channel catfish: Lysozyme activity and efficacy as plasmid DNA immunostimulant against Aeromonas hydrophila infection

    Science.gov (United States)

    The objectives of this study were: 1) to investigate whether recombinant channel catfish lysozyme g (CC-Lys-g) produced in E. coli expression system possesses any lysozyme activity; and 2) to evaluate whether channel catfish lysozyme g plasmid DNA could be used as an immunostimulant to protect chann...

  7. Enhanced antibacterial activity of lysozyme immobilized on chitin nanowhiskers.

    Science.gov (United States)

    Jiang, Suisui; Qin, Yang; Yang, Jie; Li, Man; Xiong, Liu; Sun, Qingjie

    2017-04-15

    In this paper, the contribution of chitin nanowhiskers (CHNW) to the enzymatic activity and antimicrobial activity of lysozyme adsorbed on CHNW was investigated. An activity assay showed that the lysozyme-CHNW system exhibited significant promotion potency on lysozyme activity, which was approximately 1.5-fold greater than that of free lysozyme. The molecular promotion mechanism of lysozyme immobilized by adsorption onto CHNW was investigated by ultraviolet-visible spectrophotometry, fluorescence spectroscopy, and circular dichroism spectroscopy. The results indicated that changes in the secondary structure of lysozyme adsorption onto CHNW resulted in superior enzymatic activity. Furthermore, the antimicrobial assays indicated that the antimicrobial activity of the lysozyme-CHNW system was greater than that of free lysozyme, whereas its antimicrobial effect on a gram-negative bacterium was better than that on gram-positive bacteria. This research provides a facile and promising approach for increasing the application of chitin-derived and enhancing the antibacterial efficiency on preservation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. ALTERED ENZYMATIC ACTIVITY OF LYSOZYMES BOUND TO VARIOUSLY SULFATED CHITOSANS

    Institute of Scientific and Technical Information of China (English)

    Hong-wei Wang; Lin Yuan; Tie-liang Zhao; He Huang; Hong Chen; Di Wu

    2012-01-01

    The purpose of this research is to investigate the effects of the variously sulfated chitosans on lysozyme activity and structure.It was shown that the specific enzymatic activity of lysozyme remained almost similar to the native protein after being bound to 6-O-sulfated chitosan (6S-chitosan) and 3,6-O-sulfated chitosan (3,6S-chitosan),but decreased greatly after being bound to 2-N-6-O-sulfated chitosan (2,6S-chitosan).Meanwhile,among these sulfated chitosans,2,6S-chitosan induced the greatest conformational change in lysozyme as indicated by the fluorescence spectra.These findings demonstrated that when sulfated chitosans of different structures bind to lysozyme,lysozyme undergoes conformational change of different magnitudes,which results in corresponding levels of lysozyme activity.Further study on the interaction of sulfated chitosans with lysozyme by surface plasmon resonance (SPR) suggested that their affinities might be determined by their molecular structures.

  9. Forced Desorption of Bovine Serum Albumin and Lysozyme from Graphite: Insights from Molecular Dynamics Simulation.

    Science.gov (United States)

    Mücksch, Christian; Urbassek, Herbert M

    2016-08-18

    We use molecular dynamics (MD) simulation to study the adsorption and desorption of two widely different proteins, bovine serum albumin (BSA) and lysozyme, on a graphite surface. The adsorption is modeled using accelerated MD to allow the proteins to find optimum conformations on the surface. Our results demonstrate that the "hard protein" lysozyme retains much of its secondary structure during adsorption, whereas BSA loses it almost completely. BSA has a considerably larger adsorption energy compared to that of lysozyme, which does not scale with chain length. Desorption simulations are carried out using classical steered MD. The BSA molecule becomes fully unzipped during pull-off, whereas several helices survive this process in lysozyme. The unzipping process shows up in the force-distance curve of BSA as a series of peaks, whereas only a single or few, depending on protein orientation, force peaks occur for lysozyme. The maximum desorption force is larger for BSA than for lysozyme, but only by a factor of about 2.3.

  10. Binding properties of drospirenone with human serum albumin and lysozyme in vitro

    Science.gov (United States)

    Wang, Qing; Ma, Xiangling; He, Jiawei; Sun, Qiaomei; Li, Yuanzhi; Li, Hui

    2016-01-01

    The interaction of drospirenone (DP) with human serum albumin (HSA)/lysozyme (LYZ) was investigated using different optical techniques and molecular models. Results from the emission and time resolved fluorescence studies revealed that HSA/LYZ emission quenching with DP was initiated by static quenching mechanism. The LYZ-DP system was more easily influenced by temperature than the HSA-DP system. Displacement experiments demonstrated that the DP binding site was mainly located in site 1 of HSA. Based on the docking methods, DP was mainly bound in the active site hinge region where Trp-62 and Trp-63 are located. Conformation study showed that DP had different effects on the local conformation of HSA and LYZ molecules.

  11. 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.

  12. Recombinant goose-type lysozyme in channel catfish: lysozyme activity and efficacy as plasmid DNA immunostimulant against Aeromonas hydrophila infection.

    Science.gov (United States)

    Pridgeon, Julia W; Klesius, Phillip H; Dominowski, Paul J; Yancey, Robert J; Kievit, Michele S

    2013-10-01

    The objectives of this study were: 1) to investigate whether recombinant channel catfish lysozyme-g (CC-Lys-g) produced in Escherichia coli expression system possesses any lysozyme activity; and 2) to evaluate whether channel catfish lysozyme-g plasmid DNA could be used as an immunostimulant to protect channel catfish against Aeromonas hydrophila infection. Recombinant CC-Lys-g produced in E. coli expression system exhibited significant (P recombinant channel catfish lysozyme-g (pcDNA-Lys-g) was transfected in channel catfish gill cells G1B, the over-expression of pcDNA-Lys-g offered significant (P DNA injection. Macrophages of fish injected with pcDNA-Lys-g produced significantly (P DNA injection. Taken together, our results suggest that pcDNA-Lys-g could be used as a novel immunostimulant to offer immediate protection to channel catfish against A. hydrophila infection.

  13. Characterization and function of kuruma shrimp lysozyme possessing lytic activity against Vibrio species.

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    Hikima, Sonomi; Hikima, Jun ichi; Rojtinnakorn, Jiraporn; Hirono, Ikuo; Aoki, Takashi

    2003-10-16

    Lysozyme cDNA was isolated from a kuruma shrimp, Marsupenaeus japonicus, hemocyte cDNA library. The cDNA consists of 1055 base pairs (bp) and encodes a chicken-type (c-type) lysozyme with a deduced amino acid sequence of 156 residues. The kuruma shrimp lysozyme has a high identity (79.7%) with pacific white shrimp lysozyme, and low to moderate identities (33.3-43.0%) with lysozymes of insects and vertebrates. Comparisons with other c-type lysozymes from invertebrates and vertebrates showed that the two catalytic residues (Glu58 and Asp75) and the eight cysteine residue motif were completely conserved. Two novel insertion sequences were also observed in the kuruma and pacific white shrimp lysozyme amino acid sequences. Interestingly, phylogenetic analysis revealed that the kuruma shrimp lysozyme was more closely related to vertebrate c-type lysozymes. Expression of the cDNA in insect cells, using a baculovirus expression system, yielded a recombinant lysozyme with optimum activity at pH 7.5 and 50 degrees C, as evaluated by a lysoplate assay. The kuruma shrimp lysozyme displayed lytic activities against several Vibrio species and fish pathogens, including Vibrio penaeicida (a pathogenic bacteria to the kuruma shrimp) and suggested that shrimp lysozyme affects a greater variety of pathogens.

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

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    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

  15. Mixed-mode chromatography integrated with high-performance liquid chromatography for protein analysis and separation: Using bovine serum albumin and lysozyme as the model target.

    Science.gov (United States)

    Xia, Hai-Feng; Don, Bin-Bin; Zheng, Meng-Jie

    2016-05-01

    A type of mixed-mode chromatography was integrated with high-performance liquid chromatography for protein analysis and separation. The chromatographic behavior was tested using bovine serum albumin and lysozyme as model proteins. For the mixed-mode column, the silica beads were activated with γ-(2,3-epoxypropoxy)-propytrimethoxysilane and coupled with 4-mercaptopyridine as the functional ligand. The effects of pH, salt, and the organic solvent conditions of the mobile phase on the retention behavior were studied, which provided valuable clues for separation strategy. When eluted with a suitable pH gradient, salt concentration gradient, and acetonitrile content gradient, the separation behavior of bovine serum albumin and lysozyme could be controlled by altering the conditions of the mobile phase. The results indicated this type of chromatography might be a useful method for protein analysis and separation.

  16. 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.

  17. 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

  18. 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 investiga

  19. The Plasmid-Encoded Regulator Activates Factors Conferring Lysozyme Resistance on Enteropathogenic Escherichia coli Strains▿

    Science.gov (United States)

    Salinger, Nina; Kokona, Bashkim; Fairman, Robert; Okeke, Iruka N.

    2009-01-01

    We demonstrate that enhanced lysozyme resistance of enteropathogenic Escherichia coli requires the plasmid-encoded regulator, Per, and is mediated by factors outside the locus for enterocyte effacement. EspC, a Per-activated serine protease autotransporter protein, conferred enhanced resistance on nonpathogenic E. coli, and a second Per-regulated, espC-independent lysozyme resistance mechanism was identified. PMID:18997020

  20. The plasmid-encoded regulator activates factors conferring lysozyme resistance on enteropathogenic Escherichia coli strains.

    Science.gov (United States)

    Salinger, Nina; Kokona, Bashkim; Fairman, Robert; Okeke, Iruka N

    2009-01-01

    We demonstrate that enhanced lysozyme resistance of enteropathogenic Escherichia coli requires the plasmid-encoded regulator, Per, and is mediated by factors outside the locus for enterocyte effacement. EspC, a Per-activated serine protease autotransporter protein, conferred enhanced resistance on nonpathogenic E. coli, and a second Per-regulated, espC-independent lysozyme resistance mechanism was identified.

  1. Antimicrobial activity of lysozyme with special relevance to milk

    African Journals Online (AJOL)

    STORAGESEVER

    2008-12-29

    Dec 29, 2008 ... Lysozyme is a hydrolytic enzyme which has been purified from cells, secretions and tissues of virtually ... breakdown of peptidoglycan polymers of bacterial cell ...... Small Rum. ..... myeloid leukemia: An analysis of 208 cases.

  2. Destroying activity of magnetoferritin on lysozyme amyloid fibrils

    Energy Technology Data Exchange (ETDEWEB)

    Kopcansky, Peter; Siposova, Katarina [Institute of Experimental Physics, SAS, Watsonova 47, 040 01 Kosice (Slovakia); Melnikova, Lucia, E-mail: melnikova@saske.sk [Institute of Experimental Physics, SAS, Watsonova 47, 040 01 Kosice (Slovakia); Bednarikova, Zuzana [Institute of Experimental Physics, SAS, Watsonova 47, 040 01 Kosice (Slovakia); Institute of Chemical Sciences, Faculty of Sciences, Safarik University, Kosice (Slovakia); Timko, Milan; Mitroova, Zuzana; Antosova, Andrea [Institute of Experimental Physics, SAS, Watsonova 47, 040 01 Kosice (Slovakia); Garamus, Vasil M. [Helmholtz-Zentrum Geesthacht: Centre for Materials and Coastal Research, Max-Planck-Street 1, 21502 Geesthacht (Germany); Petrenko, Viktor I. [Joint Institute for Nuclear Research, Joliot-Curie 6, Dubna, 141980 Moscow Region (Russian Federation); Kyiv Taras Shevchenko National University, Volodymyrska Street 64, Kyiv 01033 (Ukraine); Avdeev, Mikhail V. [Joint Institute for Nuclear Research, Joliot-Curie 6, Dubna, 141980 Moscow Region (Russian Federation); Gazova, Zuzana [Institute of Experimental Physics, SAS, Watsonova 47, 040 01 Kosice (Slovakia); Department of Medical and Clinical Biochemistry and LABMED, Tr. SNP 1, 040 11 Kosice (Slovakia)

    2015-03-01

    Presence of protein amyloid aggregates (oligomers, protofilaments, fibrils) is associated with many diseases as diabetes mellitus or Alzheimer's disease. The interaction between lysozyme amyloid fibrils and magnetoferritin loaded with different amount of iron atoms (168 or 532 atoms) has been investigated by small-angle X-rays scattering and thioflavin T fluorescence measurements. Results suggest that magnetoferritin caused an iron atom-concentration dependent reduction of lysozyme fibril size. - Highlights: • The interaction between lysozyme amyloid fibrils and magnetoferritin loaded with different amount of iron atoms (168 or 532 atoms) has been investigated by small-angle X-rays scattering and thioflavin T fluorescence measurements. • Results suggest that magnetoferritin caused an iron atom-concentration dependent reduction of lysozyme fibril size.

  3. Lysozyme activity and L(+)-lactic acid production in saliva in schoolchildren with high Lactobacillus counts.

    Science.gov (United States)

    Twetman, S; Dahllöf, G; Wikner, S

    1987-04-01

    Out of 374 schoolchildren, aged 13-15 yr, 42 with high counts of salivary lactobacilli (greater than or equal to 10(5] were selected for this study. Lysozyme activity in saliva and L(+)-lactic acid (LA) production after addition of glucose were determined. The mean values of lysozyme activity and LA concentration were 19.4 micrograms/ml and 1.4 mmol/l respectively. The levels of LA produced without addition of glucose were less than 0.2 mmol/l. The results showed a statistically significant (P less than 0.05) negative correlation between lysozyme activity and the levels of LA produced. The findings of this study suggest that lysozyme may be of importance in limiting acid production in saliva.

  4. 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.

  5. Investigation on the interactions of silymarin to bovine serum albumin and lysozyme by fluorescence and absorbance

    Energy Technology Data Exchange (ETDEWEB)

    Pang Bo [College of Chemistry, Changchun Normal University, Changchun 130032 (China); Bi Shuyun, E-mail: sy_bi@sina.com [College of Chemistry, Changchun Normal University, Changchun 130032 (China); Wang Yu; Yan Lili; Wang Tianjiao [College of Chemistry, Changchun Normal University, Changchun 130032 (China)

    2012-04-15

    The interactions of silymarin with bovine serum albumin (BSA) and lysozyme (LYS) were investigated in physiological buffer (pH = 7.4) by fluorescence spectroscopy and UV-vis absorption spectroscopy. The mechanism study indicated that silymarin could strongly quench the intrinsic fluorescence of BSA and LYS through static quenching procedures. At 291 K, the values of the binding constant K{sub A} were 4.20 Multiplication-Sign 10{sup 4} and 4.71 Multiplication-Sign 10{sup 4} L mol{sup -1} for silymarin-BSA and silymarin-LYS, respectively. Using thermodynamic equations, the conclusion that hydrophobic and electrostatic forces played an important role in stabilizing complex of silymarin-BSA or silymarin-LYS was obtained. The effects of Cu{sup 2+}, Mg{sup 2+}, Ca{sup 2+}, Fe{sup 2+}, and Fe{sup 3+} on the binding were also studied at 291 K. According to Foerster's nonradiative energy transfer theory, the distances r{sub 0} between donor and acceptor were calculated to be 3.36 and 2.71 nm for silymarin-BSA and silymarin-LYS, respectively. Synchronous fluorescence spectra showed that the conformation of BSA and LYS were changed by silymarin. - Highlights: Black-Right-Pointing-Triangle Quenchings of BSA and LYS fluorescence by silymarin were all static quenchings. Black-Right-Pointing-Triangle Binding constants, binding sites, and thermodynamic parameters were calculated. Black-Right-Pointing-Triangle Hydrophobic and electrostatic forces were the major forces in the two systems. Black-Right-Pointing-Triangle The binding of silymarin to BSA and LYS changed the conformation of BSA and LYS. Black-Right-Pointing-Triangle Energy transfer occurred between silymarin and protein.

  6. 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.

  7. Serum Amyloid P Component Ameliorates Neurological Damage Caused by Expressing a Lysozyme Variant in the Central Nervous System of Drosophila melanogaster.

    Directory of Open Access Journals (Sweden)

    Linda Helmfors

    Full Text Available Lysozyme amyloidosis is a hereditary disease in which mutations in the gene coding for lysozyme leads to misfolding and consequently accumulation of amyloid material. To improve understanding of the processes involved we expressed human wild type (WT lysozyme and the disease-associated variant F57I in the central nervous system (CNS of a Drosophila melanogaster model of lysozyme amyloidosis, with and without co-expression of serum amyloid p component (SAP. SAP is known to be a universal constituent of amyloid deposits and to associate with lysozyme fibrils. There are clear indications that SAP may play an important role in lysozyme amyloidosis, which requires further elucidation. We found that flies expressing the amyloidogenic variant F57I in the CNS have a shorter lifespan than flies expressing WT lysozyme. We also identified apoptotic cells in the brains of F57I flies demonstrating that the flies' neurological functions are impaired when F57I is expressed in the nerve cells. However, co-expression of SAP in the CNS prevented cell death and restored the F57I flies' lifespan. Thus, SAP has the apparent ability to protect nerve cells from damage caused by F57I. Furthermore, it was found that co-expression of SAP prevented accumulation of insoluble forms of lysozyme in both WT- and F57I-expressing flies. Our findings suggest that the F57I mutation affects the aggregation process of lysozyme resulting in the formation of cytotoxic species and that SAP is able to prevent cell death in the F57I flies by preventing accumulation of toxic F57I structures.

  8. C-terminus of TRAP in Staphylococcus can enhance the activity of lysozyme and lysostaphin

    Institute of Scientific and Technical Information of China (English)

    Guang Yang; Ningsheng Shao; Yaping Gao; Jiannan Feng; Yong Huang; Shaohua Li; Yu Liu; Chuan Liu; Ming Fan; Beifen Shen

    2008-01-01

    In Staphylococcus aureus, the target of RNAⅢ activating protein (TRAP) is a membrane-associated protein whose Cterminus can be used as a vaccine to provide protection against staphylococcal infection. Here, we show for the first time by surface plasmon resonance and enzyme-linked immunosorbent assay that TRAP can specifically bind lysozyme and iysostaphin through its C-terminus (amino acids 155-167) and enhance lysozomal activities in vitro. It was also found that the traP mutant strain is more resistant to iysostaphin than wild-type. Our previous data showed that the C-terminus of TRAP might be extracellular. So our results suggested that the C-terminus of TRAP could act as the specific targeting protein of the lysozyme/lysostaphin on the S. aureus cell wall and the biological significance of the interaction might be to facilitate lysozyme/lysostaphin-mediated cell iysis.

  9. cDNA cloning, expression and antibacterial activity of lysozyme C in the blue shrimp (Litopenaeus stylirostris)

    Institute of Scientific and Technical Information of China (English)

    Weijun Mai; Chaoqun Hu

    2009-01-01

    The gene coding for lysozyme in blue shrimp (Litopenaeus stylirostris) was cloned, sequenced and expressed in pET-32a vector. The deduced amino acid sequence of F. Merguiensi lysozyme showed 37-93% similarity with the mouse, human, chicken, and tiger prawn counterparts. The lysozyme was purified to homogeneity and observed as a band of approximately 15 kDa in 15% SDS-PAGE. Semi-quantitative RT-PCR analysis demonstrated that mRNA transcripts of lysozyme could be mainly detected in the tissues of haemocytes, gill, gonad and the lymphoid organ of unchallenged shrimps, whereas the expression of lysozyme transcripts was increased in all the tested tissues after the heat-killed Vibrio alginolyticus challenge. The temporal expression of lysozyme mRNA in haemolymph challenged by Micrococcus luteus and V. Alginolyticus was both up-regulated and reached the maximum level at 8 and 16 h post-stimulation, respec-tively, and then dropped back to the original level. Bacteriolytic activity of the lysozyme against different bacterial cultures was deter-mined by the solid phase and turbidimetric assays. The results demonstrated that the lysozyme we obtained was not only against Gram-positive and Gram-negative bacteria but also against shrimp pathogens V. Alginolyticus and V. Parahemolyticus. In addition, the study of the inhibition mechanism revealed that the antibacterial activity of the lysozyme was a result of the bactericidal effect.

  10. Lysozyme-mediated biomineralization of titanium-tungsten oxide hybrid nanoparticles with high photocatalytic activity.

    Science.gov (United States)

    Kim, Jung Kyu; Jang, Ji-ryang; Choi, Noori; Hong, Dahyun; Nam, Chang-Hoon; Yoo, Pil J; Park, Jong Hyeok; Choe, Woo-Seok

    2014-10-21

    Titanium-tungsten oxide composites with greatly enhanced photocatalytic activity were synthesized by lysozyme-mediated biomineralization. It was shown for the first time that simple control of the onset of biomineralization could enable fine tuning of the composition and crystallinity of the composites to determine their photocatalytic performance.

  11. Enhancement of nisin, lysozyme, and monolaurin antimicrobial activities by ethylenediaminetetraacetic acid and lactoferrin.

    Science.gov (United States)

    Branen, Jill K; Davidson, P Michael

    2004-01-01

    A microtiter plate assay was employed to systematically assess the interaction between ethylenediaminetetraacetic acid (EDTA) or lactoferrin and nisin, lysozyme, or monolaurin against strains of Listeria monocytogenes, Escherichia coli, Salmonella enteritidis, and Pseudomonas fluorescens. Low levels of EDTA acted synergistically with nisin and lysozyme against L. monocytogenes but EDTA and monolaurin interacted additively against this microorganism. EDTA synergistically enhanced the activity of nisin, monolaurin, and lysozyme in tryptic soy broth (TSB) against two enterohemorrhagic E. coli strains. In addition, various combinations of nisin, lysozyme, and monolaurin with EDTA were bactericidal to some gram-negative bacteria whereas none of the antimicrobials alone were bactericidal. Lactoferrin alone (2000 microg ml(-1)) did not inhibit any of the bacterial strains, but did enhance nisin activity against both L. monocytogenes strains. Lactoferrin in combination with monolaurin inhibited growth of E. coli O157:H7 but not E. coli O104:H21. While lactoferrin combined with nisin or monolaurin did not completely inhibit growth of the gram-negative bacteria, there was some growth inhibition. All combinations of EDTA or lactoferrin with antimicrobials were less effective in 2% fat UHT milk than in TSB. S. enteritidis and P. fluorescens strains were consistently more resistant to antimicrobial combinations. Resistance may be due to differences in the outer membrane and/or LPS structure.

  12. Chicken-type lysozyme in channel catfish: Expression analysis, lysozyme activity and efficacy as immunostimulant against Aeromonas hydrophila infection

    Science.gov (United States)

    To understand whether chicken-type lysozyme (Lys-c) in channel catfish was induced by infection of Aeromonas hydrophila, the transcriptional levels of Lys-c in skin, gut, liver, spleen, posterior kidney, and blood cells in healthy channel catfish was compared to that in channel catfish infected with...

  13. Involvement of PKA, PKC, and Ca2+ in LPS-activated expression of the chicken lysozyme gene.

    Science.gov (United States)

    Regenhard, P; Goethe, R; Phi-van, L

    2001-04-01

    The lysozyme gene is activated in myelomonocytic HD11 cells in response to LPS. In this study, we described the involvement of LPS-activated signal transduction pathways in activation of the lysozyme gene. Pre-treatment of HD 11 cells with H-89, H-7, TMB-8, or KN-93 resulted in inhibition of the LPS-enhanced lysozyme expression, suggesting that PKA, PKC, and Ca2+-dependent protein kinases participate in the LPS activation. CaMKII seems to be required for the processing of lysozyme transcripts. TPA and calcium ionophore A23187, when separately added to HD11 cells, stimulated the lysozyme expression effectively, and forskolin was ineffective. It is interesting that simultaneous treatment of cells with forskolin and calcium ionophore A23187 resulted in a potentiated increase in lysozyme mRNA expression, indicating a synergistic cooperation of PKA and Ca2+. This synergistic effect of PKA and Ca2+ was observed on the expression of a stably integrated CAT construct, controlled by the lysozyme promoter and the -6.1-kb enhancer containing binding sites for C/EBP and NF-kappaB/Rel. Therefore, we discussed the role of C/EBPbeta(NF-M), CREB, and NF-kappaB/Rel as possible targets for phosphorylation mediated by PKA, PKC, and Ca2+.

  14. Interactions between high pressure homogenization and antimicrobial activity of lysozyme and lactoperoxidase.

    Science.gov (United States)

    Vannini, L; Lanciotti, R; Baldi, D; Guerzoni, M E

    2004-07-15

    It was the objective of this work to evaluate the effect of high pressure homogenization on the activity of antimicrobial enzymes such as lysozyme and lactoperoxidase against a selected group of Gram positive and Gram negative species inoculated in skim milk. Lactobacillus helveticus, Lactobacillus plantarum and Listeria monocytogenes were the most pressure resistant species while Bacillus subtilis, Pseudomonas putida, Salmonella typhimurium, Staphylococcus aureus, Proteus vulgaris and Salmonella enteritidis were found to be very sensitive to the hyperbaric treatment. The enzyme addition enhanced the instantaneous pressure efficacy on almost all the considered species as indicated by their instantaneous viability loss following the treatment. Moreover, the combination of the enzyme and high pressure homogenization significantly affected the recovery and growth dynamics of several of the considered species. Although L. monocytogenes was slightly sensitive to pressure, the combination of the two stress factors induced a significant viability loss within 3 h and an extension of lag phases in skim milk during incubation at 37 degrees C. The hypothesis formulated in this work is that the interaction of high pressure homogenization and lysozyme or lactoperoxidase is associated to conformational modifications of the two proteins with a consequent enhancement of their activity. This hypothesis is supported by the experimental results also regarding the increased antimicrobial activity against L. plantarum of the previously pressurised lysozyme with respect to that of the native enzyme.

  15. Lysozyme expression in Lactococcus lactis

    NARCIS (Netherlands)

    Guchte, Maarten van de; Wal, Fimme Jan van der; Kok, Jan; Venema, Gerhardus

    1992-01-01

    Three lysozyme-encoding genes, one of eukaryotic and two of prokaryotic origin, were expressed in Lactococcus lactis subsp. lactis. Hen egg white lysozyme (HEL) could be detected in L. lactis lysates by Western blotting. No lysozyme activity was observed, however, presumably because of the absence o

  16. Antibacterial activity of hen egg white lysozyme modified by heat and enzymatic treatments against oenological lactic acid bacteria and acetic acid bacteria.

    Science.gov (United States)

    Carrillo, W; García-Ruiz, A; Recio, I; Moreno-Arribas, M V

    2014-10-01

    The antimicrobial activity of heat-denatured and hydrolyzed hen egg white lysozyme against oenological lactic acid and acetic acid bacteria was investigated. The lysozyme was denatured by heating, and native and heat-denatured lysozymes were hydrolyzed by pepsin. The lytic activity against Micrococcus lysodeikticus of heat-denatured lysozyme decreased with the temperature of the heat treatment, whereas the hydrolyzed lysozyme had no enzymatic activity. Heat-denatured and hydrolyzed lysozyme preparations showed antimicrobial activity against acetic acid bacteria. Lysozyme heated at 90°C exerted potent activity against Acetobacter aceti CIAL-106 and Gluconobacter oxydans CIAL-107 with concentrations required to obtain 50% inhibition of growth (IC50) of 0.089 and 0.013 mg/ml, respectively. This preparation also demonstrated activity against Lactobacillus casei CIAL-52 and Oenococcus oeni CIAL-91 (IC50, 1.37 and 0.45 mg/ml, respectively). The two hydrolysates from native and heat-denatured lysozyme were active against O. oeni CIAL-96 (IC50, 2.77 and 0.3 mg/ml, respectively). The results obtained suggest that thermal and enzymatic treatments increase the antibacterial spectrum of hen egg white lysozyme in relation to oenological microorganisms.

  17. 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.

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

    Institute of Scientific and Technical Information of China (English)

    Viswanathan Karthik; Thomas Ancy; Dharmaraj Ramkumar; Narayanasamy Mathivanan; Rangarajan Badri Narayanan

    2014-01-01

    Objective: To assess the multitudinal antimicrobial effects of recombinant lysozyme fromFenneropenaeus 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 testedKlebsiella 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.

  19. Correlation of Conformational Changes and Protein Degradation with Loss of Lysozyme Activity Due to Chlorine Dioxide Treatment.

    Science.gov (United States)

    Ooi, Beng Guat; Branning, Sharon Alyssa

    2016-12-13

    Chlorine dioxide (ClO2) is a potent oxidizing agent used for the treatment of drinking water and decontamination of facilities and equipment. The purpose of this research is to elucidate the manner in which ClO2 destroys proteins by studying the effects of ClO2 on lysozyme. The degree of enzyme activity lost can be correlated to the treatment time and levels of the ClO2 used. Lysozyme activity was drastically reduced to 45.3% of original enzyme activity when exposed to 4.3 mM ClO2 in the sample after 3 h. Almost all activities were lost in 3 h after exposure to higher ClO2 concentrations of up to 16.8 and 21.9 mM. Changes in protein conformation and amount as a result of ClO2 treatment were determined using the Raman spectroscopy and gel electrophoresis. Raman shifts and the alteration of spectral features observed in the ClO2-treated lysozyme samples are associated with loss of the α-helix secondary structure, tertiary structure, and disulfide bond. Progressive degradation of the denatured lysozyme by increasing levels of chlorine dioxide was also observed in gel electrophoresis. Hence, ClO2 can effectively cause protein denaturation and degradation resulting in loss of enzyme activity.

  20. Production of active lysozyme films by matrix assisted pulsed laser evaporation at 355 nm

    DEFF Research Database (Denmark)

    Purice, Andreea; Schou, Jørgen; Kingshott, P.;

    2007-01-01

    Thin lysozyme films have been produced in a dry environment by MAPLE (matrix assisted pulsed laser evaporation) from a water ice matrix irradiated by laser light at 355 nm above the absorption threshold of the protein. A significant part of the lysozyme molecules are transferred to the film without...

  1. 蛋清、重组溶菌酶酶学性质及其对小鼠肠道菌群和血清生化指标的影响%The Enzymatic Properties of Two Kinds Lysozyme and Their Effects on the Intestinal Flora and Serum Biochemical Indicators of Mice

    Institute of Scientific and Technical Information of China (English)

    田洪源; 刘明启; 戴贤君

    2012-01-01

    Objective: The enzymatic properties were studied to determine the optimum condition of activity for egg white lysozyme extracted from the eggs and recombinant lysozyme expressed from the Pichia pastoris including human lysozyme gene. The intestinal Escherichia coli., Staphylococcus aureus, Enterococcus faecalis, Bifidobacterium and Lacto-bacilliis were cultured, and serum biochemical indicators were determined for the mice orally administered with two lysozymes. Methods: Based on the turbidimetry value of micrococcus, lysozyme activity was determined under different reaction conditions (temperature, pH value, artificial gastric and bowel liquid, and common metal ions); Mice were orally administered with the lysozymes through drinking water including the egg white lysozyme and recombinant lysozyme of 2 500 and 5 000 U/mL. The serum were collected to determinate the concentration of total protein, urea nitrogen, triglyc-eride and cholesterol through picking eye ball of the mice at the experimental time of 1,7,14, 21 days, then the mice were slaughtered for the intestinal contents taken out in sterile environment to culture the Escherichia coli., Staphylococcus aureus, Enterococcus faecalis, Bifidobacterium and Lactobacillus. Results; Egg white lysozyme exhibited the optimal temperature was 40 ℃, and the optimal pH value was 6.0; Recombinant lysozyme exhibited the optimal temperature was 45 ℃, and the optimal pH value was 6.0. Both lysozyme activity would decline rapidly above 50℃, but keep high in the different pH6~9 conditions. Their activities would decline rapidly when both lysozyme were kept in artificial gastric and bowel liquid, only recombinant lysozyme would contain 44.85% activity in artificial bowel liquid after 120min; The Fe3+, Ca2+, K+, Mn2+, Zn2+, Mg2+, Al3+, Cu2+ ions could restrain the lysozyme activity which had no significant difference between two lysozymes(f>0.05). Compared with control group, both lysozyme can restrain the growth of

  2. Influence of Dietary Dinamune® on Growth Performance and Lysozyme Activity in Rainbow Trout (Oncorhynchus mykiss Fry

    Directory of Open Access Journals (Sweden)

    Sudabeh RAMEZANI

    2014-05-01

    Full Text Available This study was conducted to evaluate the effect of a commercial β-glucan based immunostimulant preparation, Dinamune®, in the form of a feed supplement, on the growth performance and lysozyme activity of rainbow trout fry (Oncorhynchus mykiss weighing 1 g. Fish were fed diets containing 0 (control, 0.5, 1 and 1.5 g Dinamune® kg-1 of dry diet for a period of 11 weeks. Body weight was generally increased in fish which were fed diets supplemented with concentrations (1.5 Dinamune®/kg feed (23.75 ± 7.48, compared with the control (21.38 ± 7.04 (p < 0.05. Specific growth rate was significantly (p < 0.05 the highest in 1.5 g Dinamune® kg-1 of dry diet (4.27 ± 0.40 and the lowest in the control group (4.13 ± 0.41. FCR value was found to be the best in the 1g kg-1 group (0.67 ± 0.17, followed by the 1.5g kg-1 group (0.73 ± 0.23, the 0.5 g kg-1 group (0.80 ± 0.36 and the control group (1.00 ± 0.25. Hepatosomatic index (HSI showed a significant decrease in 1.5 g Dinamune® kg-1 of dry diet (0.88 ± 0.18, compared with the control (1.15 ± 0.19. Condition factor (CF in the control, 0.5 g kg-1 diet, 1 g kg-1 diet and 1.5 g kg-1 diet was (1.30 ± 0.16, 1.34 ± 0.15, 1.19 ± 0.10 and 1.18 ± 0.10 respectively. Lysozyme activity of the liver, kidney, and serum were significantly stimulated in the 0.5, 1 and 1.5 g Dinamune® supplemented groups at the end of experiment, compared with the control (p < 0.05.doi:10.14456/WJST.2014.77

  3. The balance of flexibility and rigidity in the active site residues of hen egg white lysozyme

    Institute of Scientific and Technical Information of China (English)

    Qi Jian-Xun; Jiang Fan

    2011-01-01

    The crystallographic temperature factors (B factor) of individual atoms contain important information about the thermal motion of the atoms in a macromolecule. Previously the theory of flexibility of active site has been established based on the observation that the enzyme activity is sensitive to low concentration denaturing agents. It has been found that the loss of enzyme activity occurs well before the disruption of the three-dimensional structural scaffold of the enzyme. To test the theory of conformational flexibility of enzyme active site, crystal structures were perturbed by soaking in low concentration guanidine hydrochloride solutions. It was found that many lysozyme crystals tested could still diffract until the concentration of guanidine hydrochloride reached 3 M. It was also found that the B factors averaged over individually collected data sets were more accurate. Thus it suggested that accurate measurement of crystal temperature factors could be achieved for medium-high or even medium resolution crystals by averaging over multiple data sets. Furthermore, we found that the correctly predicted active sites included not only the more flexible residues, but also some more rigid residues. Both the flexible and the rigid residues in the active site played an important role in forming the active site residue network, covering the majority of the substrate binding residues. Therefore, this experimental prediction method may be useful for characterizing the binding site and the function of a protein, such as drug targeting.

  4. 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.

  5. Lysozyme-coated silver nanoparticles for differentiating bacterial strains on the basis of antibacterial activity

    Science.gov (United States)

    Ashraf, Sumaira; Chatha, Mariyam Asghar; Ejaz, Wardah; Janjua, Hussnain Ahmed; Hussain, Irshad

    2014-10-01

    Lysozyme, an antibacterial enzyme, was used as a stabilizing ligand for the synthesis of fairly uniform silver nanoparticles adopting various strategies. The synthesized particles were characterized using UV-visible spectroscopy, FTIR, dynamic light scattering (DLS), and TEM to observe their morphology and surface chemistry. The silver nanoparticles were evaluated for their antimicrobial activity against several bacterial species and various bacterial strains within the same species. The cationic silver nanoparticles were found to be more effective against Pseudomonas aeruginosa 3 compared to other bacterial species/strains investigated. Some of the bacterial strains of the same species showed variable antibacterial activity. The difference in antimicrobial activity of these particles has led to the conclusion that antimicrobial products formed from silver nanoparticles may not be equally effective against all the bacteria. This difference in the antibacterial activity of silver nanoparticles for different bacterial strains from the same species may be due to the genome islands that are acquired through horizontal gene transfer (HGT). These genome islands are expected to possess some genes that may encode enzymes to resist the antimicrobial activity of silver nanoparticles. These silver nanoparticles may thus also be used to differentiate some bacterial strains within the same species due to variable silver resistance of these variants, which may not possible by simple biochemical tests.

  6. Identification of lysozyme activity from two edible bivalves - Perna viridis (Linnaeus) and Meretrix casta (Chemnitz)

    Digital Repository Service at National Institute of Oceanography (India)

    Sharma, S.; Tanu; Chatterji, A.

    Shellfish Immunol., 11, 611–622. Data, S. (2005). Purification of lysozyme from shell liquor of eastern oysters (crassostrea virginica) and its use in antimicrobial films to Preserve smoked fish. Thesis Louisiana State University and Agricultural...

  7. Preparation and antibacterial activity of lysozyme and layered double hydroxide nanocomposites.

    Science.gov (United States)

    Yang, Qin-Zheng; Chang, Ying-Yue; Zhao, Hua-Zhang

    2013-11-01

    It is necessary to develop "green" disinfection technology which does not produce disinfection by-products. Lysozyme-layered double hydroxide nanocomposites (LYZ-LDHs) were prepared by intercalating LYZ in LDH for the first time. Their antibacterial activity was evaluated using staphylococcus aureus as a target. The bacteria removal mechanism was also studied. Characterization of LYZ-LDHs by X-ray diffraction and Fourier transform infrared spectroscopy indicated that LYZ was successfully intercalated in LDH, compressed and deformed without secondary structural change. LYZ-LDHs showed excellent bactericidal effectiveness against staphylococcus aureus. The antibacterial performance of LYZ-LDHs was found to be affected by the LYZ/LDH ratio and the pH of the bacteria-containing water. The bacteria removal efficiency of LYZ-LDHs with LYZ/LDH mass ratio of 0.8 was consistently above 94% over the pH range of 3-9. LYZ-LDHs adsorbed bacteria to their surface by LDH and then killed them by the immobilized LYZ. This new material integrated the bactericidal ability of LYZ and adsorption ability of LDH. Moreover, the antibacterial ability of LYZ-LDHs was persistent and not limited by the adsorption capacity.

  8. Yam tuber mucilage as a candidate substance for saliva substitute: in vitro study of its viscosity and influences on lysozyme and peroxidase activities.

    Science.gov (United States)

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

    2014-03-01

    To investigate the viscosity of yam tuber mucilage (YTM) and its effects on lysozyme and peroxidase activities in solution phase and on surface phase. Two kinds of YTM were extracted, one containing both protein and carbohydrate and the other containing mainly carbohydrate. Hen egg-white lysozyme and bovine lactoperoxidase were used as lysozyme and peroxidase sources, respectively. Viscosity was measured with a cone-and-plate digital viscometer. Lysozyme activity was determined using the turbidimetric method, and peroxidase activity was determined using the NbsSCN assay. Hydroxyapatite beads were used as a solid phase. The viscosity values of YTM followed a pattern of a non-Newtonian fluid. The carbohydrate concentration affected the viscosity values at all shear rates, while the protein concentration affected the viscosity values at low shear rates. It could be suggested that YTM composed of 1.0 mg/ml protein and 1.0 mg/ml carbohydrate has viscosity values similar to those of unstimulated whole saliva at shear rates present at routine oral functions. Hydroxyapatite-adsorbed YTM significantly increased the adsorption and subsequent enzymatic activities of lysozyme, but not those of peroxidase. Yam tuber mucilage has viscoelastic properties similar to those of human saliva and enhances the enzymatic activity of lysozyme on hydroxyapatite surfaces. © 2012 John Wiley & Sons A/S and The Gerodontology Association. Published by John Wiley & Sons Ltd.

  9. Antimicrobial activity and synergism of lactoferrin and lysozyme against cariogenic microorganisms.

    Science.gov (United States)

    de Andrade, Flaviana Bombarda; de Oliveira, Jair Caetano; Yoshie, Marjorie Takei; Guimarães, Bruno Martini; Gonçalves, Rafael Braga; Schwarcz, Waleska Dias

    2014-01-01

    The present study evaluated the antimicrobial in vitro effects of the salivary proteins lactoferrin and lysozyme on microorganisms involved in the carious process, obtaining their minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Streptococcus mutans (ATCC 25175) and Lactobacillus casei (ATCC 7469) were submitted to broth macrodilution of lysozyme at 80 mg/mL and lactoferrin at 200 mg/mL. The tubes were read in a spectrophotometer after they had been incubated at 37 °C for 18 h, in a carbon dioxide chamber, in order to read the MIC. A new subculture was carried on agar plates to obtain the MBC. The agar diffusion method was also tested, using BHI agar with 100 µL of the standardized microbial inocula. Filter-paper disks soaked in 10 µL of the solutions lactoferrin (200 µg/mL) and lysozyme (80 µg/mL) were placed on the agar surface. Inhibition halos were not observed on the plates, showing the absence of the antimicrobial effects of these proteins in this method. The bactericidal and bacteriostatic effects of lysozyme on L. casei were 50.3 mg/mL and 43.1 mg/mL respectively. The bactericidal and bacteriostatic effects on S. mutans were 68.5 mg/mL and 58.7 mg/mL. Lactoferrin did not induce any inhibitory effects on any microorganism, even in the concentration of 200 mg/mL. There was not a synergic antimicrobial effect of proteins, when they were tested together, even in the concentration of 42.8 mg/mL of lysozyme and 114 mg/mL of lactoferrin (the highest values evaluated). S. mutans and L. casei were only inhibited by lysozyme, not affected by lactoferrin and by the synergic use of both proteins.

  10. Enhancing the antimicrobial activity of Sus scrofa lysozyme by N-terminal fusion of a sextuple unique homologous peptide.

    Science.gov (United States)

    Zhu, Dewei; Cai, Guolin; Li, Xiaomin; Lu, Jian; Zhang, Liang

    2017-02-10

    Sus scrofa lysozyme (SSL), an important component of the pig immune system, is a potential candidate to replace antibiotics in feed. However, there is little antimicrobial activity of natural SSL against gram-negative bacteria, which limits its application. In this study, a unique peptide (A-W-V-A-W-K) with antimicrobial activity against gram-negative bacteria was discovered and purified from trypsin hydrolysate of natural SSL. This unique peptide was fused to natural SSL and the recombinant fused SSL exhibited improved activity against gram-negative bacteria. The N-terminal fusion likely increased the membrane penetrability and induced programmed bacterial cell death. The recombinant fused SSL also showed higher activity against some gram-positive bacteria with O-acetylation. By N-terminal fusion of the sextuple peptide, the anti-microbial activity, either to gram-positive or negative bacteria, of the recombinant SSL was higher than the fusion of only one copy of the peptide. This study provides a general, feasible, and highly useful strategy to enhance the antimicrobial activity of lysozyme.

  11. IL-1α-induced microvascular endothelial cells promote neutrophil killing by increasing MMP-9 concentration and lysozyme activity.

    Science.gov (United States)

    Liu, Xiaoye; Dong, Hong; Wang, Mingming; Gao, Ying; Zhang, Tao; Hu, Ge; Duan, Huiqing; Mu, Xiang

    2016-02-01

    The recruitment of neutrophils by endothelial cells during infection has been extensively studied, but little is known about the regulation of neutrophils activity by endothelial cells. To examine the role of microvascular endothelial cells in neutrophil killing, we established a transmigration model using rat intestinal microvascular endothelial cells (RIMVECs) and measured the extracellular and intracellular killing of Escherichia coli, Lactobacillus acidophilus, and Staphylococcus aureus by transendothelial neutrophils. We observed that blood neutrophils engulfed bacteria but did not kill them, and lipopolysaccharide- or hemolysin-injured RIMVECs inhibited the extracellular and intracellular bactericidal activity of transendothelial neutrophils. In comparison, interleukin-1α-induced RIMVECs promoted the extracellular and intracellular killing activity of transendothelial neutrophils and significantly increased MMP-9 concentration and lysozyme activity in transendothelial neutrophils (p neutrophils and bacterial toxin damage of endothelial cells led to reduction in bactericidal activity of transendothelial neutrophils. These findings offered new insight into the role of endothelial cells in the bactericidal activity of neutrophils.

  12. Effect of thermal pasteurisation and high-pressure processing on immunoglobulin content and lysozyme and lactoperoxidase activity in human colostrum.

    Science.gov (United States)

    Sousa, Sílvia G; Delgadillo, Ivonne; Saraiva, Jorge A

    2014-05-15

    Human milk, and particularly human colostrum, is the gold standard for newborn nourishment. Colostrum contains the highest concentration of immune factors, being the most potent immune booster known to science. In this work, we investigated Holder pasteurisation and high-pressure processing (HPP) effects on colostral IgA, IgM, IgG, lysozyme and lactoperoxidase. The amount of Igs was significantly decreased after Holder pasteurisation (20%, 51% and 23% for IgA, IgM and IgG, respectively), but fully preserved after HPP at 200 and 400 MPa. HPP at 600 MPa for 2.5 min resulted in the maintenance of IgA and losses of IgM and IgG (21% for both). The pressure treatments at 600 MPa for 15 and 30 min led to similar or higher losses than pasteurisation. D-values (min) for Igs ranged from 4941 to 452 at 400 MPa and from 235 to 40 at 600 MPa. Lysozyme activity was lost after pasteurisation (decreased 44%) and maintained after HPP. Lactoperoxidase activity was not detected. As far as the authors are aware, this is the first study evaluating HPP effects on human colostrum.

  13. A new family of lysozyme inhibitors contributing to lysozyme tolerance in gram-negative bacteria.

    Directory of Open Access Journals (Sweden)

    Lien Callewaert

    2008-03-01

    Full Text Available Lysozymes are ancient and important components of the innate immune system of animals that hydrolyze peptidoglycan, the major bacterial cell wall polymer. Bacteria engaging in commensal or pathogenic interactions with an animal host have evolved various strategies to evade this bactericidal enzyme, one recently proposed strategy being the production of lysozyme inhibitors. We here report the discovery of a novel family of bacterial lysozyme inhibitors with widespread homologs in gram-negative bacteria. First, a lysozyme inhibitor was isolated by affinity chromatography from a periplasmic extract of Salmonella Enteritidis, identified by mass spectrometry and correspondingly designated as PliC (periplasmic lysozyme inhibitor of c-type lysozyme. A pliC knock-out mutant no longer produced lysozyme inhibitory activity and showed increased lysozyme sensitivity in the presence of the outer membrane permeabilizing protein lactoferrin. PliC lacks similarity with the previously described Escherichia coli lysozyme inhibitor Ivy, but is related to a group of proteins with a common conserved COG3895 domain, some of them predicted to be lipoproteins. No function has yet been assigned to these proteins, although they are widely spread among the Proteobacteria. We demonstrate that at least two representatives of this group, MliC (membrane bound lysozyme inhibitor of c-type lysozyme of E. coli and Pseudomonas aeruginosa, also possess lysozyme inhibitory activity and confer increased lysozyme tolerance upon expression in E. coli. Interestingly, mliC of Salmonella Typhi was picked up earlier in a screen for genes induced during residence in macrophages, and knockout of mliC was shown to reduce macrophage survival of S. Typhi. Based on these observations, we suggest that the COG3895 domain is a common feature of a novel and widespread family of bacterial lysozyme inhibitors in gram-negative bacteria that may function as colonization or virulence factors in bacteria

  14. Effects of cortisol and stress on channel catfish (Ictalurus punctatus) pathogen susceptibility and lysozyme activity following exposure to Edwardsiella ictaluri.

    Science.gov (United States)

    Small, Brian C; Bilodeau, A Lelania

    2005-05-15

    Periods of stress are often associated with disease outbreaks in cultured fish, and stress is often characterized by the secretion of cortisol. Although stress and cortisol secretion are highly correlated in fish, the role of cortisol in affecting channel catfish (Ictalurus punctatus) pathogen susceptibility is unclear. The effects of short-term stress and exogenous cortisol administration on channel catfish susceptibility to Edwardsiella ictaluri, the etiologic agent of enteric septicemia of catfish (ESC), were investigated. Channel catfish were exposed to virulent E. ictaluri following a standardized 30-min low-water stress or administration of dietary cortisol (100 mg/kg feed) and compared to a pathogen-challenged control group of catfish. Pathogen susceptibility increased in stressed catfish (43.3% mortality) when compared to cortisol-fed catfish (26.7%) and controls (26.7%). A greater (Pcortisol-fed catfish (13.0%) over the course of the study, however, average levels of circulating bacteria were not different (P>0.05) among the treatments. Catfish challenged by the low-water stress event had elevated (Pcortisol 1-day post-pathogen exposure and elevated (Pcortisol-fed and control-challenged catfish. Cortisol concentrations were not correlated (P>0.05) to either lysozyme activity or bacterial levels; however, lysozyme activity was positively correlated (P=0.0197) to blood bacterial concentrations. These results implicate other stress factors or pathways, separate from or possibly in conjunction with cortisol, in the stress-associated immunosuppression of channel catfish as it relates to ESC susceptibility.

  15. Serum paraoxonase 1 activity in dogs

    DEFF Research Database (Denmark)

    Rossi, Gabriele; Giordano, Alessia; Pezzia, Francesca

    2013-01-01

    Serum activity of paraoxonase (PON1) decreases during inflammation in many species. Little information is available on paraoxon-based tests and the possible role of PON1 in dogs.......Serum activity of paraoxonase (PON1) decreases during inflammation in many species. Little information is available on paraoxon-based tests and the possible role of PON1 in dogs....

  16. A novel analytical procedure for assaying lysozyme activity using an end-blocked chitotetraose derivative as substrate.

    Science.gov (United States)

    Ogata, Makoto; Matsui, Megumi; Kono, Haruka; Matsuzaki, Yuka; Kato, Yuna; Usui, Taichi

    2017-09-23

    An end-modified β-d-galactosyl chitotetraose derivative [4(4)-O-β-d-galactosyl-β-tri-N-acetylchitotriosyl 2-acetamide-2,3-dideoxy-glucopyranose; Gal(GlcN)3D] was designed and synthesized from chitin tetrasaccharide. The derivative was chemically modified by dehydration of the reducing end GlcN and enzymatic addition of a Gal group to the non-reducing end GlcN. Hydrolysis of Gal(GlcN)3D and related compounds using hen egg-white lysozyme was then examined. Gal(GlcN)3D was specifically cleaved to Gal(GlcN)2 and GlcND. Kinetic studies and docking simulations were further conducted to elucidate its mode of binding to lysozyme. These analyses revealed the binding of Gal(GlcN)3D to lysozyme is more favorable than that of (GlcN)4D. We conclude the 4-O-substituted Gal group at the non-reducing end of Gal(GlcN)3D does not prohibit the action of lysozyme, but gives some affinity to the subsite (i.e. equivalent to GlcN). From these results, a new assay method for quantifying lysozyme was established by utilizing the Morgan-Elson reaction based on the generation of product D (2-acetamide-2,3-dideoxy-glucopyranose), which serves as a chromophore, formed from Gal(GlcN)3D by lysozyme through a conjugated reaction involving β-N-acetylhexosaminidase. The assay system gave a linear dose-response curve in the range of 2-31 μg of lysozyme during a 15 min incubation. This novel assay method for the quantification of lysozyme is highly specific, sensitive, accurate and reproducible. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. 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.

  18. Baicalin promotes the bacteriostatic activity of lysozyme on S. aureus in mammary glands and neutrophilic granulocytes in mice.

    Science.gov (United States)

    Gao, Xuejiao; Guo, Mengyao; Zhang, Zecai; Shen, Peng; Yang, Zhengtao; Zhang, Naisheng

    2017-03-21

    Staphylococcus aureus causes mastitis as a result of community-acquired or nosocomial infections. Lysozyme (LYSO) is an enzyme that is upregulated in many organisms during the innate immune response against infection by bacterial pathogens. Baicalin is a bioactive flavonoid that can bind to enzymes, often to potentiate their effect. Here we tested the effects of baicalin on the activity of LYSO using the S. aureus mastitis mouse model and neutrophilic granulocyte model of S. aureus infection. In our experiments, S. aureus counts decreased with increasing baicalin concentration. Furthermore, qPCR and western blot analyses showed that LYSO expression was unaffected by baicalin, while fluorescence quenching and UV fluorescence spectral analyses showed that baicalin binds to LYSO. To test whether this binding increased LYSO activity, we assessed LYSO-induced bacteriostasis in the presence of baicalin. Our results showed that LYSO-induced S. aureus bacteriostasis increased with increasing concentrations of baicalin, and that baicalin binding to LYSO synergistically increased the antibacterial activity of LYSO. These results demonstrate that baicalin enhances LYSO-induced bacteriostasis during the innate immune response to S. aureus. They suggest baicalin is a potentially useful therapeutic agent for the treatment of bacterial infections.

  19. Study on Antimicrobial and Antiviral Activities of Lysozyme From Marine Strain S-12-86 In Vitro

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    In this study, the in vitro antimicrobial and antiviral activities of the lysozyme from marine strain S-12-86 (LS) were investigated. The antimicrobial activity of LS was tested by minimum inhibition concentration (MIC) and minimum bactericidal concentration (MBC) method. The inhibiting effects of LS on pseudo rabies virus (PRV) in swine kidney cells (PK-15 cells) were judged by cytopathogenic effect test (CPE). The results showed LS had a broad antimicrobial spectrum against several standard strains including gram-positive bacteria, gram-negative bacteria, fungi, etc. The MIC of LS was 0.25-4.00 mg mL-1 and its MBC was 0.25-8.00 mg mL-1, respectively. Observation under the transmission electron microscope revealed that the cell wall of Candida albicans was distorted seriously, and the cytoplasm with many cavities was asymmetrical after being hydrolyzed by LS. The median cytotoxicity concentration (TC50) of LS was 100.0 μg mL-1, the median effective concentration (EC50) was 0.46 μg mL-1, and the selectivity index (TI = TC50/EC50) was 217. LS could inhibit PRV in PK-15 cells when it was added to cell culture medium at 0, 2, 4, 6, and 8 h after PK-15 cells had been infected by PRV. From the results, we concluded that LS had broad antimicrobial spectrum and good inhibiting effects on PRV.

  20. Different Analytical Approaches in Assessing Antibacterial Activity and the Purity of Commercial Lysozyme Preparations for Dairy Application

    Directory of Open Access Journals (Sweden)

    Luisa Pellegrino

    2013-05-01

    Full Text Available Hen egg-white lysozyme (LSZ is currently used in the food industry to limit the proliferation of lactic acid bacteria spoilage in the production of wine and beer, and to inhibit butyric acid fermentation in hard and extra hard cheeses (late blowing caused by the outgrowth of clostridial spores. The aim of this work was to evaluate how the enzyme activity in commercial preparations correlates to the enzyme concentration and can be affected by the presence of process-related impurities. Different analytical approaches, including turbidimetric assay, SDS-PAGE and HPLC were used to analyse 17 commercial preparations of LSZ marketed in different countries. The HPLC method adopted by ISO allowed the true LSZ concentration to be determined with accuracy. The turbidimetric assay was the most suitable method to evaluate LSZ activity, whereas SDS-PAGE allowed the presence of other egg proteins, which are potential allergens, to be detected. The analytical results showed that the purity of commercially available enzyme preparations can vary significantly, and evidenced the effectiveness of combining different analytical approaches in this type of control.

  1. Serum adenosine deaminase activity in cutaneous anthrax.

    Science.gov (United States)

    Sunnetcioglu, Mahmut; Karadas, Sevdegul; Aslan, Mehmet; Ceylan, Mehmet Resat; Demir, Halit; Oncu, Mehmet Resit; Karahocagil, Mustafa Kasım; Sunnetcioglu, Aysel; Aypak, Cenk

    2014-07-06

    Adenosine deaminase (ADA) activity has been discovered in several inflammatory conditions; however, there are no data associated with cutaneous anthrax. The aim of this study was to investigate serum ADA activity in patients with cutaneous anthrax. Sixteen patients with cutaneous anthrax and 17 healthy controls were enrolled. We measured ADA activity; peripheral blood leukocyte, lymphocyte, neutrophil, and monocyte counts; erythrocyte sedimentation rate; and C reactive protein levels. Serum ADA activity was significantly higher in patients with cutaneous anthrax than in the controls (panthrax.

  2. Recent advances for the production and recovery methods of lysozyme.

    Science.gov (United States)

    Ercan, Duygu; Demirci, Ali

    2016-12-01

    Lysozyme is an antimicrobial peptide with a high enzymatic activity and positive charges. Therefore, it has applications in food and pharmaceutical industries as an antimicrobial agent. Lysozyme is ubiquitous in both animal and plant kingdoms. Currently, egg-white lysozyme is the most commercially available form of lysozyme. The main concerns of egg-white lysozyme are high recovery cost, low activity and most importantly the immunological problems to some people. Therefore, human lysozyme production has gained importance in recent years. Scientists have developed transgenic plants, animals and microorganisms that can produce human lysozyme. Out of these, microbial production has advantages for commercial productions, because high production levels are achievable in a relatively short time. It has been reported that fermentation parameters, such as pH, temperature, aeration, are key factors to increase the effectiveness of the human lysozyme production. Moreover, purification of the lysozyme from the fermentation broth needs to be optimized for the economical production. In conclusion, this review paper covers the mechanism of lysozyme, its sources, production methods and recovery of lysozyme.

  3. 溶菌酶酶解物的抗菌活性研究%Study on the antibacterial activity of lysozyme hydrolysate

    Institute of Scientific and Technical Information of China (English)

    朱伶俐; 朱明捷; 杨严俊

    2012-01-01

    Objective:Provide the theoretical foundation for broadening the application of lysozyme in food industry through researching on the bacteriostatic ability of Lysozyme hydrolysate. Methods:Lysozyme hydrolysate(LH) was obtained by using pepsin. The antibacterial activity of LH was tested by antibacterial circle experiment; meanwhile,lysozyme and nisin were also tested. The minimum bactericide concentration of LH against common food bacteria was determined with the two-fold dilution method. Results:LH showed great inhibition effect on positive bacteria,especially staphylococcus aureus. Without heat treatment,the bacteriostatic ability of LH on staphylococcus aureus was weaker than lysozyme,but much stronger than nisin. After heat treatment,LH was more antibacterial than lysozyme and nisin. LH was promising to be a natural food preservative.%目的:考察溶菌酶酶解物的抑菌能力.为扩展溶菌酶的应用提供理论依据。方法:采用胃蛋白酶对溶菌酶进行酶解,得到溶菌酶酶解物(Lysozymehydrolysate,LH);采用二倍稀释法检测LH对几种常见食品腐败菌的抑菌能力;以金黄色葡萄球菌为受试菌种,通过抑菌圈实验探讨LH的抑菌活性,并与溶菌酶和乳酸链球菌素作比较。结果:LH对几种常见阳性菌都有很强的抑制作用,其中对金黄色葡萄球菌的抑制作用最强;未经热处理时,LH的抑菌活性略低于溶菌酶,而高于乳酸链球菌素;经热处理后,LH的抑菌活性高于溶菌酶和乳酸链球菌素。菌酶酶解物有望被开发成一种天然的食品防腐剂。

  4. 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.

  5. Effects of dimerized lysozyme (KLP-602) on the cellular and humoral defence mechanisms in sheatfish (Silurus glanis): in vitro and in vivo study.

    Science.gov (United States)

    Morand, M; Siwicki, A; Pozet, F; Klein, P; Vinaize, J C; Keck, N

    1999-01-01

    This study examined the effects of the dimerized lysozyme (KLP-602) on the immunocompetence cell activity in sheatfish (Silurus glanis) and its influence in vivo on the non-specific defence mechanisms and protection against motile aeromonad septicaemia (MAS). The in vitro study showed that the lysozyme dimer (KLP-602), at concentrations between 5 and 50 micrograms/mL of medium significantly (P < 0.05) increased the respiratory burst activity and potential killing activity of pronephric macrophages, as well as the proliferative ability of pronephric lymphocytes stimulated by ConA and LPS. The in vivo study showed that injecting lysozyme dimer (Lydium-KLP) intraperitoneally at doses of 50 micrograms/kg bw stimulated cell-mediated and humoral-mediated imunity. On day 5, after application of Lydium-KLP in vivo, a statistically higher (P < 0.05) respiratory burst activity and potential killing activity of blood and pronephros phagocytes were observed. A higher proliferative ability of blood and pronephros lymphocytes stimulated by Concanavaline A (ConA) or lipopolysaccharide (LPS) was also observed. At the same time, the myeloperoxidase activity in the PMN cells and the lysozyme activity and total Ig levels in serum were significantly higher (P < 0.05), compared to the control group. A challenge test with Aeromonas hydrophila showed that dimerized lysozyme increased the protection against MAS. Dimerized lysozyme stimulates non-specific cellular and humoral mechanisms and protection against MAS in sheatfish.

  6. CRYSTAL-STRUCTURES OF HEVAMINE, A PLANT DEFENSE PROTEIN WITH CHITINASE AND LYSOZYME ACTIVITY, AND ITS COMPLEX WITH AN INHIBITOR

    NARCIS (Netherlands)

    VANSCHELTINGA, ACT; KALK, KH; BEINTEMA, JJ; DIJKSTRA, BW

    1994-01-01

    Background: Hevamine is a member of one of several families of plant chitinases and lysozymes that are important for plant defence against pathogenic bacteria and fungi. The enzyme can hydrolyze the linear polysaccharide chains of chitin and peptidoglycan. A full understanding of the structure/funct

  7. Influence of Soil Humic and Fulvic Acid on the Activity and Stability of Lysozyme and Urease

    NARCIS (Netherlands)

    Li, Yan; Tan, WenFeng; Koopal, Luuk K.; Wang, MingXia; Liu, Fan; Norde, Willem

    2013-01-01

    Humic substances (HS), including humic acids (HA) and fulvic acids (FA), are important components of soil systems. HS form strong complexes with oppositely charged proteins, which will lead to changes in the enzyme activity. The effect of soil HS on the activity and stability of two enzymes was inve

  8. Influence of soil humic and fulvic acid on the activity and stability of lysozyme and urease

    NARCIS (Netherlands)

    Li, Y.; Tan, W.; Koopal, L.K.; Wang, M.; Liu, Fan; Norde, W.

    2013-01-01

    Humic substances (HS), including humic acids (HA) and fulvic acids (FA), are important components of soil systems. HS form strong complexes with oppositely charged proteins, which will lead to changes in the enzyme activity. The effect of soil HS on the activity and stability of two enzymes was inve

  9. A complex of equine lysozyme and oleic acid with bactericidal activity against Streptococcus pneumoniae.

    Directory of Open Access Journals (Sweden)

    Emily A Clementi

    Full Text Available HAMLET and ELOA are complexes consisting of oleic acid and two homologous, yet functionally different, proteins with cytotoxic activities against mammalian cells, with HAMLET showing higher tumor cells specificity, possibly due to the difference in propensity for oleic acid binding, as HAMLET binds 5-8 oleic acid molecules per protein molecule and ELOA binds 11-48 oleic acids. HAMLET has been shown to possess bactericidal activity against a number of bacterial species, particularly those with a respiratory tropism, with Streptococcus pneumoniae displaying the greatest degree of sensitivity. We show here that ELOA also displays bactericidal activity against pneumococci, which at lower concentrations shows mechanistic similarities to HAMLET's bactericidal activity. ELOA binds to S. pneumoniae and causes perturbations of the plasma membrane, including depolarization and subsequent rupture, and activates an influx of calcium into the cells. Selective inhibition of calcium channels and sodium/calcium exchange activity significantly diminished ELOA's bactericidal activity, similar to what we have observed with HAMLET. Finally, ELOA-induced death was also accompanied by DNA fragmentation into high molecular weight fragments - an apoptosis-like morphological phenotype that is seen during HAMLET-induced death. Thus, in contrast to different mechanisms of eukaryote cell death induced by ELOA and HAMLET, these complexes are characterized by rather similar activities towards bacteria. Although the majority of these events could be mimicked using oleic acid alone, the concentrations of oleic acid required were significantly higher than those present in the ELOA complex, and for some assays, the results were not identical between oleic acid alone and the ELOA complex. This indicates that the lipid, as a common denominator in both complexes, is an important component for the complexes' bactericidal activities, while the proteins are required both to solubilize

  10. A complex of equine lysozyme and oleic acid with bactericidal activity against Streptococcus pneumoniae.

    Science.gov (United States)

    Clementi, Emily A; Wilhelm, Kristina R; Schleucher, Jürgen; Morozova-Roche, Ludmilla A; Hakansson, Anders P

    2013-01-01

    HAMLET and ELOA are complexes consisting of oleic acid and two homologous, yet functionally different, proteins with cytotoxic activities against mammalian cells, with HAMLET showing higher tumor cells specificity, possibly due to the difference in propensity for oleic acid binding, as HAMLET binds 5-8 oleic acid molecules per protein molecule and ELOA binds 11-48 oleic acids. HAMLET has been shown to possess bactericidal activity against a number of bacterial species, particularly those with a respiratory tropism, with Streptococcus pneumoniae displaying the greatest degree of sensitivity. We show here that ELOA also displays bactericidal activity against pneumococci, which at lower concentrations shows mechanistic similarities to HAMLET's bactericidal activity. ELOA binds to S. pneumoniae and causes perturbations of the plasma membrane, including depolarization and subsequent rupture, and activates an influx of calcium into the cells. Selective inhibition of calcium channels and sodium/calcium exchange activity significantly diminished ELOA's bactericidal activity, similar to what we have observed with HAMLET. Finally, ELOA-induced death was also accompanied by DNA fragmentation into high molecular weight fragments - an apoptosis-like morphological phenotype that is seen during HAMLET-induced death. Thus, in contrast to different mechanisms of eukaryote cell death induced by ELOA and HAMLET, these complexes are characterized by rather similar activities towards bacteria. Although the majority of these events could be mimicked using oleic acid alone, the concentrations of oleic acid required were significantly higher than those present in the ELOA complex, and for some assays, the results were not identical between oleic acid alone and the ELOA complex. This indicates that the lipid, as a common denominator in both complexes, is an important component for the complexes' bactericidal activities, while the proteins are required both to solubilize and/or present the

  11. A NF-κB-dependent dual promoter-enhancer initiates the lipopolysaccharide-mediated transcriptional activation of the chicken lysozyme in macrophages.

    Directory of Open Access Journals (Sweden)

    James Witham

    Full Text Available The transcriptional activation of the chicken lysozyme gene (cLys by lipopolysaccharide (LPS in macrophages is dependent on transcription of a LPS-Inducible Non-Coding RNA (LINoCR triggering eviction of the CCCTC-binding factor (CTCF from a negative regulatory element upstream of the lysozyme transcription start site. LINoCR is transcribed from a promoter originally characterized as a hormone response enhancer in the oviduct. Herein, we report the characterization of this cis-regulatory element (CRE. In activated macrophages, a 60 bp region bound by NF-κB, AP1 and C/EBPβ controls this CRE, which is strictly dependent on NF-κB binding for its activity in luciferase assays. Moreover, the serine/threonine kinase IKKα, known to be recruited by NF-κB to NF-κB-dependent genes is found at the CRE and within the transcribing regions of both cLys and LINoCR. Such repartition suggests a simultaneous promoter and enhancer activity of this CRE, initiating cLys transcriptional activation and driving CTCF eviction. This recruitment was transient despite persistence of both cLys transcription and NF-κB binding to the CRE. Finally, comparing cLys with other LPS-inducible genes indicates that IKKα detection within transcribing regions can be correlated with the presence of the elongating form of RNA polymerase II or concentrated in the 3' end of the gene.

  12. A NF-κB-dependent dual promoter-enhancer initiates the lipopolysaccharide-mediated transcriptional activation of the chicken lysozyme in macrophages.

    Science.gov (United States)

    Witham, James; Ouboussad, Lylia; Lefevre, Pascal F

    2013-01-01

    The transcriptional activation of the chicken lysozyme gene (cLys) by lipopolysaccharide (LPS) in macrophages is dependent on transcription of a LPS-Inducible Non-Coding RNA (LINoCR) triggering eviction of the CCCTC-binding factor (CTCF) from a negative regulatory element upstream of the lysozyme transcription start site. LINoCR is transcribed from a promoter originally characterized as a hormone response enhancer in the oviduct. Herein, we report the characterization of this cis-regulatory element (CRE). In activated macrophages, a 60 bp region bound by NF-κB, AP1 and C/EBPβ controls this CRE, which is strictly dependent on NF-κB binding for its activity in luciferase assays. Moreover, the serine/threonine kinase IKKα, known to be recruited by NF-κB to NF-κB-dependent genes is found at the CRE and within the transcribing regions of both cLys and LINoCR. Such repartition suggests a simultaneous promoter and enhancer activity of this CRE, initiating cLys transcriptional activation and driving CTCF eviction. This recruitment was transient despite persistence of both cLys transcription and NF-κB binding to the CRE. Finally, comparing cLys with other LPS-inducible genes indicates that IKKα detection within transcribing regions can be correlated with the presence of the elongating form of RNA polymerase II or concentrated in the 3' end of the gene.

  13. Resistance screening essay of wine lactic acid bacteria on lysozyme: efficacy of lysozyme in unclarified grape musts.

    Science.gov (United States)

    Delfini, Claudio; Cersosimo, Manuela; Del Prete, Vincenzo; Strano, Morela; Gaetano, Giuseppe; Pagliara, Adolfo; Ambrò, Stefano

    2004-04-01

    In wine making, the bacteriolytic activity of lysozyme has primarily been used to control the malolactic fermentation in wines. The use of lysozyme in musts before settling and the beginning of the alcoholic fermentation to inhibit the growth of lactic acid bacteria could be very beneficial. In a resistance test carried out in MT/b broth, lysozyme had greater antimicrobial activity toward Oenococcus oeni than Lactobacillus species. Several strains of wine bacteria belonging to Oenococcus proved sensitive to the bacteriolytic activity of lysozyme at low concentrations in both synthetic medium (MT/b) (50 mg/L), white must, or red must made with or without the skins (100 mg/L). Lactobacillus and Pediococcus strains survived at lysozyme concentrations of 200-500 and 500 mg/L, respectively, in MT/b and musts. Suspended solids in unclarified musts may strongly bind to lysozyme thereby causing its removal by filtration or centrifugation. One hour after lysozyme was added to musts, it was quantified by HPLC and found after centrifugation to be 40-50% and only 10% in musts made with or without the skins, respectively. Although appreciable amounts of lysozyme were bound to wine components, this did not appear to be a serious hindrance to lysozyme activity.

  14. IMMOBILIZATION OF LYSOZYME IN POLYVINYL ALCOHOL CRYOGEL

    Directory of Open Access Journals (Sweden)

    S. S. Dekina

    2014-06-01

    Full Text Available The lysozyme immobilization in cryogel of polyvinyl alcohol and physico-chemical properties of obtained preparation was investigated. Hydrolytic activity of lysozyme was determined by bacteriolytic method, using Micrococcus lysodeikticus cells acetone powder as substrate. Protein content was determined by the Lowry–Hartree method. Immobilization of lysozyme was conducted by entrapment in polyvinyl alcohol gel with subsequent cycles of freezing-thawing. Antimicrobial activity was studied by standard disk-diffusional method. The hydrogel filmic coatings with antimicrobial action, insoluble at physiological conditions, with quantitative retaining of protein and hydrolytic activity of lysozyme were obtained. The product is characterized by the widened pH-profile of activity at acidic pH values, stability in acidic medium (pH 5.5 and at storage. Its antimicrobial action against Staphylococcus aureus ATCC 25923 F-49, Pseudomonas aeruginosa 415, Escherichia coli 055 K 59912/4 and Candida albicans ATCC 885-653 was noted. The proposed method of lysozyme immobilization allows to obtain stable, highly effective product with antimicrobial activity, prospective for usage in biomedical investigations.

  15. Study of lysozyme resistance in Rhodococcus equi.

    Science.gov (United States)

    Hébert, Laurent; Bidaud, Pauline; Goux, Didier; Benachour, Abdellah; Laugier, Claire; Petry, Sandrine

    2014-03-01

    Lysozyme is an important and widespread component of the innate immune response that constitutes the first line of defense against bacterial pathogens. The bactericidal effect of this enzyme relies on its capacity to hydrolyze the bacterial cell wall and also on a nonenzymatic mechanism involving its cationic antimicrobial peptide (CAMP) properties, which leads to membrane permeabilization. In this paper, we report our findings on the lysozyme resistance ability of Rhodococcus equi, a pulmonary pathogen of young foals and, more recently, of immunocompromised patients, whose pathogenic capacity is conferred by a large virulence plasmid. Our results show that (i) R. equi can be considered to be moderately resistant to lysozyme, (ii) the activity of lysozyme largely depends on its muramidase action rather than on its CAMP activity, and (iii) the virulence plasmid confers part of its lysozyme resistance capacity to R. equi. This study is the first one to demonstrate the influence of the virulence plasmid on the stress resistance capacity of R. equi and improves our understanding of the mechanisms enabling R. equi to resist the host defenses.

  16. Inhibitory activity of reuterin, nisin, lysozyme and nitrite against vegetative cells and spores of dairy-related Clostridium species.

    Science.gov (United States)

    Avila, Marta; Gómez-Torres, Natalia; Hernández, Marta; Garde, Sonia

    2014-02-17

    The butyric acid fermentation, responsible for late blowing of cheese, is caused by the outgrowth in cheese of some species of Clostridium, resulting in texture and flavor defects and economical losses. The aim of this study was to evaluate the effectiveness of different antimicrobial compounds against vegetative cells and spores of C. tyrobutyricum, C. butyricum, C. beijerinckii and C. sporogenes strains isolated from cheeses with late blowing defect. Minimal inhibitory concentration (MIC) for reuterin, nisin, lysozyme and sodium nitrite were determined against Clostridium strains in milk and modified RCM (mRCM) after 7d exposure. Although the sensitivity of Clostridium to the tested antimicrobials was strain-dependent, C. sporogenes and C. beijerinckii generally had higher MIC values than the rest of Clostridium species. The majority of Clostridium strains were more resistant to antimicrobials in milk than in mRCM, and vegetative cells exhibited higher sensitivity than spores. Reuterin (MIC values 0.51-32.5 mM) and nisin (MIC values 0.05-12.5 μg/ml) were able to inhibit the growth of vegetative cells and spores of all assayed Clostridium strains in milk and mRCM. Strains of C. tyrobutyricum exhibited the highest sensitivity to lysozyme (MIC valuesClostridium spp. spores and vegetative cells, may be the best options to control Clostridium growth in dairy products and to prevent associated spoilage, such as late blowing defect of cheese. However, further studies in cheese would be necessary to validate this hypothesis.

  17. Purification, amino acid sequence, and some properties of rabbit kidney lysozyme.

    Science.gov (United States)

    Ito, Y; Yamada, H; Nakamura, S; Imoto, T

    1990-02-01

    The lysozyme (rabbit kidney lysozyme) from the homogenate of rabbit kidney (Japanese white) was purified by repeated cation-exchange chromatography on Bio-Rex 70. The amino acid sequence was determined by automated gas-phase Edman degradation of the peptides obtained from the digestion of reduced and S-carboxymethylated rabbit lysozyme with Achromobacter protease I (lysyl endopeptidase). The sequence thus determined was KIYERCELARTLKKLGLDGYKGVSLANWMCLAKWESSYNTRATNYNPGDKSTDYGIFQ INSRYWCNDGKTPRAVNACHIPCSDLLKDDITQAVACAKRVVSDPQGIRAWVAWRNHCQ NQDLTPYIRGCGV, indicating 25 amino acid substitutions from human lysozyme. The lytic activity of rabbit lysozyme against Micrococcus lysodeikticus at pH 7, ionic strength of 0.1, and 30 degrees C was found to be 190 and 60% of those of hen and human lysozymes, respectively. The lytic activity-pH profile of rabbit lysozyme was slightly different from those of hen and human lysozymes. While hen and human lysozymes had wide optimum activities at around pH 5.5-8.5, the optimum activity of rabbit lysozyme was at around pH 5.5-7.0. The high proline content (five residues per molecule compared with two prolines per molecule in hen or human lysozyme) is one of the interesting features of rabbit lysozyme. The transition temperatures for the unfolding of rabbit, human, and hen lysozymes in 3 M guanidine hydrochloride at pH 5.5 were 51.2, 45.5, and 45.4 degrees C, respectively, indicating that rabbit lysozyme is stabler than the other two lysozymes. The high proline content may be responsible for the increased stability of rabbit lysozyme.

  18. Edwardsiella tarda Ivy, a lysozyme inhibitor that blocks the lytic effect of lysozyme and facilitates host infection in a manner that is dependent on the conserved cysteine residue.

    Science.gov (United States)

    Wang, Chong; Hu, Yong-hua; Sun, Bo-guang; Li, Jun; Sun, Li

    2013-10-01

    Edwardsiella tarda is a Gram-negative bacterial pathogen with a broad host range that includes fish and humans. In this study, we examined the activity and function of the lysozyme inhibitor Ivy (named IvyEt) identified in the pathogenic E. tarda strain TX01. IvyEt possesses the Ivy signature motif CKPHDC in the form of (82)CQPHNC(87) and contains several highly conserved residues, including a tryptophan (W55). For the purpose of virulence analysis, an isogenic TX01 mutant, TXivy, was created. TXivy bears an in-frame deletion of the ivyEt gene. A live infection study in a turbot (Scophthalmus maximus) model showed that, compared to TX01, TXivy exhibited attenuated overall virulence, reduced tissue dissemination and colonization capacity, an impaired ability to replicate in host macrophages, and decreased resistance against the bactericidal effect of host serum. To facilitate functional analysis, recombinant IvyEt (rIvy) and three mutant proteins, i.e., rIvyW55A, rIvyC82S, and rIvyH85D, which bear Ala, Ser, and Asp substitutions at W55, C82, and H85, respectively, were prepared. In vitro studies showed that rIvy, rIvyW55A, and rIvyH85D were able to block the lytic effect of lysozyme on a Gram-positive bacterium, whereas rIvyC82S could not do so. Likewise, rIvy, but not rIvyC82S, inhibited the serum-facilitated killing effect of lysozyme on E. tarda. In vivo analysis showed that rIvy, but not rIvyC82S, restored the lost pathogenicity of TXivy and enhanced the infectivity of TX01. Together these results indicate that IvyEt is a lysozyme inhibitor and a virulence factor that depends on the conserved C82 for biological activity.

  19. Adaptive functional diversification of lysozyme in insectivorous bats.

    Science.gov (United States)

    Liu, Yang; He, Guimei; Xu, Huihui; Han, Xiuqun; Jones, Gareth; Rossiter, Stephen J; Zhang, Shuyi

    2014-11-01

    The role of gene duplication in generating new genes and novel functions is well recognized and is exemplified by the digestion-related protein lysozyme. In ruminants, duplicated chicken-type lysozymes facilitate the degradation of symbiotic bacteria in the foregut. Chicken-type lysozyme has also been reported to show chitinase-like activity, yet no study has examined the molecular evolution of lysozymes in species that specialize on eating insects. Insectivorous bats number over 900 species, and lysozyme expression in the mouths of some of these species is associated with the ingestion of insect cuticle, suggesting a chitinase role. Here, we show that chicken-type lysozyme has undergone multiple duplication events in a major family of insect-eating bats (Vespertilionidae) and that new duplicates have undergone molecular adaptation. Examination of duplicates from two insectivorous bats-Pipistrellus abramus and Scotophilus kuhlii-indicated that the new copy was highly expressed in the tongue, whereas the other one was less tissue-specific. Functional assays applied to pipistrelle lysozymes confirmed that, of the two copies, the tongue duplicate was more efficient at breaking down glycol chitin, a chitin derivative. These results suggest that the evolution of lysozymes in vespertilionid bats has likely been driven in part by natural selection for insectivory.

  20. Low serum alkaline phosphatase activity in Wilson's disease.

    Science.gov (United States)

    Shaver, W A; Bhatt, H; Combes, B

    1986-01-01

    Low values for serum alkaline phosphatase activity were observed early in the course of two patients with Wilson's disease presenting with the combination of severe liver disease and Coombs' negative acute hemolytic anemia. A review of other cases of Wilson's disease revealed that 11 of 12 patients presenting with hemolytic anemia had values for serum alkaline phosphatase less than their respective sex- and age-adjusted mean values; in eight, serum alkaline phosphatase activity was less than the lower value for the normal range of the test. Low values for serum alkaline phosphatase were much less common in Wilson's disease patients with more chronic forms of presentation. Copper added in high concentration to serum in vitro did not have an important effect on serum alkaline phosphatase activity. The mechanism responsible for the decrease in serum alkaline phosphatase activity in patients is uncertain.

  1. Lysozymes in the animal kingdom

    Indian Academy of Sciences (India)

    Lien Callewaert; Chris W Michiels

    2010-03-01

    Lysozymes (EC 3.2.1.17) are hydrolytic enzymes, characterized by their ability to cleave the -(1,4)-glycosidic bond between -acetylmuramic acid and -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.

  2. Lysozymes and lysozyme-like proteins from the fall armyworm, Spodoptera frugiperda.

    Science.gov (United States)

    Chapelle, Michael; Girard, Pierre-Alain; Cousserans, François; Volkoff, Nathalie-Anne; Duvic, Bernard

    2009-12-01

    Lysozyme is an important component of the insect non-specific immune response against bacteria that is characterized by its ability to break down bacterial cell-walls. By searching an EST database from the fall armyworm, Spodoptera frugiperda (Negre et al., 2006), we identified five sequences encoding proteins of the lysozyme family. The deduced protein sequences corresponded to three classical c-type lysozymes Sf-Lys1, Sf-Lys2 and Sf-Lys3, and two lysozyme-like proteins, Sf-LLP1 and Sf-LLP2. Sf-Lys1 was purified from the hemolymph of Escherichia coli-challenged S. frugiperda larvae. The mature protein had a molecular mass of 13.975 Da with an isoelectric point of 8.77 and showed 98.3% and 96.7% identity with lysozymes from Spodoptera litura and Spodoptera exigua, respectively. As the other insect lysozymes, Sf-Lys1 was active against gram positive bacteria such as Micrococcus luteus but also induced a slight permeabilization of the inner membrane of E. coli. Genes encoding these five Sf-Lys or Sf-LLPs were differentially up-regulated in three immune-competent tissues (hemocytes, fat body and gut) after challenges with non-pathogenic bacteria, E. coli and M. luteus, or entomopathogenic bacterium, Photorhabdus luminescens. Sf-Lys1 and Sf-Lys2 were mainly induced in fat body in the presence of E. coli or P. luminescens. Sf-Lys3, which had an acidic isoelectric point, was found to be the most up-regulated of all five Sf-Lys or Sf-LLPs in hemocytes and gut after challenge with P. luminescens. More molecular data are now available to investigate differences in physiological functions of these different members of the lysozyme superfamily.

  3. 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.

  4. Differential major histocompatibility complex-related activation of idiotypic suppressor T cells. Suppressor T cells cross-reactive to two distantly related lysozymes are not induced by one of them.

    Science.gov (United States)

    Adorini, L; Harvey, M A; Rozycka-Jackson, D; Miller, A; Sercarz, E E

    1980-09-01

    B10 (H-2b) mice are genetic nonresponders to hen egg-white lysozyme (HEL) and the distantly related human lysozyme (HUL). However, anti-HEL or anti-HUL primary antibody responses in vivo or in vitro can be obtained in B10 mice by immunization with the appropriate lysozyme coupled to erythrocytes. T cells able to suppress either anti-lysozyme plaque-forming cells (PFC) response are induced in B10 mice after immunization with HEL-complete Freund's adjuvant (CFA) or HUL-CFA. This cross-reactivity of HEL and HUL in the induction and the expression of suppressive activity is in marked contrast to their very low cross-reactivity at the PFC level. These results suggest that either HEL or HUL can stimulate a suppressor T cell which recognizes a particular epitope present on both lysozymes. Suppressor cells induced by HEL or HUL bear the same predominant idiotype found on the majority of anti-HEL antibodies, and on the small proportion of anti-HUL antibodies cross-reactive with HEL. B10.Q (H-2q) mice are responders in vivo to HEL-CFA, but not to HUL-CFA. In contrast to B10, HEL-CFA priming in B10.Q micr induces helper cells whereas HUL-CFA priming induces suppressor cells. These suppressor cells are cross-reactive with HEL and are fully able to suppress HEL-specific helper cells. The presence of HEL-specific suppressor cell precursors in B10.Q mice which are not activated by HEL, seems to implicate differential choice by the antigen presenting system as a basis for Ir gene control, rather than the absence of a regulatory cell type from the T cell repertoire.

  5. 水解溶菌酶污泥减量过程中的污泥特性%Characterization of activated sludge in a sludge reduction process using lysozyme

    Institute of Scientific and Technical Information of China (English)

    宋勇; 施周; 陈世洋; 罗璐

    2012-01-01

    利用水解溶菌酶对从SBR系统中取出的部分污泥进行水解,然后再回流到SBR系统中,通过与未加水解溶菌酶的相同系统对比,研究了水解溶菌酶对SBR系统中污泥减量与污泥特性等方面的影响.结果表明,在连续50d的运行期间,水解溶菌酶作用下的SBR系统中剩余污泥减量总计达到76.29%,而该系统对COD与氨氮的降解效率与未加酶系统基本持平,分别为88.21%与68.72%,但其TP平均去除率较未加酶系统降低了17.2%;系统中由于水解溶菌酶的添加,污泥的微生物活性得到强化,比氧气吸收速率平均提高35%,ATP的平均值比对比系统提高了3.12nmol/mgMLSS.%Partial residual sludge from a sequential batch reactor (SBR), after mixed with lysozyme, was fed back to the SBR. The effects of the lysozyme on sludge reduction and sludge characteristics in the SBR system were evaluated through the comparative observation of the sludge in a reference SBR without lysozyme addition. Experimental results showed that the lysozyme could effectively reduce the production of excess sludge by 76.29% in the test SBR system during 50 days operation. The average removal efficiency of chemical oxygen demand and ammonia-N in the SBR with lysozyme addition was almost the same as that in the reference system, which were 88.12% and 69.05% respectively, but the average TP removal decreased by 17.2% in the lysozyme added system comparing with the reference one. It was further found that after treated with lysozyme, microbial activity of the sludge was enhanced, the specific oxygen uptake rate (SOUR) was elevated by 35%, and the average ATP value was improved by 3.12nmol/mgMLSS comparing with that of the reference system.

  6. [Purification and characterization of a lysozyme from a marine microorganism].

    Science.gov (United States)

    Zou, Yan-Li; Sun, Mi; Wang, Yue-Jun

    2005-05-01

    A novel lysozyme was purified from a marine microorganism and its major characteristics were studied. Cell-free supernatant was prepared by centrifugation of culture broth, ultrafiltration using a hollow fiber (molecular weight cut off, 50kD) and concentration using a hollow fiber (molecular weight cut off, 10kD). The crude lysozyme was purified 34.7 fold to electrophoretic homogeneity with a recovery of 24.1% by CM-Sepharose FF cationic-exchange and Sephadex G-100 gel chromatography. The relative molecular weight of this lysozyme was determined as about 39 kD. The optimum pH and temperature towards Micrococcus lysodleikticus were pH 8.0 and 35 degrees C respectively, and the enzyme was stable at temperature below 50 degrees C and pH 5.0 - 10.0. The lysozyme activity was slightly enhanced by Zn2+ and Cu2+ and slightly inhibited by Mn2+ and Ag+. The lysozyme showed good compatibility to many common chemical agents such as EDTA (0.1%) and KH2 PO4 (1.0%). The lysozyme had broad-spectrum against many bacteria, including a number of pathogens, which were resistant to egg-white lysozyme.

  7. The Invertebrate Lysozyme Effector ILYS-3 Is Systemically Activated in Response to Danger Signals and Confers Antimicrobial Protection in C. elegans

    Science.gov (United States)

    Gravato-Nobre, Maria João; Vaz, Filipa; Filipe, Sergio; Chalmers, Ronald; Hodgkin, Jonathan

    2016-01-01

    Little is known about the relative contributions and importance of antibacterial effectors in the nematode C. elegans, despite extensive work on the innate immune responses in this organism. We report an investigation of the expression, function and regulation of the six ilys (invertebrate-type lysozyme) genes of C. elegans. These genes exhibited a surprising variety of tissue-specific expression patterns and responses to starvation or bacterial infection. The most strongly expressed, ilys-3, was investigated in detail. ILYS-3 protein was expressed constitutively in the pharynx and coelomocytes, and dynamically in the intestine. Analysis of mutants showed that ILYS-3 was required for pharyngeal grinding (disruption of bacterial cells) during normal growth and consequently it contributes to longevity, as well as being protective against bacterial pathogens. Both starvation and challenge with Gram-positive pathogens resulted in ERK-MAPK-dependent up-regulation of ilys-3 in the intestine. The intestinal induction by pathogens, but not starvation, was found to be dependent on MPK-1 activity in the pharynx rather than in the intestine, demonstrating unexpected communication between these two tissues. The coelomocyte expression appeared to contribute little to normal growth or immunity. Recombinant ILYS-3 protein was found to exhibit appropriate lytic activity against Gram-positive cell wall material. PMID:27525822

  8. SERUM PARAOXONASE ACTIVITY AND ITS RELATION TO SERUM LIPIDS IN CORONARY ARTERY DISEASE

    Directory of Open Access Journals (Sweden)

    Rakhi S

    2015-10-01

    Full Text Available Human serum paraoxonase1 (PON1 is a calcium dependent es terase enzyme that hydrolyses lipid peroxides accumulating on low density lipoproteins. In the serum, PON1 enzyme is almost exclusively located on the high density lipoprotein (HDL. It hydrolyses oxidised lipids in low density lipoprotein (LDL and could therefore retard the development of atherosclerosis. As PON1 activity has a role in preventing atherosclerosis and coronary heart disease is common in Kerala, this study was conducted to assess the activity of the enzyme and its corelation to serum lipids. MATERIALS AND METHODS : In this case - control study, one hundred patients with coronary artery disease and one hundred healthy controls were included. Serum paraoxonase activity was meausured using phenyl acetate as substrate and lipid profile was done in a uto analyzer . RESULTS : The mean PON 1 activity {(70.21  27.62 in cases against (135.86  33.48 in controls (p value .000} was significantly low in CAD group compared to the control group. The CHD group had a significantly lower mean total cholesterol level (202.82  57.77 against 219.45  46.08 and lower mean HDL level (40.88  10.08 against 56.79  17.24. Correlation of the activity of PON1 with other variables for the combined group ( C ases and controls taken together showed that age is negatively correlated an d HDL is positively correlated with PON 1 activity. CONCLUSIONS : The activity of serum paraoxonase enzyme (PON1 is low in patients with coronary artery disease compared to healthy controls. Thus low serum PON1 enzyme activity is a risk factor for CAD. The activity of the serum PON1 enzyme increases with increase in HDL level and decreases with increase in age for the combined group.

  9. Antibacterial activities of serum from the Komodo Dragon (Varanus komodoensis

    Directory of Open Access Journals (Sweden)

    Mark Merchant

    2013-07-01

    Full Text Available Komodo dragons (Varanus komodoensis are able to feed on large prey items by injecting a dose of toxic bacteria with their bite that, over time, kills the prey by systemic infection. Dragons also suffer bites from other members of their own species during territorial disputes and feeding frenzies. However, they do not suffer the same fate as their prey, suggesting that they have developed a strong immunity to bacterial infections. This study was undertaken to determine the antibacterial activities of serum from the Komodo dragon. Bacterial cultures were treated with different volumes serum from Varanus komodoensis and the growth was monitored by optical density at 430 nm. In addition, the serum was treated with protease, chelators of divalent metal ions, or with mild heat to determine the mechanism of antibacterial activities. Treatment of bacterial cultures with serum from Komodo dragons (Varanus komodoensis resulted in a volume-dependent decrease in bacterial growth. Cultures of Escherichia coli, Staphylococcus aureus, and Klebsiella oxytoca exhibited moderate-strong growth inhibition by V. komodoensis serum, while cultures of Streptococcus epidermitis, Salmonella typhimurium, Providencia stuartii, and Shigella flexneri were nearly completely obliterated for 24 h by only 10% (v/v serum. The antibacterial activity of V. komodensis serum occurred very rapidly, as 18% of E. coli growth was inhibited by a five min exposure to serum. Furthermore, 10- and 20-min incubations of E. coli with serum from V. komodoensis resulted in 43 and 68% inhibition of bacterial growth, respectively. The bactericidal capacity of the serum against E. coli was 2,075,000 bacteria/μL serum, and was inhibited by mild heat treatment, pronase, EDTA, and phosphate, indicating that the anti-bacterial action is most probably due to the presence of a potent serum complement protein system.

  10. Non-covalent conjugation of CdTe QDs with lysozyme binding DNA for fluorescent sensing of lysozyme in complex biological sample.

    Science.gov (United States)

    Li, Shujia; Gao, Zhidan; Shao, Na

    2014-11-01

    Water-soluble cysteamine (CA) capped CdTe quantum dots (QDs) conjugated with lysozyme binding DNA (LBD) was constructed for luminescent sensing of lysozyme by forming a ternary self-assembly complex. Addition of negatively charged lysozyme binding DNA to the positively charged CA capped CdTe QDs buffer solution (Tris-HCl pH 7.4) could lead to the formation of QDs-LBD complex through electrostatic interactions. Once lysozyme was introduced into the CdTe QDs-LBD system, it could bind specifically with the QDs-LBD complex, resulting in fluorescence emission enhancement of the QDs due to the surface inert of QDs. At a given amount of LBD and CdTe QDs (LBD: QDs=2: 1), the fluorescence intensity enhancement of QDs was linear with lysozyme concentration over the range of 8.9-71.2 nM, with a detection limit of 4.3 nM. Due to the specific binding of LBD with lysozyme, this approach displayed high selectivity for lysozyme recognition. The sensing mechanism was confirmed by DLS and zeta potential measurement, and agarose gel electrophoresis experiment. Furthermore, the proposed CA-capped CdTe QDs-LBD sensor was applied to lysozyme detection in mouse serum and human morning urine samples, which showed high sensitivity and selectivity in the complex biological sample.

  11. Type IV collagen-degrading enzyme activity in human serum.

    OpenAIRE

    Hashimoto, Noriaki; Kobayashi, Michio; Watanabe,Akiharu; Higashi, Toshiro; Tsuji, Takao

    1988-01-01

    Type IV collagen-degrading enzyme activity was detected in human serum. Serum was preincubated with 4-aminophenylmercuric acetate and trypsin to activate the enzyme prior to assay. Type IV collagen, purified from human placentas and radiolabeled with [1-14C] acetic anhydride, was used as the substrate. The enzyme activity was measured at pH 7.5 and inhibited by treatment with ethylenediaminetetraacetic acid or heat. The assay of type IV collagen-degrading enzyme in human serum might be useful...

  12. Type IV collagen-degrading enzyme activity in human serum.

    Directory of Open Access Journals (Sweden)

    Hashimoto,Noriaki

    1988-02-01

    Full Text Available Type IV collagen-degrading enzyme activity was detected in human serum. Serum was preincubated with 4-aminophenylmercuric acetate and trypsin to activate the enzyme prior to assay. Type IV collagen, purified from human placentas and radiolabeled with [1-14C] acetic anhydride, was used as the substrate. The enzyme activity was measured at pH 7.5 and inhibited by treatment with ethylenediaminetetraacetic acid or heat. The assay of type IV collagen-degrading enzyme in human serum might be useful for estimating the degradation of type IV collagen.

  13. Serum Renalase Levels Correlate with Disease Activity in Lupus Nephritis.

    Directory of Open Access Journals (Sweden)

    Chaojun Qi

    Full Text Available Lupus nephritis (LN is among the most serious complications of systemic lupus erythematosus (SLE, which causes significant morbidity and mortality. Renalase is a novel, kidney-secreted cytokine-like protein that promotes cell survival. Here, we aimed to investigate the relationship of serum renalase levels with LN and its role in the disease progression of LN.For this cross-sectional study, 67 LN patients and 35 healthy controls were enrolled. Seventeen active LN patients who received standard therapies were followed up for six months. Disease activity was determined by the SLE Disease Activity-2000 (SLEDAI-2K scoring system and serum renalase amounts were determined by ELISA. Predictive value of renalase for disease activity was assessed. Furthermore, the expression of renalase in the kidneys of patients and macrophage infiltration was assessed by immunohistochemistry.Serum renalase amounts were significantly higher in LN patients than in healthy controls. Moreover, patients with proliferative LN had more elevated serum renalase levels than Class V LN patients. In proliferative LN patients, serum renalase levels were significantly higher in patients with active LN than those with inactive LN. Serum renalase levels were positively correlated with SLEDAI-2K, 24-h urine protein excretion, ds-DNA and ESR but inversely correlated with serum albumin and C3. Renalase amounts decreased significantly after six-months of standard therapy. The performance of renalase as a marker for diagnosis of active LN was 0.906 with a cutoff value of 66.67 μg/ml. We also observed that the amount of renalase was significantly higher in glomerular of proliferative LN along with the co-expression of macrophages.Serum renalase levels were correlated with disease activity in LN. Serum renalase might serve as a potential indicator for disease activity in LN. The marked increase of glomerular renalase and its association with macrophages suggest that it might play an

  14. Bioengineered lysozyme in combination therapies for Pseudomonas aeruginosa lung infections

    Science.gov (United States)

    Griswold, Karl E; Bement, Jenna L; Teneback, Charlotte C; Scanlon, Thomas C; Wargo, Matthew J; Leclair, Laurie W

    2014-01-01

    There is increasing urgency in the battle against drug-resistant bacterial pathogens, and this public health crisis has created a desperate need for novel antimicrobial agents. Recombinant human lysozyme represents one interesting candidate for treating pulmonary infections, but the wild type enzyme is subject to electrostatic mediated inhibition by anionic biopolymers that accumulate in the infected lung. We have redesigned lysozyme’s electrostatic potential field, creating a genetically engineered variant that is less susceptible to polyanion inhibition, yet retains potent bactericidal activity. A recent publication demonstrated that the engineered enzyme outperforms wild type lysozyme in a murine model of Pseudomonas aeruginosa lung infection. Here, we expand upon our initial studies and consider dual therapies that combine lysozymes with an antimicrobial peptide. Consistent with our earlier results, the charge modified lysozyme combination outperformed its wild type counterpart, yielding more than an order-of-magnitude reduction in bacterial burden following treatment with a single dose. PMID:24637705

  15. Release of lysozyme from electrospun PVA/lysozyme-gelatin scaffolds

    Institute of Scientific and Technical Information of China (English)

    Dong-zhi YANG; Yu-hua LONG; Jun NIE

    2008-01-01

    This article describes an electrospinning process in fabricating ultra fine fibers with core-shell structure. A biodegradable polymer, poly(vinyl alcohol) (PVA), was used as the shell; lysozyme was a kind of antioxidant; and gelatin were used as the core. Morphology and microstruc-ture of the ultra fine fibers were characterized by scanning electron microscope (SEM), transmission electron micro-scopy (TEM) and X-ray photoelectron spectroscopy (XPS) analysis. As a comparison, composite nanofiber PVA/lysozyme-gelatin blend was prepared by a normal electrospinning process. In vitro drug release behaviors of the nanofibrous membranes were determined in phosphate-buffered saline (PBS) solution. It was found that core-shell nanofibers PVA/lysozyme-gelatin obviously exhibit higher initial release rates compared to that of PVA/lysozyme-gelatin blend nanofibers. The current method may find wide application in controlled release of bioactive proteins and tissue engineering.

  16. Prognostic value of serum angiogenic activity in colorectal cancer patients.

    Science.gov (United States)

    Gonzalez, Francisco-Jesus; Quesada, Ana-Rodriguez; Sevilla, Isabel; Baca, Juan-Javier; Medina, Miguel-Angel; Amores, Jose; Diaz, Juan Miguel; Rius-Diaz, Francisca; Marques, Eduardo; Alba, Emilio

    2007-01-01

    Angiogenesis, resulting from an imbalance between angiogenic activator factors and inhibitors, is required for tumour growth and metastasis. The determination of the circulating concentration of all angiogenic factors (activators and inhibitors) is not feasible at present. We have evaluated diagnostic and prognostic values of the measurement of serum angiogenic activity in colorectal carcinoma (CRC) patients. Serum proliferative activity (PA) on human umbilical vein endothelial cells (HUVEC) in vitro, and serum vascular endothelial growth factor (VEGF) levels were determined by ELISA in 53 patients with primary CRC, 16 subjects with non-neoplastic gastrointestinal disease (SC) and 34 healthy individuals. Data were compared with clinical outcome of the patients. Although serum from CRC patients significantly increased the PA of HUVEC, compared to culture control (HUVEC in medium + 10% foetal bovine serum (FBS); P < 0.001); our results indicate that serum PA in CRC patients was similar to that of SC or healthy individuals. There was no correlation between serum PA and circulating VEGF concentrations. Surgery produced a decrease of PA at 8 hrs after tumour resection in CRC patients compared to pre-surgery values (186 +/- 47 versus 213 +/- 41, P < 0.001). However, an increase in serum VEGF values was observed after surgery (280 [176-450] versus 251 [160-357] pg/ml, P = 0.004). Patients with lower PA values after surgery showed a worse outcome that those with higher PA values. Therefore, this study does not support a diagnostic value for serum angiogenic activity measured by proliferative activity on HUVEC but suggests it could have a prognostic value in CRC patients.

  17. Expression of lysozymes from Erwinia amylovora phages and Erwinia genomes and inhibition by a bacterial protein.

    Science.gov (United States)

    Müller, Ina; Gernold, Marina; Schneider, Bernd; Geider, Klaus

    2012-01-01

    Genes coding for lysozyme-inhibiting proteins (Ivy) were cloned from the chromosomes of the plant pathogens Erwinia amylovora and Erwinia pyrifoliae. The product interfered not only with activity of hen egg white lysozyme, but also with an enzyme from E. amylovora phage ΦEa1h. We have expressed lysozyme genes from the genomes of three Erwinia species in Escherichia coli. The lysozymes expressed from genes of the E. amylovora phages ΦEa104 and ΦEa116, Erwinia chromosomes and Arabidopsis thaliana were not affected by Ivy. The enzyme from bacteriophage ΦEa1h was fused at the N- or C-terminus to other peptides. Compared to the intact lysozyme, a His-tag reduced its lytic activity about 10-fold and larger fusion proteins abolished activity completely. Specific protease cleavage restored lysozyme activity of a GST-fusion. The bacteriophage-encoded lysozymes were more active than the enzymes from bacterial chromosomes. Viral lyz genes were inserted into a broad-host range vector, and transfer to E. amylovora inhibited cell growth. Inserted in the yeast Pichia pastoris, the ΦEa1h-lysozyme was secreted and also inhibited by Ivy. Here we describe expression of unrelated cloned 'silent' lyz genes from Erwinia chromosomes and a novel interference of bacterial Ivy proteins with a viral lysozyme.

  18. Structure and stability of complex coacervate core micelles with lysozyme.

    Science.gov (United States)

    Lindhoud, Saskia; Vries, Renko de; Norde, Willem; Stuart, Martien A Cohen

    2007-07-01

    Encapsulation of enzymes by polymers is a promising method to influence their activity and stability. Here, we explore the use of complex coacervate core micelles for encapsulation of enzymes. The core of the micelles consists of negatively charged blocks of the diblock copolymer PAA42PAAm417 and the positively charged homopolymer PDMAEMA150. For encapsulation, part of the positively charged homopolymer was replaced by the positively charged globular protein lysozyme. We have studied the formation, structure, and stability of the resulting micelles for three different mixing ratios of homopolymer and lysozyme: a system predominantly consisting of homopolymer, a system predominantly consisting of lysozyme, and a system where the molar ratio between the two positively charged molecules was almost one. We also studied complexes made of only lysozyme and PAA42PAAm417. Complex formation and the salt-induced disintegration of the complexes were studied using dynamic light-scattering titrations. Small-angle neutron scattering was used to investigate the structures of the cores. We found that micelles predominantly consisting of homopolymer are spherical but that complex coacervate core micelles predominantly consisting of lysozyme are nonspherical. The stability of the micelles containing a larger fraction of lysozyme is lower.

  19. Serum paraoxonase activity and protein thiols in patients with hyperlipidemia

    Institute of Scientific and Technical Information of China (English)

    Mungli Prakash; Jeevan K Shetty; Sudeshna Tripathy; Pannuri Vikram; Manish Verma

    2009-01-01

    Objective: In the present study we evaluated the paraoxonase activity and protein thiols level in south Indian population with newly diagnosed hyperlipidemia. Methods: The study was conducted on 55 newly diagnosed hyperlipidemic pa-tients and 57 healthy controls. Serum paraoxonase activity and protein thiols were estimated by spectrophotometeric method and lipid profile by enzymatic kinetic assay method. Results: Serum paraoxonase activity, protein thiols and high density lipoprotein levels were low and total cholesterol, triglycerides and low density lipoprutein levels were high in patients with hyperlipidemia compared to healthy controls ( P < 0.01 ). Serum paranxonase activity correlated positively with protein thiols and high density lipoprotein (P<0.01). Conclusion: Decreased paraoxonase activity and protein thiols were found in patients with hyperlipi-demia. This may indicate the susceptibility of this population to accelerated atherogenesis and protein oxidation.

  20. EFFECT OF AQUATIC ENVIRONMENT PH ON THE LEVEL OF ECTOPARASITE INFESTATION, PROTEIN AND LYSOZYME CONTENT IN SOME CYPRINID SPECIES (CYPRINIDAE

    Directory of Open Access Journals (Sweden)

    L. Kurovskaya

    2016-03-01

    environments with a significant reduction in the number of ectoparasites, the protein and lysozyme levels in fish serum and organs were not significantly different from those of non-infected individuals that were kept in water of corresponding pH values. In Pseudorasbora parva with low level of parasite infestation, we did not detect any changes in the protein and lysozyme levels in fish serum and organs in a neutral pH environment. The effect of different pH levels of water on the studied parameters of Pseudorasbora parva showed their significant changes. Originality. For the first time, we demonstrated the change in the level of ectoparasite infestation, protein and lysozyme contents in fish serum and organs (Cyprinus carpio, Carassius auratus gibelio, Pseudorasbora parva, which were kept in the slightly acidic or slightly alkaline aquatic environments. Practical value. The concentration of hydrogen ions, or an aqueous active reaction (pH, plays an important role in the life of fish, their development and in the predisposition to various diseases. The performed studies showed that infestation of fish with parasites can be significantly decreased when changing water pH. This leads to the recovery and restoration of fish normal physiological functions, the impairment of which causes the toxic effects of parasites.

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

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    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.

  2. Lower Serum Paraoxonase-1 Activity Is Related to Higher Serum Amyloid A Levels in Metabolic Syndrome

    NARCIS (Netherlands)

    Kappelle, Paul Jan Willem Herman; Bijzet, Johan; Hazenberg, Bouke Pier; Dullaart, Robin Pieter Frank

    Background and Aims. High-density lipoproteins (HDL) contain the anti-oxidative enzyme, paraoxonase-1 (PON-1), which is important for atheroprotection. The acute phase reactant, serum amyloid A (SAA), an HDL-associated apolipoprotein, may impair PON-1 activity, whereas SAA and PON-1 are reciprocally

  3. Lower Serum Paraoxonase-1 Activity Is Related to Higher Serum Amyloid A Levels in Metabolic Syndrome

    NARCIS (Netherlands)

    Kappelle, Paul Jan Willem Herman; Bijzet, Johan; Hazenberg, Bouke Pier; Dullaart, Robin Pieter Frank

    2011-01-01

    Background and Aims. High-density lipoproteins (HDL) contain the anti-oxidative enzyme, paraoxonase-1 (PON-1), which is important for atheroprotection. The acute phase reactant, serum amyloid A (SAA), an HDL-associated apolipoprotein, may impair PON-1 activity, whereas SAA and PON-1 are reciprocally

  4. 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.

  5. Biophysical aspects of lysozyme adduct with monocrotophos.

    Science.gov (United States)

    Amaraneni, Sreenivasa Rao; Kumar, Sudhir; Gourinath, Samudrala

    2014-09-01

    The present study on in vitro formation and characterization of lysozyme adduct with monocrotophos (MP) evaluates the potential of lysozyme to be used as a sensitive biomarker to monitor exposure levels to the commonly used organophosphorus pesticide monocrotophos. Crystallization of lysozyme protein adduct with monocrotophos was also undertaken to understand the adduct formation mechanism at a molecular level. The binding of organophosphorus pesticides to lysozyme is one of the key steps in their mutagenicity. The formation and structural characterization of lysozyme adduct with monocrotophos was done using MALDI-TOFMS, fluorescence, UV/Vis spectroscopy, circular dichroism, and X-ray diffraction studies. We report the crystal structure of lysozyme adduct with monocrotophos at 1.9 Å. It crystallized in the P43 space group with two monomers in one asymmetric unit having one molecule of monocrotophos bound to each protein chain. The results proved that the fluorescence quenching of lysozyme by monocrotophos is due to binding of monocrotophos with a tryptophan residue of lysozyme. Monocrotophos interacts most strongly with the Trp-108 and Asp-52 of lysozyme. The interactions of the monocrotophos molecule with the lysozyme suggest the formation of a stable adduct. In addition, the alteration of lysozyme secondary structure in the presence of monocrotophos was confirmed by circular dichroism and fluorescence inhibition of lysozyme by increasing monocrotophos and UV/Vis spectrophotometry. The formation of lysozyme adduct with monocrotophos was confirmed by MALDI-TOFMS.

  6. 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.

  7. Protective properties of lysozyme on β-amyloid pathology: implications for Alzheimer disease.

    Science.gov (United States)

    Helmfors, Linda; Boman, Andrea; Civitelli, Livia; Nath, Sangeeta; Sandin, Linnea; Janefjord, Camilla; McCann, Heather; Zetterberg, Henrik; Blennow, Kaj; Halliday, Glenda; Brorsson, Ann-Christin; Kågedal, Katarina

    2015-11-01

    The hallmarks of Alzheimer disease are amyloid-β plaques and neurofibrillary tangles accompanied by signs of neuroinflammation. Lysozyme is a major player in the innate immune system and has recently been shown to prevent the aggregation of amyloid-β1-40 in vitro. In this study we found that patients with Alzheimer disease have increased lysozyme levels in the cerebrospinal fluid and lysozyme co-localized with amyloid-β in plaques. In Drosophila neuronal co-expression of lysozyme and amyloid-β1-42 reduced the formation of soluble and insoluble amyloid-β species, prolonged survival and improved the activity of amyloid-β1-42 transgenic flies. This suggests that lysozyme levels rise in Alzheimer disease as a compensatory response to amyloid-β increases and aggregation. In support of this, in vitro aggregation assays revealed that lysozyme associates with amyloid-β1-42 and alters its aggregation pathway to counteract the formation of toxic amyloid-β species. Overall, these studies establish a protective role for lysozyme against amyloid-β associated toxicities and identify increased lysozyme in patients with Alzheimer disease. Therefore, lysozyme has potential as a new biomarker as well as a therapeutic target for Alzheimer disease. Copyright © 2015. Published by Elsevier Inc.

  8. ISOLATION AND PURIFICATION OF LYSOZYME FROM THE HEN EGG WHITE

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    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.

  9. The Release of Egg White Lysozyme Containing EDTA from Composite Edible Film Based on Whey Protein, Konjac Flour and Lipid

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    Mulia W. Apriliyani

    2014-01-01

    Full Text Available The objectives of this research were to find out the effect of EDTA addition on antibacterial spectrum broadening of lysozyme on Gram negative bacteria and the release of lysozyme from composite edible film made of whey protein, konjac glucomannan and several lipids type and content. The research were conducted with 2 steps. Step I: The addition of EDTA on lysozyme aquaeous (Lysozyme (mg/mL: EDTA (mg/mL = 11.14:8.14; 11.14:11.14 and 11.14:14.14 using Randomyzed Block Design, the variables were, antibacterial of lysozyme on Micrococcus lysodeikticus and Escherichia coli. Step II: Lipid content (5 and 10% and kind of lipid (butter, margarine, palm oil and beeswax using nested Randomyzed Block Design, the variables were lysozyme release, Water Vapor Permeability (WVP, protein solublity and microstructure of composite edible film. The results were, step I: the treatment didn’t gave significantly effect (p>0.05 on lysozyme activity. EDTA decrease cell membrane stabilization and lysozyme made lysis of cell membrane. EDTA chelate Ca2+ and Mg2+ salts as bridge between Lypopolysachcharide (LPS in outer membrane so LPS released from cell wall of Gram negative bacteria. Step II: The treatment didn’t gave significantly effect (p>0.05 on release of lysozyme and water vapour permeability, but gave significantly effect (p<0.05 on protein solubility. The release of lysozyme from composite edible film gave the best lysozyme release from beeswax 10% addition.

  10. Eating a healthy lunch improves serum alanine aminotransferase activity.

    Science.gov (United States)

    Iwamoto, Masako; Yagi, Kaori; Yazumi, Kayoko; Komine, Airi; Shirouchi, Bungo; Sato, Masao

    2013-09-14

    Nutritional guidance and diet control play important roles in the treatment of obesity and non-alcoholic fatty liver. However, in Japan, nutritional guidance is difficult to provide in practice. Therefore, we evaluated the effects of providing the 'once-a-day' intervention of a healthy lunch on various metabolic parameters. For a 1-month preparatory period, 10 subjects generally consumed the lunches that were provided by the worksite cafeteria. This was followed by a 1-week washout period, after which, the subjects consumed healthy, low-calorie, well-balanced lunches for a 1-month test period. After the preparatory and test periods, blood samples were obtained from all subjects. The serum levels of indices relevant to metabolic syndrome and fatty liver were measured. Serum alanine aminotransferase activity significantly decreased by 20.3% after the healthy intervention. However, the indices of metabolic syndrome did not significantly change. Analysis of the relationship between serum alanine aminotransferase activity and nutrient content indicated that the improvement of serum alanine aminotransferase status was due to the higher vegetable content and lower animal-source protein of the meals provided. In summary, the 'once-a-day' intervention of providing a healthy lunch improved serum alanine aminotransferase status. A diet high in vegetables and low in animal-based protein is important in maintaining a healthy condition.

  11. 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.

  12. Automated conductimetric assay of human serum cholinesterase activity.

    Science.gov (United States)

    Duffy, P; Wallach, J M

    1989-01-01

    Serum cholinesterase activity was determined by conductimetry using samples in the microliter range. Butyrylcholine iodide was demonstrated to be a convenient substrate for the conductimetric assay. Validation of the microassay was made by using either purified enzyme or control serum. In the range of 0-60 U/l, a linear relationship was demonstrated. Correlation with a reference spectrophotometric method was obtained with a slope of 1.18. An explanation of this value is proposed, as different hydrolysis rates were obtained with human sera, depending on the substrate used (butyrylthio- or butyryl-choline ester).

  13. Mapping the solid-state properties of crystalline lysozyme during pharmaceutical unit-operations.

    Science.gov (United States)

    Mohammad, Mohammad Amin; Grimsey, Ian M; Forbes, Robert T

    2015-10-10

    Bulk crystallisation of protein therapeutic molecules towards their controlled drug delivery is of interest to the biopharmaceutical industry. The complexity of biotherapeutic molecules is likely to lead to complex material properties of crystals in the solid state and to complex transitions. This complexity is explored using batch crystallised lysozyme as a model. The effects of drying and milling on the solid-state transformations of lysozyme crystals were monitored using differential scanning calorimetry (DSC), X-ray powder diffraction (XRPD), FT-Raman, and enzymatic assay. XRPD was used to characterise crystallinity and these data supported those of crystalline lysozyme which gave a distinctive DSC thermogram. The apparent denaturation temperature (Tm) of the amorphous lysozyme was ∼201 °C, while the Tm of the crystalline form was ∼187 °C. Raman spectra supported a more α-helix rich structure of crystalline lysozyme. This structure is consistent with reduced cooperative unit sizes compared to the amorphous lysozyme and is consistent with a reduction in the Tm of the crystalline form. Evidence was obtained that milling also induced denaturation in the solid-state, with the denatured lysozyme showing no thermal transition. The denaturation of the crystalline lysozyme occurred mainly through its amorphous form. Interestingly, the mechanical denaturation of lysozyme did not affect its biological activity on dissolution. Lysozyme crystals on drying did not become amorphous, while milling-time played a crucial role in the crystalline-amorphous-denatured transformations of lysozyme crystals. DSC is shown to be a key tool to monitor quantitatively these transformations.

  14. Lysozyme in water-acetonitrile mixtures: Preferential solvation at the inner edge of excess hydration

    Science.gov (United States)

    Sirotkin, Vladimir A.; Kuchierskaya, Alexandra A.

    2017-06-01

    Preferential solvation/hydration is an effective way for regulating the mechanism of the protein destabilization/stabilization. Organic solvent/water sorption and residual enzyme activity measurements were performed to monitor the preferential solvation/hydration of hen egg-white lysozyme at high and low water content in acetonitrile at 25 °C. The obtained results show that the protein destabilization/stabilization depends essentially on the initial hydration level of lysozyme and the water content in acetonitrile. There are three composition regimes for the dried lysozyme. At high water content, the lysozyme has a higher affinity for water than for acetonitrile. The residual enzyme activity values are close to 100%. At the intermediate water content, the dehydrated lysozyme has a higher affinity for acetonitrile than for water. A minimum on the residual enzyme activity curve was observed in this concentration range. At the lowest water content, the organic solvent molecules are preferentially excluded from the dried lysozyme, resulting in the preferential hydration. The residual catalytic activity is ˜80%, compared with that observed after incubation in pure water. Two distinct schemes are operative for the hydrated lysozyme. At high and intermediate water content, lysozyme is preferentially hydrated. However, in contrast to the dried protein, at the intermediate water content, the initially hydrated lysozyme has the increased preferential hydration parameters. At low water content, the preferential binding of the acetonitrile molecules to the initially hydrated lysozyme was detected. No residual enzyme activity was observed in the water-poor acetonitrile. Our data clearly show that the initial hydration level of the protein macromolecules is one of the key factors that govern the stability of the protein-water-organic solvent systems.

  15. Serum enzymes activities in Plasmodium falciparum infection in Southern Pakistan

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    Koay Yen Chin

    2011-05-01

    Full Text Available Objective: Serum levels of lactate dehydrogenase (LDH,aspartate aminotranferase (AST, alanine aminotransferase(ALT, and alkaline phosphatase (ALP were assessed todetermine the liver functions of patients infected withPlasmodium falciparum. The enzyme activities were assessedin 60 malarial patients and a control group of 44 people.Materials and Methods: The data for the study was collectedfrom the survey conducted from Liaquat University of medicaland health sciences Hospital, Hyderabad, Pakaistan. Sample of60 patients aged between 20 and 50 years were collected. Acontrol group of 44 healthy individual adults was also assessedfor comparative purposes. All the malaria patients who visitedthe OPD during the study period enrolled in the study.Results: The LDH activity in male patients was found to be674.89 ± 33.354 IU/L. This is above the control LDH activity of296.59 ± 14.476 IU/L. Similarly, in female patients, the serumLDH activity of 580.25 ± 24.507 IU/L is over twice the controlfemale serum LDH activity of 302.18 ± 18.082 IU/L. Furtherone-way anova test was performed to find any significance ininfected and control male and female.Conclusion: Hepatic dysfunction was found to be associated toP. falciparum malaria infection.

  16. Silica Exposure and Serum Angiotensin Converting Enzyme Activity

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    YK Sharma

    2010-01-01

    Full Text Available Background: Silicosis is known in industrial workers for centuries. Till recently, the mainstay of its diagnosis and progress was clinical examination of the respiratory system, pulmonary function test and chest radiography. Several biomarkers such as serum angiotensin converting enzyme (ACE activity have been examined to determine the extent of silicosis. Objective: To elucidate the effect of age, gender, duration of exposure to silica dust, smoking habit, and pulmonary function status on the serum ACE activity among quartz stone workers without disease.Methods: A cross-sectional study was carried out on 134 (111 men and 14 women workers of quartz stone crushing units were studied. Standard diagnostic criteria were used for diagnosing silicosis and tuberculosis. Pulmonary functions of the participants were also assessed.Results: The mean±SD age for participants was 26.1±6.3 years (26.6±6.3 for men and 21.9±4.3 for women. The mean±SD duration of exposure was 1.1±1.9 years. In the present study, only one case of silicosis and eight cases of tuberculosis were found. The mean±SD serum ACE levels for those with and without respiratory disease were 68.44±11.61, and 66.9±14.4 IU/L, respectively (p>0.05.Conclusion: We could not observe any association between serum ACE activity and age, gender, duration of exposure, smoking habits and pulmonary function status. However, elevated levels of serum ACE was found in a solitary case of silicosis.

  17. Normal serum alanine aminotransferase activity in uncomplicated obesity

    Institute of Scientific and Technical Information of China (English)

    Gianluca Iacobellis; Antonio Moschetta; Maria Cristina Ribaudo; Alessandra Zappaterreno; Concetta Valeria Iannucci; Frida Leonetti

    2005-01-01

    AIM: To evaluate serum alanine aminotransferase (ALT)activity in a well-characterized group of uncomplicated obese subjects and its correlation with insulin resistance,plasma adiponectin, and leptin concentrations.METHODS: One hundred and five uncomplicatedobese subjects (87 women, 18 men, age 34.3±9.6 years,BMI 39.9±8.3 kg/m2)were studied. Serum ALT activity was evaluated. Insulin sensitivity was assessed by euglycemic hyperinsulinemic clamp (M index) and fasting insulin. Plasma leptin and adiponectin levels were also measured.RESULTS: Serum ALT concentration in the whole group of uncomplicated obese subjects was 17.73±6.33 U/L with none of the subjects presenting ALT levels greater than 43 U/L and only 9 (11%) women and 3 (19%) men showed ALT levels >19 and >30 U/L for women and men,respectively. No significant difference was detected in serum ALT levels between severe obese subjects (BMI >40 kg/m2) and those with BMI <40 kg/m2 (18.63±6.25 vs 17.26±6.02 U/L). ALT was significantly correlated with fasting insulin (r = 0.485, P = 0.02) and triglycerides (r= 0.358, P= 0.03).CONCLUSION: Serum ALT activity is practically normal in uncomplicated obese subjects, independently of their obesity degree. These findings suggest the role of obesityrelated comorbidities and not of BMI as main risk factors for elevated ALT levels in obese subjects.

  18. Study on Reaction Mechanism and Their Structure -Activity Relationship between Three Amphetamines and Lysozyme%3种苯丙胺类药物与溶菌酶的作用机制及构效关系研究

    Institute of Scientific and Technical Information of China (English)

    张爱平; 黄茜; 郝娟; 文雯; 高晓亚

    2011-01-01

    在模拟人体生理条件下,采用荧光光谱法研究了3种不同结构的苯丙胺类药物(麻黄碱、伪麻黄碱和甲基麻黄碱)与溶菌酶之间的相互作用,计算了其结合常数、结合位点数和热力学参数,并探讨了3种药物对溶菌酶构象的影响.研究发现,三者可对溶菌酶内源性荧光产生强烈的猝灭作用,其猝灭过程均为静态猝灭.麻黄碱、伪麻黄碱和甲基麻黄碱与溶菌酶均形成1 :1复合物,在308 K温度下的结合常数K分别为5.11×103、4.04×103、2.80×103 L·mol-1,结合距离r分别为0.241、0.350、0.422 nm,与溶菌酶结合的焓变分别为-123、-126、-52.1 kJ·mol-1,熵变分别为-329、-339、-103 J·mol-1·K-1.研究结果表明,苯丙胺类药物的构型和取代基对其与溶菌酶的相互作用有重要影响,3种苯丙胺类药物与溶菌酶的作用力顺序为麻黄碱>伪麻黄碱>甲基麻黄碱,体系的主要作用力为氢键和范德华力.溶菌酶与3种药物的同步荧光光谱也表明,溶菌酶的构象在作用前后基本不变.%The interactions of lysozyme with ephedrine, pseudoephedrine and methylephedrine, as well as their structure -activity relationship were investigated by fluorescence spectrometry under simulative physiological conditions. The binding constant, the number of binding sites and the thermodynamic parameters were calculated and the effects of ephedrine, pseudoephedrine and methylephedrine on the conformation of lysozyme were studied. The results showed that the endogenous fluorescence of lysozyme was significantly quenched by ephedrine, pseudoephedrine and methylephedrine.The mechanism of fluorescence quenching was static quenching. The 1 : 1 complex was formed between each amphetamine and lysozyme. The binding constants K of ephedrine, pseudoephedrine and methylephedrine were 5.11×103, 4. 04 × 103 , 2. 80 × 103 L · mol-1, respectively. According to the theory of Fǒster dipole -dipole non-radiation energy

  19. Serum fucosyl transferase activity and serum fucose levels as diagnostic tools in malignancy.

    Directory of Open Access Journals (Sweden)

    Sen,Umi

    1983-12-01

    Full Text Available Glycoproteins play a significant role in neoplastic transformations. Both the levels of fucose and the activity of fucosyl transferase, which mediates the assembly of the oligosaccharide moieties of the glycoprotein chains, have been found to be elevated in neoplastic conditions. Since these elevations are common features of a variety of neoplastic cells, these two have been designated as non-specific markers of malignancy. In the present study, the fucose level and fucosyl transferase activity were determined in the sera of cancer patients and an attempt was made to establish a relationship between the two. It was found that both the fucose levels and fucosyl transferase activities showed considerable elevation in the five cancer groups studied, establishing them as useful diagnostic parameters. However, it was also observed that the rate of increased fucosyl transferase activity was not fully reflected in the resulting serum fucose levels in a few cases.

  20. Serum melatonin in juvenile rheumatoid arthritis: correlation with disease activity.

    Science.gov (United States)

    El-Awady, Hanaa Mahmoud; El-Wakkad, Amany Salah El-Dien; Saleh, Maysa Tawheed; Muhammad, Saadia Ibraheem; Ghaniema, Eiman Mahmoud

    2007-05-01

    The study was conducted to investigate the abnormalities in early morning serum melatonin among patients with Juvenile Rheumatoid Arthritis (JRA) and to outline its relation to disease activity and severity. Twenty one patients with JRA and twenty healthy age and sex matched controls were enrolled in the study. Fifteen patients had polyarticular JRA, 3 had oligoarticular and 3 had systemic onset JRA. Evaluation was carried out clinically, functionally and radiologically by using disease activity score, Juvenile Arthritis Functional Assessment Report for Children (JAFAR-C score) and modified Larsen score, respectively. Laboratory investigations included Complete Blood Picture (CBC), The Erythrocyte Sedimentation Rate (ESR), C-Reactive Protein (CRP), classic IgM Rheumatoid Factor (RF), Anti-nuclear Antibodies (ANA) and melatonin estimation in serum. The serum levels of melatonin were significantly increased in JRA patients (mean +/- SD = 13.9 +/- 8 pg mL(-1)) as compared to healthy controls (mean +/- SD = 8.1 +/- 2.7 pg mL(-1), p 0.05). Hence the study conclude that the elevated melatonin levels among JRA patients with active synovitis and its close relation to disease activity rather than disease severity suggests that melatonin might play a promoting role in rheumatoid arthritis. Hence, inhibition of its synthesis and/or action by specific antagonists may be of therapeutic value.

  1. 天然药物活性成分与溶菌酶相互作用的研究进展%Advance of studies on interaction between active components of natural medicines and lysozymes

    Institute of Scientific and Technical Information of China (English)

    陈晨; 张洪峰; 王乐; 黄芸; 魏亚超; 郭金明; 郭莹

    2012-01-01

    溶菌酶是一种广泛存在于生物体内的小分子碱性蛋白,具有抗菌消炎、抗病毒、增强免疫力等多种药理作用.能够和许多外源和内源性物质结合,运载多种药物.该文综述了不同种类的天然药物活性成分与溶菌酶的相互作用,对于全面认识天然药物的药理机制,了解药物在生物体内的转运和代谢过程以及优化药物分子结构、提高天然药物的生物利用率和生物效应等都具有重要的意义.%Lysozyme (LYSO) , as an alkalescent protein micromolecule in living organisms, exhibits important pharmacological actions such as antibiosis, anti-inflammatory, antivirus and enhancing immunity. LYSO can combine with many exogenous and endogenous substances and carry many drugs. This essay summarizes interaction between different kincis of active components of natural medicines and lysozymes, which is significant to comprehensively understand pharmacological mechanism of natural drugs and their transfer and metabolic process in organisms, optimize molecule structures of drugs and increase bioavailability and biological effects of natural drugs.

  2. Serum Albumin Binding and Esterase Activity: Mechanistic Interactions with Organophosphates

    Directory of Open Access Journals (Sweden)

    Nikolay V. Goncharov

    2017-07-01

    Full Text Available The albumin molecule, in contrast to many other plasma proteins, is not covered with a carbohydrate moiety and can bind and transport various molecules of endogenous and exogenous origin. The enzymatic activity of albumin, the existence of which many scientists perceive skeptically, is much less studied. In toxicology, understanding the mechanistic interactions of organophosphates with albumin is a special problem, and its solution could help in the development of new types of antidotes. In the present work, the history of the issue is briefly examined, then our in silico data on the interaction of human serum albumin with soman, as well as comparative in silico data of human and bovine serum albumin activities in relation to paraoxon, are presented. Information is given on the substrate specificity of albumin and we consider the possibility of its affiliation to certain classes in the nomenclature of enzymes.

  3. Listeria monocytogenes is resistant to lysozyme through the regulation, not the acquisition, of cell wall-modifying enzymes.

    Science.gov (United States)

    Burke, Thomas P; Loukitcheva, Anastasia; Zemansky, Jason; Wheeler, Richard; Boneca, Ivo G; Portnoy, Daniel A

    2014-11-01

    Listeria monocytogenes is a Gram-positive facultative intracellular pathogen that is highly resistant to lysozyme, a ubiquitous enzyme of the innate immune system that degrades cell wall peptidoglycan. Two peptidoglycan-modifying enzymes, PgdA and OatA, confer lysozyme resistance on L. monocytogenes; however, these enzymes are also conserved among lysozyme-sensitive nonpathogens. We sought to identify additional factors responsible for lysozyme resistance in L. monocytogenes. A forward genetic screen for lysozyme-sensitive mutants led to the identification of 174 transposon insertion mutations that mapped to 13 individual genes. Four mutants were killed exclusively by lysozyme and not other cell wall-targeting molecules, including the peptidoglycan deacetylase encoded by pgdA, the putative carboxypeptidase encoded by pbpX, the orphan response regulator encoded by degU, and the highly abundant noncoding RNA encoded by rli31. Both degU and rli31 mutants had reduced expression of pbpX and pgdA, yet DegU and Rli31 did not regulate each other. Since pbpX and pgdA are also present in lysozyme-sensitive bacteria, this suggested that the acquisition of novel enzymes was not responsible for lysozyme resistance, but rather, the regulation of conserved enzymes by DegU and Rli31 conferred high lysozyme resistance. Each lysozyme-sensitive mutant exhibited attenuated virulence in mice, and a time course of infection revealed that the most lysozyme-sensitive strain was killed within 30 min of intravenous infection, a phenotype that was recapitulated in purified blood. Collectively, these data indicate that the genes required for lysozyme resistance are highly upregulated determinants of L. monocytogenes pathogenesis that are required for avoiding the enzymatic activity of lysozyme in the blood. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  4. Serum paraoxonase 1 activity status in patients with liver disorders

    Directory of Open Access Journals (Sweden)

    Kedage Vivekananda

    2010-01-01

    Full Text Available Background/Aim: Paraoxonase 1 (PON1 is an esterase, exclusively synthesized by liver. The present study has two objectives: to determine the PON1 activity status in various disorders associated with hepatocellular damage and to correlate the changes of PON1 activity with the standard liver function and fasting lipid profile tests in these disorders. Patients and Methods: The study groups consisted of 95 patients with liver diseases including acute viral hepatitis (14, cirrhosis with portal hypertension (33, leptospirosis (14, sepsis and multi organ failure (15, left ventricular failure (9, and falciparum malaria (10; and 53 healthy controls. Serum PON1 activity was measured manually using spectrophotometer. Liver function test parameters and fasting lipid profile were performed in clinical chemistry auto analyzer (Hitachi 912. Results: The serum PON1 activity in patients with acute viral hepatitis and sepsis decreased significantly ( P < 0.001 and moderately in falciparum malaria ( P < 0.05. However, in patients with cirrhosis, leptospirosis and left ventricular patients, its activity did not change significantly. On applying Pearson correlation, serum PON1 activity correlated positively with high-density lipoprotein-cholesterol (HDL-C in patients with sepsis (r=0.633, P < 0.05, left ventricular failure patients (r=0.814, P < 0.05 and negatively with acute viral hepatitis patients (r=- 0.528, P < 0.05. Conclusion: PON1 activity has decreased significantly in acute viral hepatitis, sepsis with multi organ failure and falciparum malaria patients. Determination of PON1 activity may serve as a useful additional test in assessing these conditions.

  5. Complex coacervate core micelles with a lysozyme-modified corona

    NARCIS (Netherlands)

    Danial, M.; Klok, H.A.; Norde, W.; Cohen Stuart, M.A.

    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.

  6. Comparative evaluation of multi-purpose solutions in the stabilization of tear lysozyme.

    Science.gov (United States)

    Barniak, Vicki L; Burke, Susan E; Venkatesh, Srini

    2010-12-01

    The range and extent of tear proteins removed by various multi-purpose solutions has been investigated, but there is little information in the literature about their ability to prevent denaturation of tear proteins, particularly lysozyme. The purpose of this study was to determine the ability of Bausch+Lomb Biotrue™ multi-purpose solution and other care solutions to affect denaturation of lysozyme using a lysozyme activity assay. The test solutions used were: Biotrue multi-purpose solution, Bausch+Lomb renu(®) fresh™, formerly ReNu MultiPlus(®), Alcon OPTI-FREE RepleniSH, Alcon OPTI-FREE EXPRESS, CIBA VISION AQuify, and AMO COMPLETE Multi-Purpose Solution Easy Rub Formula. A phosphate-buffered saline (PBS) solution served as a control. The test and control solutions containing lysozyme were exposed to sodium dodecyl sulfate (SDS), a known denaturant of the enzyme. The assay was based on digestion of the cell wall of Micrococcus luteus in a suspension, a substrate sensitive to active lysozyme. Enzymatic activity against M. luteus was used to assess activity of lysozyme. The decrease in the turbidity of the cell wall suspension, a measure of relative enzyme activity, was determined by following the decrease in absorbance (at 450nm) over time using a spectrophotometer. Statistically significant greater stabilization of lysozyme was observed with Biotrue multi-purpose solution and renu fresh than with OPTI-FREE RepleniSH, OPTI-FREE EXPRESS, AQuify, COMPLETE Multi-Purpose Solution Easy Rub Formula, and a PBS control. The lysozyme activity assay revealed that Biotrue multi-purpose solution and renu fresh have the ability to stabilize lysozyme under conditions that typically denature the protein.

  7. Complex coacervates of hyaluronic acid and lysozyme

    DEFF Research Database (Denmark)

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

    2014-01-01

    by differential scanning calorimetry. Furthermore, the protein stability of lysozyme was found to be improved upon complexation during a 12-weeks storage study at room temperature, as shown by a significant increase in recovered protein when complexed (94 ± 2% and 102 ± 5% depending on the polymer-protein weight...... 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...

  8. 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.

  9. Interaction between lysozyme and procyanidin: multilevel structural nature and effect of carbohydrates.

    Science.gov (United States)

    Liang, Miao; Liu, Rui; Qi, Wei; Su, Rongxin; Yu, Yanjun; Wang, Libing; He, Zhimin

    2013-06-01

    The interaction of procyanidins with proteins has aroused extensive attention due to its important relationship with the bioavailability and astringent property of polyphenols. In the present work, we have investigated the interactions of lysozyme with procyanidin dimer (B3) using various biophysical approaches, which aims to provide insights into the mechanism of protein/polyphenol aggregation. Procyanidin B3 spontaneously binds lysozyme, inducing the multilevel structural changes in lysozyme and the formation of insoluble complexes. The relationship between lysozyme aggregation and the loss of enzymatic activity was monitored using dynamic light scattering and fluorescence quenching. The influences of two carbohydrates (gum arabic and sucrose) on lysozyme/B3 aggregation were also studied. Gum arabic effectively inhibited the formation of insoluble aggregates, but was unable to restore the fluorescence and activity of lysozyme. However, sucrose concomitantly decreased the aggregate size with the recovery of fluorescence and lysozyme activity. These results proposed two probable mechanisms by which these two carbohydrates inhibit protein/polyphenol aggregation.

  10. Evolution of the mammalian lysozyme gene family

    Directory of Open Access Journals (Sweden)

    Biegel Jason M

    2011-06-01

    Full Text Available Abstract Background Lysozyme c (chicken-type lysozyme has an important role in host defense, and has been extensively studied as a model in molecular biology, enzymology, protein chemistry, and crystallography. Traditionally, lysozyme c has been considered to be part of a small family that includes genes for two other proteins, lactalbumin, which is found only in mammals, and calcium-binding lysozyme, which is found in only a few species of birds and mammals. More recently, additional testes-expressed members of this family have been identified in human and mouse, suggesting that the mammalian lysozyme gene family is larger than previously known. Results Here we characterize the extent and diversity of the lysozyme gene family in the genomes of phylogenetically diverse mammals, and show that this family contains at least eight different genes that likely duplicated prior to the diversification of extant mammals. These duplicated genes have largely been maintained, both in intron-exon structure and in genomic context, throughout mammalian evolution. Conclusions The mammalian lysozyme gene family is much larger than previously appreciated and consists of at least eight distinct genes scattered around the genome. Since the lysozyme c and lactalbumin proteins have acquired very different functions during evolution, it is likely that many of the other members of the lysozyme-like family will also have diverse and unexpected biological properties.

  11. Smoking Discriminately Changes the Serum Active and Non-Active Forms of Vitamin B12.

    Science.gov (United States)

    Shekoohi, Niloofar; Javanbakht, Mohammad Hassan; Sohrabi, Marjan; Zarei, Mahnaz; Mohammadi, Hamed; Djalali, Mahmoud

    2017-06-01

    Smoking may modify the appetite, and consequently affect nutrient intake and serum micronutrients. The effect of smoking on vitamin B12 status has been considered in several studies. The research proposed that organic nitrites, nitro oxide, cyanides, and isocyanides of cigarette smoke interfere with vitamin B12 metabolism, and convert it to inactive forms. This research was carried out to determine the serum level of active and inactive forms of vitamin B12 in male smokers in comparison with male nonsmokers. This is a case-control study, in which the participants were 85 male smokers and 85 male nonsmokers. The serum levels of total and active form of vitamin B12 were measured. Dietary intake was recorded by a quantitative food frequency questionnaire and one-day 24-hour dietary recall method. Independent two sample T test was used to compare quantitative variables between the case and control groups. The serum level of total vitamin B12 was not significantly different between two groups, but serum level of active form of vitamin B12 in the smoking group was significantly lower than non-smoking group (Pvitamin B12 in smokers in the Iranian community. The results of this study identified that serum level of total vitamin B12 might be not different between smoking and non-smoking people, but the function of this vitamin is disturbed in the body of smokers through the reduction of serum level of active form of vitamin B12.

  12. 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

  13. Covalent immobilization of lysozyme on ethylene vinyl alcohol films for nonmigrating antimicrobial packaging applications.

    Science.gov (United States)

    Muriel-Galet, V; Talbert, J N; Hernandez-Munoz, P; Gavara, R; Goddard, J M

    2013-07-10

    The objective of this study was to develop a new antimicrobial film, in which lysozyme was covalently attached onto two different ethylene vinyl alcohol copolymers (EVOH 29 and EVOH 44). The EVOH surface was modified with UV irradiation treatment to generate carboxylic acid groups, and lysozyme was covalently attached to the functionalized polymer surface. Surface characterization of control and modified films was performed using attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and dye assay. The value of protein loading after attachment on the surface was 8.49 μg protein/cm(2) and 5.74 μg protein/cm(2) for EVOH 29 and EVOH 44, respectively, after 10 min UV irradiation and bioconjugation. The efficacy of the EVOH-lysozyme films was assessed using Micrococcus lysodeikticus. The antimicrobial activity of the films was tested against Listeria monocytogenes and was similar to an equivalent amount of free enzyme. The reduction was 1.08 log for EVOH 29-lysozyme, 0.95 log for EVOH 44-lysozyme, and 1.34 log for free lysozyme. This work confirmed the successful use of lysozyme immobilization on the EVOH surface for antimicrobial packaging.

  14. 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 stro

  15. Complex coacervation of lysozyme and heparin

    DEFF Research Database (Denmark)

    van de Weert, Marco; Andersen, Mia Bendix; Frokjaer, Sven

    2004-01-01

    To characterize complex coacervates/flocculates of lysozyme and heparin in terms of binding stoichiometry and to determine the effect of complexation on protein structure and stability.......To characterize complex coacervates/flocculates of lysozyme and heparin in terms of binding stoichiometry and to determine the effect of complexation on protein structure and stability....

  16. The Lysozyme from Insect (Manduca sexta) is a Cold-Adapted Enzyme

    Energy Technology Data Exchange (ETDEWEB)

    Sotelo-Mundo,R.; Lopez-Zavala, A.; Garcia-Orozco, K.; Arvizu-Flores, A.; Velazquez-Contreras, E.; Valenzuela-Soto, E.; Rojo-Dominguez, A.; Kanost, M.

    2007-01-01

    Enzymatic activity is dependent on temperature, although some proteins have evolved to retain activity at low temperatures at the expense of stability. Cold adapted enzymes are present in a variety of organisms and there is ample interest in their structure-function relationships. Lysozyme (E.C. 3.2.1.17) is one of the most studied enzymes due to its antibacterial activity against Gram positive bacteria and is also a cold adapted protein. In this work the characterization of lysozyme from the insect Manduca sexta and its activity at low temperatures is presented. Both M. sexta lysozymes natural and recombinant showed a higher content of {alpha}-helix secondary structure compared to that of hen egg white lysozyme and a higher specific enzymatic activity in the range of 5-30 {sup o}C. These results together with measured thermodynamic activation parameters support the designation of M. sexta lysozyme as a cold adapted enzyme. Therefore, the insect recombinant lysozyme is feasible as a model for structure-function studies for cold-adapted proteins.

  17. Salivary lysozyme in smoking alcohol dependent persons.

    Science.gov (United States)

    Waszkiewicz, Napoleon; Zalewska-Szajda, Beata; Zalewska, Anna; Waszkiewicz, Magdalena; Szajda, Slawomir Dariusz; Repka, Bernadeta; Szulc, Agata; Kepka, Alina; Minarowska, Alina; Ladny, Jerzy Robert; Zwierz, Krzysztof

    2012-01-01

    The purpose of this study was to evaluate the effect of chronic alcohol intoxication and smoking on the concentration and output of salivary lysozyme. Thirty seven men participated in the study, including 17 male smoking alcohol-dependent patients after chronic alcohol intoxication (AS), and 20 control non-smoking male social drinkers (CNS) with no history of alcohol abuse or smoking. The level of lysozyme was assessed by the radial immunodiffusion method. Significantly lower lysozyme output in the AS group compared to the CNS group was found. Moreover, gingival index was significantly higher in AS than in the CNS group. It appeared that the reduced salivary lysozyme output was more likely the result of ethanol action than smoking. In conclusion, persons addicted to alcohol and nicotine have a poorer periodontal status than non-smoking social drinkers, which may partially be due to the diminished protective effects of lysozyme present in the saliva.

  18. Control of electrostatic interactions between F-actin and genetically modified lysozyme in aqueous media

    Energy Technology Data Exchange (ETDEWEB)

    Sanders, Lori K.; Xian, Wujing; Guaqueta, Camilo; Strohman, Michael J.; Vrasich, Chuck R.; Luijten, Erik; Wong, Gerard C.L. (UIUC)

    2008-07-11

    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.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2009-06-04

    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.

  20. Serum inflammatory mediators as markers of human Lyme disease activity.

    Directory of Open Access Journals (Sweden)

    Mark J Soloski

    Full Text Available Chemokines and cytokines are key signaling molecules that orchestrate the trafficking of immune cells, direct them to sites of tissue injury and inflammation and modulate their states of activation and effector cell function. We have measured, using a multiplex-based approach, the levels of 58 immune mediators and 7 acute phase markers in sera derived from of a cohort of patients diagnosed with acute Lyme disease and matched controls. This analysis identified a cytokine signature associated with the early stages of infection and allowed us to identify two subsets (mediator-high and mediator-low of acute Lyme patients with distinct cytokine signatures that also differed significantly (p<0.0005 in symptom presentation. In particular, the T cell chemokines CXCL9 (MIG, CXCL10 (IP-10 and CCL19 (MIP3B were coordinately increased in the mediator-high group and levels of these chemokines could be associated with seroconversion status and elevated liver function tests (p = 0.027 and p = 0.021 respectively. There was also upregulation of acute phase proteins including CRP and serum amyloid A. Consistent with the role of CXCL9/CXCL10 in attracting immune cells to the site of infection, CXCR3+ CD4 T cells are reduced in the blood of early acute Lyme disease (p = 0.01 and the decrease correlates with chemokine levels (p = 0.0375. The levels of CXCL9/10 did not relate to the size or number of skin lesions but elevated levels of serum CXCL9/CXCL10 were associated with elevated liver enzymes levels. Collectively these results indicate that the levels of serum chemokines and the levels of expression of their respective chemokine receptors on T cell subsets may prove to be informative biomarkers for Lyme disease and related to specific disease manifestations.

  1. Serum inflammatory mediators as markers of human Lyme disease activity.

    Science.gov (United States)

    Soloski, Mark J; Crowder, Lauren A; Lahey, Lauren J; Wagner, Catriona A; Robinson, William H; Aucott, John N

    2014-01-01

    Chemokines and cytokines are key signaling molecules that orchestrate the trafficking of immune cells, direct them to sites of tissue injury and inflammation and modulate their states of activation and effector cell function. We have measured, using a multiplex-based approach, the levels of 58 immune mediators and 7 acute phase markers in sera derived from of a cohort of patients diagnosed with acute Lyme disease and matched controls. This analysis identified a cytokine signature associated with the early stages of infection and allowed us to identify two subsets (mediator-high and mediator-low) of acute Lyme patients with distinct cytokine signatures that also differed significantly (p<0.0005) in symptom presentation. In particular, the T cell chemokines CXCL9 (MIG), CXCL10 (IP-10) and CCL19 (MIP3B) were coordinately increased in the mediator-high group and levels of these chemokines could be associated with seroconversion status and elevated liver function tests (p = 0.027 and p = 0.021 respectively). There was also upregulation of acute phase proteins including CRP and serum amyloid A. Consistent with the role of CXCL9/CXCL10 in attracting immune cells to the site of infection, CXCR3+ CD4 T cells are reduced in the blood of early acute Lyme disease (p = 0.01) and the decrease correlates with chemokine levels (p = 0.0375). The levels of CXCL9/10 did not relate to the size or number of skin lesions but elevated levels of serum CXCL9/CXCL10 were associated with elevated liver enzymes levels. Collectively these results indicate that the levels of serum chemokines and the levels of expression of their respective chemokine receptors on T cell subsets may prove to be informative biomarkers for Lyme disease and related to specific disease manifestations.

  2. Resistance to mucosal lysozyme compensates for the fitness deficit of peptidoglycan modifications by Streptococcus pneumoniae.

    Directory of Open Access Journals (Sweden)

    Kimberly M Davis

    2008-12-01

    Full Text Available The abundance of lysozyme on mucosal surfaces suggests that successful colonizers must be able to evade its antimicrobial effects. Lysozyme has a muramidase activity that hydrolyzes bacterial peptidoglycan and a non-muramidase activity attributable to its function as a cationic antimicrobial peptide. Two enzymes (PgdA, a N-acetylglucosamine deacetylase, and Adr, an O-acetyl transferase that modify different sites on the peptidoglycan of Streptococcus pneumoniae have been implicated in its resistance to lysozyme in vitro. Here we show that the antimicrobial effect of human lysozyme is due to its muramidase activity and that both peptidoglycan modifications are required for full resistance by pneumococci. To examine the contribution of lysozyme and peptidoglycan modifications during colonization of the upper respiratory tract, competition experiments were performed with wild-type and pgdAadr mutant pneumococci in lysozyme M-sufficient (LysM(+/+ and -deficient (LysM(-/- mice. The wild-type strain out-competed the double mutant in LysM(+/+, but not LysM(-/- mice, indicating the importance of resistance to the muramidase activity of lysozyme during mucosal colonization. In contrast, strains containing single mutations in either pgdA or adr prevailed over the wild-type strain in both LysM(+/+ and LysM(-/- mice. Our findings demonstrate that individual peptidoglycan modifications diminish fitness during colonization. The competitive advantage of wild-type pneumococci in LysM(+/+ but not LysM(-/- mice suggests that the combination of peptidoglycan modifications reduces overall fitness, but that this is outweighed by the benefits of resistance to the peptidoglycan degrading activity of lysozyme.

  3. Serum Inflammatory Mediators as Markers of Human Lyme Disease Activity

    Science.gov (United States)

    Soloski, Mark J.; Crowder, Lauren A.; Lahey, Lauren J.; Wagner, Catriona A.

    2014-01-01

    Chemokines and cytokines are key signaling molecules that orchestrate the trafficking of immune cells, direct them to sites of tissue injury and inflammation and modulate their states of activation and effector cell function. We have measured, using a multiplex-based approach, the levels of 58 immune mediators and 7 acute phase markers in sera derived from of a cohort of patients diagnosed with acute Lyme disease and matched controls. This analysis identified a cytokine signature associated with the early stages of infection and allowed us to identify two subsets (mediator-high and mediator-low) of acute Lyme patients with distinct cytokine signatures that also differed significantly (pLyme disease (p = 0.01) and the decrease correlates with chemokine levels (p = 0.0375). The levels of CXCL9/10 did not relate to the size or number of skin lesions but elevated levels of serum CXCL9/CXCL10 were associated with elevated liver enzymes levels. Collectively these results indicate that the levels of serum chemokines and the levels of expression of their respective chemokine receptors on T cell subsets may prove to be informative biomarkers for Lyme disease and related to specific disease manifestations. PMID:24740099

  4. Stomach lysozymes of the three-toed sloth (Bradypus variegatus), an arboreal folivore from the Neotropics.

    Science.gov (United States)

    Pacheco, M Andreína; Concepción, Juan Luís; Rangel, José David Rosales; Ruiz, Marie Christine; Michelangeli, Fabián; Domínguez-Bello, María G

    2007-07-01

    Lysozymes are antimicrobial defences that act as digestive enzymes when expressed in the stomach of herbivores with pre-gastric fermentation. We studied this enzyme in the complex stomach of the three-toed sloth (Bradypus variegatus), a folivore with pre-gastric fermentation. Lysozymes were identified by SDS-PAGE and immunoblotting in all portions: diverticulum, pouch, glandular and muscular prepyloric area with 14.3 kDa of molecular mass. Purified lysozymes from all areas but the diverticulum were characterized by MALDI-TOF, optimal pH, optimal ionic strength, and specific activity. The differences observed suggested at least three isoforms. The optimal pHs were similar to the pH of the stomach portion where the enzymes were isolated. The lysozyme from the pouch (fermentation chamber) exhibited higher specific activity and concentration than the others. The specific activity of the enzyme from the acid muscular prepyloric portion was comparable to that reported in the cow abomasums; however, its concentration was lower than that observed in cow. This distinctive pattern of secretion/specific activity and overall low concentration suggests different roles for the lysozymes in this herbivore compared to Artiodactyla. We postulate that sloth stomach lysozymes may still be antimicrobial defences by protecting the microbial flora of the fermentation chamber against foreign bacteria.

  5. Serum arylesterase and paraoxonase activity in patients with chronic hepatitis

    Institute of Scientific and Technical Information of China (English)

    Suleyman Sirri Kilic; Suleyman Aydin; Nermin Kilic; Fazilet Erman; Suna Aydin; (I)lhami Celik

    2005-01-01

    AIM: To investigate the relationship between serum paraoxonase (PON1), AST, ALT, GGT, and arylesterase (AE) activity alterations and the degree of liver damage in patients with chronic hepatitis.METHODS: We studied 34 chronic hepatitis patients and 32 control subjects, aged between 35 and 65 years,in the Department of Infection and Clinical Microbiology at the Firat University School of Medicine. Blood samples were collected from subjects between 8:00 and 10:00 a.m. following a 12-h fast. Baseline and salt-stimulated PON1 activities were measured by the hydrolysis of paraoxon. Phenyl acetate was used as the substrate and formed phenol was measured spectrophotometrically at 270 nm after the addition of a 10-fold diluted serum sample in AE activity measurements.RESULTS: The results of this investigation revealed that the levels of AE activity decreased from 132±52 to 94±36 (29%), baseline PON1 activity from 452±112 to 164±67 (64%), salt-stimulated PON1 activity from 746±394 to 294±220 (61%), HDL from 58.4±5.1 to 47.2±5.6(20%), triglyceride from 133±51.2 to 86±34.0 (35%),while a slight increase in the level of LDL (from 163±54.1 to 177.3±56.0; 9%) and significant increases in the levels of AST (from 29±9.3 to 98±44), ALP (from 57.2±13.1 to 91±38.1), ALT (from 27.9±3.32 to 89±19.1), GGT (from 24.3±2.10 to 94±48.2), total bilirubin (from 0.74±0.02 to 1.36±0.06; 84%) and direct bilirubin (from 0.18±0.01 to 0.42±0.04; 133%) were detected.However, the levels of albumin, total protein, cholesterol,and uric acid were almost the same in chronic hepatitis and the control subjects.CONCLUSION: Low PON1 and AE activity may contribute to the increased liver dysfunction in chronic hepatitis patients by reducing the ability of HDL to retard LDL oxidation and might be clinically useful for monitoring the disease of chronic hepatitis.

  6. Serum YKL-40 concentrations in patients with rheumatoid arthritis: relation to disease activity

    DEFF Research Database (Denmark)

    Johansen, J S; Stoltenberg, M; Hansen, M

    1999-01-01

    OBJECTIVE: YKL-40, also called human cartilage glycoprotein-39, is secreted by chondrocytes, synovial cells, macrophages and neutrophils. Studies have shown that YKL-40 is an autoantigen in rheumatoid arthritis (RA). We evaluated whether serum YKL-40 was related to disease activity in patients...... with RA. METHODS: Serum YKL-40 was determined by radioimmunoassay in 156 patients with RA during a 1 yr longitudinal study. RESULTS: Serum YKL-40 was increased in 54% of the patients with clinically active disease. Patients with clinically active disease initially who became inactive after 12 months had...... a significant decrease in serum YKL-40 (-30%, P patients who changed from inactive to active disease had an increase in serum YKL-40. Patients who remained active had unchanged serum YKL-40 during the study. Serum YKL-40 decreased rapidly (-24% after 7 days, P

  7. Evaluation of Serum Lactate Dehydrogenase Activity in a Virtual Environment

    Directory of Open Access Journals (Sweden)

    V.M.T. Trindade

    2013-05-01

    Full Text Available Introduction: Lactate dehydrogenase is a citosolic enzyme involved in reversible transformation of pyruvate to lactate. It participates in anaerobic glycolysis of skeletal muscle and red blood cells, in liver gluconeogenesis and in aerobic metabolism of heart muscle. The determination of its activity helps in the diagnosis of various diseases, because it is increased in serum of patients suffering from myocardial infarction, acute hepatitis, muscular dystrophy and cancer. This paper presents a learning object, mediated by computer, which contains the simulation of the laboratory determination serum lactate dehydrogenase activity measured by the spectrophotometric method, based in the decrease of absorbance at 340 nm. Materials and Methods: Initially, pictures and videos were obtained recording the procedure of the methodology. The most representative images were selected, edited and inserted into an animation developed with the aid of the tool Adobe ® Flash ® CS3. The validation of the object was performed by the students of Biochemistry I (Pharmacy-UFRGS from the second semester of 2009 and both of 2010. Results and Discussion: The analysis of students' answers revealed that 80% attributed the excellence of the navigation program, the display format and to aid in learning. Conclusion: Therefore, this software can be considered an adequate teaching resource as well as an innovative support in the construction of theoretical and practical knowledge of Biochemistry. Available at: http://www6.ufrgs.br/gcoeb/LDH

  8. Effects of Chitosan, Lysozyme and Oregano Oil on Antimicrobial Activity of Soy Protein Isolate Film%壳聚糖、溶菌酶和牛至油对大豆分离蛋白膜抑菌效果的影响

    Institute of Scientific and Technical Information of China (English)

    杜会云; 赵寿经; 王新伟; 马中苏; 吴晓艳; 张艳平

    2011-01-01

    采用双倍稀释法研究壳聚糖、溶菌酶和牛至油对大肠杆菌和金黄色葡萄球菌的最小抑菌浓度.采用抑菌圈法研究添加天然抑菌剂壳聚糖、溶菌酶和牛至油的大豆分离蛋白膜对大肠杆菌、金黄色葡萄球菌、酿酒酵母和黑曲霉的抑菌效果.结果表明,添加壳聚糖、溶菌酶和牛至油的大豆分离蛋白膜对大肠杆菌、金黄色葡萄球菌、酿酒酵母和黑曲霉均有抑制作用.对大肠杆菌和金黄色葡萄球菌抑制效果为:牛至油>壳聚糖>溶菌酶.对酿酒酵母的抑制效果为:壳聚糖>牛至油>溶菌酶;对黑曲零的抑制效果为:牛至油>壳聚糖>溶菌酶.因此,添加壳聚糖、溶菌酶和牛至油的大豆分离蛋白膜具有较好的抑菌效果和应用前景.%Double dilution method was used to determine minimum inhibitory concentration of chitosan, lysozyme and oregano oil against Escherichia coli and Staphyloccocus aureus.Inhibition zone method was used to determine antimicrobial effect of soy protein isolate films enriched with chitosan, lysozyme and oregano oil against E.coli, S.aureus,Saccharomyces cerevisiae and Aspergillus niger.Results indicated that soy protein isolate films enriched with chitosan,lysozyme and oregano oil could inhibit the growth of E.coli, S.aureus, S.cerevisiae and A.niger.The inhibitory effect on E.coli and S.aureus: oregano oil > chitosan > lysozyme.The inhibitory effect on S.cerevisiae: chitosan > oregano oil > lysozyme; the inhibitory effect on A.niger: oregano oil > chitosan > lysozyme.This study showed the films enriched with chitosan, lysozyme and oregano oil have potential to be used as active biodegradable films with strong antimicrobial effects.

  9. 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.

  10. Dramatic Changes of Matrix Metalloproteinases-7 and Lysozyme in the Ulcerative Colitis of Mice Induced by Dextran Sulfate Sodium

    Institute of Scientific and Technical Information of China (English)

    KANG Jing-jing; YANG Yu-rong; LIANG Hong-de; ZHAO De-ming; TENG Ke-dao; JIAO Xi-lan; WANG Ping-li; SUN Zhe; NI Pei-pei; WANG Zhi-feng; ZHANG Rui

    2014-01-01

    Ulcerative colitis (UC) is a lifelong illness with profound emotional and social impacts, and could cause serious damage to large intestine, especially in colon. However, the pathogenesis of UC remained unclear. The present study attempts to ifnd out the role of matrix metalloproteinases-7 (MMP-7) and lysozyme in the pathogenesis of UC through a mice model induced by dextran sulfate sodium (DSS). The UC model was evaluated both by disease activity index (DAI) and the intestinal histopathology. The results show that there is a high correlation between the DAI score and the pathological changes of colon. Interleukin-6 (IL-6) serum levels and large intestinal lfuids levels in UC mice are always higher than that of the control groups, which might be associated with the degree of the inlfammation damage in the colon. The change tendency of the MMP-7 mRNA and protein expressions are both up-regulated ifrstly and then down-regulated from 1 to 5 d in the colon, but only the MMP-7 protein is up-regulated at 7 d again. The up-regulated MMP-7 levels in the early stage of UC may play a protective role through the activated defensins, while the down-regulated levels in the mid-later stage of UC may be connected with the severe lesions in the colon. However, the up-regulated MMP-7 levels in the later stage of UC in the colon may also contribute to the tissue repair or be served as a marker to CRC (colorectal cancer). The distribution of lysozyme protein indicates that there may be Paneth-like cells in the colon. Both the changes of MMP-7 and lysozyme in the small intestine may play a protective role for the safe environment of the whole gut, especially to the colon of UC.

  11. Binding of copper to lysozyme: Spectroscopic, isothermal titration calorimetry and molecular docking studies

    Science.gov (United States)

    Jing, Mingyang; Song, Wei; Liu, Rutao

    2016-07-01

    Although copper is essential to all living organisms, its potential toxicity to human health have aroused wide concerns. Previous studies have reported copper could alter physical properties of lysozyme. The direct binding of copper with lysozyme might induce the conformational and functional changes of lysozyme and then influence the body's resistance to bacterial attack. To better understand the potential toxicity and toxic mechanisms of copper, the interaction of copper with lysozyme was investigated by biophysical methods including multi-spectroscopic measurements, isothermal titration calorimetry (ITC), molecular docking study and enzyme activity assay. Multi-spectroscopic measurements proved that copper quenched the intrinsic fluorescence of lysozyme in a static process accompanied by complex formation and conformational changes. The ITC results indicated that the binding interaction was a spontaneous process with approximately three thermodynamical binding sites at 298 K and the hydrophobic force is the predominant driven force. The enzyme activity was obviously inhibited by the addition of copper with catalytic residues Glu 35 and Asp 52 locating at the binding sites. This study helps to elucidate the molecular mechanism of the interaction between copper and lysozyme and provides reference for toxicological studies of copper.

  12. Serum cryoglobulins and disease activity in systematic lupus erythematosus

    Directory of Open Access Journals (Sweden)

    Mansoor Karimifar

    2013-01-01

    Full Text Available Background: To determine the prevalence of cryoglobulins in Iranian patients with systemic lupus erythematosus (SLE and evaluate the correlation of cryoglobulins with disease activity in these patients. Materials and Methods: In a cross-sectional study, we investigated 80 consecutive women who fulfilled the 1982 revised criteria of the American College of Rheumatology for the classification of SLE. All the patients had undergone a medical interview and general physical examination by a rheumatologist for clinical and serologic characteristics of SLE. For the determination of cryoglobulins, sera were collected by a standard protocol at 37°C, and after incubation at 4°C for seven days, the level of cryoglobulins was estimated for each patient. Results: Cryoglobulins were detected in the sera of 39 (48.8% patients. All of these patients had cryocrit over 5%. Disease was active in 30 patients [SLEDAI ≥6 (DAI: disease activity index] and inactive in 50 (SLEDAI <6. There was no significant difference between active and inactive patients for the presence of serum cryoglobulins (r = 0.086, P = 0.56. A significant positive correlation was observed between antinuclear antibody (ANA, anti-dsDNA (dsDNA: Double-stranded deoxyribonucleic acid, CH50 (CH50: total hemolytic complement assay, and C-reactive protein (CRP (r = 0.21, P = 0.004, r = 0.65, P = 0.001, r = 0.45, P = 0.023, r = 0.38, P = 0.036, respectively. Hepatitis C virus (HCV infection was not detected in any of the SLE patients. Conclusion: Although the presence of cryoglobulins in the SLE patients correlated with positive anti-ds DNA and low CH50, it could not be predict activity of the disease.

  13. A comparison of the physical properties of ultrasonically synthesized lysozyme- and BSA-shelled microbubbles.

    Science.gov (United States)

    Vong, Fiona; Son, Younggyu; Bhuiyan, Sadia; Zhou, Meifang; Cavalieri, Francesca; Ashokkumar, Muthupandian

    2014-01-01

    Ultrasonic technique has been used for synthesising protein microspheres possessing specific physical and functional properties. Various proteins have been used as shell materials under different experimental conditions. In previous studies, thermal or chemical denaturation of the proteins was used to obtain stable bovine-serum albumin (BSA) and lysozyme microbubbles (MBs), respectively. It is ideal to establish a generic procedure to synthesise microspheres irrespective of the nature of the protein. In order to see if a generic procedure can be established, ultrasonic synthesis of lysozyme and BSA MBs was carried out under similar experimental conditions and their properties were evaluated. The size, size distribution and the stability of the MBs were significantly different for the lysozyme and BSA MBs. The size and size distribution of the lysozyme coated MBs were larger than BSA bubbles. The mechanical strength of MBs against the shear forces, generated when irradiated by high frequency ultrasound, was studied using pulsed-sonoluminescence (SL). This study indicated that lysozyme MBs were significantly more stable than BSA MBs. An increase in mechanical strength of the MBs may lead to an increase in their storage lifetime and stability against gas diffusion. Possible reasons for such observations have been discussed.

  14. Effect of lysozyme on "flor" velum yeasts in the biological aging of sherry wines.

    Science.gov (United States)

    Roldán, Ana; Lasanta, Cristina; Caro, Ildefonso; Palacios, Víctor

    2012-05-01

    Biological aging is a key step in the production of Sherry wine classified as "fine". During this stage, a film of yeast referred to as "flor velum" covers the surface of the wine and substantially alters its characteristics. Other microorganisms may coexist with flor yeasts, such as lactic acid bacteria and non-Saccharomyces yeasts, whose growth may be favored under certain conditions, causing organoleptic deviations and deterioration of the wine. To prevent the development of lactic bacteria, lysozyme usage has been introduced. Lysozyme is a hydrolytic enzyme with muramidase activity that can lyse gram-positive bacteria; its use in winemaking was approved by the OIV in 1997 (resolution OENO 10/97). Thus far, the use of lysozyme during the production of Sherry wines is not widespread despite its effectiveness in controlling lactic acid bacteria. However, there have been no studies on the effect of lysozyme on flor velum. The aim of this study was to determine the influence of lysozyme on yeast growth and the formation, development and metabolism of flor velum during the biological aging process of Sherry wine. The results indicate that lysozyme does not affect the flor yeast during the fermentative stage or biofilm stage. However, if yeast inoculation is carried out under submerged culture conditions during biological aging, low doses of lysozyme (≥12.5 g/hL) affect cell multiplication and the membrane hydrophobicity of the yeast, inhibiting their aggregation and flotation and the subsequent development of flor velum. Thus, the yeast inoculation protocol and the methodology used for the addition of lysozyme influence velum development, its metabolism and the wine characteristics.

  15. Hexafluoroisopropanol-induced catanionic-surfactants-based coacervate extraction for analysis of lysozyme.

    Science.gov (United States)

    Xu, Jia; Niu, Manli; Xiao, Yuxiu

    2017-02-01

    A coacervate extraction method, based on hexafluoroisopropanol (HFIP)-induced catanionic surfactants and coupled with a back-extraction procedure, was developed for separation and purification of proteins, using sodium dodecyl sulfate (SDS) and dodecyltrimethyl ammonium bromide (DTAB) as representative catanionic surfactants and lysozyme as a model protein. After the coacervate extraction and back extraction, the obtained lysozyme solutions were examined in terms of quantitative analysis by capillary electrophoresis, bacteriolytic activity, and circular dichroism (CD). The effects of several parameters including back-extraction solvent, HFIP content, total surfactant concentration, and SDS/DTAB molar ratio were investigated in detail on the extraction efficiency and activity of lysozyme. Under the optimized extraction conditions (66 mM KH2PO4 buffer with pH 6.2 as back-extraction solvent, SDS/DTAB molar ratio = 1:1 mol/mol, total surfactant concentration = 30 mM, HIFP concentration = 8 % v/v), the extraction recovery was 89.8 % (±4.7, n = 3), limit of detection was 2.2 (±0.3, n = 3) μg mL(-1), and meanwhile nearly 65 % of native lysozyme activity was retained. In addition, the activity and CD assays showed that SDS/DTAB molar ratio had a significant influence on the activity and structure of lysozyme after extraction. The DTAB-rich extraction systems, in which the DTAB mole fraction was equal to or larger than 70 %, could keep the activity and structure of lysozyme almost in the native state. Graphical Abstract Procedure of HFIP-induced SDS/DTAB coacervate extraction and back extraction of lysozyme.

  16. Effect of Fe{sub 3}O{sub 4} magnetic nanoparticles on lysozyme amyloid aggregation

    Energy Technology Data Exchange (ETDEWEB)

    Bellova, Andrea; Koneracka, Martina; Kopcansky, Peter; Tomasovicova, Natalia; Timko, Milan; Bagelova, Jaroslava; Gazova, Zuzana [Department of Biophysics, Department of Magnetism, Institute of Experimental Physics, Slovak Academy of Science, Watsonova 47, 04001 Kosice (Slovakia); Bystrenova, Eva; Valle, Francesco; Biscarini, Fabio, E-mail: gazova@saske.sk [CNR-Instituto per lo Studio dei Materiali Nanostrutturati, via Gobetti 101, I-40129 Bologna (Italy)

    2010-02-10

    Peptide amyloid aggregation is a hallmark of several human pathologies termed amyloid diseases. We have investigated the effect of electrostatically stabilized magnetic nanoparticles of Fe{sub 3}O{sub 4} on the amyloid aggregation of lysozyme, as a prototypical amyloidogenic protein. Thioflavin T fluorescence assay and atomic force microscopy were used for monitoring the inhibiting and disassembly activity of magnetic nanoparticles of Fe{sub 3}O{sub 4}. We have found that magnetic Fe{sub 3}O{sub 4} nanoparticles are able to interact with lysozyme amyloids in vitro leading to a reduction of the amyloid aggregates, thus promoting depolymerization; the studied nanoparticles also inhibit lysozyme amyloid aggregation. The ability to inhibit lysozyme amyloid formation and promote lysozyme amyloid disassembly exhibit concentration-dependent characteristics with IC50 = 0.65 mg ml{sup -1} and DC50 = 0.16 mg ml{sup -1} indicating that nanoparticles interfere with lysozyme aggregation already at stoichiometric concentrations. These features make Fe{sub 3}O{sub 4} nanoparticles of potential interest as therapeutic agents against amyloid diseases and their non-risk exploitation in nanomedicine and nanodiagnostics.

  17. 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.

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

    Science.gov (United States)

    Kong, Hailong; Lv, Min; Mao, Nian; Wang, Cheng; Cheng, Yunxia; Zhang, Lei; Jiang, Xingfu; Luo, Lizhi

    2016-01-01

    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. PMID:27575006

  19. Serum thymidine kinase activity as a useful marker for bovine leukosis.

    Science.gov (United States)

    Sakamoto, Leo; Ohbayashi, Tetsu; Matsumoto, Kotaro; Kobayashi, Yoshiyasu; Inokuma, Hisashi

    2009-11-01

    Serum thymidine kinase (TK) activity has recently been evaluated as a serum marker for human and canine hematopoietic neoplasms. The purpose of the current study was to establish the significance of serum TK activity in the diagnosis of bovine leukosis. The discrimination value for TK activity was set at 5.4 U/l based on the receiver operating characteristic curve. In the group of clinically healthy cows, only 2 out of 83 cows (2.4%) had serum TK activity above the discrimination value. In contrast, 19 out of 20 cows (95.0%) with bovine leukosis showed serum TK activity above the discrimination value, although only 7 of 79 (8.9%) cows diagnosed with diseases other than bovine leukosis showed elevated serum TK activity. Thymidine kinase activities of all Bovine leukemia virus-positive cows with or without lymphocytosis were below the discrimination value. Sensitivity and specificity of measuring serum TK activity as a diagnostic tool for bovine leukosis was 95.0% and 95.9%, respectively. Results indicate that serum TK activity may be a marker for bovine leukosis.

  20. Serum YKL-40, a potential new marker of disease activity in patients with inflammatory bowel disease

    DEFF Research Database (Denmark)

    Vind, Ida; Johansen, J S; Price, P A

    2003-01-01

    BACKGROUND: YKL-40 is secreted by macrophages and neutrophils and is a growth factor for vascular endothelial cells and fibroblasts. Elevated serum concentrations of YKL-40 are found in patients with diseases characterized by inflammation or ongoing fibrosis. The aim of this study was to seek...... association between serum YKL-40 in patients with ulcerative colitis (UC) and Crohn disease (CD) and clinical disease activity. METHODS: One-hundred-and-sixty-four patients with UC and 173 patients with CD were studied. The Simple Clinical Colitis Activity Index (SCCAI) and the Harvey-Bradshaw (H-B) score...... were used to assess disease activity. Serum YKL-40 (determined by ELISA) was related to C-reactive protein (CRP) and disease activity. RESULTS: In patients with UC, the median serum YKL-40 rose with increasing disease activity, and patients with severe active disease had higher serum YKL-40 (median 59...

  1. On the adsorption of magnetite nanoparticles on lysozyme amyloid fibrils.

    Science.gov (United States)

    Majorosova, Jozefina; Petrenko, Viktor I; Siposova, Katarina; Timko, Milan; Tomasovicova, Natalia; Garamus, Vasil M; Koralewski, Marceli; Avdeev, Mikhail V; Leszczynski, Błażej; Jurga, Stefan; Gazova, Zuzana; Hayryan, Shura; Hu, Chin-Kun; Kopcansky, Peter

    2016-10-01

    An adsorption of magnetic nanoparticles (MNP) from electrostatically stabilized aqueous ferrofluids on amyloid fibrils of hen egg white lysozyme (HEWL) in 2mg/mL acidic dispersions have been detected for the MNP concentration range of 0.01-0.1vol.%. The association of the MNP with amyloid fibrils has been characterized by transmission electron microscopy (TEM), small-angle X-ray scattering (SAXS) and magneto-optical measurements. It has been observed that the extent of adsorption is determined by the MNP concentration. When increasing the MNP concentration the formed aggregates of magnetic particles repeat the general rod-like structure of the fibrils. The effect is not observed when MNP are mixed with the solution of lysozyme monomers. The adsorption has been investigated with the aim to clarify previously found disaggregation activity of MNP in amyloid fibrils dispersions and to get deeper insight into interaction processes between amyloids and MNP. The observed effect is also discussed with respect to potential applications for ordering lysozyme amyloid fibrils in a liquid crystal phase under external magnetic fields.

  2. Serum creatine kinase and lactate dehydrogenase activities in ...

    African Journals Online (AJOL)

    hypothyroidism is defined by the finding of elevated serum. TSH concentrations ... Access this article online. Quick Response ... in thyroid disorders condition is higher in women than men and increases with ..... thyroid diseases and pregnancy.

  3. Platelet antibodies, activated platelets and serum leptin in childhood immune thrombocytopenic purpura.

    Science.gov (United States)

    Badrawy, Hosny; Elsayh, Khalid I; Zahran, Asmaa M; El-Ghazali, Mohamad Hamdy

    2013-01-01

    The aim of this study was to evaluate the levels of platelet-associated antibodies (PAIgG and PAIgM), activated platelets and serum leptin in children with acute immune thrombocytopenic purpura (ITP). The study included 40 patients with ITP and 40 healthy age- and sex-matched controls. PAIgG, PAIgM and activated platelet levels were estimated by flow cytometry, and serum leptin levels were estimated by ELISA. Activated platelets and serum leptin were significantly higher in the ITP patients than in the controls. The percentage and mean fluorescence intensity of PAIgG and PAIgM staining were significantly higher in the patients than in the controls. Serum leptin and activated platelet levels in patients with thrombocytopenia of brief duration were significantly lower than those in patients with thrombocytopenia of prolonged duration. The levels of activated platelets, serum leptin and PAIgG were positively correlated, and the levels of serum leptin, activated platelets and platelet counts were negatively correlated. The increased levels of activated platelets, serum leptin and platelet-associated antibodies in children with acute ITP suggest that these factors could play a role in ITP pathogenesis. Additionally, activated platelets and serum leptin could have prognostic significance in paediatric acute ITP. Copyright © 2013 S. Karger AG, Basel.

  4. Determination of alternative pathway of complement activity in mouse serum using rabbit erythrocytes

    NARCIS (Netherlands)

    Dijk, H. van; Rademaker, P.M.; Willers, J.M.N

    1980-01-01

    Rabbit, mouse and sheep erythrocytes expressing different concentrations of membrane sialic acid were used to study possible modes of activation of the alternative complement (C) pathway in mouse, human and guinea pig serum. Mouse erythrocytes activated only human serum, whereas rabbit erythrocytes

  5. Inhibition of lysozyme amyloidogenesis by phospholipids. Focus on long-chain dimyristoylphosphocholine.

    Science.gov (United States)

    Ponikova, Slavomira; Kubackova, Jana; Bednarikova, Zuzana; Marek, Jozef; Demjen, Erna; Antosova, Andrea; Musatov, Andrey; Gazova, Zuzana

    2017-11-01

    Protein amyloid aggregation is an important pathological feature of a group of different degenerative human diseases called amyloidosis. We tested effect of two phospholipids, 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC) on amyloid aggregation of hen egg white (HEW) lysozyme in vitro. Effect of phospholipids was investigated using spectroscopic techniques (fluorescence and CD spectroscopy), atomic force microscopy and image analysis. Phospholipids DMPC and DHPC are able dose-dependently inhibit lysozyme fibril formation. The length of the phospholipid tails and different structural arrangement of the phospholipid molecules affect inhibitory activity; long-chain DMPC inhibits fibrillization more efficiently. Interestingly, interference of DMPC with lysozyme amyloid fibrils has no effect on their morphology or amount. Phospholipid molecules have significant effect on lysozyme amyloid fibrillization. We suggest that inhibitory activity is due to the interference of phospholipids with lysozyme leading to the blocking of the intermolecular protein interactions important for formation of the cross-β structure within the core of the fibrils. The higher inhibitory activity of DMPC is probably due to adsorption of protein molecules on the liposome surfaces which caused decrease of species needed for fibrillization. Interaction of the phospholipids with formed fibrils is not sufficient enough to interrupt the bonds in β-sheets which are required for destroying of amyloid fibrils. The obtained results contribute to a better understanding of the effect of phospholipids on amyloid fibrillization of the lysozyme. The data suggest that DMPC and DHPC phospholipids represent agents able to modulate lysozyme amyloid aggregation. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Intermittent Photoperiod Schedule does not Influence Brain and Serum Melatonin and Selected Serum Antioxidant Enzymes Activity in Broiler Chickens

    Directory of Open Access Journals (Sweden)

    Mosleh N

    2016-07-01

    Full Text Available This study evaluated the effect of the intermittent light program on serum and brain melatonin concentrations, antioxidant enzyme activities, and homocysteine concentration in broiler chickens. A total of 60 one-day-old broiler chickens (Cobb 500 were distributed in three light-proof controlled rooms (20 chicks per room. All birds were reared in continuous light until 3 days of age. Then, chicks were treated as follows: 1 intermittent lighting program (1L: 3D cycles, 2 continuous lighting program (24L, and 3 nonintermittent restricted lighting program (8L: 16D. At day 42 of age, sera and brains were collected from all chicks of each group (at the end of the dark phase for groups 1 and 3. Serum and brain melatonin concentrations were not affected by the different photoperiod schedules. Serum glutathione reductase, glucose-6-phosphate dehydrogenase, superoxide dismutase, and glutathione peroxidase activity as well as homocysteine concentration were also similar between different treatment groups. In conclusion, intermittent lighting program during the rearing period does not affect serum and brain melatonin levels as well as antioxidant status at the end of the dark phase in broilers.

  7. IgG, IgA, and lysozyme in Martina Franca donkey jennies and their foals.

    Science.gov (United States)

    Veronesi, Maria C; Dall'Ara, Paola; Gloria, Alessia; Servida, Francesco; Sala, Elisabetta; Robbe, Domenico

    2014-04-01

    Because immune transfer from jenny to donkey foal is mostly unknown, the aim of the present study was to evaluate, from 5 days before to 10 days after foaling, immunoglobulin (Ig)G, IgA, and lysozyme peripartal concentrations in serum and mammary secretions of 10 healthy, spontaneously foaling Martina Franca jennies and in serum of their mature, viable, healthy foals, in the first 10 days after birth. The results showed that, in jennies, mammary secretion of IgG levels (ranging between 16 and 75 mg/mL) and IgA (0.9-2 mg/mL), and IgG (6.8-13.5 mg/mL) and IgA (0.5-2.4 mg/mL) serum concentrations were not different along the time of study. Also, IgG concentrations in serum of foals did not show significant differences although a high level was observed at 12 hours after birth (8 mg/mL), and IgA concentrations in serum of foals did not show any significant difference, although a high level was observed at 12 hours after birth (1.2 mg/mL). Lysozyme increased significantly at Day 2 after parturition in mammary secretions of jennies (551.9 μg/mL) and at 12 hours in serum of foals (25.9 μg/mL). The study demonstrated that the pattern of passive immune transfer in donkey foals seems to be similar to that reported for the horse foal, with IgG predominating IgA in serum and mammary secretions of the jenny and also in serum of foals. The most significant early increase in foals' serum concerns lysozyme, which probably plays an important role in the innate immunity of the donkey foal in the first challenging hours after birth.

  8. Acetylated Lysozyme as Impurity in Lysozyme Crystals: Constant Distribution Coefficient

    Science.gov (United States)

    Thomas, B. R.; Chernov, A. A.

    2000-01-01

    Hen egg white lysozyme (HEWL) was acetylated to modify molecular charge keeping the molecular size and weight nearly constant. Two derivatives, A and B, more and less acetylated, respectively, were obtained, separated, purified and added to the solution from which crystals of tetragonal HEWL crystals were grown. Amounts of the A or B impurities added were 0.76, 0.38 and 0.1 milligram per millimeter while HEWL concentration were 20, 30 and 40 milligram per milliliter. The crystals grown in 18 experiments for each impurity were dissolved and quantities of A or B additives in these crystals were analyzed by cation exchange high performance liquid chromatography. All the data for each set of 18 samples with the different impurity and regular HEWL concentrations is well described by one distribution coefficient K = 2.15 plus or minus 0.13 for A and K = 3.42 plus or minus 0.25 for B. The observed independence of the distribution coefficient on both the impurity concentration and supersaturation is explained by the dilution model described in this paper. It shows that impurity adsorption and incorporation rate is proportional to the impurity concentration and that the growth rate is proportional to the crystallizing protein in solution. With the kinetic coefficient for crystallization, beta = 5.10(exp -7) centimeters per second, the frequency at which an impurity molecule near the growing interface irreversibly joins a molecular site on the crystal was found to be 3 1 per second, much higher than the average frequency for crystal molecules. For best quality protein crystals it is better to have low microheterogeneous protein impurity concentration and high supers aturation.

  9. Changes in serum angiotensin I converting enzyme activity due to carbon disulfide exposure

    Energy Technology Data Exchange (ETDEWEB)

    Filipovic, N.; Bilalbegovic, Z.; Sefic, M.; Djuric, D.

    1984-05-01

    The activity of serum angiotensin I converting enzyme (ACE) was determined in 50 workers from a viscose factory in Banja Luka, Yugoslavia, and in 50 control subjects. Activity of serum ACE was significantly lower in workers exposed to carbon disulfide than in the control group. No correlation was found between a decrease of serum ACE in exposed workers and duration of exposure. These findings indicate that the serum ACE may be influenced by carbon disulfide, but the mechanism of these changes remains to be elucidated in this case.

  10. Changes in serum angiotensin I converting enzyme activity due to carbon disulfide exposure.

    Science.gov (United States)

    Filipović, N; Bilalbegović, Z; Sefić, M; Djurić, D

    1984-01-01

    The activity of serum angiotensin I converting enzyme (ACE) was determined in 50 workers from a viscose factory in Banja Luka, Yugoslavia, and in 50 control subjects. Activity of serum ACE was significantly lower in workers exposed to carbon disulfide than in the control group. No correlation was found between a decrease of serum ACE in exposed workers and duration of exposure. These findings indicate that the serum ACE may be influenced by carbon disulfide, but the mechanism of these changes remains to be elucidated in this case.

  11. Effect of surface energy of solid surfaces on the micro- and macroscopic properties of adsorbed BSA and lysozyme.

    Science.gov (United States)

    Sharma, Indu; Pattanayek, Sudip K

    2017-07-01

    The surface energy, a macroscopic property, depends on the chemical functionality and micro- and macroscopic roughness of the surface. The adsorption of two widely used proteins bovine serum albumin (BSA) and lysozyme on surfaces of four different chemical functionalities were done to find out the interrelation between macroscopic and microscopic properties. We have observed the secondary structure of protein after its adsorption. In addition, we observed the variation of surface energy of proteins due to variation in adsorption time, change in protein concentration and effect of a mixture of proteins. Surfaces of three different chemical functionalities namely, amine, hydroxyl and octyl were obtained through self-assembled monolayer on silica surfaces and were tested for responses towards adsorption of lysozyme and BSA. The adsorbed lysozyme has higher surface energy than the adsorbed BSA on amine and octyl surfaces. On hydroxyl functional surface, the surface energy due to the adsorbed lysozyme or BSA increases slowly with time. The surface energy of the adsorbed protein increases gradually with increasing protein concentration on hydrophobic surfaces. On hydrophilic surfaces, with increasing BSA concentration in bulk solution, the surface energy of the adsorbed protein on GPTMS and amine surfaces is maximum at 1μM concentration. During the adsorption from a mixture of BSA and lysozyme on octyl surface, first lysozyme adsorbs and subsequent BSA adsorption leads to a high surface energy. Copyright © 2016. Published by Elsevier B.V.

  12. Neutron structure of the T26H mutant of T4 phage lysozyme provides insight into the catalytic activity of the mutant enzyme and how it differs from that of wild type.

    Science.gov (United States)

    Hiromoto, Takeshi; Meilleur, Flora; Shimizu, Rumi; Shibazaki, Chie; Adachi, Motoyasu; Tamada, Taro; Kuroki, Ryota

    2017-07-13

    T4 phage lysozyme is an inverting glycoside hydrolase that degrades the murein of bacterial cell walls by cleaving the β-1,4-glycosidic bond. The substitution of the catalytic Thr26 residue to a histidine converts the wild type from an inverting to a retaining enzyme, which implies that the original general acid Glu11 can also act as an acid/base catalyst in the hydrolysis. Here, we have determined the neutron structure of the perdeuterated T26H mutant to clarify the protonation states of Glu11 and the substituted His26, which are key in the retaining reaction. The 2.09-Å resolution structure shows that the imidazole group of His26 is in its singly protonated form in the active site, suggesting that the deprotonated Nɛ2 atom of His26 can attack the anomeric carbon of bound substrate as a nucleophile. The carboxyl group of Glu11 is partially protonated and interacts with the unusual neutral state of the guanidine moiety of Arg145, as well as two heavy water molecules. Considering that one of the water-binding sites has the potential to be occupied by a hydronium ion, the bulk solvent could be the source for the protonation of Glu11. The respective protonation states of Glu11 and His26 are consistent with the bond lengths determined by an unrestrained refinement of the high-resolution X-ray structure of T26H at 1.04-Å resolution. The detail structural information, including the coordinates of the deuterium atoms in the active site, provides insight into the distinctively different catalytic activities of the mutant and wild type enzymes. © 2017 The Authors Protein Science published by Wiley Periodicals, Inc. on behalf of The Protein Society.

  13. Serum interleukin-18 levels are associated with physical activity in Japanese men.

    Directory of Open Access Journals (Sweden)

    Kanae Oda

    Full Text Available OBJECTIVE: To investigate the link between serum interleukin-18 (IL-18 levels and physical activity in Japanese men. METHODS: A total of 81 men (45.7±17.6 years old was enrolled in this cross-sectional investigation study. We assessed anthropometric and body composition parameters. Serum IL-18 levels, physical activity by uniaxial accelerometers, peak oxygen uptake and metabolic risk parameters were also evaluated. RESULTS: Serum IL-18 levels were 179.4±84.7 pg/mL. Physical activity evaluated by Σ[metabolic equivalents × h per week (METs⋅h/w]was significantly and negatively correlated with serum IL-18 levels (r = -0.252, p = 0.0235. These associations remained even after adjusting for age, peak oxygen uptake and other confounding factors. CONCLUSION: Serum IL-18 levels were closely associated with physical activity independent of peak oxygen uptake in Japanese men.

  14. Influence of some formulation and process parameters on the stability of lysozyme incorporated in corn flour- or corn starch-based extruded materials prepared by melt blending processing.

    Science.gov (United States)

    Jbilou, Fouzia; Galland, Sophie; Telliez, Camille; Akkari, Zied; Roux, Roselyne; Oulahal, Nadia; Dole, Patrice; Joly, Catherine; Degraeve, Pascal

    2014-12-01

    In order to obtain an antimicrobial biodegradable material, corn flour was extruded with 1% of lysozyme. Since the limited stability of natural preservatives such as lysozyme is a common bottleneck to the elaboration of active biomaterials by melt blending processes, the influence of formulation and of extrusion processing temperature on its residual enzymatic activity was investigated. To assess the contribution of process parameters such as temperature, shear stress and of related formulation parameters such as glycerol and moisture contents, the stability of lysozyme following its extrusion or its thermoforming with plasticized corn starch or thermal treatments in aqueous glycerol solutions was also studied. Increasing glycerol content from 25% to 30% significantly limited inactivation of lysozyme during extrusion, while increasing initial moisture content of the mixture from 14.5% to 28.5% had the opposite effect. These observations open the possibility to prepare active materials retaining more than 60±7% of initial lysozyme activity.

  15. Serum paraoxonase activity and lipid hydroperoxide levels in adult ...

    African Journals Online (AJOL)

    EB

    2013-09-03

    Sep 3, 2013 ... different substrates.16 First, the rate of hydrolysis of paraoxon was ... when p value was less than 0.05. ... parameters, and serum LOOH levels, paraoxonase .... inhibition of cell growth. ... Hambrecht R, Wolf A, Gielen S, et al.

  16. 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.

  17. Hydrophobic nano-carrier for lysozyme adsorption

    Indian Academy of Sciences (India)

    CANAN ALTUNBAS; FULDEN ZEYNEP URAL; MURAT UYGUN; NESIBE AVCIBASI; UGUR AVCIBASI; DENIZ AKTAS UYGUN; SINAN AKGÖL

    2016-04-01

    In this work, poly(HEMA–APH) nanoparticles were synthesized by surfactant-free emulsion polymerization technique.Magnetic behaviour was introduced by simple addition of Fe$_3$O$_4$ into the polymerization medium.Characterization of the nanoparticle was carried out by FTIR, ESR, SEM, AFM and EDX analyses. These synthesized magnetic nanoparticles were used for adsorption of lysozyme. For this purpose, adsorption conditions wereoptimized and maximum lysozyme binding capacity was found to be 278.8 mg g$^{−1}$ polymer in pH 7.0 phosphate buffer at 25$^{\\circ}$C. Desorption and reusability properties of the nanoparticles were investigated and lysozyme adsorption efficiency did not change significantly at the end of the 10 successive reuses.

  18. Serum markers associated with disease activity in giant cell arteritis and polymyalgia rheumatica

    NARCIS (Netherlands)

    van der Geest, Kornelis S. M.; Abdulahad, Wayel H.; Rutgers, Abraham; Horst, Gerda; Bijzet, Johan; Arends, Suzanne; Roffel, Mirjam P.; Boots, Annemieke M. H.; Brouwer, Elisabeth

    Objective. To compare multiple serum markers for their ability to detect active disease in patients with GCA and in those with PMR. Methods. Twenty-six markers related to immune cells that may be involved in GCA and PMR were determined by ELISA and multiplex assay in the serum of 24 newly diagnosed,

  19. Serum markers associated with disease activity in giant cell arteritis and polymyalgia rheumatica

    NARCIS (Netherlands)

    van der Geest, Kornelis S. M.; Abdulahad, Wayel H.; Rutgers, Abraham; Horst, Gerda; Bijzet, Johan; Arends, Suzanne; Roffel, Mirjam P.; Boots, Annemieke M. H.; Brouwer, Elisabeth

    2015-01-01

    Objective. To compare multiple serum markers for their ability to detect active disease in patients with GCA and in those with PMR. Methods. Twenty-six markers related to immune cells that may be involved in GCA and PMR were determined by ELISA and multiplex assay in the serum of 24 newly diagnosed,

  20. Serum markers associated with disease activity in giant cell arteritis and polymyalgia rheumatica

    NARCIS (Netherlands)

    van der Geest, Kornelis S. M.; Abdulahad, Wayel H.; Rutgers, Abraham; Horst, Gerda; Bijzet, Johan; Arends, Suzanne; Roffel, Mirjam P.; Boots, Annemieke M. H.; Brouwer, Elisabeth

    2015-01-01

    Objective. To compare multiple serum markers for their ability to detect active disease in patients with GCA and in those with PMR. Methods. Twenty-six markers related to immune cells that may be involved in GCA and PMR were determined by ELISA and multiplex assay in the serum of 24 newly diagnosed,

  1. Interaction of fullerenol with lysozyme investigated by experimental and computational approaches

    Energy Technology Data Exchange (ETDEWEB)

    Yang Shengtao; Wang Haifang; Guo Lin; Gao Yang; Liu Yuanfang [Beijing National Laboratory for Molecular Sciences, Department of Chemical Biology, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871 (China); Cao Aoneng [Institute of Nanochemistry and Nanobiology, Shanghai University, Shanghai 200444 (China)], E-mail: haifangw@pku.edu.cn, E-mail: ancao@shu.edu.cn

    2008-10-01

    The potential biomedical applications of fullerenol C{sub 60}(OH){sub x} (x{approx}24) have been extensively studied. However, the structural information of the interaction of fullerenol with the bio-system at the molecular level, which is essential for understanding its bioactivity and toxicity, is still missing. In this study, lysozyme was selected as a model protein to investigate the interaction between fullerenol and biomolecules. A strong induced circular dichroism (CD) signal of achiral fullerenol was observed after binding with lysozyme. Activity assay shows that lysozyme activity is inhibited significantly by fullerenol. No heat capacity difference between the folded and unfolded states of lysozyme was measured by differential scanning calorimetry (DSC) in the presence of fullerenol, indicating that fullerenol prefers to bind with the hydrophobic residues. Both experimental and Autodock computational results suggest that the binding site on lysozyme for fullerenol is close to Trp 62, and a {pi}-{pi} stacking interaction might play an important role in binding.

  2. High-level soluble expression of the functional peptide derived from the C-terminal domain of the sea cucumber lysozyme and analysis of its antimicrobial activity

    Directory of Open Access Journals (Sweden)

    Lina Cong

    2014-11-01

    Conclusion: These results indicate that the expressed rSjLys-C is a highly soluble product and has a strong antimicrobial activity. Therefore, gaining a large quantity of biologically active rSjLys-C will be used for further biochemical and structural studies and provide a potential use in aquaculture and medicine.

  3. Judging disease activity in rheumatoid arthritis by serum free kappa and lambda light chain levels.

    Science.gov (United States)

    Ye, Yun; Li, Su-Liang; Xie, Ming; Jiang, Ping; Liu, Kai-Ge; Li, Ya-Jun

    2013-10-01

    The study aimed to evaluate the levels of serum free kappa (κ) and lambda (λ) light chains in patients with rheumatoid arthritis (RA) as well as exploring the association between serum free κ and λ light chains and activity of RA. For this purpose, healthy individuals and patients with active RA and RA in remission were enrolled, and their serum levels of free κ and λ light chains were measured using rate nephelometry. The diagnostic accuracy of serum free κ and λ light chains was evaluated by receiver operating characteristic curves and 95% confidence intervals for areas under the curve (AUC). The results obtained indicated that the levels of serum free κ and λ light chains in patients with active RA were significantly higher than those of patients in remission and of healthy controls (p light chain and 0.781 for free λ light chain. When the optimal cut-off point for serum κ light chain was 8.02 g/L, the maximum sensitivity and specificity were 82.5% and 82.5%, respectively, and when the optimal cut-off point for serum λ light chain was 3.57 g/L, the maximum sensitivity and specificity were 80% and 82.5%, respectively. It was thus found that serum levels of free κ and λ light chains were positively correlated with disease activity in RA, the Disease Activity Score 28 (DAS28), and values for C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), platelet count (PLT), rheumatoid factor (RF), and anticitrullinated protein antibody (ACPA) (p light chains in patients with active RA are closely correlated with disease activity parameters including DAS28, CRP, ESR, PLT, RF, and ACPA. Thus, the above-mentioned levels of serum free κ and λ light chains may be used as important indicators of activity of RA.

  4. Lysozyme immobilization onto PVC catheters grafted with NVCL and HEMA for reduction of bacterial adhesion

    Science.gov (United States)

    Guadarrama-Zempoalteca, Yesica; Díaz-Gómez, Luis; Meléndez-Ortiz, H. Iván; Concheiro, Angel; Alvarez-Lorenzo, Carmen; Bucio, Emilio

    2016-09-01

    The aim of the present work was to functionalize poly(vinyl chloride) (PVC) urinary catheters with grafted copolymers that can improve the biocompatibility and serve as binding points of lysozyme. PVC catheters were modified by grafting a mixture of N-vinylcaprolactam (NVCL) and 2-hydroxyethylmethacrylate (HEMA) applying a gamma-ray pre-irradiation method. The effect of absorbed dose, monomer concentration, temperature, and reaction time on the grafting percentage was evaluated. The grafted catheters were characterized regarding surface composition (FTIR-ATR spectroscopy), thermal properties (DSC and TGA) and swelling in aqueous medium. Lysozyme was directly coupled onto PVC-g-(NVCL/HEMA) previously activated using carbonyldiimidazole. Antimicrobial lytic activity of the modified catheters over time was tested against Micrococcus lysodeikticus. Lysozyme diminished the adhesion of Staphylococcus aureus onto the functionalized catheters, which may be suitable to prevent biofilm formation.

  5. Viscometric study of lysozyme solution with sugar and urea at various temperatures

    Directory of Open Access Journals (Sweden)

    Jamal Akhter Siddique

    2016-11-01

    at temperatures (293.15, 303.15, 313.13 and 323.15 K at various concentrations of glucose, maltose and urea. Change in entropy (ΔH, enthalpy (ΔS and free energy of activation (ΔG have also been evaluated for these systems. Value of B-coefficient of d (− glucose, maltose and urea has also been calculated from viscosity data in aqueous lysozyme solution. Viscosity B-coefficients of glucose and maltose in aqueous lysozyme solution are positive while that of the urea–lysozyme water system it is negative due to the structure breaking effect of urea. The values of entropy of activation are negative due to attainment of transition state for viscous flow, which is accompanied by bond formation and increase in order.

  6. Expression, Characterization and Antimicrobial Ability of a Variant T4 Lysozyme in Pichia pastoris

    Institute of Scientific and Technical Information of China (English)

    Ning SUN; Sanfeng CHEN; Xiangming XIE; Yueju WANG; Gangqiang LI; Nan WANG; Dehu LIU

    2014-01-01

    T4 lysozyme was engineered with disulfide bonds and expressed in Pichia pastoris. The secreted proteins were purified and made into powder by lyophiliza-tion. Recombinant protein purity was more than 70% measured by HPLC. The lytic activity of variant T4-lysozyme was measured by the lysis of the cel wal of Xan-thomonas oryzae pv. oryzae, X. oryzae pv. oryzicola, Ralstonia solanacearum comb. nov, Clavibacter michiganensis subsp. michiganensis, X. campestris pv. mal-vacearum, Fusarium oxysporium sp. vasinfectum, Verticil ium dahliae kleb. Inhibition zone assay showed that variant T4 lysozyme significantly inhibited X. o. oryzicola and X. c. malvacearum. The antifungal activities of this protein against F. o. vasin-fectum and V. d. kleb were also analyzed.

  7. Effects of breed and zeranol implantation on serum insulin, somatomedin-like activity and fibroblast proliferative activity.

    Science.gov (United States)

    Wangsness, P J; Olsen, R F; Martin, R J

    1981-01-01

    Twenty-eight Suffolk-sired (Sx) and 28 Finnsheep-sired (Fx) lambs were implanted with either 0 or 12 mg zeranol. Zeranol significantly increased average daily gain over that of controls. Serum taken at biweekly intervals for 6 weeks was assayed for insulin, somatomedin-like activity (Sm) and fibroblast proliferative activity (FPA). Insulin appeared to increase with time, but there were no consistent time changes for FPA or Sm. Serum insulin concentration was higher (P less than .05) in implanted lambs than in controls (33.4 vs 25.6 microU/ml). Unlike insulin, serum Sm and FPA were not affected by zeranol implantation, and, thus, these serum factors appeared not to be involved in zeranol-stimulated growth. Sm was higher in the faster growing Sx lambs than in the slower growing Fx lambs. Thus, serum Sm activity may be involved in normal regulation of growth.

  8. THE EFFECT OF SLEEP DEPRIVATION ON SERUM IGG RESPONSES TO AEROBIC ACTIVITY IN COLLEGE STUDENT ATHLETES

    Directory of Open Access Journals (Sweden)

    Saeed Jamshidi Far

    2014-11-01

    Full Text Available Background & Aims: Sleep is a restorative process for the immune system. There are many situations in which sleep is disturbed prior to an athletic event. However, the effect of sleep deprivation on immune indices in response to exercise remains unknown. The aim of this study was to investigate the effects of sleep deprivation on serum IgG responses to aerobic activity. Materials & Methods: In this quasi-experimental study, 10 male physical education students were voluntarily participated. Study was performed in two separate occasions; control and experimental within two weeks. In the control occasion, normal sleep and aerobic activity and in the experimental occasion, sleep deprivation and aerobic activity was applied. Aerobic activity was performed on bicycle ergometer for 30 minutes at intensity of 70 to 75 percent of maximum heart rate. Changes in serum IgG concentrations in pre-test, before and after aerobic activity in both occasions were analyzed by the two repeated measures ANOVA and dependent T-test using SPSS software. Results: The results showed that sleep deprivation not significantly effect on Serum IgG response to aerobic activity (p=0.130. Also, aerobic activity not significantly effect on Serum IgG concentration (p=0.357. But sleep deprivation caused a significantly increase in serum IgG concentration (p=0.035. Conclusion: No significant effect of sleep deprivation on serum IgG concentrations response to aerobic activity.

  9. Properties of lysozyme/sodium alginate complexes for the development of antimicrobial films.

    Science.gov (United States)

    Amara, Chedia Ben; Eghbal, Noushin; Oulahal, Nadia; Degraeve, Pascal; Gharsallaoui, Adem

    2016-11-01

    Complexation study of lysozyme (0.714g/L) by sodium alginate at pH7 showed that aggregates formation was a two-phase process. The first phase (from 0 to 0.1g/L sodium alginate) corresponded to the combination of individual complexes to form aggregates which caused an increase of turbidity and average size and a rapid sedimentation. Charge neutralization estimated by ζ-potential measurements occurred at 0.1g/L sodium alginate concentration. The second phase (from 0.1 to 4g/L of sodium alginate) was characterized by the formation of aggregates having a less dense structure with higher average size despite the drop in turbidity and the high dispersion in the medium. Lysozyme enzymatic activity decreased upon complexation with sodium alginate but was fully recovered after calcium chloride addition. In order to check whether lysozyme reversible inactivation was only due to substrate diffusion limitation or to conformational changes upon complexation, fluorescence and UV-Vis absorption measurements were performed. Moreover, lysozyme/sodium alginate complexes were used to manufacture an edible antimicrobial film to target lysozyme sensitive microorganisms. Copyright © 2016 Elsevier Ltd. All rights reserved.

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

    Energy Technology Data Exchange (ETDEWEB)

    Hughes, Ashley J. [Institute of Integrative Biology, University of Liverpool, Liverpool L69 7ZB (United Kingdom); Diamond Light Source Ltd., Diamond House, Didcot, Oxfordshire OX11 0DE (United Kingdom); Hussain, Rohanah [Diamond Light Source Ltd., Diamond House, Didcot, Oxfordshire OX11 0DE (United Kingdom); Cosentino, Cesare; Guerrini, Marco [Istituto di Chimica e Biochimica ' G. Ronzoni' , Via G. Colombo 81, Milano 20133 (Italy); Siligardi, Giuliano [Diamond Light Source Ltd., Diamond House, Didcot, Oxfordshire OX11 0DE (United Kingdom); Yates, Edwin A., E-mail: eayates@liv.ac.uk [Institute of Integrative Biology, University of Liverpool, Liverpool L69 7ZB (United Kingdom); Rudd, Timothy R., E-mail: trudd@liv.ac.uk [Institute of Integrative Biology, University of Liverpool, Liverpool L69 7ZB (United Kingdom); Istituto di Chimica e Biochimica ' G. Ronzoni' , Via G. Colombo 81, Milano 20133 (Italy)

    2012-09-07

    Highlights: Black-Right-Pointing-Pointer Zinc-heparan sulfate complex destabilises lysozyme, a model amyloid protein. Black-Right-Pointing-Pointer Addition of zinc, without heparan sulfate, stabilises lysozyme. Black-Right-Pointing-Pointer 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 {beta}-rich amyloid by far UV circular dichroism (increased {beta}-sheet: +25%), with a significantly reduced melting temperature (from 68 to 58 Degree-Sign 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.

  11. Clinical value of serum amylase, serum trypsinogen-2, IL-10 and platelet activating factor detection in diagnosis of post-ERCP pancreatitis

    Institute of Scientific and Technical Information of China (English)

    Di Wang; Xue-Song Li; Peng-Peng Cai

    2016-01-01

    Objective:To explore the clinical value of serum amylase, serum trypsinogen-2, IL-10 and platelet activating factor detection in the diagnosis of post-ERCP pancreatitis.Methods:A total of 256 patients with ERCP were selected in our hospital of the digestive department of internal medicine from January 2015 to December 2015. According to different symptoms, signs, serum amylase and pancreatic CT examination results after ERCP, the subjects were divided into three groups: control group, hyperlipidemia group and PEP group. Serum trypsinogen-2, IL-10, platelet activating factor were detected.Results:There were statistically significant differences in serum trypsinogen-2, IL-10, platelet activating factor among control group, hyperlipidemia group and PEP group patients with serum amylase (Fgroup = 269.578, 139.492, 145.637, 126.891, allP = 0.000), and the value of PEP group was the highest. There were statistically significant differences in serum trypsinogen-2, IL-10, platelet activating factor at different time point (Ftime = 602.506, 494.375, 462.512, 239.564, allP = 0.000), the serum amylase level in the patients with hyperlipidemia group reached the highest value after 4 h, but the 24 h was lower than the normal level after operation, the serum amylase in PEP group increased rapidly after 4 h, and the 24 h was still increased after operation; The serum trypsinogen-2, IL-10, platelet activating factor were all highest 4 h after the operation in the hyperlipidemia group and PEP group, and the 24 h after the operation were all decreased. In PEP group, serum amylase, serum trypsinogen-2, IL-10, platelet activating factor 4 h after the operation and 24 h after the operation were significantly higher than those before operation (Finteractive = 28.492, 22.614, 16.573, 7.819, allP = 0.000). In PEP group, the serum amylase had positive correlations with serum trypsinogen-2, IL-10, platelet activating factor (r=0.591, 0.662, 0.681, allP<0.05).Conclusions: Serum amylase

  12. Antimicrobial Activity of Ceftriaxone Compared with Cefotaxime in the Presence of Serum Albumin

    Directory of Open Access Journals (Sweden)

    Swapan K Nath

    1995-01-01

    Full Text Available The effect of serum albumin on the antimicrobial activity of ceftriaxone, cefotaxime, and a 1:1 ratio of cefotaxime and its desacetyl metabolite against nonpseudomonal Gram-negative bacilli was determined. Antimicrobial activity of drugs was evaluated by measuring minimum inhibitory (mic and bactericidal (mbc concentrations in broth with and without human serum albumin. The analysis of logarithmically transformed mean mics and mbcs showed that there was a highly significant interaction between drug and serum albumin (P<0.0001. The inhibitory and bactericidal activities were greatest for cefotaxime followed by cefotaxime/desacetylcefotaxime and ceftriaxone (P<0.01. Time-kill kinetics demonstrated that ceftriaxone was less bactericidal than cefotaxime in broth with albumin. On the basis of these results it was concluded that the in vitro antimicrobial activity of ceftriaxone compared with that of cefotaxime was significantly diminished in the presence of serum albumin.

  13. Serum bactericidal activity as indicator of innate immunity in pacu Piaractus mesopotamicus (Holmberg, 1887

    Directory of Open Access Journals (Sweden)

    J.D. Biller-Takahashi

    2013-12-01

    Full Text Available The immune system of teleost fish has mechanisms responsible for the defense against bacteria through protective proteins in several tissues. The protein action can be evaluated by serum bactericidal activity and this is an important tool to analyze the immune system. Pacu, Piaractus mesopotamicus, is one of the most important fish in national aquaculture. However there is a lack of studies on its immune responses. In order to standardize and assess the accuracy of the serum bactericidal activity assay, fish were briefly challenged with Aeromonas hydrophila and sampled one week after the challenge. The bacterial infection increased the concentration of protective proteins, resulting in a decrease of colony-forming unit values expressed as well as an enhanced serum bactericidal activity. The protocol showed a reliable assay, appropriate to determine the serum bactericidal activity of pacu in the present experimental conditions.

  14. Lysozyme uptake by oxidized starch polymer microgels

    NARCIS (Netherlands)

    Li, Y.; Vries, R.D.; Kleijn, M.; Slaghek, T.; Timmermans, J.; Stuart, M.C.; Norde, W.

    2010-01-01

    With the aim of determining suitable conditions for uptake and release of globular proteins on microgels, we studied the interaction between phosphated, highly cross-linked, negatively charged oxidized potato starch polymer (OPSP) microgel particles and lysozyme from hen eggs. Our microgel shows a

  15. Complex coacervates of hyaluronic acid and lysozyme

    DEFF Research Database (Denmark)

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

    2014-01-01

    Complex coacervates of hyaluronic acid and lysozyme, a model protein, were formed by ionic interaction using bulk mixing and were characterized in terms of binding stoichiometry and protein structure and stability. The complexes were formed at pH 7.2 at low ionic strength (6 mM) and the binding s...

  16. Relationship between changed alveolar-capillary permeability and angiotensin converting enzyme activity in serum in sarcoidosis.

    OpenAIRE

    Eklund, A; Blaschke, E

    1986-01-01

    The effect of altered alveolar-capillary permeability on angiotensin converting enzyme (ACE) activity in serum (SACE) was studied in 45 patients with sarcoidosis and 21 healthy controls. In sarcoidosis increased albumin concentrations in the bronchoalveolar lavage fluid (L albumin) and increased ratios of L albumin to albumin in serum (S albumin) indicated an increased permeability of the alveolar-capillary membrane. ACE activity in the lavage fluid (LACE) was correlated with the number of al...

  17. BINDING OF THERMO-SENSITIVE AND pH-SENSITIVE BUTYLATED POLY(ALLYLAMINE)S WITH LYSOZYME

    Institute of Scientific and Technical Information of China (English)

    Jing-jing Liu; Yun-feng Yan; Ping Yao

    2011-01-01

    Butyl modified poly(allylamine)s with butyl substitution degrees of 15% to 70% were prepared. The polymers show pH sensitive property and lower critical solution temperature (LCST) behavior. The LCST appears at lower temperature, lower pH and lower polymer concentration for the polymer with higher butylated degree. The binding of native lysozyme with the polymers depends on the hydrophobicity of the polymers at the pH range that the protein and the polymer carry the same positive charges. The increase of polymer hydrophobicity can increase the binding with lysozyme, but the self-aggregation of the polymer decreases the binding. The bound lysozyme molecules can recover their native activity completely after the dissociation of the complexes. Compared with native lysozyme, the denatured one which exposes the hydrophobic residues can increase the binding with the polymer and form stable complex nanoparticles.

  18. 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.

  19. [Serum gamma-glutamyl transpeptidase activity in children with chronic hepatitis B].

    Science.gov (United States)

    Lebensztejn, Dariusz Marek; Skiba, Elzbieta; Sobaniec-Lotowska, Maria Elzbieta; Kaczmarski, Maciej

    2005-03-01

    THE AIM OF THE STUDY was evaluation of serum activity of chosen enzymes (ALT, AST, GGT and ALP) in assessment of fibrosis degree in children with chronic hepatitis B. We determined serum activity of liver enzymes in 47 children aged 4-16 with biopsy-verified chronic hepatitis B, prior to interferon alpha treatment. Fibrosis stage was assessed in a blinded fashion according to Batts and Ludwig. We defined advanced fibrosis as a score =3. Receiver operating characteristics (ROC) analysis was used to calculate the power of the assays to detect advanced liver fibrosis (AccuROC, Canada). Serum GGT activity of 19 IU/I had a sensitivity of 50% and a specificity of 90% (AUC=0.7324, p=0.0391) to predict advanced fibrosis. All other enzymes did not allow a useful prediction. GGT is suitable serum marker to predict advanced liver fibrosis in children with chronic hepatitis B.

  20. The protective effect of salicylic acid on lysozyme against riboflavin-mediated photooxidation

    Science.gov (United States)

    Li, Kun; Wang, Hongbao; Cheng, Lingli; Zhu, Hui; Wang, Mei; Wang, Shi-Long

    2011-06-01

    As a metabolite of aspirin in vivo, salicylic acid was proved to protect lysozyme from riboflavin-mediated photooxidation in this study. The antioxidative properties of salicylic acid were further studied by using time-resolved laser flash photolysis of 355 nm. It can quench the triplet state of riboflavin via electron transfer from salicylic acid to the triplet state of riboflavin with a reaction constant of 2.25 × 10 9 M -1 s -1. Mechanism of antioxidant activities of salicylic acid on lysozyme oxidation was discussed. Salicylic acid can serve as a potential antioxidant to quench the triplet state of riboflavin and reduce oxidative pressure.

  1. Characterization of Serum Phospholipase A2 Activity in Three Diverse Species of West African Crocodiles

    Directory of Open Access Journals (Sweden)

    Mark Merchant

    2011-01-01

    Full Text Available Secretory phospholipase A2, an enzyme that exhibits substantial immunological activity, was measured in the serum of three species of diverse West African crocodiles. Incubation of different volumes of crocodile serum with bacteria labeled with a fluorescent fatty acid in the sn-2 position of membrane lipids resulted in a volume-dependent liberation of fluorescent probe. Serum from the Nile crocodile (Crocodylus niloticus exhibited slightly higher activity than that of the slender-snouted crocodile (Mecistops cataphractus and the African dwarf crocodile (Osteolaemus tetraspis. Product formation was inhibited by BPB, a specific PLA2 inhibitor, confirming that the activity was a direct result of the presence of serum PLA2. Kinetic analysis showed that C. niloticus serum produced product more rapidly than M. cataphractus or O. tetraspis. Serum from all three species exhibited temperature-dependent PLA2 activities but with slightly different thermal profiles. All three crocodilian species showed high levels of activity against eight different species of bacteria.

  2. Tear Film and Serum Prolidase Activity and Oxidative Stress in Patients With Keratoconus.

    Science.gov (United States)

    Göncü, Tuğba; Akal, Ali; Adıbelli, Fatih Mehmet; Çakmak, Sevim; Sezen, Hatice; Yılmaz, Ömer Faruk

    2015-09-01

    To determine and compare the serum and tear film prolidase activity (PA) between patients with keratoconus and healthy subjects. Also, we aimed to evaluate the serum oxidative stress level and the correlation with serum PA in patients with keratoconus. This prospective, comparative clinical study included 31 patients with keratoconus and 33 age-matched and sex-matched control subjects. All participants underwent a detailed ophthalmologic examination. Serum and tear samples were obtained from all participants. Tears and serum PA and serum oxidative stress markers were measured. No significant differences in demographic characteristics were detected between groups (P > 0.05). The serum PA was significantly lower in the keratoconus group than in the control group (895.6 ± 198.7 vs. 1145.9 ± 285.4 U/L, P keratoconus group than in the control group; however, this difference was not significant (3075.4 ± 672.2 vs. 3225.8 ± 903.2 U·L⁻¹·g⁻¹ protein, P = 0.45). Oxidative stress markers, such as total oxidant status and oxidative stress index, were found to be significantly higher in the keratoconus group (P keratoconus than in the controls. Additionally, serum oxidative stress markers were found to be higher than those of the controls. Thus, prolidase and systemic oxidative stress may have a role in the pathogenesis of keratoconus.

  3. Serum paraoxonase activity, total thiols levels, and oxidative status in patients with acute brucellosis.

    Science.gov (United States)

    Esen, Ramazan; Aslan, Mehmet; Kucukoglu, Mehmet Emin; Cıkman, Aytekin; Yakan, Umit; Sunnetcioglu, Mahmut; Selek, Sahbettin

    2015-06-01

    It is well known that paraoxonase-1 (PON1) activity may decrease during the course of infection and inflammation. The aim of this study was to investigate serum PON1 activity, oxidative status, and thiols levels in patients with acute brucellosis. In addition, we investigated the PON1 phenotype in patients with acute brucellosis. Thirty patients with acute brucellosis and 35 healthy controls were enrolled. Serum paraoxonase and arylesterase activities, thiols levels, lipid hydroperoxide levels, total antioxidant capacity (TAC), total oxidant status (TOS) and oxidative stress index (OSI) were determined. Serum basal and salt-stimulated paraoxonase-arylesterase activities, TAC levels and thiols levels were significantly lower in patients with acute brucellosis than controls (for all, p brucellosis. These results indicate that lower PON1 activity is associated with oxidant-antioxidant imbalance.

  4. SPE-HPLC purification of endocrine disrupting compounds from human serum for assessment of xenoestrogenic activity

    DEFF Research Database (Denmark)

    Hjelmborg, P.S.; Ghisari, Mandana; Bonefeld-Jørgensen, Eva

    2006-01-01

    Assessment of xenoestrogenic activity in human serum samples requires the removal of endogenous sex hormones to assure that the activity measured originates from xenobiotic compounds only. Serum samples representing high, medium and lower accumulation of persistent organic pollutants (POPs) were...... extracted using solid-phase extraction (SPE) followed by normal-phase high-performance liquid chromatography (NP-HPLC) for separation of POPs from endogenous hormones. The recovery of polychlorinated biphenyl (PCB) congeners in spiked serum samples was up to 86 %, making the extraction method suitable...... measured by ERE-CALUX was validated and considered to be a valuable tool to assess the combined ER effect of lipophilic serum POPs where additive/synergistic and agonistic/antagonistic effects are integrated giving an overall estimate of exposure and bioactivity....

  5. Serum cysteine proteases and their inhibitors in rheumatoid arthritis: relation to disease activity and radiographic progression

    DEFF Research Database (Denmark)

    Kos, Janko; Krašovec, Marta; Troelsen, Lone

    2011-01-01

    This study aims to investigate the serum levels of cysteine proteases cathepsins B and H and their inhibitors stefin A, stefin B, and cystatin C, as well as traditional inflammatory markers such as C-reactive protein in patients with rheumatoid arthritis and to correlate these markers with scores...... of disease activity and radiographic progression. Seventy-two patients with rheumatoid arthritis were included from two previously described cohorts of patients with chronic polyarthritis. At inclusion, disease activity was assessed by a 28-joint count, patient global assessment, and serum C-reactive protein....... Erosive status of hands and wrists was expressed by the Larsen score and recorded at inclusion and after 1 year. Serum levels of cathepsin B, cathepsin H, stefin A, stefin B, and cystatin C were determined by enzyme-linked immunosorbent assay. Neither cathepsin B nor cathepsin H serum levels were...

  6. Relationship of serum somatomedin-like activity and fibroblast proliferative activity with age and growth in the rat.

    Science.gov (United States)

    Olsen, R F; Wangsness, P J; Patton, W H; Martin, R J

    1980-03-01

    An assay for fibroblast proliferative activity (FPA) using human lung fibroblasts, WI-38, was described. The assay was responsive to varying rat serum levels and was not influenced by direct growth hormone (GH) addition. The relationship of serum growth factors to age and body weight was examined in the rat. In study 1, serum was obtained from lean Zucker rats at 3, 5, 7, 9, 11 and 30 wk of age. Six samples were taken at each age and serum samples were analyzed for somatomedin-like activity (Sm) and fibroblast proliferative activity (FPA). Serum Sm was not different at any of the sampling ages. FPA was low at 3 wk, but was higher and constant from 5 wk to 30 wk. In study 2, 73 lean Zucker rats (7 wk of age) were maintained on laboratory chow and water ad libitum for 4 wk, and then serum was obtained by decapitation. The rats were ranked according to each of four different criteria: average daily gain (ADG) for the duration of the study, ADG for the fourth week, total body protein and total body fat. Serum Sm, FPA and insulin concentrations on the top 10 and bottom 10 rats of each ranking were compared. Neither FPA nor insulin was significantly different for any ranking. Serum Sm was significantly higher in the top 10 rats ranked by ADG for the duration of the study. Sm was not significantly different in rats ranked by body weight, total body protein or total body fat. The data suggest that serum somatomedin-like activity (Sm) may be important in the earlier stages of growth in rats.

  7. Increased serum YKL-40 in patients with pulmonary sarcoidosis--a potential marker of disease activity?

    DEFF Research Database (Denmark)

    Johansen, Julia S; Milman, Nils; Hansen, Michael

    2005-01-01

    YKL-40, a growth factor for fibroblasts and vascular endothelial cells, is secreted by macrophages and neutrophils. Elevated serum YKL-40 is found in patients with diseases characterized by inflammation, tissue remodelling and ongoing fibrosis. The aim was to evaluate whether macrophages and gian...... cells in the granulomatous sarcoid lesions of patients with pulmonary sarcoidosis produce YKL-40 and to determine whether serum YKL-40 in these patients were associated with disease activity....

  8. Antimicrobial lysozyme-containing starch microgel to target and inhibit amylase-producing micro-organisms

    NARCIS (Netherlands)

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

    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

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

    NARCIS (Netherlands)

    Li, Yuan; Kadam, Sachin; Abee, Tjakko; Slaghek, Ted M.; Timmermans, Johan W.; Stuart, Martien A. Cohen; Norde, Willem; Kleijn, Mieke 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

  10. Antimicrobial lysozyme-containing starch microgel to target and inhibit amylase-producing micro-organisms

    NARCIS (Netherlands)

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

    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

  11. 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

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

    NARCIS (Netherlands)

    Li, Yuan; Kadam, Sachin; Abee, Tjakko; Slaghek, Ted M.; Timmermans, Johan W.; Stuart, Martien A. Cohen; Norde, Willem; Kleijn, Mieke J.

    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

  13. [Cancer procoagulant and cathepsin D activity in blood serum in patients with bladder cancer].

    Science.gov (United States)

    Szajda, Sławomir Dariusz; Darewicz, Barbara; Kudelski, Jacek; Chlabicz, Marcin; Domel, Tomasz; Chabielska, Ewa; Skrzydlewski, Zdzisław

    2005-06-01

    The increasing morbidity and mortality rates of bladder cancer forced the scientists to search for new unfailing diagnostic and therapeutic methods that will improve treatment effects. There are biochemical cancer markers as cancer procoagulant (CP) and cathepsin D which may be used to this end. The aim of the study was to evaluate the activity of the cancer procoagulant and cathepsin D in the blood serum in patients with superficial bladder cancer. The venous blood samples were from 15 patients with microscopically proved superficial bladder carcinoma (i.e. study group) and 15 normal volunteers as a control group. The serum blood CP activity was determined by the Gordon-Benson's coagulation method and expressed by the clotting time in seconds (s) while the cathepsin D activity was determined by the Folin-Ciocalteau's method and expressed by a quantity of released tyrosine in nmol/ml per 4 hours. The CP activity in serum of patients with superficial bladder cancer was increased in statistically way as compared to the non-cancer controls (p<0.0001). The cathepsin D activity in blood serum of the study group was also enhanced as compared to the control group and the said values differed statistically (p<0.0351). It appears to be justifiable to apply the determination of the CP and cathepsin D activity in blood serum for the diagnostics of superficial bladder cancer.

  14. Serum B-cell activating factor in myeloperoxiase-antineutrophil cytoplasmic antibodies-associated vasculitis.

    Science.gov (United States)

    Xin, Gang; Chen, Min; Su, Yun; Xu, Li-Xia; Zhao, Ming-Hui; Li, Kang-Sheng

    2014-07-01

    In this study, the serum B-cell activating factor belonging to tumor necrosis factor family (BAFF) levels in patients with myeloperoxidase (MPO)-antineutrophil cytoplasmic antibodies (ANCA)-associated vasculitis (AAV) were measured, and their clinical significance was further analyzed. One hundred twenty-one patients with MPO-AAV were enrolled in this study. Eighty-three patients had active vasculitis and 38 were in remission. Fifty-five healthy individuals were used as healthy controls. The levels of serum BAFF were assessed using commercial available enzyme-linked immunosorbent assay kits. The correlations between serum BAFF and Birmingham Vasculitis Activity Score, erythrocyte sedimentation rate and MPO-ANCA were further evaluated. The levels of serum BAFF of patients with MPO-AAV in both active (6.06±5.02 ng/mL) and remission phases (3.60±3.83 ng/mL) were significantly higher than those in healthy controls (0.87±0.31 ng/mL) (Pvasculitis were significantly higher than those in remission (PVasculitis Activity Score (r=0.320, Pvasculitis were 1.58 and 4.20 ng/mL, respectively. The levels of serum BAFF were elevated in patients with MPO-AAV and associated with disease activity, but they were not related with the levels of MPO-ANCA.

  15. Identification and characterization of serum complement activity in the American crocodile (Crocodylus acutus).

    Science.gov (United States)

    Merchant, Mark; McFatter, Justin; Mead, Stephanie; McAdon, Charles; Wasilewski, Joe

    2010-02-15

    Incubation of unsensitized sheep red blood cells with serum from the American crocodile (Crocodylus acutus) resulted in a concentration-dependent hemolysis. The hemolytic activity was heat-sensitive, and inhibited by EDTA in a concentration-dependent manner. The EDTA-inhibited SRBC hemolysis could be restored by the addition of excess Ca2+ or Mg2+, but not Ba2+ or Cu2+, revealing the specificity of this activity for these two divalent cations. The hemolytic activity of crocodile serum was titer-dependent, with 329 microL producing 50% of maximal SRBC hemolysis. The complement activity was also temperature-dependent, with decreased activity at lower temperatures (5-15 degrees C) and maximal activity occurred at 30-40 degrees C. The hemolysis occurred relatively slowly, with near zero activity after 10 min, 40% of activity observed within 15 min of exposure to SRBCs, and maximal activity at 30 min.

  16. Preparation of lysozyme molecularly imprinted polymers and purification of lysozyme from egg white.

    Science.gov (United States)

    Wang, Xuejiao; Dong, Shaohua; Bai, Quan

    2014-06-01

    Molecular imprinting as a promising and facile separation technique has received much attention because of its high selectivity for target molecules. In this study, lysozyme molecularly imprinted polymers (Lys-MIPs) were successfully prepared by the entrapment method with lysozyme as the template molecule, acrylamide as the functional monomer and N,N-methylenebisacrylamide as the cross-linker. The removal of the template lysozyme from the molecularly imprinted polymers was investigated in detail by two methods. The synthesized Lys-MIPs were characterized by scanning electron microscopy and Fourier transform-infrared, and the adsorption capacity, selectivity and reproducibility of the Lys-MIPs were also evaluated. The maximum adsorption capacity reached 94.8 mg/g, which is twice that of nonmolecularly imprinted polymers, and satisfactory selectivity and reproducibility were achieved. Using the Lys-MIP column, lysozyme could be separated completely from egg white, with purity close to 100% and mass recovery of 98.2%. This illustrated that the synthesized Lys-MIPs had high specific recognition and selectivity to the template lysozyme when they were applied to a mixture of protein standards and a real sample.

  17. Cyclophosphamide-induced changes of serum angiotensin converting enzyme activity and pulmonary microvessels ultrastructure.

    Science.gov (United States)

    Musiatowicz, B; Terlikowski, S; Sulik, M; Famulski, W; Giedrojć, J; Jakubowski, A; Sobaniec-Lotowska, M; Pasztaleniec, L; Baltaziak, M; Jabłońska, E

    1997-01-01

    The effect of cyclophosphamide (CP) on the ultrastructure of the lung tissue and the activity of angiotensin converting enzyme (ACE) in serum was evaluated in rats. The animals were given cyclophosphamide (CP) in a single intraperitoneal dose of 150 mg/kg b.w. ACE activity was evaluated in the blood serum collected from the left ventricle of the heart using the spectrophometric method. In all time subgroups, the CP-receiving animals showed a decrease in ACE activity. Ultrastructural examinations of CP-treated animals revealed increased adhesion of neutrophiles and monocytes to the damage endothelium of the alveolar septa vessels and focally accumulation of the platelets.

  18. An Evaluation of the Correlation between Hepcidin Serum Levels and Disease Activity in Inflammatory Bowel Disease

    Directory of Open Access Journals (Sweden)

    Zehra Betül Paköz

    2015-01-01

    Full Text Available Aim. While there are many well-defined serological markers for inflammatory bowel disease (IBD, there is limited evidence that they positively affect clinical outcomes. This study aimed to evaluate the correlation between hepcidin serum levels and disease activity in IBD. Materials and Methods. Eighty-five consecutive IBD patients were enrolled in the study. Hepcidin serum levels were assessed using an enzyme-linked immunosorbent assay (ELISA and were compared with disease activity as well as the interleukin-6 (IL-6 and C-reactive protein (CRP levels. Results. The mean hepcidin serum levels in Crohn’s disease (CD patients in remission and in the active phase were 3837±1436 and 3752±1274 pg/mL, respectively P=0.613. The mean hepcidin serum levels in ulcerative colitis (UC patients in remission and in the active phase were 4285±8623 and 3727±1176 pg/mL, respectively P=0.241. Correlation analysis between inflammatory markers and hepcidin serum levels indicated that there was no correlation between hepcidin levels and IL-6 P=0.582 or CRP P=0.783. Conclusion. As an acute-phase protein, hepcidin seems to have a lower efficacy than other parameters in the detection of activation in IBD.

  19. Serum paraoxonase 1 activity in patients with iron deficiency anemia

    Science.gov (United States)

    Gedikbasi, Asuman; Akalin, Nilgul; Gunaldi, Meral; Yilmaz, Deniz; Mert, Meral; Harmankaya, Ozlem; Soylu, Aliye; Karakaya, Pinar; Kumbasar, Abdulbaki

    2016-01-01

    Introduction In this study we aimed to detect paraoxonase 1 (PON-1) activity in iron deficiency anemia (IDA) and to compare it with healthy controls by observing the change after iron therapy. Material and methods In this study, 50 adult patients with IDA and 40 healthy subjects were enrolled. All patients were analyzed at the beginning and after treatment according to laboratory assessments. Results Mean paraoxonase and arylesterase activities in the iron deficiency anemia group were significantly lower than mean activities of the control group (102.4 ±19.2 U/l and 163.3 ±13.68 U/l, respectively and 157.3 ±26.4 U/l and 256.1 ±24.6 U/l, respectively; p = 0.0001 for both). Paraoxonase and arylesterase activities significantly increased after treatment for IDA (143.2 ±13.9 and 197.6 ±27.9 U/l, respectively, p = 0.0001). Mean activities after treatment with iron were significantly lower than mean activities in the control group (p = 0.002; p = 0.0001 respectively). Conclusions Paraoxonase and arylesterase activities in patients with IDA significantly increased after treatment with iron therapy. In adults IDA may also be one of the factors associated with increased risk of atherosclerosis. PMID:27478448

  20. Elevated serum interleukin-23 levels in ankylosing spondylitis patients and the relationship with disease activity.

    Science.gov (United States)

    Ugur, Mahir; Baygutalp, Nurcan Kilic; Melikoglu, Meltem Alkan; Baygutalp, Fatih; Altas, Elif Umay; Seferoglu, Buminhan

    2015-11-01

    This study was aimed to evaluate the relationship between serum interleukin-23 (IL-23) levels and ankylosing spondylitis (AS).Twenty male patients diagnosed with ankylosing spondylitis according to the 1984 modified New York criteria for AS and twenty male healthy controls were included in this study.The demographic characteristics, clinical and laboratory findings of the patients were recorded. Serum IL-23 levels, C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) were measured in both the AS and control groups. The Bath ankylosing spondylitis disease activity ındex (BASDAI), the Bath ankylosing spondylitis functional index (BASFI), and the Bath ankylosing spondylitis metrology index (BASMI) were evaluated as disease activity parameters. The AS patients were divided into two subgroups as active and inactive in respect of CRP, ESR levels and BASDAI scores. The mean serum IL-23 levels of the AS and control groups were 334.45±176.54 pg/ml and 166.49±177.50 pg/ml respectively, and there was a significant difference between the groups. Correlation analysis of serum IL-23 levels with clinical and laboratory parameters showed that there were positive correlations between serum IL-23 levels and the BASDAI, BASFI scores in total, active and inactive patients and the BASMI scores in total and inactive patients and negative correlations between serum IL-23 levels and ESR in inactive patients. It was shown that altered serum IL-23 levels were related to AS disease activity. Further studies in large patient series are necessary to investigate the role of IL-23 protein in etiopathogenesis of AS.

  1. Expression of recombinant human lysozyme in the milk of transgenic mice

    Institute of Scientific and Technical Information of China (English)

    YU Zhengquan; FAN Baoliang; DAI Yunping; ZHENG Ming; NIU Huiling; WANG Meili; WANG Lili; FEI Jing; LI Ning

    2003-01-01

    Human lysozyme is a 130-aa (amino acid) alkaline polypeptide, and has both anti-bacterial and anti-viral properties which make it an important component of human natural immunity system. As a first step toward the ultimate goal ofimproving the anti-bacterial properties of bovine and ovine milk, a transgenic mouse that contains the genomic DNA sequence of the human lysozme gene has been generated for the first time. From 83 mice generated by microinjection, a total of 6 positive transgenic mice were identified by PCR and Southern blot. F1 mice positive for transgene in lines were also detected by PCR. This shows that transgene could be transmitted from founder transgenic mice to their offspring. Recombinant human lysozyme (rHlys) was found in the whey of 3 female positive transgenic mice by Western blot. The highest concentration of rHlys for transgenic micewas 0.2 mg/mL. The antibacterial activity of the whey for transgenic mice was highly enhanced up to 0.4 times as much as that of human, while that of non-transgenic mouse was very low. Although the lysozyme activity of transgenic mice is still lower than that of human, the rHlys exhibits the same specific activity as that of human lysozyme. It provides a strong basis for further studies into the possible application of rHlys express in mammary gland.

  2. Serum paraoxonase 1 activity status in patients with liver disorders

    OpenAIRE

    Kedage Vivekananda; Muttigi Manjunatha; Shetty Mahesh; Suvarna Renuka; Rao Soumya; Joshi Chitralekha; Prakash Mungli

    2010-01-01

    Background/Aim: Paraoxonase 1 (PON1) is an esterase, exclusively synthesized by liver. The present study has two objectives: to determine the PON1 activity status in various disorders associated with hepatocellular damage and to correlate the changes of PON1 activity with the standard liver function and fasting lipid profile tests in these disorders. Patients and Methods: The study groups consisted of 95 patients with liver diseases including acute viral hepatitis (14), cirrhosis with port...

  3. POTENT INVITRO ANTI-HUMAN IMMUNODEFICIENCY VIRUS-1 ACTIVITY OF MODIFIED HUMAN SERUM ALBUMINS

    NARCIS (Netherlands)

    JANSEN, RW; MOLEMA, G; PAUWELS, R; SCHOLS, D; DECLERCQ, E; MEIJER, DKF

    1991-01-01

    A series of neoglycoproteins was synthesized by coupling of thiophosgene-activated p-aminophenyl derivatives [Biol. Cell. 47:95-110 (1983); J. Histochem. Cytochem. 32:1091-1094 (1984)] of various sugars to human serum albumin. The compounds were evaluated for their in vitro activity against human im

  4. A correlation of pregnancy term, disease activity, serum female hormones, and cytokines in uveitis

    OpenAIRE

    Chan, C-C; Reed, G F; Kim, Y.; Agrón, E; Buggage, R R

    2004-01-01

    Background/aims: Pregnancy and the postpartum period are associated with the activity of autoimmune diseases including uveitis. Although the exact mechanism is unknown, hormones are reported to alter inflammatory cytokines and influence disease activity. The authors studied ocular inflammation, female hormones, and serum cytokine levels during and after pregnancy.

  5. IL-18 Serum Level in Adult Onset Still’s Disease: A Marker of Disease Activity

    Directory of Open Access Journals (Sweden)

    Serena Colafrancesco

    2012-01-01

    Conclusions. Higher levels of IL-18 are detected in active AODS patients and correlate with disease activity and inflammatory laboratory features. ROC-AUC analysis of the serum concentration of IL-18 suggests that it can be considered a diagnostic marker of AOSD. This paper supports the targeting of this cytokine as a possible therapeutic option in AOSD.

  6. Basal Secretion of Lysozyme from Human Airways in Vitro

    Directory of Open Access Journals (Sweden)

    Patricia Roger

    1999-01-01

    Full Text Available The aim of this study was to examine the basal release of lysozyme from isolated human lung tissues. Measurements of lysozyme in the fluids derived from lung preparations were performed using a rate-of-lysis assay subsequent to acidification of the biological samples. Lysozyme released from bronchial preparations into fluids was greater than that observed for parenchymal tissues. The lysozyme quantities detected in bronchial fluids were not modified by removal of the surface epithelium. Furthermore, the quantities of lysozyme in bronchial fluids was correlated with the size of the bronchial preparations. These results suggest that the lysozyme was principally secreted by the human bronchi (submucosal layer rather than by parenchyma tissues and that a greater release was observed in the proximal airways.

  7. Serum boron concentration in rheumatoid arthritis: correlation with disease activity, functional class, and rheumatoid factor

    Directory of Open Access Journals (Sweden)

    Fadhil Muhsin

    2013-02-01

    Full Text Available Objectives: Rheumatoid arthritis (RA is a common chronic inflammatory arthropathy of unknown etiology. Trace elements have a great role in a number of biological processes. The aim of this study was to assess the serum element boron in a sample of Iraqi patients with RA and to evaluate its relationship if present with disease activity, functional class of the disease, and rheumatoid factor (RF. Methods: A cross sectional study enrolled 107 RA patients and 214 controls matched in age and sex. The American College of Rheumatology 1987 revised criteria was used for diagnosis of RA. Disease Activity Score index of 28 joints (DAS28, functional class of RA patients, RF, erythrocyte sedimentation rate (ESR were measured in patients’ group; serum boron levels were measured using a flame atomic absorption spectrophotometer in both patients and controls groups. Results: RA patients had significantly lower serum boron level than controls (P 0.05. Also, RF titer was a significant predictor of low serum boron level (P = 0.023, OR = –0.07, 95%CI –0.13-(–0.01. Conclusions: There was a significant low serum boron level in RA patients. RF titer was significant predictor of low serum boron level. This may suggest that boron element may play a role in pathophysiology of RA and its severity. Supplementation with boron element and diets rich in fruits, vegetables, nuts, and pulses may be useful. [J Exp Integr Med 2013; 3(1: 9-15

  8. Proteolytic activity of IgGs from blood serum of wistar rats at experimental rheumatoid arthritis

    Directory of Open Access Journals (Sweden)

    Yu. Ya. Kit

    2014-10-01

    Full Text Available The aim of this work was to study the proteolytic activity of IgGs purified from blood serum of Wistar rats at experimental rheumatoid arthritis (ERA induced by an injection of bovine collagen of type II. Twenty rats were immunized with a preparation of bovine collagen II (Sigma-Aldrich, USA in the presence of complete Freund’s adjuvant. ERA development was determined by inflammation in limbs of treated animals. IgG preparations were isolated from blood serum of immunized and non-immunized animals by precipitation of antibodies with 33% ammonium sulfate followed by chromatography on the Protein G-Sepharose column. Human histone H1, bovine collagen II, calf thymus histones, myelin basic protein (MBP, bovine serum albumin (BSA, and bovine casein were used as substrates of the proteolytic activity of IgGs. It was found that IgG preparations from blood serum of rats with ERA were capable of cleaving histone H1 and MBP, however, they were catalytically inactive towards collagen II, casein, BSA, and core histones. IgGs from blood serum of non-immunized rats were proteolytically inactive towards all used protein substrates. Thus, we demonstrated that immunization of rats with bovine collagen II induced IgG-antibodies possessing the proteolytic activity towards histone H1 and MBP. This activity might be associated with the development of inflammatory processes in the immunized rats.

  9. Serum B cell-activating factor (BAFF) level in connective tissue disease associated interstitial lung disease.

    Science.gov (United States)

    Hamada, Tsutomu; Samukawa, Takuya; Kumamoto, Tomohiro; Hatanaka, Kazuhito; Tsukuya, Go; Yamamoto, Masuki; Machida, Kentaro; Watanabe, Masaki; Mizuno, Keiko; Higashimoto, Ikkou; Inoue, Yoshikazu; Inoue, Hiromasa

    2015-09-30

    Interstitial lung diseases (ILDs) are common in patients with connective tissue diseases (CTDs). Although the diagnosis of an underlying CTD in ILD (CTD-ILD) affects both prognosis and treatment, it is sometimes difficult to distinguish CTD-ILD from chronic fibrosing interstitial pneumonia (CFIP). B cell-activating factor belonging to the tumour necrosis factor family (BAFF) plays a crucial role in B cell development, survival, and antibody production. We examined serum levels of BAFF, surfactant protein D (SP-D), and Krebs von den Lungen-6 (KL-6) in 33 patients with CTD-ILD, 16 patients with undifferentiated CTD-ILD, 19 patients with CFIP, and 26 healthy volunteers. And we analysed the relationship between serum BAFF levels and pulmonary function, as well as the expression of BAFF in the lung tissue of patients with CTD-ILD. Serum levels of BAFF were significantly higher in CTD-ILD patients compared to healthy subjects and CFIP patients. However, there were no significant differences in serum levels of SP-D and KL-6. Furthermore, serum BAFF levels in CTD-ILD patients were inversely correlated with pulmonary function. BAFF was strongly expressed in the lungs of CTD-ILD patients, but weakly in normal lungs. This is the first study to demonstrate that serum BAFF levels were significantly higher in CTD-ILD patients compared to healthy subjects and CFIP patients. Furthermore, serum BAFF levels were correlated with pulmonary function. We consider that serum BAFF levels in patients with CTD-ILD reflect the presence of ILDs disease activity and severity. These finding suggest that BAFF may be a useful marker for distinguishing CTD-ILD from CFIP.

  10. Human Serum-Specific Activation of Alternative Sigma Factors, the Stress Responders in Aggregatibacter actinomycetemcomitans

    Science.gov (United States)

    Tang-Siegel, Gaoyan; Bumgarner, Roger; Ruiz, Teresa; Kittichotirat, Weerayuth; Chen, Weizhen; Chen, Casey

    2016-01-01

    Aggregatibacter actinomycetemcomitans, a known pathogen causing periodontal disease and infective endocarditis, is a survivor in the periodontal pocket and blood stream; both environments contain serum as a nutrient source. To screen for unknown virulence factors associated with this microorganism, A. actinomycetemcomitans was grown in serum-based media to simulate its in vivo environment. Different strains of A. actinomycetemcomitans showed distinct growth phenotypes only in the presence of human serum, and they were grouped into high- and low-responder groups. High-responders comprised mainly serotype c strains, and showed an unusual growth phenomenon, featuring a second, rapid increase in turbidity after 9-h incubation that reached a final optical density 2- to 7-fold higher than low-responders. Upon further investigation, the second increase in turbidity was not caused by cell multiplication, but by cell death. Whole transcriptomic analysis via RNA-seq identified 35 genes that were up-regulated by human serum, but not horse serum, in high-responders but not in low-responders, including prominently an alternative sigma factor rpoE (σE). A lacZ reporter construct driven by the 132-bp rpoE promoter sequence of A. actinomycetemcomitans responded dramatically to human serum within 90 min of incubation only when the construct was carried by a high responder strain. The rpoE promoter is 100% identical among high- and low-responder strains. Proteomic investigation showed potential interactions between human serum protein, e.g. apolipoprotein A1 (ApoA1) and A. actinomycetemcomitans. The data clearly indicated a different activation process for rpoE in high- versus low-responder strains. This differential human serum-specific activation of rpoE, a putative extra-cytoplasmic stress responder and global regulator, suggests distinct in vivo adaptations among different strains of A. actinomycetemcomitans. PMID:27490177

  11. Evaluation of Serum Superoxide Dismutase Activity, Malondialdehyde, and Zinc and Copper Levels in Patients With Keratoconus.

    Science.gov (United States)

    Kılıç, Raşit; Bayraktar, Aslhan Cavunt; Bayraktar, Serdar; Kurt, Ali; Kavutçu, Mustafa

    2016-12-01

    The aim of this study was to evaluate the relationship between antioxidant superoxide dismutase (SOD) enzyme activity, malondialdehyde (MDA) as a lipid peroxidation marker, and some trace elements such as zinc (Zn) and copper (Cu) levels in patients with keratoconus. A total of 58 patients with keratoconus and 53 control subjects with similar age and sex were evaluated in this study. The modified Krumeich keratoconus classification was used to divide the patients into 4 stages. Serum SOD activity, MDA, and zinc and copper levels were compared between the patient and control groups. The median serum SOD activity, MDA, and Zn and Cu levels were 27.2 (42.4-13.7) U/mL, 10.2 (11.9-8.5) nmol/mL, 87.9 (104.6-76.5) μmol/L, and 103.2 (117.9-90.3) μmol/L in the keratoconus group and 26.2 (32.5-14.4) U/mL, 8.8 (11.4-7.1) nmol/mL, 100.5 (121.1-81.8) μmol/L, and 98.4 (120.3-83.4) μmol/L in the control group, respectively. There was a statistically significant difference between the MDA and Zn levels of the keratoconus group and control subjects but not between the respective SOD activities or Cu levels (P = 0.016, P = 0.031, P = 0.440, and P = 0.376, respectively). We found no significant difference between the keratoconus group stages for serum SOD activity, serum MDA, and Zn and Cu levels (P > 0.05), and there was also no significant correlation between the keratoconus group stages and serum SOD activity, serum MDA, and Zn and Cu levels (P > 0.05). There is imbalance in the systemic oxidant/antioxidant status where Zn deficiency also plays a role in patients with keratoconus.

  12. Serum YKL-40, a potential new marker of disease activity in patients with inflammatory bowel disease

    DEFF Research Database (Denmark)

    Vind, Ida; Johansen, J S; Price, P A;

    2003-01-01

    BACKGROUND: YKL-40 is secreted by macrophages and neutrophils and is a growth factor for vascular endothelial cells and fibroblasts. Elevated serum concentrations of YKL-40 are found in patients with diseases characterized by inflammation or ongoing fibrosis. The aim of this study was to seek...... association between serum YKL-40 in patients with ulcerative colitis (UC) and Crohn disease (CD) and clinical disease activity. METHODS: One-hundred-and-sixty-four patients with UC and 173 patients with CD were studied. The Simple Clinical Colitis Activity Index (SCCAI) and the Harvey-Bradshaw (H-B) score...

  13. Serum Xenohormone activity and DNA integrity of Europeans and Inuits

    DEFF Research Database (Denmark)

    Bonefeld-Jørgensen, Eva Cecilie

      Background. The toxicological assessment of the lipophilic persistent organic pollutants (POPs), including PCBs, pesticides and dioxins, is complicated since individuals are exposed to a complex mixture of contaminants. Therefore we developed ex vivo cell systems to determine the actual...... integrated level of  xenobiotic activity in the human serum fraction containing the POPs. Sperm DNA integrity is essential for the accurate transmission of genetic information and sperm DNA damage can cause decreased male fertility.    The AIM was to determine the integrated serum xenobiotic activity of POPs...

  14. [Method of determining tissue renin activity using heterologous serum].

    Science.gov (United States)

    Orbetsova, V Ts; Kiprov, D

    1979-01-01

    The authors described a method for determination of tissue renin activity with heterologous substrate. The preparation of the substrate was performed at several stages: salting with amonium sulfate; dialisis of the precipitate till complete separation of amonium sulfate molecules; distruction of angiotensinases by interchangeble souring and alcalization of the medium; lyophylization of the pure substrate. The obtained renin-substrate was preserved in ampules and its usage had a series of advantages--duration, economic, a possibility for standartization of the determination, etc., which were described in details in the article. The described in details also the quantitative determination of the renin activity in the tissues (renal and cerebral) with the help of the obtained substrate as the moments, modiied by the authors, were indicated.

  15. Serum microRNAs as biomarkers of human lymphocyte activation in health and disease.

    Directory of Open Access Journals (Sweden)

    Paola ede Candia

    2014-02-01

    Full Text Available Induction of the adaptive immune system is evaluated mostly by assessment of serum antibody titers and T lymphocyte responses in peripheral blood, although T and B cell activation occurs in lymphoid tissues. In recent years, the release of microRNAs (miRNAs in the extra-cellular environment has been exploited to assess cell functions at distance via measurement of serum miRNAs. Also activated lymphocytes release a large amount of nano-sized vesicles (exosomes, containing miRNA, but there are still few data on whether this phenomenon is reflected in modulation of serum miRNAs. Interestingly, miRNA signatures of CD4+ T cell-derived exosomes are substantially different from intracellular miRNA signatures of the same cells. We have recently identified serum circulating miR-150 as a sensor of general lymphocyte activation and we strongly believe that the identification of miRNAs differentially released by specific CD4+ effector T cell subsets (Th1, Th2, Th17 and Treg may work as serum biomarkers of their elicitation in lymphoid tissues but also in damaged tissues, thus providing pivotal information about the nature of immune responses occurring in health and disease.

  16. In vitro evaluation of synergistic activity between ciprofloxacin and broad snouted caiman serum against Escherichia coli.

    Science.gov (United States)

    Siroski, P A; Russi, N B; Ortega, H H; Formentini, E A

    2015-02-01

    The in vitro synergistic activity between ciprofloxacin and serum of broad snouted caiman on Escherichia coli was studied. The estimated MIC value of ciprofloxacin was 0.0188 µg/ml, and two assays of kill curve during 5 hours were performed: the first one in a standard culture medium and the second one in the presence of caiman serum. Different concentrations of ciprofloxacin were tested. Ciprofloxacin showed higher values of bacterial elimination rate in the presence of caiman serum in all concentrations tested. The combined activity of sub-inhibitory concentrations of ciprofloxacin and the humoral immune factors present in caiman serum determined an increase in the bacterial elimination observed in this assay. We suggest that the antibacterial activity of complement and natural antibodies present in caiman serum, which can bind to both Gram-negative and Gram-positive bacteria and acting through the classical complement pathway, can inhibit bacterial growth of Escherichia coli by lysis. Copyright © 2014 Elsevier Ltd. All rights reserved.

  17. Effect of Physical Activity on Serum Homocysteine Levels in Obese and Overweight Women

    Directory of Open Access Journals (Sweden)

    R. Soori

    2016-09-01

    Full Text Available Aims: Recently, homocysteine has been noticed as the major pathogenesis factor of the cardiovascular diseases. The aim of the study was to investigate the effects of physical activities on the serum homocysteine levels, as well as other cardiovascular risk factors in either obese or overweight women. Materials & Methods: In the controlled pretest-posttest semi-experimental study, 18 women referred to the Alzahra sport complexes in districts 3 and 4 of Tehran were studied in 2015. The subjects were selected via random sampling method and randomly divided into two groups; physical activity and control groups. And the intervention program was conducted in the former, while the latter received no intervention. The exercise protocol consisted of 10-week (5 sessions a week stretching exercises and aerobic activities (60 to 75% of the maximum heart beat. The serum homocystein level and lipids were measured both at the start and 48 hours after the exercises. Data was analyzed by SPSS 16 software using paired T and independent T tests. Findings: After the exercises, the mean serum homocysteine level in physical activity group significantly decreased than control group (p=0.001. Nevertheless, the difference between the lipid levels of physical activity and control groups was not significant (p>0.05. Conclusion: Reducing the serum homocysteine concentration, 10-week physical activity might also reduce the risk factors of cardiovascular diseases in either obese or overweight women.

  18. Serum lipoproteins attenuate macrophage activation and Toll-Like Receptor stimulation by bacterial lipoproteins

    Directory of Open Access Journals (Sweden)

    James Richard W

    2010-09-01

    Full Text Available Abstract Background Chlamydia trachomatis was previously shown to express a lipoprotein, the macrophage infectivity potentiator (Mip, exposed at the bacterial surface, and able to stimulate human primary monocytes/macrophages through Toll Like Receptor (TLR2/TLR1/TLR6, and CD14. In PMA-differentiated THP-1 cells the proinflammatory activity of Mip was significantly higher in the absence than in the presence of serum. The present study aims to investigate the ability of different serum factors to attenuate Mip proinflammatory activity in PMA-differentiated THP-1 cells and in primary human differentiated macrophages. The study was also extend to another lipoprotein, the Borrelia burgdorferi outer surface protein (OspA. The proinflammatory activity was studied through Tumor Necrosis Factor alpha (TNF-α and Interleukin (IL-8 release. Finally, TLR1/2 human embryonic kidney-293 (HEK-293 transfected cells were used to test the ability of the serum factors to inhibit Mip and OspA proinflammatory activity. Results In the absence of any serum and in the presence of 10% delipidated FBS, production of Mip-induced TNF-α and IL-8 in PMA-differentiated THP-1 cells were similar whereas they were significantly decreased in the presence of 10% FBS suggesting an inhibiting role of lipids present in FBS. In the presence of 10% human serum, the concentrations of TNF-α and IL-8 were 2 to 5 times lower than in the presence of 10% FBS suggesting the presence of more potent inhibitor(s in human serum than in FBS. Similar results were obtained in primary human differentiated macrophages. Different lipid components of human serum were then tested (total lipoproteins, HDL, LDL, VLDL, triglyceride emulsion, apolipoprotein (apoA-I, B, E2, and E3. The most efficient inhibitors were LDL, VLDL, and apoB that reduced the mean concentration of TNF-α release in Mip-induced macrophages to 24, 20, and 2%, respectively (p Conclusions These results demonstrated the ability of

  19. [Modifying effect of the blood serum from patients with soft-tissue sarcomas on interleukin-2 production and activity].

    Science.gov (United States)

    Goretskiĭ, B A; Berezhnaia, N M; Palivets, A Iu; Konovalenko, V F

    1989-01-01

    Interleukin-2 (IL-2) production by lymphocytes of a peripheral blood (LPB) in patients with soft tissue sarcomas is considerably lower in comparison with that in donors. Autological serum had either suppressive or stimulative effect on LPB IL-2 production in the observed patients. The investigated serum affected LPB of donors in the same way. Serum of patients with soft tissue sarcomas had the same dual effect on the ability of the active drug IL-2 to proliferate human T-lymphoblasts. Moreover serum-inhibitors of production acted as IL-2 activity stimulators, while serum-stimulators of IL-2 production inhibited the mediator effects.

  20. Serum cytokine and periodontal profiles in relation to disease activity of rheumatoid arthritis in Japanese adults.

    Science.gov (United States)

    Kobayashi, Tetsuo; Murasawa, Akira; Komatsu, Yasutaka; Yokoyama, Tomoko; Ishida, Kohei; Abe, Asami; Yamamoto, Kouji; Yoshie, Hiromasa

    2010-05-01

    Rheumatoid arthritis (RA) and periodontitis are common chronic inflammatory conditions and share many pathologic features. A similar profile of cytokines is involved in the pathogenesis of the two diseases. The relationship between the disease activity of RA and the periodontal condition remains unclear. This study examines whether the disease activity of RA affects serum cytokine and periodontal profiles. The study subjects consisted of 84 Japanese adults with RA and 22 race-matched control individuals. After periodontal and rheumatologic examination, the disease activity of RA was determined with the Disease Activity Score including 28 joints using C-reactive protein (DAS28-CRP). Serum levels of cytokines including interleukin (IL)-1beta, IL-6, IL-12, IL-12 p40, IL-18, and tumor necrosis factor-alpha (TNF-alpha) were determined by an enzyme-linked immunosorbent assay. High-sensitive CRP was also measured with a latex particle-enhanced nephelometric method. Of 84 patients with RA, 28 and 56 patients exhibited low and moderate to high disease activity, respectively. Serum levels of IL-6, TNF-alpha, and CRP were significantly different between the two groups (P <0.05). Additionally, a significant correlation was observed between DAS28-CRP and percentage of sites with bleeding on probing (BOP) (P = 0.008) and between serum TNF-alpha levels and percentage of sites with BOP (P = 0.01) in 56 patients with RA with moderate to high activity. These results suggest that the disease activity of RA correlated with serum levels of IL-6, TNF-alpha, and CRP, and it might influence BOP in the patients with moderate to high disease activity.

  1. Serum Adenosine deaminase activity and C-reactive protein levels in unstable angina

    Directory of Open Access Journals (Sweden)

    Rani Surekha

    2003-01-01

    Full Text Available In unstable angina (USA patients, immunological responses contributing to inflammation play a vital role in plaque rupture and thrombosis causing stroke. In the present study an attempt is made to estimate the levels of adenosine deaminase activity, an immunoenzyme marker and C-reactive protein, a marker of inflammation in USA patients. 45 patients presenting USA and 50 age and sex matched healthy controls were included in the study. Serum ADA activity was measured spectrophotometrically at 630nm and serum C-reactive protein was detected using Avitex CRP kit, which is a rapid latex agglutination test. The Mean ADA levels were 41.15 ± 11.04 in patients and 20.71±5.63 in controls and 66.6% of patients and none of the controls were positive to CRP. The present study observed the importance of ADA as a serum marker in addition to CRP for assessing the immune response in USA patients.

  2. Detecting endotoxin activity in bovine serum using an automated testing system.

    Science.gov (United States)

    Suzuki, Kazuyuki; Shimamori, Toshio; Sato, Ayano; Tsukano, Kenji; Tsuchiya, Masakazu; Lakritz, Jeffrey

    2015-08-01

    The aim of the present study was to compare the ability of the commercially available portable test system (PTS(TM)) to detect endotoxin activity in bovine serum, with that of the traditional LAL-kinetic turbidimetric (KT) and chromogenic (KC) assays. Prior to testing, serum samples, which were obtained from endotoxin-challenged cattle, were diluted 1:20 in endotoxin-free water and heated to 80°C for 10 min. The performance of the PTS(TM) was not significantly different from that of the traditional LAL-based assays. The results using PTS(TM) correlated with those using KT (r(2)=0.963, PPTS(TM) could be applied as a simplified system to assess endotoxin activity in bovine serum.

  3. EFFECT OF ACUPUNCTURE ON SERUM TNF-α ACTIVITY IN ALLERGIC BRONCHIAL ASTHMA PATIENTS

    Institute of Scientific and Technical Information of China (English)

    HUANG Tiejun; ZHANG Ji

    2002-01-01

    In this study, the therapeutic effect of acupuncture and its effect on serum tumor necrosis factor apha (TNF-α) level was observed in 25 cases of allergic bronchial asthma patients. Acupoints used were Dazhui (GV 14),Feishu (BL 13), Dingchuan (EX-B 1), Pishu (BL 20), Tanzhong (CV 17), Shenshu (BL 23) and Fengchi (GB 20),supplemented with other acupoints according to syndrome differentiation. After 15 sessions of treatment, results showed that the total effective rate was 96 %. Before treatment, serum TNF-α activity was significantly higher than that of healthy subjects (25 cases, P < 0.01 ). After treatment, TNF-α level reduced considerably in comparison with that of pre-treatment (P< 0.05). These findings indicate that acupuncture can significantly improve allergic asthma patients' clinical symptoms and lower serum TNF-α activity.

  4. Evaluation of serum antioxidant activity in patients with liver pathology by the chemiluminescent method.

    Science.gov (United States)

    Titov, V N; Sazhina, N N

    2014-12-01

    Total antioxidant activity of the serum in patients with liver pathology was assessed by two chemiluminescent methods based on different models of free radical oxidation: Hb-H2O2-luminol and ABAP-luminol. Comparative analysis showed a significant, but not high correlation of the results (r=0.798), which can be explained by different mechanisms of induction of free radicals and effects of various serum components (proteins and bilirubin) on the initiation process. The influence of aphysiological concentration of analyzed values manifested in the Hb-H2O2 model. Disagreement between the results of measurements was more pronounced in patients with abnormally high serum bilirubin content. The results suggested that ABAP-luminol chemiluminescent model is more preferable for evaluation of antioxidant activity in clinical practice.

  5. Lead exposure is associated with decreased serum paraoxonase 1 (PON1) activity and genotypes.

    Science.gov (United States)

    Li, Wan-Fen; Pan, Mei-Hung; Chung, Meng-Chu; Ho, Chi-Kung; Chuang, Hung-Yi

    2006-08-01

    Lead exposure causes cardiac and vascular damage in experimental animals. However, there is considerable debate regarding the causal relationship between lead exposure and cardiovascular dysfunction in humans. Paraoxonase 1 (PON1), a high-density lipoprotein-associated antioxidant enzyme, is capable of hydrolyzing oxidized lipids and thus protects against atherosclerosis. Previous studies have shown that lead and several other metal ions are able to inhibit PON1 activity in vitro. To investigate whether lead exposure has influence on serum PON1 activity, we conducted a cross-sectional study of workers from a lead battery manufactory and lead recycling plant. Blood samples were analyzed for whole-blood lead levels, serum PON1 activity, and three common PON1 polymorphisms (Q192R, L55M, -108C/T). The mean blood lead level (+/-SD) of this cohort was 27.1+/-15 microg/dL. Multiple linear regression analysis showed that blood lead levels were significantly associated with decreased serum PON1 activity (p<0.001) in lead workers. This negative correlation was more evident for workers who carry the R192 allele, which has been suggested to be a risk factor for coronary heart disease. Taken together, our results suggest that the decrease in serum PON1 activity due to lead exposure may render individuals more susceptible to atherosclerosis, particularly subjects who are homozygous for the R192 allele.

  6. [Serum calcium and phosphorus concentration and alkaline phosphatase activity in healthy children during growth and development].

    Science.gov (United States)

    Savić, Ljiljana; Savić, Dejan

    2008-01-01

    Many changes happen during growth and development in an organism as a result of important hormon changes, especially biohumoral ones. These changes make a problem when interpreting biochemical results in pediatric population. The most important changes are intensive calcium and phosphorus metabolic turnover in bone tissue with changes in alkaline phosphatase activity as a result of osteoblast activity. The aim of this study was to follow the serum calcium and phosphorus concentration and alkaline phosphatase activity in children 1-15 years old in different growth and development period and of different sexes and to fortify the influence of growth and development dynamics on biohumoral status in healthy male and female children. We evaluated 117 healthy children of both sexes from 1-15 years of age and divided them into three age groups: 1-5, 6-10 and 11-15 years. We followed the serum calcium and phosphorus concentration and alkaline phosphatase activity in different groups and in different sexes. Our investigation found significantly higher values of serum calcium in boys than in girls with no important changes between the age groups and significantly higher values of serum phosphorus in the youngest age group in all children and in different sexes with no important sex differences. Alkaline phosphatase activity followed the growth spurt and was the biggest in 6-10 years group in girls and in 11-15 years group in boys.

  7. Serum protease activity in chronic kidney disease patients: The GANI_MED renal cohort.

    Science.gov (United States)

    Wolke, Carmen; Teumer, Alexander; Endlich, Karlhans; Endlich, Nicole; Rettig, Rainer; Stracke, Sylvia; Fiene, Beate; Aymanns, Simone; Felix, Stephan B; Hannemann, Anke; Lendeckel, Uwe

    2017-03-01

    Serum or plasma proteases have been associated with various diseases including cancer, inflammation, or reno-cardiovascular diseases. We aimed to investigate whether the enzymatic activities of serum proteases are associated with the estimated glomerular filtration rate (eGFR) in patients with different stages of chronic kidney disease (CKD). Our study population comprised 268 participants of the "Greifswald Approach to Individualized Medicine" (GANI_MED) cohort. Enzymatic activity of aminopeptidase A, aminopeptidase B, alanyl (membrane) aminopeptidase, insulin-regulated aminopeptidase, puromycin-sensitive aminopeptidase, leucine aminopeptidase 3, prolyl-endopeptidase (PEP), dipeptidyl peptidase 4 (DPP4), angiotensin I-converting enzyme, and angiotensin I-converting enzyme 2 (ACE2) proteases was measured in serum. Linear regression of the respective protease was performed on kidney function adjusted for age and sex. Kidney function was modeled either by the continuous Modification of Diet in Renal Disease (MDRD)-based eGFR or dichotomized by eGFR serum protease activities showed no associations with age or sex. Our data indicate that ACE2 and DPP4 enzymatic activity are associated with the eGFR in patients with CKD. This finding distinguishes ACE2 and DPP4 from other serum peptidases analyzed and clearly indicates that further analyses are warranted to identify the precise role of these serum ectopeptidases in the pathogenesis of CKD and to fully elucidate underlying molecular mechanisms. Impact statement • Renal and cardiac diseases are very common and often occur concomitantly, resulting in increased morbidity and mortality. Understanding of molecular mechanisms linking both diseases is limited, available fragmentary data point to a role of the renin-angiotensin system (RAS) and, in particular, Ras-related peptidases. • Here, a comprehensive analysis of serum peptidase activities in patients with different stages of chronic kidney disease (CKD) is

  8. Human serum activates CIDEB-mediated lipid droplet enlargement in hepatoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Singaravelu, Ragunath [Department of Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, Ontario K1N 6N5 (Canada); National Research Council of Canada, Ottawa, Ontario K1A 0R6 (Canada); Lyn, Rodney K. [Department of Chemistry, University of Ottawa, Ottawa, Ontario K1N 6N5 (Canada); National Research Council of Canada, Ottawa, Ontario K1A 0R6 (Canada); Srinivasan, Prashanth [National Research Council of Canada, Ottawa, Ontario K1A 0R6 (Canada); Delcorde, Julie [Department of Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, Ontario K1N 6N5 (Canada); National Research Council of Canada, Ottawa, Ontario K1A 0R6 (Canada); Steenbergen, Rineke H.; Tyrrell, D. Lorne [Department of Medical Microbiology and Immunology, University of Alberta (Canada); Li Ka Shing Institute of Virology, Katz Centre for Pharmacy and Health Research, Edmonton, Alberta T6G 2S2 (Canada); Pezacki, John P., E-mail: John.Pezacki@nrc-cnrc.gc.ca [Department of Chemistry, University of Ottawa, Ottawa, Ontario K1N 6N5 (Canada); National Research Council of Canada, Ottawa, Ontario K1A 0R6 (Canada)

    2013-11-15

    Highlights: •Human serum induced differentiation of hepatoma cells increases cellular lipid droplet (LD) size. •The observed increase in LD size correlates with increased PGC-1α and CIDEB expression. •Induction of CIDEB expression correlates with rescue of VLDL secretion and loss of ADRP. •siRNA knockdown of CIDEB impairs the human serum mediated increase in LD size. •This system represents a cost-efficient model to study CIDEB’s role in lipid biology. -- Abstract: Human hepatocytes constitutively express the lipid droplet (LD) associated protein cell death-inducing DFFA-like effector B (CIDEB). CIDEB mediates LD fusion, as well as very-low-density lipoprotein (VLDL) maturation. However, there are limited cell culture models readily available to study CIDEB’s role in these biological processes, as hepatoma cell lines express negligible levels of CIDEB. Recent work has highlighted the ability of human serum to differentiate hepatoma cells. Herein, we demonstrate that culturing Huh7.5 cells in media supplemented with human serum activates CIDEB expression. This activation occurs through the induced expression of PGC-1α, a positive transcriptional regulator of CIDEB. Coherent anti-Stokes Raman scattering (CARS) microscopy revealed a correlation between CIDEB levels and LD size in human serum treated Huh7.5 cells. Human serum treatment also resulted in a rapid decrease in the levels of adipose differentiation-related protein (ADRP). Furthermore, individual overexpression of CIDEB was sufficient to down-regulate ADRP protein levels. siRNA knockdown of CIDEB revealed that the human serum mediated increase in LD size was CIDEB-dependent. Overall, our work highlights CIDEB’s role in LD fusion, and presents a new model system to study the PGC-1α/CIDEB pathway’s role in LD dynamics and the VLDL pathway.

  9. Serum prolidase enzyme activity in obese subjects and its relationship with oxidative stress markers.

    Science.gov (United States)

    Aslan, Mehmet; Duzenli, Ufuk; Esen, Ramazan; Soyoral, Yasemin Usul

    2017-10-01

    The relationship between increased serum enzyme activity of prolidase and increased rate of collagen turnover in the arterial wall has been asserted in previous studies. Collagen reflects much of the strength to the connective tissue involved in the arterial wall. Atherosclerosis is very common vessel disease and oxidative stress plays a pivotal role in the etiopathogenesis. Our objective was to examine the serum enzyme activity of prolidase and its possible relationships with oxidative stress parameters in obese subjects. Our present study was conducted 27 obese subjects and 26 age-matched healthy control subjects. The serum enzyme activity of prolidase in all study population was evaluated spectrophotometrically. Oxidative stress levels in obese subjects were analyzed with total antioxidant capacity (TAC) and total oxidant status (TOS) as well as oxidative stress index (OSI). Obese subjects have higher serum TOS and OSI indicators as well as prolidase activity than those in control subjects (for all; p<0.001). Moreover, obese subjects have lower levels of TAC than in those in healthy subjects (p<0.001). In the Pearson's correlation analysis, enzyme activity of prolidase was positively related with TOS (p<0.001, r=0.529) and OSI (p<0.001, r=0.519) as well as BMI (p<0.001, r=0.692) and inversely related with TAC (p<0.05, r=-0.405) in obese subjects. Increased serum prolidase activity and decreased antioxidant levels are likely to be a results of increased of oxidative stress levels in obese subjects. The significantly correlation between increased oxidative stress and increased prolidase activity may play a pivotal role in etiopathogenesis of atherosclerotic cardiovascular diseases in obese subjects. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Quinopeptide formation associated with the disruptive effect of epigallocatechin-gallate on lysozyme fibrils.

    Science.gov (United States)

    Cao, Na; Zhang, Yu-Jie; Feng, Shuang; Zeng, Cheng-Ming

    2015-01-01

    Numerous studies demonstrate that natural polyphenols can inhibit amyloid formation and disrupt preformed amyloid fibrils. In the present study, the fibril-disruptive effects of epigallocatechin-3-gallate (EGCG) were examined using lysozyme as a model protein. The results indicated that EGCG dose dependently inhibited lysozyme fibrillation and modified the peptide chains with quinonoid moieties under acidic conditions, as measured by ThT fluorescence, transmission electron microscopy, and an NBT-staining assay. Moreover, EGCG transformed the preformed lysozyme fibrils to amorphous aggregates through quinopeptide formation. The thiol blocker, N-ethylmaleimide, inhibited the disruptive effect of EGCG on preformed fibrils, suggesting that thiol groups are the binding sites for EGCG. We propose that the formation of quinone intermediates via oxidation and subsequent binding to lysozyme chains are the main processes driving the inhibition of amyloid formation and disruption of preformed fibrils by EGCG. The information presented in this study may provide fresh insight into the link between the antioxidant capacity and anti-amyloid activity of polyphenols. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Biparametric multicommutated flow analysis system for determination of human serum phosphoesterase activity.

    Science.gov (United States)

    Tymecki, Łukasz; Strzelak, Kamil; Koncki, Robert

    2013-10-03

    A multicommutated flow analysis (MCFA) system constructed of microsolenoid valves and pumps offering simultaneous determination of activity of acid phosphatase (ACP) and alkaline phosphatase (ALP) in human serum samples has been developed. The MCFA system is based on optoelectronic flow-through detector made of two light emitting diodes and operating according to paired emitter detector diode (PEDD) principle. This photometric PEDD device has been dedicated for detection of p-nitrophenol (NP) generated in the course of enzymatic hydrolysis of p-nitrophenyl phosphate and optimized for the determination of NP in human serum samples. The developed PEDD-based MCFA system allows independent optimization of conditions for reaction and detection steps of photometric ACP and ALP bioassays. Moreover, it allows elimination of photometric interferences from serum matrix components according to two-points kinetic mode of measurement. The single measurement cycle takes 12 min, consists of four measurements (two for each phosphoesterase) and enables determination of serum ACP and ALP activities at physiological and pathological levels. The real analytical utility of the developed MCFA system has been confirmed by analysis of control sera as well as real human serum samples from healthy persons and oncological patients. Copyright © 2013 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    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.

  13. The activity of lysosomal exoglycosidases in serum of alcohol-dependent men supplemented with borage oil enriched with vitamin E.

    Science.gov (United States)

    Zaniewska, Agnieszka; Borzym-Kluczyk, Malgorzata; Szajda, Slawomir D; Romatowski, Jacek; Gil, Andrzej; Knas, Malgorzata; Dobryniewski, Jacek; Zwierz, Krzysztof

    2009-08-01

    The aim of this study was to determine the activity of the lysosomal exoglycosidases: alpha-mannosidase (MAN), alpha-fucosidase (FUC), and beta-glucuronidase (GLUCUR) in serum of alcohol-dependent men supplemented and not supplemented with borage oil enriched with vitamin E. Serum was collected from eight social drinkers and 16 alcohol-dependent men after a drinking period. The activity of exoglycosidases and the concentration of protein in serum were determined. The increase in specific activity of MAN and GLUCUR was significant in serum of alcohol-dependent men both not supplemented and supplemented with borage oil enriched with vitamin E, in comparison with the specific activity in serum of social drinkers. In serum of alcohol-dependent men treated with borage oil enriched with vitamin E, specific activity of MAN and GLUCUR fluctuated in comparison with alcohol-dependent men not supplemented. Specific activity of FUC in serum of alcohol-dependent men both not supplemented and supplemented with borage oil enriched with vitamin E showed a tendency to increase, in comparison with social drinkers. Specific activity of FUC had a tendency to decrease in serum of alcohol-dependent men supplemented with borage oil enriched with vitamin E, in comparison with alcohol-dependent men not supplemented. Thus, supplementation of alcohol-dependent men after a long-lasting drinking period with borage oil and vitamin E did not change the rate of catabolism of the oligosaccharide chains of glycoconjugates, as evaluated by serum activity of exoglycosidases.

  14. Characterization of serum complement activity of saltwater (Crocodylus porosus) and freshwater (Crocodylus johnstoni) crocodiles.

    Science.gov (United States)

    Merchant, Mark; Britton, Adam

    2006-04-01

    We employed a spectroscopic assay, based on the hemolysis of sheep red blood cells (SRBCs), to assess the innate immune function of saltwater and freshwater crocodiles in vitro. Incubation of serum from freshwater and saltwater crocodiles with SRBCs resulted in concentration-dependent increases in SRBC hemolysis. The hemolytic activity occurred rapidly, with detectable activity within 2 min and maximum activity at 20 min. These activities, in both crocodilian species, were heat sensitive, unaffected by 20 mM methylamine, and completely inhibited by low concentrations of EDTA, suggesting that the alternative serum complement cascade is responsible for the observed effects. The hemolytic activities of the sera were inhibited by other chelators of divalent metal ions, such as phosphate and citrate. The inhibition of SRBC hemolysis by EDTA could be completely restored by the addition of 10 mM Ca2+ or Mg2+, but not Ba2+, Cu2+ or Fe2+, indicating specificity for these metal ions. The serum complement activities of both crocodilians were temperature-dependent, with peak activities occurring at 25-30 degrees C and reduced activities below 25 degrees C and above 35 degrees C.

  15. Lysozyme Photochemistry as a Function of Temperature. The Protective Effect of Nanoparticles on Lysozyme Photostability

    Science.gov (United States)

    Oliveira Silva, Catarina; Petersen, Steffen B.; Pinto Reis, Catarina; Rijo, Patrícia; Molpeceres, Jesús; Vorum, Henrik; Neves-Petersen, Maria Teresa

    2015-01-01

    The presence of aromatic residues and their close spatial proximity to disulphide bridges makes hen egg white lysozyme labile to UV excitation. UVB induced photo-oxidation of tryptophan and tyrosine residues leads to photochemical products, such as, kynurenine, N–formylkynurenine and dityrosine and to the disruption of disulphide bridges in proteins. We here report that lysozyme UV induced photochemistry is modulated by temperature, excitation power, illumination time, excitation wavelength and by the presence of plasmonic quencher surfaces, such as gold, and by the presence of natural fluorescence quenchers, such as hyaluronic acid and oleic acid. We show evidence that the photo-oxidation effects triggered by 295 nm at 20°C are reversible and non-reversible at 10°C, 25°C and 30°C. This paper provides evidence that the 295 nm damage threshold of lysozyme lies between 0.1 μW and 0.3 μW. Protein conformational changes induced by temperature and UV light have been detected upon monitoring changes in the fluorescence emission spectra of lysozyme tryptophan residues and SYPRO® Orange. Lysozyme has been conjugated onto gold nanoparticles, coated with hyaluronic acid and oleic acid (HAOA). Steady state and time resolved fluorescence studies of free and conjugated lysozyme onto HAOA gold nanoparticles reveals that the presence of the polymer decreased the rate of the observed photochemical reactions and induced a preference for short fluorescence decay lifetimes. Size and surface charge of the HAOA gold nanoparticles have been determined by dynamic light scattering and zeta potential measurements. TEM analysis of the particles confirms the presence of a gold core surrounded by a HAOA matrix. We conclude that HAOA gold nanoparticles may efficiently protect lysozyme from the photochemical effects of UVB light and this nanocarrier could be potentially applied to other proteins with clinical relevance. In addition, this study confirms that the temperature plays a

  16. Study on the relationship between serum interleukins, platelet activation indexes and cerebral infarction

    Institute of Scientific and Technical Information of China (English)

    Bo Wu

    2015-01-01

    Objective:To study and investigate the relationship between serum interleukins, platelet activation indexes and cerebral infarction.Methods:58 patients with cerebral infarction in our hospital from March 2013 to September 2014 were selected as observation group; meanwhile, 58 healthy persons at the same period were selected as control group, then the serum interleukins and platelet activation indexes of two groups were detected and compared, then the detection results of observation group with different stages and severity of cerebral infarction were compared too, and the relationship between those blood detection indexes and cerebral infarction were analyzed by the Logistic analysis.Results:The serum interleukins and platelet activation indexes of observation group were obviously higher than those of control group, and the detection levels of observation group with cerebral infarction at early and severe stage were obviously higher than those of patients at other stages and light, moderate, and those blood indexes all had close relationship to the cerebral infarction by the Logistic analysis,P<0.05. Conclusion:The serum interleukins and platelet activation indexes all have close relationship to cerebral infarction, and they can be as the important monitoring indexes of the disease.

  17. Serum 5'nucleotidase activity in rats: a method for automated analysis and criteria for interpretation.

    Science.gov (United States)

    Carakostas, Michael C.; Power, Richard J.; Banerjee, Asit K.

    1990-01-01

    A manual kit for determining serum 5'nucleotidase (5'NT, EC 3.1.3.5) activity was adapted for use with rat samples on a large discrete clinical chemistry analyzer. The precision of the method was good (within-run C.V. = 2.14%; between-run C.V. = 5.5%). A comparison of the new automated method with a manual and semi-automated method gave regression statistics of y = 1.18X -3.66 (Sy. x = 4.54), and y = 0.733X + 1.97 (Sy. x = 1.69), respectively. Temperature conversion factors provided by the kit manufacturer for human samples were determined to be inaccurate for converting results from rat samples. Analysis of components contributing to normal variation in rat serum 5'NT activity showed age and sex to be major factors. Increased serum 5'NT activity was observed in female rats when compared to male rats beginning at about 5 to 6 weeks of age. An analysis of variance of serum 5'NT, alkaline phosphatase, and GGT activities observed over a 9-week period in normal rats suggests several advantages for 5'NT as a predictor of biliary lesions in rats.

  18. Liver and extrahepatic contributions to postheparin serum lipase activity of the rat

    NARCIS (Netherlands)

    H. Jansen (Hans); W.C. Hülsmann (William)

    1974-01-01

    textabstractThe influence of the amount of heparin injected on the contributions of liver and of extrahepatic tissues to the lipase activity of postheparin serum of the rat was studied. It was found that when high doses of heparin (20 I.U./100 g bodyweight) were injected, the liver contributes for 6

  19. Serum matrix metalloproteinase 3 in early rheumatoid arthritis is correlated with disease activity and radiological progression

    NARCIS (Netherlands)

    Posthumus, MD; Limburg, PC; Westra, J; van Leeuwen, MA; van Rijswijk, MH

    2000-01-01

    Objective. To analyze the clinical significance of serial measurements of serum matrix metalloproteinase 3 (MMP-3) levels in relation to markers of disease activity and radiological progression in early rheumatoid arthritis (RA). Methods. In a 3 year prospective study of 33 patients with early RA (s

  20. Serum Amyloid A Induces Inflammation, Proliferation and Cell Death in Activated Hepatic Stellate Cells.

    Directory of Open Access Journals (Sweden)

    Sören V Siegmund

    Full Text Available Serum amyloid A (SAA is an evolutionary highly conserved acute phase protein that is predominantly secreted by hepatocytes. However, its role in liver injury and fibrogenesis has not been elucidated so far. In this study, we determined the effects of SAA on hepatic stellate cells (HSCs, the main fibrogenic cell type of the liver. Serum amyloid A potently activated IκB kinase, c-Jun N-terminal kinase (JNK, Erk and Akt and enhanced NF-κB-dependent luciferase activity in primary human and rat HSCs. Serum amyloid A induced the transcription of MCP-1, RANTES and MMP9 in an NF-κB- and JNK-dependent manner. Blockade of NF-κB revealed cytotoxic effects of SAA in primary HSCs with signs of apoptosis such as caspase 3 and PARP cleavage and Annexin V staining. Serum amyloid A induced HSC proliferation, which depended on JNK, Erk and Akt activity. In primary hepatocytes, SAA also activated MAP kinases, but did not induce relevant cell death after NF-κB inhibition. In two models of hepatic fibrogenesis, CCl4 treatment and bile duct ligation, hepatic mRNA levels of SAA1 and SAA3 were strongly increased. In conclusion, SAA may modulate fibrogenic responses in the liver in a positive and negative fashion by inducing inflammation, proliferation and cell death in HSCs.

  1. Liver and extrahepatic contributions to postheparin serum lipase activity of the rat

    NARCIS (Netherlands)

    H. Jansen (Hans); W.C. Hülsmann (William)

    1974-01-01

    textabstractThe influence of the amount of heparin injected on the contributions of liver and of extrahepatic tissues to the lipase activity of postheparin serum of the rat was studied. It was found that when high doses of heparin (20 I.U./100 g bodyweight) were injected, the liver contributes for

  2. A highly amyloidogenic region of hen lysozyme.

    Science.gov (United States)

    Frare, Erica; Polverino De Laureto, Patrizia; Zurdo, Jesús; Dobson, Christopher M; Fontana, Angelo

    2004-07-23

    Amyloid fibrils obtained after incubating hen egg-white lysozyme (HEWL) at pH 2.0 and 65 degrees C for extended periods of time have been found to consist predominantly of fragments of the protein corresponding to residues 49-100, 49-101, 53-100 and 53-101, derived largely from the partial acid hydrolysis of Asp-X peptide bonds. These internal fragments of HEWL encompass part of the beta-domain and all the residues forming the C-helix in the native protein, and contain two internal disulfide bridges Cys64-Cys80 and Cys76-Cys94. The complementary protein fragments, including helices A, B and D of the native protein, are not significantly incorporated into the network of fibrils, but remain largely soluble, in agreement with their predicted lower propensities to aggregate. Further analysis of the properties of different regions of HEWL to form amyloid fibrils was carried out by studying fragments produced by limited proteolysis of the protein by pepsin. Here, we show that only fragment 57-107, but not fragment 1-38/108-129, is able to generate well-defined amyloid fibrils under the conditions used. This finding is of particular importance, as the beta-domain and C-helix of the highly homologous human lysozyme have been shown to unfold locally in the amyloidogenic variant D67H, which is associated with the familial cases of systemic amyloidosis linked to lysozyme deposition. The identification of the highly amyloidogenic character of this region of the polypeptide chain provides strong support for the involvement of partially unfolded species in the initiation of the aggregation events that lead to amyloid deposition in clinical disease.

  3. Decreased levels of serum platelet-activating factor acetylhydrolase in patients with rheumatic diseases

    Directory of Open Access Journals (Sweden)

    P. Vergne

    1997-01-01

    Full Text Available PAF is a potent inflammatory compound known to stimulate the release of various cytokines involved in rheumatic diseases. Elevated blood PAF levels are reported in these patients. We report that serum PAF acetylhydrolase activity (AHA levels are decreased in patients with rheumatoid arthritis or osteoarthritis as compared to healthy controls. Serum and synovial fluid AHA levels were correlated in these patients. The present study suggests the potential role of AHA in controling systemic and/or local PAF levels in patients with rheumatic diseases.

  4. VARIATIONS IN THE COMPLEMENT ACTIVITY AND FIXABILITY OF GUINEA PIG SERUM.

    Science.gov (United States)

    Noguchi, H; Bronfenbrenner, J

    1911-01-01

    The following conclusions may be drawn from the foregoing series of experiments. The complementary activity varies within a definite limit in different specimens of guinea pig serum. With sera which stood in contact with the clot for twenty hours, the strongest and weakest were in the ratio of 0.015 cubic centimeter to 0.04 cubic centimeter. The former was 2.66 times stronger than the latter. The variation observed with the same series of sera after forty-six hours was still more striking. The strongest was 0.013 cubic centimeter, and the weakest, 0.06 cubic centimeter, that is, the former was 4.6 times stronger than the latter. These findings agree with those made by Massol and Grysez. The variations were not so marked with the majority of sera. It is noteworthy that a large number of the sera gained in the complementary activity when remaining in contact with the clot for forty-six hours, while some sera became weakened during the same length of time. The amount of serum fixed by given constant quantities of syphilitic serum and antigen varies much more markedly than the variations in their complementary activity. One serum failed altogether to be fixed. On the other hand, one sample of serum was so easily fixable that 0.24 cubic centimeter (corresponding to 9.6 complement units of this specimen) disappeared, while the average quantity fixed was only 0.098 cubic centimeter (corresponding to 4.64 complement units). The normal standard of fixability was shown in about 50 per cent. of the specimens examined. If the zone of normal fixability is enlarged in both directions by one unit, the percentage of normal fixability would become 65.8. There is no definite relationship between the complementary activity and the fixability of a given specimen of guinea pig serum. The facts derived from our present experiments, especially in regard to the exceptions in the fixative quality of this serum, demand the utmost precaution from those intending to employ it for diagnostic

  5. Action of egg white lysozyme on Clostridium tyrobutyricum.

    Science.gov (United States)

    Wasserfall, F; Teuber, M

    1979-08-01

    A 500-U ml-1 portion of egg white lysozyme was able to kill 99% of 5 X 10(5) resting vegetative cells of Clostridium tyrobutyricum within 24 h of incubation at 25 degrees C. Spores were completely resistant to lysozyme. Proliferating vegetative cells were severely inhibited, although lysozyme-resistant cells developed in growing cultures in the presence of lysozyme. Whereas early stages of spore germination (loss of optical refractility and heat resistance) were not inhibited by lysozyme, the overall outgrowth of spore cells into vegetative cells was delayed by 1 day in the presence of 500 U of lysosyme ml-1. This delay was independent of the lysozyme sensitivity or resistance of the mother culture of the used spores. It is suggested that this inhibition by lysozyme of the outgrowth of spore cells into vegetative cells of the lactate-fermenting C. tyrobutyricum is the basis for the observation that lysozyme can substitute for nitrate in preventing the "late gas" defect of Edam- and Gouda-type cheeses.

  6. Co-option of bacteriophage lysozyme genes by bivalve genomes

    Science.gov (United States)

    Wang, Chunyang; Jin, Min; Lan, Jiangfeng; Ye, Ting; Hui, Kaimin; Tan, Jingmin; Wang, Zheng; Wang, Wen; Han, Guan-Zhu

    2017-01-01

    Eukaryotes have occasionally acquired genetic material through horizontal gene transfer (HGT). However, little is known about the evolutionary and functional significance of such acquisitions. Lysozymes are ubiquitous enzymes that degrade bacterial cell walls. Here, we provide evidence that two subclasses of bivalves (Heterodonta and Palaeoheterodonta) acquired a lysozyme gene via HGT, building on earlier findings. Phylogenetic analyses place the bivalve lysozyme genes within the clade of bacteriophage lysozyme genes, indicating that the bivalves acquired the phage-type lysozyme genes from bacteriophages, either directly or through intermediate hosts. These bivalve lysozyme genes underwent dramatic structural changes after their co-option, including intron gain and fusion with other genes. Moreover, evidence suggests that recurrent gene duplication occurred in the bivalve lysozyme genes. Finally, we show the co-opted lysozymes exhibit a capacity for antibacterial action, potentially augmenting the immune function of related bivalves. This represents an intriguing evolutionary strategy in the eukaryote–microbe arms race, in which the genetic materials of bacteriophages are co-opted by eukaryotes, and then used by eukaryotes to combat bacteria, using a shared weapon against a common enemy. PMID:28100665

  7. Serum hepatic enzyme activity in relation to semen quality and serum reproductive hormone levels among Estonian fertile Men.

    Science.gov (United States)

    Ehala-Aleksejev, K; Punab, M

    2016-01-01

    The aim of this study was to investigate the relations of basic semen parameters and reproductive hormones with alanine aminotransferase (ALT) and gamma-glutamyl transferase (GGT). In addition, to examine possible interaction between adiposity, alcohol consumption, and liver tests in relation to male reproductive health, standard semen analysis was performed and serum levels of reproductive hormones and liver tests were measured in 245 male partners of pregnant women at a University Hospital Andrology Centres in Estonia. Quartile analysis revealed that after adjustment for covariates GGT was negatively related to sperm concentration and total sperm count. These significant changes appeared from a GGT >35.5 U/L. Next to these changes ALT was not related to sperm parameters. Both enzymes, GGT and ALT, were not related to reproductive hormones. Alcohol consumption was positively related to GGT and in cases with elevated GGT alcohol use was negatively related to sperm concentration and total sperm count. Alcohol consumption was positively related to body mass index (BMI) and waist circumference (WC). Our findings also confirm results of previous studies that BMI and WC are associated positively with ALT and GGT. According to the study, increased GGT activity might represent a possible connection between adiposity, alcohol consumption, and semen quality.

  8. Chronic Cadmium Exposure Lead to Inhibition of Serum and Hepatic Alkaline Phosphatase Activity in Wistar Rats.

    Science.gov (United States)

    Treviño, Samuel; Andrade-García, Alejandra; Herrera Camacho, Irma; León-Chavez, Bertha Alicia; Aguilar-Alonso, Patricia; Flores, Gonzalo; Brambila, Eduardo

    2015-12-01

    Alkaline phosphatase (ALP) activity in the serum and liver from rats administered with cadmium (Cd) in drinking water was studied. After metal administration, Cd showed a time-dependent accumulation in the liver, meanwhile metallothionein had a maximum increase at 1 month, remaining in this level until the end of the study. On the other hand, serum and liver ALP activity was decreased after 3 months exposure. To determine if Cd produced an inhibition on enzyme, apo-ALP prepared from both nonexposed and exposed rats was reactivated with Zn, showing 60% more activity as compared with the enzyme isolated from nonexposed rats. In vitro assays showed that Cd-ALP was partially reactivated with Zn; however, in the presence of cadmium, Zn-ALP was completely inhibited. Kinetic studies indicate a noncompetitive inhibition by Cd; these results suggest that Cd can substitute Zn, and/or Cd can interact with nucleophilic ligands essential for the enzymatic activity.

  9. Serum Malondialdehyde Concentration and Glutathione Peroxidase Activity in a Longitudinal Study of Gestational Diabetes

    Science.gov (United States)

    Miranda, María; Muriach, María; Romero, Francisco J.; Villar, Vincent M.

    2016-01-01

    Aims The main goal of this study was to evaluate the presence of oxidative damage and to quantify its level in gestational diabetes. Methods Thirty-six healthy women and thirty-six women with gestational diabetes were studied in the three trimesters of pregnancy regarding their levels of oxidative stress markers. These women were diagnosed with diabetes in the second trimester of pregnancy. Blood glucose levels after 100g glucose tolerance test were higher than 190, 165 or 145 mg/dl, 1, 2 or 3 hours after glucose intake. Results The group of women with gestational diabetes had higher serum malondialdehyde levels, with significant differences between groups in the first and second trimester. The mean values of serum glutathione peroxidase activity in the diabetic women were significantly lower in the first trimester. In the group of women with gestational diabetes there was a negative linear correlation between serum malondialdehyde concentration and glutathione peroxidase activity in the second and third trimester. Conclusions In this observational and longitudinal study in pregnant women, the alterations attributable to oxidative stress were present before the biochemical detection of the HbA1c increase. Usual recommendations once GD is detected (adequate metabolic control, as well as any other normally proposed to these patients) lowered the concentration of malondialdehyde at the end of pregnancy to the same levels of the healthy controls. Serum glutathione peroxidase activity in women with gestational diabetes increased during the gestational period. PMID:27228087

  10. Effects of single-walled carbon nanotubes on lysozyme gelation.

    Science.gov (United States)

    Tardani, Franco; La Mesa, Camillo

    2014-09-01

    The possibility to disperse carbon nanotubes in biocompatible matrices has got substantial interest from the scientific community. Along this research line, the inclusion of single walled carbon nanotubes in lysozyme-based hydrogels was investigated. Experiments were performed at different nanotube/lysozyme weight ratios. Carbon nanotubes were dispersed in protein solutions, in conditions suitable for thermal gelation. The state of the dispersions was determined before and after thermal treatment. Rheology, dynamic light scattering and different microscopies investigated the effect that carbon nanotubes exert on gelation. The gelation kinetics and changes in gelation temperature were determined. The effect of carbon and lysozyme content on the gel properties was, therefore, determined. At fixed lysozyme content, moderate amounts of carbon nanotubes do not disturb the properties of hydrogel composites. At moderately high volume fractions in carbon nanotubes, the gels become continuous in both lysozyme and nanotubes. This is because percolating networks are presumably formed. Support to the above statements comes by rheology.

  11. Transaminases activity in the sand lizard’s serum under influence of industrial pollution

    Directory of Open Access Journals (Sweden)

    O. Y. Klymenko

    2009-04-01

    Full Text Available Influence of the environmental pollution on the alanine aminotransferase and aspartate aminotransferase activity in the blood serum of the sand lizard has been studied. Aminotransferases (ALT and AST are similar by the mechanism of action. These enzymes take part in the amino acids metabolism. The increase of the transaminases activities under conditions of the pollution is found. It may be a proof of a damage of relevant organs: namely, the liver.

  12. Transaminases activity in the sand lizard’s serum under influence of industrial pollution

    OpenAIRE

    O. Y. Klymenko; V. Y. Gasso

    2009-01-01

    Influence of the environmental pollution on the alanine aminotransferase and aspartate aminotransferase activity in the blood serum of the sand lizard has been studied. Aminotransferases (ALT and AST) are similar by the mechanism of action. These enzymes take part in the amino acids metabolism. The increase of the transaminases activities under conditions of the pollution is found. It may be a proof of a damage of relevant organs: namely, the liver.

  13. Observing lysozyme's closing and opening motions by high-resolution single-molecule enzymology.

    Science.gov (United States)

    Akhterov, Maxim V; Choi, Yongki; Olsen, Tivoli J; Sims, Patrick C; Iftikhar, Mariam; Gul, O Tolga; Corso, Brad L; Weiss, Gregory A; Collins, Philip G

    2015-06-19

    Single-molecule techniques can monitor the kinetics of transitions between enzyme open and closed conformations, but such methods usually lack the resolution to observe the underlying transition pathway or intermediate conformational dynamics. We have used a 1 MHz bandwidth carbon nanotube transistor to electronically monitor single molecules of the enzyme T4 lysozyme as it processes substrate. An experimental resolution of 2 μs allowed the direct recording of lysozyme's opening and closing transitions. Unexpectedly, both motions required 37 μs, on average. The distribution of transition durations was also independent of the enzyme's state: either catalytic or nonproductive. The observation of smooth, continuous transitions suggests a concerted mechanism for glycoside hydrolysis with lysozyme's two domains closing upon the polysaccharide substrate in its active site. We distinguish these smooth motions from a nonconcerted mechanism, observed in approximately 10% of lysozyme openings and closings, in which the enzyme pauses for an additional 40-140 μs in an intermediate, partially closed conformation. During intermediate forming events, the number of rate-limiting steps observed increases to four, consistent with four steps required in the stepwise, arrow-pushing mechanism. The formation of such intermediate conformations was again independent of the enzyme's state. Taken together, the results suggest lysozyme operates as a Brownian motor. In this model, the enzyme traces a single pathway for closing and the reverse pathway for enzyme opening, regardless of its instantaneous catalytic productivity. The observed symmetry in enzyme opening and closing thus suggests that substrate translocation occurs while the enzyme is closed.

  14. Unravelling the structure of the pneumococcal autolytic lysozyme.

    Science.gov (United States)

    Monterroso, Begoña; López-Zumel, Consuelo; García, José L; Sáiz, José L; García, Pedro; Campillo, Nuria E; Menéndez, Margarita

    2005-10-01

    The LytC lysozyme of Streptococcus pneumoniae forms part of the autolytic system of this important pathogen. This enzyme is composed of a C-terminal CM (catalytic module), belonging to the GH25 family of glycosyl hydrolases, and an N-terminal CBM (choline-binding module), made of eleven homologous repeats, that specifically recognizes the choline residues that are present in pneumococcal teichoic and lipoteichoic acids. This arrangement inverts the general assembly pattern of the major pneumococcal autolysin, LytA, and the lytic enzymes encoded by pneumococcal bacteriophages that place the CBM (made of six repeats) at the C-terminus. In the present paper, a three-dimensional model of LytC built by homology modelling of each module and consistent with spectroscopic and hydrodynamic studies is shown. In addition, the putative catalytic-pair residues are identified. Despite the inversion in the modular arrangement, LytC and the bacteriophage-encoded Cpl-1 lysozyme most probably adopt a similar global fold. However, the distinct choline-binding ability and their substrate-binding surfaces may reflect a divergent evolution directed by the different roles played by them in the host (LytC) or in the bacteriophage (Cpl-1). The tight binding of LytC to the pneumococcal envelope, mediated by the acquisition of additional choline-binding repeats, could facilitate the regulation of the potentially suicidal activity of this autolysin. In contrast, a looser attachment of Cpl-1 to the cell wall and the establishment of more favourable interactions between its highly negatively charged catalytic surface and the positively charged chains of pneumococcal murein could enhance the lytic activity of the parasite-encoded enzyme and therefore liberation of the phage progeny.

  15. Relationship between serum leptin level and disease activity in patients with systemic sclerosis.

    Science.gov (United States)

    Budulgan, Mahmut; Dilek, Banu; Dağ, Şevin Buluttekin; Batmaz, Ibrahim; Yıldız, İsmail; Sarıyıldız, Mustafa Akif; Çevik, Remzi; Nas, Kemal

    2014-03-01

    To determine the relationship between serum leptin levels and disease activity in systemic sclerosis (SSc). A total of 60 subjects (30 controls and 30 patients) were included. The inflammatory markers and leptin levels were evaluated and body mass index (BMI) was measured for both groups. The assessment of the skin involvement was performed based on the modified Rodnan skin score (mRSS). Disease activity was evaluated according to the Valentini scleroderma disease activity index. There was a significant difference between the patient and control groups in terms of BMI (p 0.05). Valentini scores and mRSS were determined to be significantly higher in active patients (n = 14) than in inactive patients (n = 16) (p 0.05). However, leptin levels were significantly lower in active patients than in inactive patients (p < 0.05). We found a significant positive correlation between serum leptin and BMI (p < 0.05), and leptin and serum C3 levels (p < 0.05); no relationship was detected between leptin and other parameters. Leptin can be used as an activity marker in SSc. Further studies, including larger series, should be carried out to clarify this relationship.

  16. Detection of gelatinase B activity in serum of gastric cancer patients

    Institute of Scientific and Technical Information of China (English)

    Vesna V Dragutinovi(c); Neboj(s)a S Radovanovi(c); Lidija T Izrael-(Z)ivkovi(c); Miroslav M Vrvi(c)

    2006-01-01

    AIM: To determine the proteolytic activity and expression of gelatinase B in serum of gastric cancer patients and their correlation with the stage of the tumor.METHODS: Sera from 23 patients who underwent surgery for primary gastric cancer as the experimental group and from 11 as the control group were used to determine the proteolytic activity and its inhibition by EDTA and 1,10-phenanthroline. Gelatinase B activity was detected by SDS polyacrylamide gel electrophoresis (SDS-PAGE) and SDS-PAGE zymography.RESULTS: Proteolytic enzyme activity was increased in gastric cancer patients when compared to the control group (P<0.05). The proteinases were determined to be metalloproteinases upon inhibition test with specific metalloproteinase inhibitors 1,10-phenanthroline (P<0.05) and EDTA (P<0.01). SDS-PAGE and SDS-PAGE zymography revealed gelatinase B (proMMP-9) activity and its molecular mass of 92 ku.CONCLUSION: Proteinase activity is overexpressed in serum of gastric cancer patients. Gelatinase B in serum plays an important role in the progression of gastric cancer. ProMMP-9 can be used as a marker for invasiveness of gastric cancer.

  17. Serum Paraoxonase 1 Activity Is Associated with Fatty Acid Composition of High Density Lipoprotein

    Directory of Open Access Journals (Sweden)

    Maryam Boshtam

    2013-01-01

    Full Text Available Introduction. Cardioprotective effect of high density lipoprotein (HDL is, in part, dependent on its related enzyme, paraoxonase 1 (PON1. Fatty acid composition of HDL could affect its size and structure. On the other hand, PON1 activity is directly related to the structure of HDL. This study was designed to investigate the association between serum PON1 activity and fatty acid composition of HDL in healthy men. Methods. One hundred and forty healthy men participated in this research. HDL was separated by sequential ultracentrifugation, and its fatty acid composition was analyzed by gas chromatography. PON1 activity was measured spectrophotometrically using paraxon as substrate. Results. Serum PON1 activity was directly correlated with the amount of stearic acid and dihomo-gamma-linolenic acid (DGLA. PON1/HDL-C was directly correlated with the amount of miristic acid, stearic acid, and DGLA and was inversely correlated with total amount of ω6 fatty acids of HDL. Conclusion. The fatty acid composition of HDL could affect the activity of its associated enzyme, PON1. As dietary fats are the major determinants of serum lipids and lipoprotein composition, consuming some special dietary fatty acids may improve the activity of PON1 and thereby have beneficial effects on health.

  18. Serum hepatic enzyme activity in relation to semen quality and serum reproductive hormone levels among Estonian fertile Men

    National Research Council Canada - National Science Library

    Ehala‐Aleksejev, K; Punab, M

    2016-01-01

    ...‐glutamyl transferase (GGT). In addition, to examine possible interaction between adiposity, alcohol consumption, and liver tests in relation to male reproductive health, standard semen analysis was performed and serum levels...

  19. Continuous recording of long-chain acyl-coenzyme A synthetase activity using fluorescently labeled bovine serum albumin

    DEFF Research Database (Denmark)

    Demant, Erland J.F.; Nystrøm, Birthe T.

    2001-01-01

    acyl-Coenzyme A, synthetase, activity assay, fluorescence recording, fatty acid probe, serum albumin, hydroxycoumarin, detergent, micelles, Pseudomonas fragi, rat liver microsomes......acyl-Coenzyme A, synthetase, activity assay, fluorescence recording, fatty acid probe, serum albumin, hydroxycoumarin, detergent, micelles, Pseudomonas fragi, rat liver microsomes...

  20. A NEW METHOD TO ASSESS FUNCTIONAL ACTIVITY OF SERUM COMPLEMENT SYSTEM

    Directory of Open Access Journals (Sweden)

    E. G. Cheremnykh

    2015-01-01

    Full Text Available Complement system is an important component of innate immunity, providing primary protection against pathogens invading the body. In addition, it was shown that the complement system is associated with many diseases, not only autoimmune and infectious, but also mental disorders. In this regard, it is necessary to develop affordable and fast method of measuring activity of the complement system in real-time mode. We present a new semi-automated method for assessment of serum complement activity. The assay is based on cytolytic action of complement system upon the ciliate organism Tetrahymena pyriformis. This method consists in repeated counting of live Tetrahymena motile cells by means of specially developed Biolat device, which consists of two video cameras, light sources, and movable round plate. The plate has two rows of holes. The device also includes microprocessor control unit based on AutoCiliata software, intended for control of operation module and counting the surviving cell. The calculations are based on fixation of two sequential video-frames, with subsequent software image processing. Cell death events were observed upon incubation in triethanolamine (TEA buffer containing 5% of blood serum. We have also compared complement activity in different buffers, i.e., standard medium for culturing of ciliates, Veronal-Medinalum buffer, and the TEA buffer. TEA buffer was found superior to the Veronal buffer when applied in the test system. The time of cell death in the TEA-buffered medium containing 5% serum was < 15 minutes for all the sera studied. The parameters denoting serum complement activity were as follows: a half-life time for the moving cells (TLD50, and a similar value for 100% cell inactivation (1/TLD50, functional activity of the complement system, ACS. The sensitivity of this assay was calculated from dependencies between TLD50 and ACS, and actual serum concentrations. We have suggested an opportunity for evaluation of an

  1. Relationship of serum somatomedin-like activity and fibroblast proliferative activity with age and body weight gain in sheep.

    Science.gov (United States)

    Olsen, R F; Wangsness, P J; Patton, W H; Martin, R J

    1981-01-01

    The relationship between serum growth factors and body weight gain was examined in five Dorset lambs. The lambs were weighed and bled by jugular puncture at 2-week intervals between 2 and 18 weeks of age. Somatomedin-like activity (Sm) declined from initially high concentrations at 2 weeks to fairly constant concentrations between 6 and 18 weeks. Relative weight gain--i.e., gain expressed as a percentage of body weight--declined in a manner similar to that of Sm. Mean relative weight gain and mean Sm for the eight 2-week intervals were significantly related (r = .84). Absolute body weight gain--i.e., gain expressed in kilograms--remained fairly constant throughout the study and was not significantly correlated to Sm (r = .15). Serum fibroblast proliferative activity (FPA) was measured as a possible indicator of collective activities of serum growth factors. FPA initially followed a pattern similar to that of Sm, decreasing between 2 and 6 weeks and plateauing until 12 weeks. After 12 weeks, FPA increased to concentrations similar to those observed at 2 weeks. The increase in FPA after 12 weeks was apparently due to an increase in a non-Sm growth factor and had no obvious relationship to body weight changes. Results of the in vitro cell assay system might have been more meaningful if cell type(s) other than WI-38 fibroblasts (e.g., myogenic cells) had been used for estimating collective activities of serum mitogenic factors. The data suggest that serum Sm-like activity may be important in the regulation of growth in sheep.

  2. Low Serum Lysosomal Acid Lipase Activity Correlates with Advanced Liver Disease

    Directory of Open Access Journals (Sweden)

    Eyal Shteyer

    2016-02-01

    Full Text Available Fatty liver has become the most common liver disorder and is recognized as a major health burden in the Western world. The causes for disease progression are not fully elucidated but lysosomal impairment is suggested. Here we evaluate a possible role for lysosomal acid lipase (LAL activity in liver disease. To study LAL levels in patients with microvesicular, idiopathic cirrhosis and nonalcoholic fatty liver disease (NAFLD. Medical records of patients with microvesicular steatosis, cryptogenic cirrhosis and NAFLD, diagnosed on the basis of liver biopsies, were included in the study. Measured serum LAL activity was correlated to clinical, laboratory, imaging and pathological data. No patient exhibited LAL activity compatible with genetic LAL deficiency. However, serum LAL activity inversely predicted liver disease severity. A LAL level of 0.5 was the most sensitive for detecting both histologic and noninvasive markers for disease severity, including lower white blood cell count and calcium, and elevated γ-glutamyltransferase, creatinine, glucose, glycated hemoglobin, uric acid and coagulation function. Serum LAL activity <0.5 indicates severe liver injury in patients with fatty liver and cirrhosis. Further studies should define the direct role of LAL in liver disease severity and consider the possibility of replacement therapy.

  3. Effect of psoriasis activity and topical treatment on serum lipocalin-2 levels.

    Science.gov (United States)

    Baran, A; Świderska, M; Myśliwiec, H; Flisiak, I

    2017-03-01

    Psoriasis has been considered as systemic disorder. Lipocalin-2 might be a link between psoriasis and its comorbidities. Aim of the study was to investigate the associations between serum lipocalin-2 levels and the disease activity, markers of inflammation or metabolic disturbances and changes after topical treatment in psoriatic patients. Thirty-seven individuals with active plaque-type psoriasis and 15 healthy controls were recruited. Blood samples were collected before and after 14 days of therapy. Serum lipocalin-2 concentrations were examined by enzyme-linked immunosorbent assay. The results were correlated with Psoriasis Area and Severity Index (PASI), body mass index (BMI), inflammatory and biochemical markers, lipid profile and with effectiveness of topical treatment. Lipocalin-2 serum levels were significantly increased in psoriatic patients in comparison to the controls (p = 0.023). No significant correlations with indicators of inflammation, nor BMI or PASI were noted. A statistical association between lipocalin-2 and low-density lipoprotein-cholesterol was shown. After topical treatment serum lipocalin-2 level did not significantly change (p = 0.9), still remaining higher than in the controls, despite clinical improvement. Lipocalin-2 might be a marker of psoriasis and convey cardiovascular or metabolic risk in psoriatic patients, but may not be a reliable indicator of inflammation, severity of psoriasis nor efficacy of antipsoriatic treatment.

  4. 动物源溶菌酶研究进展%Advance in Studies of Animal-borne Lysozyme

    Institute of Scientific and Technical Information of China (English)

    张鹏; 江明锋; 王永

    2012-01-01

    动物源溶菌酶是一种动物体内广泛存在的酶类,它可以水解细菌细胞壁肽聚糖中的β-1,4糖苷键,具有消化分解细菌、抑制外源微生物生长、增强机体免疫力的作用.目前溶菌酶已被用作研究蛋白功能、性质以及分子进化的模型.首先介绍了溶菌酶及其分子的晶体结构,溶菌酶基因及其蛋白研究进展,其次介绍了动物源溶菌酶的功能,包括溶菌酶生物学功能和重组蛋白功能活性,重点介绍了溶菌酶基因在转基因工程中的应用研究,最后对动物源溶菌酶研究进行了展望.研究动物源溶菌酶对于基础科学,并应用其转变成现实生产力具有重要的指导意义.%Lysozyme is a kind of muramidases that widespread in the animal in vivo, and can catalyze the hydrolysis of β-1, 4-glycosidic bonds between the N-acetylglucosamine and N-acetylmuramic acid in the peptidoglycan layer of bacterial cell walls. It has functions of the digestion and decomposition of bacteria, and inhibition of exogenous microbial growth, and enhancing immunity; and lysozyme has been a model protein for research in the function and the nature of the enzyme, and molecular evolution. Firstly, the lysozyme and its crystal structure and the advance in studies of lysozyme gene and its protein were introduced. Secondly, the functions of animal-borne lysozyme, including the biological functions of lysozyme and functional activities of recombinant proteins were introduced too,then the applied researches in lysozyme gene in transgenic engineering were focused on. Finally, the perspectives of animal-borne lysozyme were suggested. It' s very significant to research the animal-borne lysozyme because it' s helpful to understand the basic knowledge and to use it in the production.

  5. A simplified method for detecting macroamylasemia by measuring serum amylase activity at different reaction temperatures.

    Science.gov (United States)

    Koda, T; Kuratsune, H; Kurahori, T

    1983-06-01

    Amylase activity in serum and urine, and isoamylase, were measured in 300 patients with abdominal pain to detect cases of macroamylasemia. Of these patients, 9 had hyperamylasemia and 2 were diagnosed as cases of macroamylasemia on the basis of their amylase/creatinine clearance ratio, the gel filtration pattern of their amylase on a dextran column, and results of immunological analysis. Amylase activity in macroamylasemia is reported to show an anomalous response to increase in reaction-temperature. In this report, measurements of the temperature-activity relationships of serum amylase confirmed that the ratio of serum amylase activity at 50 degrees C to that at 25 degrees C (AMY-50 degrees C/AMY-25 degrees C ratio) in patients with macroamylasemia was higher than that in normal subjects or patients with pancreatitis. Moreover, when macromolecular amylase in the sera of patients with macroamylasemia was separated from amylase of normal molecular weight by dextran gel chromatography, it showed a significantly higher AMY-50 degrees C/AMY-25 degrees C ratio than the latter. Measurement of this AMY-50 degrees C/AMY-25 degrees C ratio seems to be a convenient and useful method for differential diagnosis of hyperamylasemia.

  6. NRF2 activation is involved in ozonated human serum upregulation of HO-1 in endothelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Pecorelli, Alessandra [Department of Molecular and Developmental Medicine, University of Siena (Italy); Child Neuropsychiatry Unit, University Hospital, AOUS, Siena (Italy); Bocci, Velio [Department of Physiology, University of Siena (Italy); Acquaviva, Alessandra [Department of Molecular and Developmental Medicine, University of Siena (Italy); Belmonte, Giuseppe [Department of Biomedical Sciences, University of Siena (Italy); Gardi, Concetta [Department of Molecular and Developmental Medicine, University of Siena (Italy); Virgili, Fabio [INRAN, Rome (Italy); Ciccoli, Lucia [Department of Molecular and Developmental Medicine, University of Siena (Italy); Valacchi, Giuseppe, E-mail: giuseppe.valacchi@unife.it [Department of Life Sciences and Biotechnology, University of Ferrara (Italy); Department of Food and Nutrition, Kyung Hee University, Seoul (Korea, Republic of)

    2013-02-15

    During the last decade, it has been shown that the activation of NRF2 and the binding to electrophile-responsive element (EpREs), stimulates the expression of a great number of genes responsible for the synthesis of phase I and phase II proteins, including antioxidants enzymes and heme oxygenase-1 (HO-1). This critical cell response occurs in cardiovascular, degenerative and chronic infective diseases aggravated by a chronic oxidative stress. In our previous reports we have shown that ozonated plasma is able to up-regulate HO-1 expression in endothelial cells. In the present work we investigated a candidate mechanism involved in this process. After treatment with increasing doses of ozonated serum (20, 40 and 80 μg/mL O{sub 3} per mL of serum), a clear dose dependent activation of NRF2 and the subsequent induction of HO-1 and NAD(P)H quinone oxidoreductase 1(NQO1) was observed. This effect was also present when cells were treated with serum and hydrogen peroxide (H{sub 2}O{sub 2}) or serum and 4-hydroxynonenal (4HNE). Moreover, the treatment with ozonated serum was associated with a dose-dependent activation of extracellular-signal-regulated kinases (ERK1/2) and p38 MAP kinases (p38), not directly involved in NRF2 activation. These data, provide a new insight on the mechanism responsible for the induction of HO-1 expression by ozonated serum in the endothelium, and have a practical importance as an expedient approach to the treatment of patients with both effective orthodox drugs and ozonated autohemotherapy, targeted to the restoration of redox homeostasis. - Highlights: ► Endothelial HO1 is upregulated by ozonated plasma ► This activation is induced by NRF2 and it is ERK independent. ► 4HNE and H{sub 2}O{sub 2} are the main molecules involved in this process. ► Ozonated plasma induced a hormetic effect ► Combination of orthodox medicine and ozonated plasma can be a useful treatment.

  7. Identification of a novel virulence determinant with serum opacification activity in Streptococcus suis.

    Science.gov (United States)

    Baums, Christoph G; Kaim, Ute; Fulde, Marcus; Ramachandran, Girish; Goethe, Ralph; Valentin-Weigand, Peter

    2006-11-01

    Streptococcus suis serotype 2 is a porcine and human pathogen with adhesive and invasive properties. In other streptococci, large surface-associated proteins (>100 kDa) of the MSCRAMM family (microbial surface components recognizing adhesive matrix molecules) are key players in interactions with host tissue. In this study, we identified a novel opacity factor of S. suis (OFS) with structural homology to members of the MSCRAMM family. The N-terminal region of OFS is homologous to the respective regions of fibronectin-binding protein A (FnBA) of Streptococcus dysgalactiae and the serum opacity factor (SOF) of Streptococcus pyogenes. Similar to these two proteins, the N-terminal domain of OFS opacified horse serum. Serum opacification activity was detectable in sodium dodecyl sulfate extracts of wild-type S. suis but not in extracts of isogenic ofs knockout mutants. Heterologous expression of OFS in Lactococcus lactis demonstrated that a high level of expression of OFS is sufficient to provide surface-associated serum opacification activity. Furthermore, serum opacification could be inhibited by an antiserum against recombinant OFS. The C-terminal repetitive sequence elements of OFS differed significantly from the respective repeat regions of FnBA and SOF as well as from the consensus sequence of the fibronectin-binding repeats of MSCRAMMs. Accordingly, fibronectin binding was not detectable in recombinant OFS. To investigate the putative function of OFS in the pathogenesis of invasive S. suis diseases, piglets were experimentally infected with an isogenic mutant strain in which the ofs gene had been knocked out by an in-frame deletion. The mutant was severely attenuated in virulence but not in colonization, demonstrating that OFS represents a novel virulence determinant of S. suis.

  8. Direct biomolecule binding on nonfouling surfaces via newly discovered supramolecular self-assembly of lysozyme under physiological conditions.

    Science.gov (United States)

    Yang, Peng

    2012-08-01

    When lysozyme is dissolved in a neutral HEPES buffer solution (pH = 7.4) with 0.001-0.050 M TCEP added, a fast phase transition process occurs and the resulting novel fiber-like hierarchical supramolecular assemblies made by primary spherical-particle aggregation can function as a "superglue" that binds strongly and quickly onto non-fouling coatings. This binding is highly selective towards lysozyme, and excludes synthetic, chemical/physical activation/deactivation (blocking) steps. By using biotinylated lysozyme, such a phase transition quickly creates a perfect biotinylated surface on non-fouling surfaces for avidin binding, showing great potential for the development of low-cost and practical biochips.

  9. Dimer formation in radiation-irradiated aqueous solution of lysozyme studied by light-scattering-intensity measurement

    Energy Technology Data Exchange (ETDEWEB)

    Hashimoto, S.; Masuda, T.; Kondo, M. (Institute of Physical and Chemical Research, Wako, Saitama (Japan); Tokyo Metropolitan Univ. (Japan). Faculty of Science); Seki, H.; Imamura, M. (Institute of Physical and Chemical Research, Wako, Saitama (Japan))

    1981-07-01

    The reaction of lysozyme with OH radical, Br/sub 2/ anion radical, and e/sup -/sub(aq), produced in an aqueous solution by pulsed electrons and ..gamma..-rays, were investigated. Irradiated enzymes showed an increase in the light scattering intensity (LSI) which is proportional to the absorbed dose. Results obtained from SDS gel electrophoresis confirm dimerization of lysozyme, which is considered to be responsible for the increase in LSI. It was found that the rate constant of the dimerization of protein radicals produced in the reaction with OH radical is 2k = (1.0+-0.3) x 10/sup 6/M/sup -1/s/sup -1/ and the yield of the dimerization is 0.6 in G. The enzymatic activity of the dimer is shown to be reduced to about 30 per cent of that of the intact enzyme. It is concluded that the radiation-induced inactivation of lysozyme is largely due to dimerization.

  10. Dimer formation in radiation-irradiated aqueous solution of lysozyme studied by light-scattering-intensity measurement.

    Science.gov (United States)

    Hashimoto, S; Seki, H; Masuda, T; Imamura, M; Kondo, M

    1981-07-01

    The reaction of lysozyme with OH., Br.-2 and e-aq, produced in an aqueous solution by pulsed electrons and gamma-rays, were investigated. Irradiated enzymes showed an increase in the light scattering intensity (LSI) which is proportional to the absorbed dose. Results obtained from SDS gel electrophoresis confirm dimerization of lysozyme, which is considered to be responsible for the increase in LSI. It was found that the rate constant of the dimerization of protein radicals produced in the reaction with OH. is 2K=(1.0 +/- 0.3) X 10(6)M-1 s-1 and the yield of the dimerization is 0.6 in G. The enzymatic activity of the dimer is shown to be reduced to about 30 per cent of that of the intact enzyme. It is concluded that the radiation-induced inactivation of lysozyme is largely due to dimerization.

  11. AhR transcriptional activity in serum of Inuits across Greenlandic districts

    Directory of Open Access Journals (Sweden)

    Bonefeld-Jorgensen Eva C

    2007-10-01

    Full Text Available Abstract Background Human exposure to lipophilic persistent organic pollutants (POPs including polychlorinated dibenzo-p-dioxins/furans (PCDDs/PCDFs, polychlorinated biphenyls (PCBs and organochlorine pesticide is ubiquitous. The individual is exposed to a complex mixture of POPs being life-long beginning during critical developmental windows. Exposure to POPs elicits a number of species- and tissue-specific toxic responses, many of which involve the aryl hydrocarbon receptor (AhR. The aim of this study was to compare the actual level of integrated AhR transcriptional activity in the lipophilic serum fraction containing the actual POP mixture among Inuits from different districts in Greenland, and to evaluate whether the AhR transactivity is correlated to the bio-accumulated POPs and/or lifestyle factors. Methods The study included 357 serum samples from the Greenlandic districts: Nuuk and Sisimiut (South West Coast, Qaanaaq (North Coast and Tasiilaq (East Coast. The bio-accumulated serum POPs were extracted by ethanol: hexane and clean-up on Florisil columns. Effects of the serum extract on the AhR transactivity was determined using the Hepa 1.12cR mouse hepatoma cell line carrying an AhR-luciferase reporter gene, and the data was evaluated for possible association to the serum levels of 14 PCB congeners, 10 organochlorine pesticide residues and/or lifestyle factors. Results In total 85% of the Inuit samples elicited agonistic AhR transactivity in a district dependent pattern. The median level of the AhR-TCDD equivalent (AhR-TEQ of the separate genders was similar in the different districts. For the combined data the order of the median AhR-TEQ was Tasiilaq > Nuuk ≥ Sisimiut > Qaanaaq possibly being related to the different composition of POPs. In overall, the AhR transactivity was inversely correlated to the levels of sum POPs, age and/or intake of marine food. Conclusion i We observed that the proportion of dioxin like (DL compounds in the

  12. Impact of polychlorinated biphenyls contamination on estrogenic activity in human male serum.

    Science.gov (United States)

    Plísková, Martina; Vondrácek, Jan; Canton, Rocio Fernandez; Nera, Jirí; Kocan, Anton; Petrík, Ján; Trnovec, Tomás; Sanderson, Thomas; van den Berg, Martin; Machala, Miroslav

    2005-10-01

    Polychlorinated biphenyls (PCBs) are thought to cause numerous adverse health effects, but their impact on estrogen signaling is still not fully understood. In the present study, we used the ER-CALUX bioassay to determine estrogenic/antiestrogenic activities of the prevalent PCB congeners and PCB mixtures isolated from human male serum. The samples were collected from residents of an area with an extensive environmental contamination from a former PCB production site as well as from a neighboring background region in eastern Slovakia. We found that the lower-chlorinated PCBs were estrogenic, whereas the prevalent higher-chlorinated PCB congeners 138, 153, 170, 180, 187, 194, 199, and 203, as well as major PCB metabolites, behaved as antiestrogens. Coplanar PCBs had no direct effect on estrogen receptor (ER) activation in this in vitro model. In human male serum samples, high levels of PCBs were associated with a decreased ER-mediated activity and an increased dioxin-like activity, as determined by the DR-CALUX assay. 17beta-Estradiol (E2) was responsible for a major part of estrogenic activity identified in total serum extracts. Significant negative correlations were found between dioxin-like activity, as well as mRNA levels of cytochromes P450 1A1 and 1B1 in lymphocytes, and total estrogenic activity. For sample fractions containing only persistent organic pollutants (POPs), the increased frequency of antiestrogenic samples was associated with a higher sum of PCBs. This suggests that the prevalent non-dioxin-like PCBs were responsible for the weak antiestrogenic activity of some POPs fractions. Our data also suggest that it might be important to pay attention to direct effects of PCBs on steroid hormone levels in heavily exposed subjects.

  13. Relationship between elevated serum gamma-glutamyltransferase activity and slow coronary flow

    DEFF Research Database (Denmark)

    Sen, Nihat; Ozlü, Mehmet F; Basar, Nurcan

    2009-01-01

    . The results were compared with those of a control group consisting of 86 age- and sex-matched patients who had normal coronary arteries and normal coronary flow. RESULTS: The three groups were similar with respect to body mass index, presence of hypertension and diabetes mellitus, lipid profiles, and fasting...... was found as the only independent predictor of the mean TFC (beta=0.309; pphysiopathologic role of serum GGT activity in SCF....

  14. Impact of Polychlorinated Biphenyls Contamination on Estrogenic Activity in Human Male Serum

    OpenAIRE

    Plíšková, Martina; Vondráček, Jan; Canton, Rocio Fernandez; Nera, Jiřií; Kočan, Anton; Petrík, Ján; Trnovec, Tomáš; Sanderson, Thomas; van den Berg, Martin; Machala, Miroslav

    2005-01-01

    Polychlorinated biphenyls (PCBs) are thought to cause numerous adverse health effects, but their impact on estrogen signaling is still not fully understood. In the present study, we used the ER-CALUX bioassay to determine estrogenic/antiestrogenic activities of the prevalent PCB congeners and PCB mixtures isolated from human male serum. The samples were collected from residents of an area with an extensive environmental contamination from a former PCB production site as well as from a neighbo...

  15. Reentrant condensation of lysozyme: Implications for studying dynamics of lysozyme in aqueous solutions of lithium chloride

    Energy Technology Data Exchange (ETDEWEB)

    Mamontov, Eugene [ORNL; O' Neill, Hugh Michael [ORNL

    2014-01-01

    Recent studies have outlined the use of eutectic solution of lithium chloride in water to study microscopic dynamics of lysozyme in an aqueous solvent that is remarkably similar to pure water in many respects, yet allows experiments over a wide temperature range without the solvent crystallization. The eutectic point in (H2O)R(LiCl) system corresponds to R 7.3, and it is of interest to investigate whether less concentrated aqueous solutions of LiCl could be employed in low-temperature studies of a solvated protein. We have investigated a range of concentrations of lysozyme and LiCl in aqueous solutions to identify systems that do not show phase separation and avoid solvent crystallization on cooling down. Compared to the lysozyme concentration in solution, the concentration of LiCl in the aqueous solvent plays the major role in determining systems suitable for low-temperature studies. We have observed interesting and rich phase behavior reminiscent of reentrant condensation of proteins.

  16. Estrous sheep serum enables in vitro capacitation of ram spermatozoa while preventing caspase activation.

    Science.gov (United States)

    Del Olmo, E; García-Álvarez, O; Maroto-Morales, A; Ramón, M; Jiménez-Rabadán, P; Iniesta-Cuerda, M; Anel-Lopez, L; Martinez-Pastor, F; Soler, A J; Garde, J J; Fernández-Santos, M R

    2016-01-15

    Estrous sheep serum (ESS) is considered the most efficient agent for in vitro capacitation of ram spermatozoa. We have explored the relationship between caspase activation and capacitation in ram. Semen samples from 17 rams were cryopreserved. In vivo fertility was evaluated after intrauterine artificial insemination. Samples were submitted to four treatments: control, ESS (10%), caspase inhibitor (Z-VAD-FMK), and estrous ewe serum plus caspase inhibitor (I + E). Sperm samples were incubated for 30 minutes at 38.5 °C and 5% CO2 and analyzed with flow cytometry for mitochondrial membrane potential (MitoTracker deep red), sperm viability and apoptosis-like changes (YO-PRO-1/propidium iodide), acrosomal status (peanut agglutinin-fluorescein isothiocyanate), membrane fluidity (merocyanine 540), and caspase activity (Vybrant FAM kits for polycaspases, caspase-8, and caspases 3-7). Estrous sheep serum induced changes compatible with capacitation, doubling the proportion of viable spermatozoa with increased merocyanine 540 and increasing YO-PRO-1(+) and acrosome-reacted spermatozoa (P ram spermatozoa by using ESS suggests a downregulation in apoptotic pathways. However, males with the lowest fertility showed parameters similar to high-fertility males, suggesting that other factors were involved apart from capacitation and/or caspase activation.

  17. Serum enzyme activities following long-distance running: comparison between Ethiopian and white athletes.

    Science.gov (United States)

    Eliakim, A; Nemet, D; Shenkman, L

    1995-11-01

    Ethiopian runners are famous for their achievements in long-distance running. The recent immigration of Ethiopians to Israel provided an opportunity to compare some physiological variables between elite Ethiopian and white Israeli runners. Six Ethiopian and five white Israeli runners, aged 20 to 40 years, were studied before and after an 11 km race. Venous blood was sampled from each runner prior to the race, and 1 h, 48 h, and 5 days following the race. The activities of creatine kinase (CK), lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) were measured, and levels of serum electrolytes, urea, creatinine, phosphorus, albumin, cholesterol, and alkaline phosphatase were also determined. CK activity rose 2.6-fold with a peak 5 days after the race. LDH and AST levels rose as well (1.4-1.3-fold, respectively). Significant elevations also occurred in serum phosphorus, uric acid, and creatinine concentrations 1 h after the race. In contrast to previous studies in which higher enzyme activities were reported in blacks, we did not detect any difference in serum enzyme values between black and white runners.

  18. [Molecular cloning, recombinant expression and characterization of lysozyme from Chinese shrimp Fenneropenaeus chinensis].

    Science.gov (United States)

    Bu, Xingjiang; Du, Xinjun; Zhou, Wenjie; Zhao, Xiaofan; Wang, Jinxing

    2008-05-01

    Lysozyme hydrolyses bacterial cell walls and acts as a nonspecific innate immunity molecule against the invasion of bacterial pathogens. We cloned the cDNA of lysozyme from Fenneropenaeus chinensis and named Fc-lysozyme (FcLyz in short). The full length of the gene was of 709 bp, and the open reading frame (477 bp) encoded 158 amino acids. The predicted protein had a signal peptide (-1--18 residue) and molecular weight of the mature protein (residue 1-140) was of 16.2 kD. A Lyz 1 domain (residue 1-130) in the lysozyme was found by SMART analysis. The results of semiquantity RT-PCR showed that FcLyz was constitutively expressed in tested tissues in a low level in normal shrimp, and up-regulated in hemocytes, heart, hepatopancreas and gill of bacterial challenged shrimp. The DNA fragment of mature Fc-Lys was subcloned to pET-30a (+) expression vector, the recombinant plasmid was transformed into Escherichia coli BL21 (DE3) and then induced by isopropylthio-beta-D-galactoside (IPTG). The antibacterial activity of the purified recombinant FcLys was analyzed and minimal inhibitory concentration (MIC) was assayed. The recombinant protein showed high antibacterial activity against some Gram-positive bacteria, and MIC reached 3.43 micromol/L, and relatively low activity against Gram-negative bacteria. All together, the Fc-Lys was regulated by pathogen infection and had antibacterial activity. This suggested that the FcLyz may be one of the important molecules against pathogens in innate immunity of the shrimp.

  19. [Cancer procoagulant activity in serum and neoplastic tissue in cases of cervical and uterine carcinoma].

    Science.gov (United States)

    Szajda, Sławomir Dariusz; Jóźwik, Maciej; Jóźwik, Marcin; Zalewska, Beata; Panek, Grzegorz; Sulkowski, Stanisław; Skrzydlewski, Zdzisław

    2004-09-01

    Cancer procoagulant (CP) is a sulfhydryl proteinase thought to be synthesized mainly by neoplastic cells. Consequently, increased CP activity in blood serum was interpreted as being associated with the presence of a proliferative process in the host's body. To date, CP activity has not been systematically studied in cases of genital carcinoma. The present study is aimed at evaluation of CP activity in women with genital carcinoma. A case-controlled study backed up by histopathological examination. Peripheral blood was sampled preoperatively in a sterile manner from an antecubital vein, from 16 women with cervical carcinoma and 15 women with uterine carcinoma. Blood for the reference group of 12 healthy women was obtained in an identical manner after an overnight fast. The CP activity in serum was determined using the coagulative method according to Gordon and Benson, and was expressed as coagulation time in seconds (s). The CP activity in 10% tissue homogenates (in saline) of genital cancer was determined by the chromogenic method according to Colucci et al. The mean CP activity in serum of women with cervical carcinoma (78.28 +/- 15.25 s) and of women with uterine carcinoma (79.63 +/- 12.02 s) was significantly different (P < 0.0001) from the respective values found in healthy women (281.33 +/- 43.19 s). The CP activity in neoplastic tissue was 28.50 +/- 6.40 nmol pNa/mL for cervical carcinoma, and 28.31 +/- 3.92 nmol pNa/mL for uterine carcinoma, both values being significantly higher (P < 0.0009) than the activity found in the normal tissues. There was no established relationship between neoplastic CP activity and FIGO staging of the disease. This is the first study to demonstrate the concomitant presence of CP activity in serum and neoplastic tissue of women with genital carcinoma. These patients have decreased coagulation time and thus are likely to develop coagulation disturbances in the course of their cancer. There may be a role for CP as a tumor marker of

  20. Decreased serum DNase1-activity in patients with autoimmune liver diseases.

    Science.gov (United States)

    Gatselis, Nikolaos K; Vakrakou, Aigli G; Zachou, Kalliopi; Androutsakos, Theodoros; Azariadis, Kalliopi; Hatzis, Gregorios; Manoussakis, Menelaos N; Dalekos, George N

    2017-03-01

    Deoxyribonuclease1 (DNase1) is involved in chromatin degradation of apoptotic cells. Its deficiency results in accumulation of self-DNA, which in turn may induce inflammation and autoimmunity. We assessed for the first time serum DNase1-activity in a large consecutive cohort of treatment-naïve patients with autoimmune liver diseases (ALD). DNase1-activity was determined by single radial enzyme-diffusion (SRED) at diagnosis of 224 patients with autoimmune hepatitis (AIH), 249 with primary biliary cirrhosis (PBC) and 36 with primary sclerosing cholangitis (PSC). Sera from 146 patients with chronic hepatitis B or C, 140 with nonalcoholic fatty liver disease/nonalcoholic steatohepatitis (NAFLD/NASH) and 114 healthy individuals served as disease and healthy controls. Available serum samples during remission from 50 AIH and 39 PBC patients were also investigated by paired analyzes. DNase1-activity was significantly lower in AIH, PBC and PSC compared to viral hepatitis (p 1400 mg/dl; p < 0.05); in PBC, with AST (p < 0.01), alanine aminotransferase (ALT) (p < 0.03) and anti-mitochondrial antibodies (AMA) (p = 0.008). In PSC, DNase1-activity was inversely associated with alkaline phosphatase (ALP) (p < 0.05). In AIH, complete responders were characterized by increased baseline DNase1-activity compared to partial responders, relapsers and non-responders (p < 0.02), whereas it was significantly increased after achievement of remission (p < 0.001). Serum DNase1-activity is significantly decreased in ALD patients, indicating its potential implication in their pathogenesis. Furthermore, DNase1-activity could be used as a new surrogate biomarker for predicting response to AIH treatment.

  1. Agaricus bisporus powder improved cutaneous mucosal and serum immune parameters and up-regulated intestinal cytokines gene expression in common carp (Cyprinus carpio) fingerlings.

    Science.gov (United States)

    Khodadadian Zou, Hassan; Hoseinifar, Seyed Hossein; Kolangi Miandare, Hamed; Hajimoradloo, Abdolmajid

    2016-11-01

    The aim of the present study was to investigate immunomodulatory effects of Agaricus bisporus, white bottom mushroom powder (WBMP) on common carp (Cyprinus carpio) fingerlings. Carps were fed on different levels of WBMP (0, 0.5, 1 and 2%) for 8 weeks and at the end of feeding trial, skin mucus immune parameters (total Ig, lysozyme and protease activity), cytokines gene expression (TNF-alpha, IL1b, IL8) in intestine as well as serum non-specific immune parameters (total Ig, lysozyme and ACH50) were measured. The results showed significant dose dependent increase of skin mucus immune parameters in carps fed WBMP (P  0.05). In case of serum non-specific immune parameters, except lysozyme activity, other parameters (Ig total and ACH50) were significantly affected by dietary inclusion of WBMP (P  0.05). Furthermore, feeding on WBMP supplemented diet significantly improved growth performance (P < 0.05). These results indicated that WBMP can be considered as a promising immunostimulants in early stage of common carp culture.

  2. ANTICOMPLEMENTARY ACTIVITY IN HUMAN SERUM OF LUNG CANCER PATIENTS AND ITS POSSIBLE CLINICAL SIGNIFICANCE

    Institute of Scientific and Technical Information of China (English)

    Liu Huirong; Liang Feng; Zhang Weifang; Zheng Zhenqun

    1998-01-01

    Objective: To identify whether the level of anticomplementary activity in serum is different between lung cancer patients and normal subjects. Method: With a sensitive immune haemolytic assay, the anticomplementary activity in the sera of 50 normal subjects and 61lung cancer patients were demonstrated. Results: It was found that all samples contained complement-inhibition activity, although some were of low degree. Increased anticomplementary activity was found to be associated significantly with lung cancer. With the progression of cancer the anticomplementary activity in sera increased in different lung cancer patients. Conclusion: Such a higher anticomplementary activity in sera of lung cancer patients might be one of the immunosuppressive contents induced by the tumor cells.

  3. Carboxymethyl chitosan-poly(amidoamine) dendrimer core-shell nanoparticles for intracellular lysozyme delivery.

    Science.gov (United States)

    Zhang, Xiaoyang; Zhao, Jun; Wen, Yan; Zhu, Chuanshun; Yang, Jun; Yao, Fanglian

    2013-11-06

    Intracellular delivery of native, active proteins is challenging due to the fragility of most proteins. Herein, a novel polymer/protein polyion complex (PIC) nanoparticle with core-shell structure was prepared. Carboxymethyl chitosan-grafted-terminal carboxyl group-poly(amidoamine) (CM-chitosan-PAMAM) dendrimers were synthesized by amidation and saponification reactions. (1)H NMR was used to characterize CM-chitosan-PAMAM dendrimers. The TEM images and results of lysozyme loading efficiency indicated that CM-chitosan-PAMAM dendrimers could self-assemble into core-shell nanoparticles, and lysozyme was efficiently encapsulated inside the core of CM-chitosan-PAMAM dendrimer nanoparticles. Activity of lysozyme was completely inhibited by CM-chitosan-PAMAM Dendrimers at physiological pH, whereas it was released into the medium and exhibited a significant enzymatic activity in an acidic intracellular environment. Moreover, the CM-chitosan-PAMAM dendrimer nanoparticles did not exhibit significant cytotoxicity in the range of concentrations below 3.16 mg/ml. The results indicated that these CM-chitosan-PAMAM dendrimers have excellent properties as highly potent and non-toxic intracellular protein carriers, which would create opportunities for novel applications in protein delivery.

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

    Science.gov (United States)

    Wu, Hanyu; Cao, Dainan; Liu, Tongxin; Zhao, Jianmin; Hu, Xiaoxiang; Li, Ning

    2015-01-01

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  6. STUDY OF SERUM HAPTOGLOBIN LEVEL AND ITS RELATION TO ERYTHROPOIETIC ACTIVITY IN BETA THALASSEMIA CHILDREN .

    Directory of Open Access Journals (Sweden)

    Seham Ragab

    2015-02-01

    Full Text Available Background  :Serum haptoglobin (Hp is a reliable marker for hemolysis regardless the inflammatory state.  Objective: We investigated the possible relation between Hp depletion and hemolysis severity, hepatitis C virus (HCV infection and iron load in β-thalassemia children. Methods: Twenty  two β-thalassemia major (TM ,20 β-thalassemia  intermedia (TI children with 20 age and sex matched healthy controls were involved. Pre-transfusion hemoglobin level was considered . Serum ferritin , Hp  and transferrin receptor  levels (sTfR  (by ELISA , alanine aminotransferase (ALT and  aspartate aminotransferase (AST  (by colorimetric method were assayed. Markers of hepatitis C virus  (HCV  were done by PCR. Results:  The mean Hp levels among the studied groups were as follows; 8.02 ± 0.93 (mg/dl , 8.6 ±0.72 (mg/dl  and 122  ± 18.5(mg/dl   for TM ,TI and the controls respectively . Both patient groups had significantly lower Hp level compared to the controls (P<0.0001  with significant lower level in TM compared to TI  children ( P= 0.034  .Significant inverse correlations were  found between serum Hp and sTfR levels in thalassemia children combined and in each group (TM and TI as well as among HCV infected children. STfR   was the only significant independent predictor for  serum Hp level (t= -5.585 , P<0.0001 . Among  HCV infected patients , no significant correlation was found between serum Hp and serum transaminases  .Conclusion:  Serum Hp depletion in thalassemia had significant relation to disease severity and correlated   well with their erythropoietic activity, as assessed by the measurement of  sTfR without significant relation  HCV infection . Large sample  multicenter studies are  recommended.

  7. The Serum Complement System: A Simplified Laboratory Exercise to Measure the Activity of an Important Component of the Immune System

    Science.gov (United States)

    Inglis, Jordan E.; Radziwon, Kimberly A.; Maniero, Gregory D.

    2008-01-01

    The immune system is a vital physiological component that affords animals protection from disease and is composed of innate and adaptive mechanisms that rely on cellular and dissolved components. The serum complement system is a series of dissolved proteins that protect against a variety of pathogens. The activity of complement in serum can be…

  8. The Serum Complement System: A Simplified Laboratory Exercise to Measure the Activity of an Important Component of the Immune System

    Science.gov (United States)

    Inglis, Jordan E.; Radziwon, Kimberly A.; Maniero, Gregory D.

    2008-01-01

    The immune system is a vital physiological component that affords animals protection from disease and is composed of innate and adaptive mechanisms that rely on cellular and dissolved components. The serum complement system is a series of dissolved proteins that protect against a variety of pathogens. The activity of complement in serum can be…

  9. Serum prolidase activity, oxidative stress, and antioxidant enzyme levels in patients with renal cell carcinoma.

    Science.gov (United States)

    Pirinççi, Necip; Kaba, Mehmet; Geçit, İlhan; Güneş, Mustafa; Yüksel, Mehmet Bilgehan; Tanık, Serhat; Arslan, Ayşe; Demir, Halit

    2016-02-01

    Prolidase is a member of the matrix metalloproteinase family. It plays a vital role in collagen turnover, matrix remodeling, and cell growth. Reactive oxygen species (ROS) have been implicated in the pathogenesis of various diseases, including cancers. Oxidative stress can cause tumor angiogenesis and may be carcinogenic. However, the relationship between antioxidant capacity and various cancers has been researched in several clinical trials. In our study, we aimed to identify serum prolidase activity, oxidative stress, and antioxidant enzyme levels in patients with renal tumors and to evaluate their relationships with each other. A total of 37 male patients with renal cell cancer and with a mean age of 56.28 ± 3.1 were included in the study. The control group comprising 36 male patients (mean age 56.31 ± 2.9) was randomly selected among the volunteers. Serum samples for measurement of superoxide dismutase (SOD), glutathione peroxidase (GSHPx), glutathione-S-transferase (GST), malondialdehyde (MDA), glutathione (GSH), and prolidase levels were kept at -20°C until they were used. Serum prolidase activity and MDA levels were significantly higher in renal cancer patients than in controls (all, p levels were significantly lower (p antioxidant levels in renal cancer. © The Author(s) 2013.

  10. Deciphering systemic lupus erythematosus-associated serum biomarkers reflecting apoptosis and disease activity.

    Science.gov (United States)

    Delfani, P; Sturfelt, G; Gullstrand, B; Carlsson, A; Kassandra, M; Borrebaeck, C A K; Bengtsson, A A; Wingren, C

    2017-04-01

    Systemic lupus erythematosus (SLE) is a severe chronic inflammatory autoimmune connective tissue disease. Despite major efforts, SLE remains a poorly understood disease with unpredictable course, unknown etiology and complex pathogenesis. Apoptosis combined with deficiency in clearing apoptotic cells is an important etiopathogenic event in SLE, which could contribute to the increased load of potential autoantigen(s); however, the lack of disease-specific protein signatures deciphering SLE and the underlying biological processes is striking and represents a key limitation. In this retrospective pilot study, we explored the immune system as a specific sensor for disease, in order to advance our understanding of SLE. To this end, we determined multiplexed serum protein expression profiles of crude SLE serum samples, using antibody microarrays. The aim was to identify differential immunoprofiles, or snapshots of the immune response modulated by the disease, reflecting apoptosis, a key process in the etiology of SLE and disease activity. The results showed that multiplexed panels of SLE-associated serum biomarkers could be decoded, in particular reflecting disease activity, but potentially the apoptosis process as well. While the former biomarkers could display a potential future use for prognosis, the latter biomarkers might help shed further light on the apoptosis process taking place in SLE.

  11. Relationship between elevated serum gamma-glutamyltransferase activity and slow coronary flow

    DEFF Research Database (Denmark)

    Sen, Nihat; Ozlü, Mehmet F; Basar, Nurcan;

    2009-01-01

    OBJECTIVES: We evaluated the relationship between coronary blood flow and serum gamma-glutamyltransferase (GGT) activity in patients with slow coronary flow (SCF). STUDY DESIGN: The study included 90 patients (47 men, 43 women; mean age 50.8+/-9.4 years) with SCF and 88 patients (45 men, 43 women......; mean age 51.4+/-8.8 years) with coronary artery disease (CAD), whose diagnoses were made by coronary angiography. Patients with CAD had normal coronary flow. Coronary flow was quantified using the corrected TIMI frame count (TFC) method and serum levels of gamma-glutamyltransferase were measured....... The results were compared with those of a control group consisting of 86 age- and sex-matched patients who had normal coronary arteries and normal coronary flow. RESULTS: The three groups were similar with respect to body mass index, presence of hypertension and diabetes mellitus, lipid profiles, and fasting...

  12. AhR transcription serum activity of Inuit´s across Greenlandic districts

    DEFF Research Database (Denmark)

    Long, Manhai; Deutch, Bente; Bonefeld-Jørgensen, Eva Cecilie

    2007-01-01

    Background: Human exposure to lipophilic persistent organic pollutants (POPs) including Human exposure to lipophilic persistent organic pollutants (POPs) including polychlorinated dibenzo-p-dioxins/furans (PCDDs/PCDFs), polychlorinated biphenyls (PCBs) and organochlorine pesticide is ubiquitous...... food. Conclusions: i) We observed that the proportion of dioxin like (DL) compounds in the POP i) We observed that the proportion of dioxin like (DL) compounds in the POP mixture was the dominating factor affecting the level of serum AhR transcriptional activity even at very high level of non DL......-PCBs; ii) The inverse association between the integrated serum AhR transactivity and sum of POPs might be explained by the higher level of compounds antagonizing the AhR function probably due to selective POP bioaccumulation in the food chain....

  13. Extra-Articular Symptoms in Constellation with Selected Serum Cytokines and Disease Activity in Spondyloarthritis

    Directory of Open Access Journals (Sweden)

    Hanna Przepiera-Będzak

    2016-01-01

    Full Text Available Objectives. In this study, we assessed the extra-articular symptoms in constellation with selected serum cytokines and disease activity in spondyloarthritis (SpA. Patients and Methods. We studied 287 SpA patients: 131 had AS, 110 had PsA, and 46 had SAPHO. We assessed extra-articular symptoms in all cases. In 191 SpA patients, we measured serum interleukin-6 (IL-6, interleukin-18 (IL-18, interleukin-23 (IL-23, endothelin-1 (ET-1, vascular endothelial growth factor (VEGF, and epidermal growth factor (EGF. Results. Patients with acute anterior uveitis (AAU had higher VAS (P=0.0008, BADSDAI (P=0.0001, ASDAS-ESR (P=0.04, CRP (P=0.006, IL-6 (P=0.02, and IL-18 (P=0.03 levels. Patients with inflammatory bowel disease (IBD had higher VAS (P=0.03, CRP (P=0.0009, and IL-6 (P=0.0003 levels. Patients with skin psoriasis had lower VAS (P=0.001 and BASDAI (P=0.00007 levels. Patients with psoriatic onycholysis had lower VAS (P=0.006, BASDAI (P=0.00001, and CRP (P=0.02 and higher IL-23 (P=0.04 levels. Patients with PPP had lower BASDAI (P=0.04 and higher ET-1 (P=0.001 levels. Conclusions. SpA patients with increased serum IL-18 and decreased serum ET-1 had an increased risk of extra-articular symptoms. In SpA patients, increased disease activity was associated with an increased risk of AAU and IBD and a decreased risk of skin psoriasis, psoriatic onycholysis, and PPP.

  14. Serum activities of liver enzymes in workers exposed to sub-TLV levels of dimethylformamide

    Directory of Open Access Journals (Sweden)

    Jinjiang He

    2015-04-01

    Full Text Available Objectives: The aim of this study has been to investigate serum activities of liver enzymes in workers exposed to sub-TLV levels of dimethylformamide (DMF. Material and Methods: Seventy-two workers and 72 healthy controls participated in the study. All subjects underwent complete physical examinations and abdominal ultrasound examination. Serum aspartate aminotransferase (AST, alanine aminotransferase (ALT, and c-glutamyl transpeptidase (c-GT were determined by an auto-chemistry analyzer. The data of airborne concentrations of DMF was obtained from the local Center of Disease Control and Prevention. The level of urine N-acetyl-S-(N-methylcarbamoylcysteine (AMCC was measured by means of high-performance liquid chromatography. Results: Time weighted average (TWA concentration of the DMF in workplace was 18.6 (range: 9.8–36.2 mg/m3. The concentration of the AMCC in workers’ urine was 28.32 (range: 1.8–58.6 mg/l and 9 workers’ AMCC exceeded the biological exposure index (40 mg/l. Thirty-one workers reported gastrointestinal symptoms (abdominal pain, nausea, anorexia and 10 workers reported headache, dizziness and/or palpitation in the exposed group. Serum analysis revealed that both the mean of serum activities of liver enzymes (ALT, AST and c-GT and the percentage of workers with abnormal liver function were significantly higher in the exposed group as compared to the controls. Conclusions: Dimethylformamide can cause liver damage even if air concentration is in the sub-threshold limit value (sub-TLV level. The protection of skin contact against the exposure to the DMF might be a critical issue as far as the occupational health is concerned.

  15. Destruction of single-species biofilms of Escherichia coli or Klebsiella pneumoniae subsp. pneumoniae by dextranase, lactoferrin, and lysozyme

    Science.gov (United States)

    The activity of dextranase, lactoferrin, lysozyme, and nisin against biofilms composed of either Klebsiella pneumonia or Escherichia coli was examined using the MBEC Assay™. Mature biofilms were treated and then sonicated to remove the adherent biofilm. This material was quantified using a lumines...

  16. Active fragments of the antihemorrhagic protein HSF from serum of habu (Trimeresurus flavoviridis).

    Science.gov (United States)

    Aoki, Narumi; Deshimaru, Masanobu; Terada, Shigeyuki

    2007-04-01

    Certain snakes have antihemorrhagic proteins in their sera. Habu serum factor (HSF), an antihemorrhagic protein isolated from the serum of the Japanese habu snake (Trimeresurus flavoviridis) is composed of two cystatin-like domains (D1 and D2) and a His-rich domain, and it inhibits several snake venom metalloproteinases (SVMPs). The activity of HSF can be abolished by trinitrophenylation of Lys residues with 2,4,6-trinitrobenzene sulphonic acid. Upon complex formation of HSF with SVMP, however, the loss of its inhibitory activity by the chemical modification was suppressed, and Lys(15), Lys(41), and Lys(103) residues in HSF were not trinitrophenylated. In order to identify the domain that is critical to the inhibitory activity on SVMPs, native HSF was digested with papain followed by cleavage with cyanogen bromide, yielding a low-molecular mass fragment that was composed of two peptide chains (residues 5-89 and 312-317) linked by a disulfide bond. This fragment inhibited several SVMPs and showed significant antihemorrhagic activity. This indicates that the N-terminal half of D1 is indispensable for the antihemorrhagic activity of HSF. Furthermore, a three-dimensional model of two cystatin-like domains constructed by the homology modeling has indicated that three Lys residues (15, 41, and 103) are exposed to the same surface of HSF molecule.

  17. Lysozyme Catalyzes the Formation of Antimicrobial Silver Nanoparticles (POSTPRINT)

    Science.gov (United States)

    2009-04-01

    aseptics and therapeutic use in the future. KEYWORDS: antimicrobial · lysozyme · silver · nanoparticle · biocompatibility · biomineralization A RT IC LE VOL...protein that will adsorb to ionic and hydro- phobic surfaces, including metal surfaces.2124 After synthesis in methanol, lysozyme-stabilized nanoparti- cle...the strong ionic interactions be- tween metal nanoparticles normally make it difficult to achieve high concentrations of monodispersed and stable

  18. Effects of reduced levels of sulfite in wine production using mixtures with lysozyme and dimethyl dicarbonate on levels of volatile and biogenic amines.

    Science.gov (United States)

    Ancín-Azpilicueta, Carmen; Jiménez-Moreno, Nerea; Moler, José Antonio; Nieto-Rojo, Rodrigo; Urmeneta, Henar

    2016-10-01

    Sulphur dioxide (SO2) is an important preservative for wine, but its presence in foods can cause allergies and this has given impetus to the research for alternatives. The aim of this study was to reduce levels of sulfite in wine production using mixtures with lysozyme and dimethyl dicarbonate and examine the influence on levels of volatile and biogenic amines. To do so, vinifications were carried out using lysozyme, dimethyl dicarbonate (DMDC) and mixtures of these with SO2 in different concentrations (25 and 50 mg l(-1)). Results were compared with a control vinification with only SO2 (50 mg l(-1)). Mixing low concentrations of SO2 with lysozyme and DMDC reduced the concentration of biogenic amines (histamine, tyramine, putrescine, cadaverine, phenylethylamine + spermidine and spermine). In general, the total concentration of volatile amines (dimethylamine, isopropylamine, isobutylamine, pyrrolidine, ethylamine, diethylamine, amylamine and hexylamine) was higher in the sample fermented only with SO2. The concentrations of amines with secondary amino groups (dimethylamine, diethylamine, pyrrolidine) were higher in the sample only fermented with SO2 than those fermented with DMDC and lysozyme or with a mixture of preservatives. When SO2 was the only preservative in wine, total amine concentration (biogenic and volatile amines) was higher than for the rest of the treatments. Lysozyme by itself, and lysozyme mixed with SO2, both reduced the formation of biogenic amines but given the antioxidant activity of SO2 the use of the preservative mixture seems more advisable.

  19. Aging-Related Correlation between Serum Sirtuin 1 Activities and Basal Metabolic Rate in Women, but not in Men

    Science.gov (United States)

    2017-01-01

    Sirtuin (SIRT) is a main regulator of metabolism and lifespan, and its importance has been implicated in the prevention against aging-related diseases. The purpose of this study was to identify the pattern of serum SIRT1 activity according to age and sex, and to investigate how serum SIRT1 activity is correlated with other metabolic parameters in Korean adults. The Biobank of Jeju National University Hospital, a member of the Korea Biobank Network, provided serum samples from 250 healthy adults. Aging- and metabolism-related factors were analyzed in serum, and the data were compared by the stratification of age and sex. Basal metabolic rate (BMR) decreased with age and was significantly lower in men in their fifties and older and in women in their forties and older compared with twenties in men and women, respectively. SIRT1 activities were altered by age and sex. Especially, women in their thirties showed the highest SIRT1 activities. Correlation analysis displayed that SIRT1 activity is positively correlated with serum triglyceride (TG) in men, and with waist circumference, systolic blood pressure, diastolic blood pressure, and serum TG in women. And, SIRT1 activity was negatively correlated with aspartate aminotransferase/alanine aminotransferase ratio in women (r = −0.183, p = 0.039). Positive correlation was observed between SIRT1 activity and BMR in women (r = 0.222, p = 0.027), but not in men. Taken together, these findings suggest the possibility that serum SIRT1 activities may be utilized as a biomarker of aging. In addition, positive correlation between SIRT1 activity and BMR in women suggests that serum SIRT1 activity may reflect energy expenditure well in human. PMID:28168178

  20. The serum of rabbitfish (Siganus oramin) has antimicrobial activity to some pathogenic organisms and a novel serum L-amino acid oxidase is isolated.

    Science.gov (United States)

    Wang, Fanghua; Li, Ruijun; Xie, Mingquan; Li, Anxing

    2011-01-01

    The serum of rabbitfish (Siganus oramin) has been confirmed previously to have killing effect to Cryptocaryon irritans, an important marine ciliate protozoan that causes a disease referred to as "marine white spot disease". Herein, we find the serum of the rabbitfish also shows antibacterial activity against both gram-positive and gram-negative bacteria and has killing effect on two other parasites: Trypanosoma brucei brucei, Ichthyophthirius multifiliis. Results of scanning electron microscopy indicated that after treating with rabbitfish serum, the surface of the Staphylococcus aureus was wrinkled and pores were formed on the surface of Escherichia coli. Serum of the rabbitfish possesses a strong killing effect to Ichthyophthirius multifiliis in vitro, causing a similar effect as to C. irritans. The serum of rabbitfish also showed strong killing effect to T. b. brucei in vitro, with the minimus trypanocidal titre (MTT) only to be 1.5% in 1 h. Results of laser confocal fluorescence microscopy indicated that rabbitfish serum could also induce cell rupture of T. b. brucei. A novel antimicrobial protein (SR-LAAO) was isolated from the serum of rabbitfish by using ultrafiltration, reversed phase high performance liquid chromatography (RP-HPLC) and Native polyacrylamide gel electrophoresis (Native-PAGE). Results of gel overlay assay showed that the protein could act alone to inhibit the growth of S. aureus and E. coli. Results of western blot and automated Edman degradation showed that it was the same as the antiparasitic protein (APP) reported before to have killing effect on C. irritans. Full length cDNA sequence of the SR-LAAO was cloned. BLAST research suggested that the cDNA of SR-LAAO has a close similarity with a number of L-amino acid oxidases (LAAOs) and possesses two conserved motifs that exist in LAAOs. Combined, these results demonstrate that this protein which has antimicrobial activity to some pathogenic organisms was a novel LAAO found in the serum of

  1. Dioxin-like activities in serum across European and Inuit populations

    Directory of Open Access Journals (Sweden)

    Zvyezday Valentyna

    2006-05-01

    Full Text Available Abstract Background Persistent organic pollutants (POPs such as polychlorinated dibenzo-p-dioxins/furans, polychlorinated biphenyls (PCBs and organochlorine pesticides can cause a series of adverse effects on e.g. reproduction in animals and humans, many of which involve the aryl hydrocarbon receptor (AhR. The aim of the present study was to compare the integrated serum level of AhR mediated activity among European and Inuit populations, and evaluate whether the activity was associated to the selected POP markers, 2,2',4,4',5,5'-hexachlorobiphenyl (CB-153 and 1,1-dichloro-2,2-bis(p-chlorophenyl-ethylene (p,p'-DDE. Methods The study included 338 males from Greenland (Inuit's, Sweden, Warsaw (Poland and Kharkiv (Ukraine. The AhR transactivity of serum extracts alone (AhRag and competitive AhR activity (AhRcomp upon co-exposure with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD were determined in the lipophilic serum fraction containing the POPs using the AhR mediated luciferase reporter Hepa1.12cR cell assay. Results The European groups showed higher median level of AhR-TEQ (TCDD toxic equivalents compared to the Inuit's, whereas higher incidence of Inuits sample further induced AhRcomp activity. Neither AhRag nor AhR-TEQ were correlated to CB-153 or p,p'-DDE for any of the study groups. Multiple regressions showed a significant heterogeneity of association between the CB-153 and the AhRcomp across the study groups, and accordingly a negative association between AhRcomp and CB-153 was found for the Kharkiv group. Conclusion No consistent correlation between AhR activities and two POP markers was found. Although the difference of AhRag between European and Inuit men could not be explained by CB-153 or p,p'-DDE levels alone, we believe that the variation of AhR serum activity reflects different pattern of POP exposure, genetics and/or life style factors.

  2. Catalytically active bovine serum amine oxidase bound to fluorescent and magnetically drivable nanoparticles

    Directory of Open Access Journals (Sweden)

    Bidollari E

    2012-05-01

    Full Text Available Giulietta Sinigaglia1, Massimiliano Magro1, Giovanni Miotto1, Sara Cardillo1, Enzo Agostinelli2,3, Radek Zboril4, Eris Bidollari2,3, Fabio Vianello11Department of Biological Chemistry, University of Padua, Padua, Italy; 2Istituto Pasteur-Fondazione Cenci Bolognetti, Department of Biochemical Sciences "A. Rossi Fanelli", SAPIENZA University of Rome, Rome, Italy; 3CNR, Institute Biology and Molecular Pathology, Rome, Italy; 4Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry, Palacky University, Olomouc, Czech RepublicAbstract: Novel superparamagnetic surface-active maghemite nanoparticles (SAMNs characterized by a diameter of 10 ± 2 nm were modified with bovine serum amine oxidase, which used rhodamine B isothiocyanate (RITC adduct as a fluorescent spacer-arm. A fluorescent and magnetically drivable adduct comprised of bovine serum copper-containing amine oxidase (SAMN–RITC–BSAO that immobilized on the surface of specifically functionalized magnetic nanoparticles was developed. The multifunctional nanomaterial was characterized using transmission electron microscopy, infrared spectroscopy, mass spectrometry, and activity measurements. The results of this study demonstrated that bare magnetic nanoparticles form stable colloidal suspensions in aqueous solutions. The maximum binding capacity of bovine serum amine oxidase was approximately 6.4 mg g-1 nanoparticles. The immobilization procedure reduced the catalytic activity of the native enzyme to 30% ± 10% and the Michaelis constant was increased by a factor of 2. We suggest that the SAMN–RITC–BSAO complex, characterized by a specific activity of 0.81 IU g-1, could be used in the presence of polyamines to create a fluorescent magnetically drivable H2O2 and aldehydes-producing system. Selective tumor cell destruction is suggested as a potential future application of this system.Keywords: amine oxidase, hydrogen peroxide production, superparamagnetic

  3. Serum and blister fluid pharmacokinetics and bactericidal activities of ampicillin-sulbactam, cefotetan, cefoxitin, ceftizoxime, and ticarcillin-clavulanate.

    Science.gov (United States)

    Jaresko, G S; Barriere, S L; Johnson, B L

    1992-01-01

    Ampicillin-sulbactam, ticarcillin-clavulanate, cefoxitin, cefotetan, and ceftizoxime are promoted for the treatment of mixed aerobic-anaerobic bacterial infections. Their activities have been compared in vitro but not in vivo. In order to assess the in vivo activities of these agents in serum and interstitial fluid, we administered single, intravenous doses of these antimicrobial agents to healthy subjects. Concentrations of the antimicrobial agents in serum and suction-induced blister fluid and bactericidal activity were measured by high-pressure liquid chromatography and the standard methodology of the National Committee for Clinical Laboratory Standards, respectively. The organisms used for bactericidal activity tests were one isolate each of Staphylococcus aureus, Klebsiella pneumoniae, and Bacteroides fragilis. Pharmacokinetic parameters in serum and blister fluid were similar to those derived in other investigations. Of note were the high and prolonged concentrations of ticarcillin and cefotetan in blister fluid, despite high-level serum protein binding. The bactericidal activities in serum and blister fluid reflected the relative in vitro activities and kinetic dispositions of the various antimicrobial agents except for the bactericidal activity of cefotetan, which was substantially lower in blister fluid than serum, despite a blister fluid:serum area under the concentration-time curve ratio of 1.5. Similarly, the activity of ticarcillin-clavulanate in blister fluid was also substantially less than would have been predicted by the blister fluid:serum ratio of the area under the concentration-time curve of 1.1, possibly because of the low concentrations of clavulanate in blister fluid. The rankings of the in vivo bactericidal activities of the five drugs were as follows: for S. aureus, ampicillin-sulbactam > ticarcillin-clavulanate > ceftizoxime > cefoxitin > cefotetan; for K. pneumoniae, ceftizoxime > cefotetan > ampicillin-sulbactam = ticarcillin

  4. Serum des-R prothrombin activation peptide fragment 2: a novel prognostic marker for disseminated intravascular coagulation.

    Science.gov (United States)

    Chung, Soie; Kim, Ji-Eun; Kim, Hyun Kyung; Yeon, Eun Hee; Shin, Yong Sung; Kim, Chul Woo

    2013-06-01

    Disseminated intravascular coagulation (DIC) is diagnosed based on the combination of predisposing underlying conditions and laboratory tests for plasma coagulation markers. Because the collection of blood plasma samples is a fastidious procedure, the serum sample method may be preferred for measurement of coagulation markers when feasible. The novel serum marker des-R prothrombin activation peptide fragment 2 (des-R F2) was measured using a sandwich enzyme-linked immunosorbent assay in 181 patients suspected of having DIC. Thrombin generation potential was estimated with a calibrated automated thrombogram. Serum des-R F2 was generated with an in vitro clotting process within a serum separation tube after blood collection. Carboxypeptidase inhibitor inhibited the formation of des-R F2 during in vitro clotting. Low levels of prothrombin and thrombin generation potential resulted in low serum des-R F2 levels. Serum des-R F2 was significantly decreased in overt DIC. Levels of des-R F2 correlated with DIC severity and other coagulation markers. Of note, the decrease in serum des-R F2 levels was a significant marker for predicting mortality. The serum marker, des-R F2, can be used for the investigation of DIC severity and prognosis. It should be considered a useful marker, especially when only serum samples are available. Copyright © 2013 Elsevier Ltd. All rights reserved.

  5. Modification of serum, pancreatic, and microbial lipase activities by phorbol diesters.

    Science.gov (United States)

    Zhang, Z C; Cabot, M C

    1987-01-01

    The influence of phorbol diesters on the in vitro hydrolysis of diacylglycerols was examined using enzymes from rat serum, porcine pancreas, and Rhizopus delemar. Two main phenomena were observed: 12-O-tetradecanoylphorbol-13-acetate (TPA), when added to the enzyme assay system, stimulated 2- to 3-fold the hydrolysis of [9,10-3H]dioleoylglycerol by serum lipase. The hydrolysis of dioleoylglycerol by either pancreatic or R. delemar lipase was, on the other hand, inhibited by TPA. A 50% inhibition of the pancreatic and R. delemar enzymes was attained with 10 and 2.0 microM TPA, respectively. The pattern of enzyme stimulation (rat serum), with regard to increasing TPA concentrations, was hyperbolic. Stimulation was not influenced by Triton X-100, but it was highly dependent on the structure of the phorbol ester: TPA greater than phorbol didecanoate greater than tetradecanoylphorbol. Phorbol dibutyrate, phorbol acetate, myristic acid, and mezerein were without influence. Lipase activity was inhibited most strongly by TPA and the nonpromoter 4-O-methyl-TPA; the weaker promoter, phorbol dibutyrate, was relatively inactive. The inhibition of R. delemar lipase by TPA was reversible. Collectively, these data show that phorbol diesters can interact with enzymes other than protein kinase C. It is believed, by virtue of their structural similarity to diacylglycerols, that phorbol diesters may serve directly as intracellular regulators of lipid metabolism. In such a manner phorbol esters could sustain or attenuate the second messenger signal by modifying diacylglycerol metabolism, a manifestation of the pleiotropic action.

  6. Evaluation of serum undercarboxylated osteocalcin in premenopausal rheumatoid arthritis patients: its correlation with disease activity and bone mineral density

    Directory of Open Access Journals (Sweden)

    Heba A Esaily

    2016-01-01

    Conclusion Serum level of ucOC (which is a mirror of vitamin K deficiency was found to be higher in premenopausal RA patients than controls and correlated positively with disease activity and inversely with BMD measurement.

  7. Adenosine deaminase activity in serum, erythrocytes and lymphocytes of rats infected with Leptospira icterohaemorrhagiae.

    Science.gov (United States)

    Tonin, Alexandre A; Pimentel, Victor C; da Silva, Aleksandro S; de Azevedo, Maria Isabel; Souza, Viviane C G; Wolkmer, Patrícia; Rezer, João F P; Badke, Manoel R T; Leal, Daniela B R; Schetinger, Maria Rosa C; Monteiro, Silvia G; Lopes, Sonia T A

    2012-04-01

    Leptospirosis is a systemic disease of humans and domestic animals, mainly dogs, cattle and swine. The course of human leptospirosis varies from mild to severe fatal forms and the most severe form of human leptospirosis is principally caused by Leptospira interrogans serovar icterohaemorrhagiae (L. icterohaemorrhagiae). The enzyme adenosine deaminase (ADA) plays an important role in the production and differentiation of blood cells. The aim of this study was to evaluate the activity of ADA in serum, erythrocytes and lymphocytes of rats infected with L. icterohaemorrhagiae, as compared with non-infected rats. Twenty-four adult rats, divided into two uniform groups (A and B) were used for the enzymatic assays. The animals in Group B were inoculated intraperitoneally with 2×10(8) leptospires/rat, and the rodents in Group A (control) were not-inoculated. Blood collection was performed on days 5 and 15 post-infection (PI) and the blood used to assess the ADA activity. The infection by L.icterohaemorrhagiae altered erythrocyte count, hemoglobin concentration and hematocrit, causing a decrease in all these parameters on day 15 PI. Lymphocytes decreased significantly on day 15 PI, and ADA activity in serum was inhibited in infected rats on days 5 and 15 PI and its activity in erythrocytes were increased on day 5 PI. On day 5 PI, we found an increase in ADA activity in erythrocytes of infected rats. No correlation was observed between hematocrit and erythrocyte ADA activity on days 5 and 15 PI. The ADA activity was inhibited in rats infected on day 15 PI. A positive correlation (r(2)=60) was also observed between the number of lymphocytes and ADA activity in lymphocytes on day 15 PI (Perythrocytes in experimental infection by L.icterohaemorrhagiae in rats, concomitantly with hematological parameters.

  8. Hydrogen sulfide reduces serum triglyceride by activating liver autophagy via the AMPK-mTOR pathway.

    Science.gov (United States)

    Sun, Li; Zhang, Song; Yu, Chengyuan; Pan, Zhenwei; Liu, Yang; Zhao, Jing; Wang, Xiaoyu; Yun, Fengxiang; Zhao, Hongwei; Yan, Sen; Yuan, Yue; Wang, Dingyu; Ding, Xue; Liu, Guangzhong; Li, Wenpeng; Zhao, Xuezhu; Liu, Zhaorui; Li, Yue

    2015-12-01

    Autophagy plays an important role in liver triglyceride (TG) metabolism. Inhibition of autophagy could reduce the clearance of TG in the liver. Hydrogen sulfide (H2S) is a potent stimulator of autophagic flux. Recent studies showed H2S is protective against hypertriglyceridemia (HTG) and noalcoholic fatty liver disease (NAFLD), while the mechanism remains to be explored. Here, we tested the hypothesis that H2S reduces serum TG level and ameliorates NAFLD by stimulating liver autophagic flux by the AMPK-mTOR pathway. The level of serum H2S in patients with HTG was lower than that of control subjects. Sodium hydrosulfide (NaHS, H2S donor) markedly reduced serum TG levels of male C57BL/6 mice fed a high-fat diet (HFD), which was abolished by coadministration of chloroquine (CQ), an inhibitor of autophagic flux. In HFD mice, administration of NaSH increased the LC3BII-to-LC3BI ratio and decreased the p62 protein level. Meanwhile, NaSH increased the phosphorylation of AMPK and thus reduced the phosphorylation of mTOR in a Western blot study. In cultured LO2 cells, high-fat treatment reduced the ratio of LC3BII to LC3BI and the phosphorylation of AMPK, which were reversed by the coadministration of NaSH. Knockdown of AMPK by siRNA in LO2 cells blocked the autophagic enhancing effects of NaSH. The same qualitative effect was observed in AMPKα2(-/-) mice. These results for the first time demonstrated that H2S could reduce serum TG level and ameliorate NAFLD by activating liver autophagy via the AMPK-mTOR pathway.

  9. The role of serum cholinesterase activity and S100B protein in the evaluation of organophosphate poisoning.

    Science.gov (United States)

    Yardan, T; Baydin, A; Acar, E; Ulger, F; Aygun, D; Duzgun, A; Nar, R

    2013-10-01

    The aim of this study was to investigate the role of serum cholinesterase (SChE) activity and S100B protein in the evaluation of patients with acute organophosphate (OP) poisoning. Patients with acute OP poisoning admitted to the emergency department were included in this cross-sectional study. Twenty healthy volunteers served as controls. The SChE activity and serum S100B were determined on admission. Patients were divided into two groups (low severity and high severity). Thirty-six patients diagnosed with acute OP poisoning were enrolled. Serum S100B concentrations were higher in patients than in the control group (p poisoning.

  10. Formulation of antiacne serum based on lime peel essential oil and in vitro antibacterial activity test against Propionibacterium acnes

    Science.gov (United States)

    Fitri, Noor; Fatimah, Ifat; Chabib, Lutfi; Fajarwati, Febi Indah

    2017-03-01

    Propionibacterium acnes are a normal bacterium in human skin but it can become primary pathogens that can cause inflammation on the skin. Research about new antibacterial compounds is important because resistance of bacteria acne to antibiotics. Some of Essential oils have antibacterial properties. Lime peel essential oil and patchouli essential oil have some terpenoids that act as antibacterial compounds such as Linalool and Seychellene. The purpose of this research was to formulate anti acne serum based on lime peel essential oil and patchouli oil and to determine the zone of inhibition against of Propionibacterium acnes. This study made 21 variations of formulation of anti acne serum, consisted of lime peel essential oil, patchouli oil and olive oil. Anti acne serum was evaluated i.e. in vitro antibacterial activity test against Propionibacterium acnes for 5 days, organoleptic, stability test, pH test, viscosity test and GC-MS analysis. Nine serum formulations had been selected, which based on their most favorite order. Those favorite serums had antibacterial inhibitory against Propionibacterium acnes between 20.80 - 26.12 mm, whereas control positive only 12.47 mm and control negative 5.78 mm. The most favorite serum with the best antibacterial activity was serum formula A. The composition of serum A consist of lime peel essential oil: patchouli oil: olive oil (11:1:18).

  11. Angiotensin Converting Enzyme Activity in the Serum, Lung, Liver and Kidney in Streptozotocin -Induced Diabetic Rats and Diabetic Nephropathy

    OpenAIRE

    Üstündağ, Bilal

    2014-01-01

    To clarify the relationship between the alterations of the levels of angiotensin converting enzyme (ACE) and diabetic nephropathy, ACE activity in the lung, liver, kid-ney and serum were investigated in streptozotocin (STZ)-induced diabetic rats. The levels of serum ACE activity unchanged 3 days post STZ treatment but it was significantly an increase 12 and 30 days post STZ treatment in diabetic rats (p

  12. Serum Vitamin D Level and Rheumatoid Arthritis Disease Activity: Review and Meta-Analysis

    Science.gov (United States)

    Lin, Jin; Liu, Jian; Davies, Michael L.; Chen, Weiqian

    2016-01-01

    Background The evidence from epidemiological studies concerning the relationship between serum vitamin D concentrations and rheumatoid arthritis (RA) is inconsistent. This meta-analysis is aimed at determining the magnitude of the correlation between this common autoimmune disease and vitamin D, an important nutrient known to dampen adaptive immune responses. Methods Through multiple search strategies, relevant literature was identified and evaluated for quality before May 16 2015. Data extracted from eligible studies was synthesized to calculate pooled correlation coefficient (r), mean difference (MD) and odds ratio (OR). The Venice criteria were applied to assess the credibility of the evidence for each statistically significant association. Results A total of 24 reports involving 3489 patients were selected for analysis. RA patients had lower vitamin D levels than healthy controls (MD:-16.52 nmol/L, 95% confidence intervals [CI]:-18.85 to -14.19 nmol/L). There existed a negative relationship between serum 25-hydroxyvitamin D (25OHD) level and disease activity index, e.g. 25OHD vs. Disease Activity Score in 28 joints (DAS28): r = -0.13, 95% CI -0.16 to -0.09; 25OHD vs. C-reactive protein: r = -0.12, 95% CI -0.23 to -0.00. Additionally, latitude-stratified subgroup analysis yielded a relatively stronger negative correlation between 25OHD and DAS28 in low-latitude areas. This inverse relationship also appeared more significant in developing countries than in developed countries. No publication bias was detected. Conclusion RA patients had lower vitamin D values than healthy controls. There was a negative association between serum vitamin D and RA disease activity. However, more strictly controlled studies are needed to validate these findings. PMID:26751969

  13. Structural relationships in the lysozyme superfamily: significant evidence for glycoside hydrolase signature motifs.

    Directory of Open Access Journals (Sweden)

    Alexandre Wohlkönig

    Full Text Available BACKGROUND: Chitin is a polysaccharide that forms the hard, outer shell of arthropods and the cell walls of fungi and some algae. Peptidoglycan is a polymer of sugars and amino acids constituting the cell walls of most bacteria. Enzymes that are able to hydrolyze these cell membrane polymers generally play important roles for protecting plants and animals against infection with insects and pathogens. A particular group of such glycoside hydrolase enzymes share some common features in their three-dimensional structure and in their molecular mechanism, forming the lysozyme superfamily. RESULTS: Besides having a similar fold, all known catalytic domains of glycoside hydrolase proteins of lysozyme superfamily (families and subfamilies GH19, GH22, GH23, GH24 and GH46 share in common two structural elements: the central helix of the all-α domain, which invariably contains the catalytic glutamate residue acting as general-acid catalyst, and a β-hairpin pointed towards the substrate binding cleft. The invariant β-hairpin structure is interestingly found to display the highest amino acid conservation in aligned sequences of a given family, thereby allowing to define signature motifs for each GH family. Most of such signature motifs are found to have promising performances for searching sequence databases. Our structural analysis further indicates that the GH motifs participate in enzymatic catalysis essentially by containing the catalytic water positioning residue of inverting mechanism. CONCLUSIONS: The seven families and subfamilies of the lysozyme superfamily all have in common a β-hairpin structure which displays a family-specific sequence motif. These GH β-hairpin motifs contain potentially important residues for the catalytic activity, thereby suggesting the participation of the GH motif to catalysis and also revealing a common catalytic scheme utilized by enzymes of the lysozyme superfamily.

  14. Effect of sulfoxides on the thermal denaturation of hen lysozyme: A calorimetric and Raman study

    Science.gov (United States)

    Torreggiani, A.; Di Foggia, M.; Manco, I.; De Maio, A.; Markarian, S. A.; Bonora, S.

    2008-11-01

    A multidisciplinary study of the thermal denaturation of lysozyme in the presence of three sulfoxides with different length in hydrocarbon chain (DMSO, DESO, and DPSO) was carried out by means of DSC, Raman spectroscopy, and SDS-PAGE techniques. In particular, the Td and Δ H values obtained from the calorimetric measurements showed that lysozyme is partially unfolded by sulfoxides but most of the conformation holds native state. The sulfoxide denaturing ability increases in the order DPSO > DESO > DMSO. Moreover, only DMSO and DESO have a real effect in preventing the heat-induced inactivation of the protein and their maximum heat-protective ability is reached when the DMSO and DESO amount is ⩾25% w/w. The sulfoxide ability to act as effective protective agents against the heat-induced inactivation was confirmed by the protein analysis. The enzymatic activity, as well as the SDS-PAGE analysis, suggested that DESO, having a low hydrophobic character and a great ability to stabilise the three-dimensional water structure, is the most heat-protective sulfoxide. An accurate evaluation of the heat-induced conformational changes of the lysozyme structure before and after sulfoxide addition was obtained by the analysis of the Raman spectra. The addition of DMSO or DESO in low concentration resulted to sensitively decrease the heat-induced structural modifications of the protein.

  15. Effects of protein and phosphate buffer concentrations on thermal denaturation of lysozyme analyzed by isoconversional method.

    Science.gov (United States)

    Cao, X M; Tian, Y; Wang, Z Y; Liu, Y W; Wang, C X

    2016-07-03

    Thermal denaturation of lysozymes was studied as a function of protein concentration, phosphate buffer concentration, and scan rate using differential scanning calorimetry (DSC), which was then analyzed by the isoconversional method. The results showed that lysozyme thermal denaturation was only slightly affected by the protein concentration and scan rate. When the protein concentration and scan rate increased, the denaturation temperature (Tm) also increased accordingly. On the contrary, the Tm decreased with the increase of phosphate buffer concentration. The denaturation process of lysozymes was accelatated and the thermal stability was reduced with the increase of phosphate concentration. One part of degeneration process was not reversible where the aggregation occurred. The other part was reversible. The apparent activation energy (Ea) was computed by the isoconversional method. It decreased with the increase of the conversion ratio (α). The observed denaturation process could not be described by a simple reaction mechanism. It was not a process involving 2 standard reversible states, but a multi-step process. The new opportunities for investigating the kinetics process of protein denaturation can be supplied by this novel isoconversional method.

  16. Comparison of two codon optimization strategies enhancing recombinant Sus scrofa lysozyme production in Pichia pastoris.

    Science.gov (United States)

    Zhu, D; Cai, G; Wu, D; Lu, J

    2015-05-16

    Lysozyme has played an important role in animal feed additive industry, food additive industry and biological engineering. For improving expression efficiency of recombinant lysozyme from Sus scrofa, two genes respectively designed by the most used codon optimization strategies, "one amino acid one codon" and "codon randomization", were synthesized and expressed in Pichia pastoris X—33. At shaking flask level, Sus scrofa lysozyme (SSL) under two conditions had a highest activity of 153.33±10.41 and 538.33±15.18 U/mL after a 5 days induction of 1% methanol, with secreted protein concentration 80.03±1.94 and 239.60±4.16 mg/L, respectively. Compared with the original SSL gene, the expression of optimized SSL gene by the second strategy showed a 2.6 fold higher level, while the first method had no obvious improvement in production. In total secreted protein, the proportions of recombinant SSL encoded by the original gene, first method optimized gene and the second—strategy optimized one were 75.06±0.25%, 74.56±0.14% and 79.00±0.14%, respectively, with the same molecular weight about 18 kDa, optimum acidity pH 6.0 and optimum temperature 35degC.

  17. Introduction of Ca(2+)-binding amino-acid sequence into the T4 lysozyme.

    Science.gov (United States)

    Leontiev, V V; Uversky, V N; Permyakov, E A; Murzin, A G

    1993-03-05

    The 51-62 loop of T4 phage lysozyme was altered by site-directed mutagenesis to obtain maximal homology with the typical EF-hand motif. A Ca(2+)-binding site was designed and created by replacing both Gly-51 and Asn-53 with aspartic acid. The mutant T4 lysozyme (G51D/N53D) was expressed in Escherichia coli. The activity of the G51D/N53D-mutant was about 60% of that of the wild-type protein. This mutant can bind Ca2+ ions specifically, while the effective dissociation constant was essentially greater than that of the EF-hand proteins. Stability of the G51D/N53D-mutant apo-form to urea- or temperature-induced denaturation was the same as that of the wild-type protein. In the presence of Ca2+ ions in solution the stability of the mutant T4 phage lysozyme was less than that of the wild-type protein. It is suggested that the binding of Ca2+ by the mutant is accompanied by the considerable conformational changes in the 'corrected' loop, which can lead to the Ca(2+)-induced destabilization of the protein.

  18. Serum cholesterol concentration associated with aspirin esterase activity in older people: preliminary data

    Directory of Open Access Journals (Sweden)

    Kazuhiko Kotani, Russell Caccavello, Ricardo Hermo, Toshiyuki Yamada, Nobuyuki Taniguchi, Alejandro Gugliucci

    2010-01-01

    Full Text Available OBJECTIVE: Metabolism of aspirin (acetylsalicylic acid, commonly used in older people for the prevention of cardiovascular disease, is important to the effectiveness of this drug. Whereas part of aspirin hydrolysis occurs in blood, there is a paucity of information in regards to circulating aspirin esterase activity in various physiological and pathological conditions. High aspirin esterase activity, corresponding to faster aspirin hydrolysis (thus aspirin non-responsiveness, may occur in cardiovascular disease-prone states. The objective of this study was to investigate the effects of cardio-metabolic variables such as cholesterol on serum aspirin esterase activity in older people who participated in an intervention study on physical activity. METHODS: A total of 18 non-medicated subjects (7 men/11 women, mean age 67.8 years, body mass index = 23.4 ± 3.3 kg/m2, who completed a 3-month interventional program for a mild-to-moderate increase in physical activity, were analyzed. The body mass index, plasma glucose, serum total cholesterol and aspirin esterase activity were measured in the pre- and post-interventional phases of the study. RESULTS: During the interventional period, the changes in aspirin esterase activity correlated significantly and positively with those of total cholesterol concentrations (r = 0.542, P = 0.020; β = 0.609, P = 0.035 in a multiple linear regression analysis after adjusting for all the measured variables. CONCLUSION: The results suggest that cholesterol metabolism alterations may be associated with aspirin metabolism in older people.

  19. Interaction of Surface-active Fluorescence Probes with Bovine Serum Albumin

    Institute of Scientific and Technical Information of China (English)

    Tong Kuan XU; Xing Hai SHEN; Na LI; Hong Cheng GAO

    2005-01-01

    The binding between three surface-active substituted 3H-indole fluorescence probes and bovine serum albumin (BSA) in aqueous solution was studied using fluorescence quenching. The binding constants of 3H-indole molecules with BSA were obtained. According to the Forster resonance energy transfer theory, the distances between 3H-indole molecules and tryptophan of BSA were calculated. The results show that the oligoethyloxyethylene chain of 3H-indole molecules is longer, the binding between them is stronger, the energy transfer efficiency is higher,and the distance between tryptophan and 3H-indole is nearer.

  20. [Bactericidal activity of serum and chemotherapy in sensitive and resistant exciter (author's transl)].

    Science.gov (United States)

    Eyer, H; Metz, H; Preac-Mursic, V

    1975-11-21

    Comparing examinations with Ampicillin sensitive and resistant bacteria-strains show that the bactericidal activity of serum is dependent on the bacteria-strains, on the Ampicillin sensitivity of the particular exciter and on the number of bacteria/ml (germ count). Bactericide effect could always be obtained with sensitive strains as a result of additional chemotherapy. With several resistant strains a bactericide effect could not be obtained in this case the continuous optimal Ampicillin addition was the decisive factor. Because of the extremely complicated process of the bactericide one should not make general conclusions from the individual experimental results.

  1. Antioxidant activity of bovine serum albumin binding amino acid Schiff-bases metal complexes

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Glutamic acid-salicylaldehyde Schiff-base metal complexes are bound into bovine serum albumin (BSA), which afforded BSA binding Schiff-base metal complexes (BSA-SalGluM, M=Cu, Co, Ni, Zn). The BSA binding metal complexes were characterized by UV-vis spectra and Native PAGE. It showed that the protein structures of BSA kept after coordinating amino acid Schiff-bases metal complexes. The effect of the antioxidant activity was investigated. The results indicate that the antioxidant capacity of BSA increased more than 10 times after binding Schiff-base metal complexes.

  2. Effects of the Drug(BSZGC)--containing Serum on Proliferation of Rat's Osteoclasts and TRACP Activity in vitro

    Institute of Scientific and Technical Information of China (English)

    Shi Jingli; Zhao Yonghua; Wu Weikang

    2008-01-01

    Objective:To observe effects of the drug-containing serum of Bu Shen Zhuang Gu Capsule(BSZGC 补肾壮骨胶囊 Capsule for Tonilying the Kidney to Strengthen the Bones)on proliferation of the rat's osteoclasts and tartrate-resistant acid phosphatase (TRACP)activity in vitro SO as to delve into the mechanisms of its preventive and therapeutic actions on osteoporosis.Methods:Forty female Sprague.Dawley rats aged three months were randomly divided into high dosage BSZGC group,medium dosage BSZGC group,low dosage BSZGC group,and the control group.BSZGC was orally administered into the rats of high,medium,and low dosage groups at difierent dosages for 12 days.and isometric normal saline was orally administered to the rats of the Control group.The drug-containing serum and control serum were prepared.Osteoclasts isolated mechanically from the femur and tibia of Sprague-Dawley rats aged one week were cultured wim medium added with different drug-containing sera and control serum.The growth of osteoclasts was observed under an inverted phase-contrast microscope,and optic density(OD)value of osteoclasts was determined by MTT colorimetric assay.TRACP activity was measured by the diazol method.Results:OD value of osteoclasts in the high dosage drug-containing serum group,medium dosage drug-containing serum group,and low dosage drug-containing serum group was significantly lower than that in the control serum group(P<0.05)with a dose-effect correlation.TRACP activity in high dosage drug-containing serum group,medium dosage drug-containing serum group,low dosage drug-containing serum group was significantly lower than that of the control serum group(P<0.01),and no significant differences in TRACP activity were not found among the difierent dosages drug-containing serum groups.Conclusions:BSZGC can inhibit the proliferation of the osteoclasts and reduce TRACP activity,which may be one of the mechanisms of its preventive and therapeutic actions on osteoporosis.

  3. Intracellular modulation, extracellular disposal and serum increase of MiR-150 mark lymphocyte activation.

    Directory of Open Access Journals (Sweden)

    Paola de Candia

    Full Text Available Activated lymphocytes release nano-sized vesicles (exosomes containing microRNAs that can be monitored in the bloodstream. We asked whether elicitation of immune responses is followed by release of lymphocyte-specific microRNAs. We found that, upon activation in vitro, human and mouse lymphocytes down-modulate intracellular miR-150 and accumulate it in exosomes. In vivo, miR-150 levels increased significantly in serum of humans immunized with flu vaccines and in mice immunized with ovalbumin, and this increase correlated with elevation of antibody titers. Immunization of immune-deficient mice, lacking MHCII, resulted neither in antibody production nor in elevation of circulating miR-150. This study provides proof of concept that serum microRNAs can be detected, with minimally invasive procedure, as biomarkers of vaccination and more in general of adaptive immune responses. Furthermore, the prompt reduction of intracellular level of miR-150, a key regulator of mRNAs critical for lymphocyte differentiation and functions, linked to its release in the external milieu suggests that the selective extracellular disposal of microRNAs can be a rapid way to regulate gene expression during lymphocyte activation.

  4. Blood and colostrum/milk serum gamma-glutamyltransferase activity as a predictor of passive transfer status in lambs.

    Science.gov (United States)

    Maden, M; Altunok, V; Birdane, F M; Aslan, V; Nizamlioglu, M

    2003-04-01

    The importance of blood and colostrum/milk serum gamma-glutamyl transferase (gamma-GT) enzyme activity was evaluated to assess passive transfer status in healthy lambs. Thirty Akkaraman sheep (3-6 years old) were used which had normal pregnancy period and the same conditions, and the age of the lambs ranged between 0 and 15 days. Blood and colostrum/milk samples were collected from sheep and lambs after birth, before suckling (0) and after on 1st, 3rd, 7th and 15th days. Serum immunoglobulin G (IgG) concentration was determined by the use of Single Radial Immunodiffusion method. Serum gamma-GT activity was measured, using a commercially available kit in blood and colostrum/milk samples. Correlations were carried out between immunoglobulin and gamma-GT levels. Regression models (simple and multiple) were calculated with significant data. Linear correlation was determined between colostrum/milk gamma-GT activity and IgG concentrations and between serum gamma-GT activity and IgG concentrations in lambs on the 0 day. (r: 0.607, P: 0.001), 1st (r: 0.768, P: 0.001) and the 3rd (r: 0.603, P: 0.001) days and on the 1st (r: 0.637, P: 0.001) and 3rd (r: 0.478, P: 0.012) days in the experiment, respectively. Multivariate regression models were developed to estimate sample IgG concentration. Serum and colostrum/milk IgG concentration could be predicted using the formula: lamb serum IgG = 825 + 0.688 (lamb gamma-GT) + 52 (days); colostrum/milk IgG = 832 + 0.505 (colostrum/milk gamma-GT) - 167 (days). The regression models were moderately accurate in predicting serum IgG concentration (R2 = 0.51) and colostrum/milk IgG concentration (R2 = 0.55). Test sensitivity and positive predictive values for serum gamma-GT enzyme activity were found to be 96 and 100% and for colostrum/milk gamma-GT enzyme activity were found to be 100 and 68% to prediction IgG concentration. Serum and colostrum/milk gamma-GT activity can be used to assess passive transfer status of lambs. Along with this

  5. The Effects of Physical Activity on Serum Visfatin Level: A Literature Review

    Directory of Open Access Journals (Sweden)

    Mohsen Ghanbarzadeh

    2017-06-01

    Full Text Available Higher concentration of plasma visfatin in obese and diabetic subjects compared with their healthy counterparts shows visfatin relationship to obesity and overweight. This article reviewed the studies on contradictory and different notions regarding the role of physical activity in visfatin response following aerobic and resistance exercises. Recent reports on the impact of physical activity and exercise on visfatin concentration is contradictory. Some studies have identified that exercise can reduce visfatin concentration depending on the duration of physical activity and calorie expenditure, while others have not reported any changes in visfatin concentration. The present review indicated that a balanced diet, low in fat, and physical exercise (aerobic and aerobic-resistance exercises can reduce blood visfatin levels depending on the severity and duration, while resistance training alone exerts no significant effects on serum visfatin level.

  6. Direct Biomolecules Binding on Nonfouling Surface via Newly Discovered Supramolecular Self-assembly of Lysozyme under Physiological Condition

    Science.gov (United States)

    Yang, Peng

    2013-01-01

    A major challenge in the development of low cost and practical strategies for biomolecules immobilization on solid supports is that the multi-step chemical/physical activating and following deactivating procedures on nonfouling substrates often increase the cost and complexity of surface functional group types as well as deteriorate the surface integrity. Herein, we show a novel phase transition of lysozyme could be used to constitute a major step to address the above problem. It is found that when lysozyme is dissolved in a neutral buffer solution of 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, pH 7.4) with 1–50 mM tris(2-carboxyethyl)phosphine (TCEP) added, a fast phase transition process occurs and the resulting novel fibra-like hierarchical supramolecular assemblies made by primary spherical particles aggregation would function as a “superglue” that strongly and quickly bind onto non-fouling coatings. This binding is highly selective towards lysozyme, and excludes completely tedious synthetical, chemical/physical activation/deactivation (blocking) steps. When biotin is conjugated with lysozyme, such phase transition quickly constructs a perfect biotinylated surface on nonfouling surface for avidin binding, showing great potential for the development of low-cost and practical biochips. PMID:22707360

  7. Laser ablation of the protein lysozyme

    DEFF Research Database (Denmark)

    Schou, Jørgen; Canulescu, Stela; Amoruso, Salvatore

    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...... 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...... 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...

  8. Terahertz absorption of lysozyme in solution

    Science.gov (United States)

    Martin, Daniel R.; Matyushov, Dmitry V.

    2017-08-01

    Absorption of radiation by solution is described by its frequency-dependent dielectric function and can be viewed as a specific application of the dielectric theory of solutions. For ideal solutions, the dielectric boundary-value problem separates the polar response into the polarization of the void in the liquid, created by the solute, and the response of the solute dipole. In the case of a protein as a solute, protein nuclear dynamics do not project on significant fluctuations of the dipole moment in the terahertz domain of frequencies and the protein dipole can be viewed as dynamically frozen. Absorption of radiation then reflects the interfacial polarization. Here we apply an analytical theory and computer simulations to absorption of radiation by an ideal solution of lysozyme. Comparison with the experiment shows that Maxwell electrostatics fails to describe the polarization of the protein-water interface and the "Lorentz void," which does not anticipate polarization of the interface by the external field (no surface charges), better represents the data. An analytical theory for the slope of the solution absorption against the volume fraction of the solute is formulated in terms of the cavity field response function. It is calculated from molecular dynamics simulations in good agreement with the experiment. The protein hydration shell emerges as a separate sub-ensemble, which, collectively, is not described by the standard electrostatics of dielectrics.

  9. 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.

  10. Replacement of dietary saturated fat with trans fat reduces serum paraoxonase activity in healthy men and women.

    Science.gov (United States)

    de Roos, Nicole M; Schouten, Evert G; Scheek, Leo M; van Tol, Arie; Katan, Martijn B

    2002-12-01

    A high intake of saturated fat and of trans isomers of unsaturated fat is associated with increased risk of cardiovascular disease. Recently, we found that replacement of saturated fat by trans fat in a dietary controlled study with 32 men and women decreased serum high-density lipoprotein (HDL)-cholesterol and impaired endothelial function, suggesting that trans fats have stronger adverse effects than saturated fats. To investigate this further, we measured the activity of serum paraoxonase (PON1) in serum samples of the same volunteers after consumption of both diets. PON1 protects lipoproteins from oxidative damage, and higher PON1 activity appears to be related to lower cardiovascular disease risk. PON1 activity (mean +/- SD) was 195.9 +/- 108.9 U/L after 4 weeks of consuming a diet with 22.9% of energy (en%) from saturated fat and 184.5 +/- 99.3 U/L when 9.3 en% from saturated fat was replaced by trans fat (P =.006). Thus, replacement of dietary saturated fat by trans fat not only decreased serum HDL-cholesterol and impaired endothelial function, but also decreased the activity of serum paraoxonase. Whether the changes in serum paraoxonase activity caused the changes in endothelial function needs to be further investigated.

  11. The mechanisms of complement activation in normal bovine serum and normal horse serum against Yersinia enterocolitica O:9 strains with different outer membrane proteins content.

    Science.gov (United States)

    Miętka, K; Brzostek, K; Guz-Regner, K; Bugla-Płoskońska, G

    2016-01-01

    Yersinia enterocolitica is a common zoonotic pathogen and facultative intracellular bacterium which can survive within blood cells. Cattle and horses are considered a reservoir of Y. enterocolitica which often causes several serious syndromes associated with yersiniosis such as abortions, premature births or infertility. The aim of our investigation was to determine the vitality of Y. enterocolitica O:9 strains (Ye9) in bovine and horse sera (NBS and NHrS) and explain the role of outer membrane proteins (OMPs) in serum resistance of these bacteria. Our previous studies demonstrated moderate human serum (NHS) resistance of the wild type Ye9 strain, whereas mutants lacking YadA, Ail or OmpC remained sensitive to the bactericidal activity of NHS. The present study showed that the wild type of Ye9 strain was resistant to the bactericidal activity of both NHrS and NBS, while Ye9 mutants lacking the YadA, Ail and OmpC proteins were sensitive to NHrS and NBS as well as to NHS. The mechanisms of complement activation against Ye9 strains lacking Ail and YadA were distinguished, i.e. activation of the classical/lectin pathways decisive in the bactericidal mechanism of complement activation of NBS, parallel activation of the classical/lectin and alternative pathways of NHrS. In this research the mechanism of independent activation of the classical/lectin or the alternative pathway of NBS and NHrS against Ye9 lacking OmpC porin was also established. The results indicate that serum resistance of Ye9 is multifactorial, in which extracellular structures, i.e. outer membrane proteins (OMPs) such as Ail, OmpC or YadA, play the main role.

  12. 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

  13. [Lipase activity in the calf serum under different types of feeding].

    Science.gov (United States)

    Surynek, J

    1975-05-01

    The development of lipase activity in serum (S-lipase activity) was studied in two groups of the calves of the Bohemian Red-Pied breed in the post-natal period up to the age of nine weeks from birth. From the eighth day of age, the two groups differed from each other in their lactic nutrition. Group A calves were given whole milk (obtained at primary production) and group B calves were fed the Laktosan mixture instead of milk. The different type of nutrition did not influence the character of the development of S-lipase activity with increasing age. A difference was found between the two groups as to the level of the activity of the enzyme under study. In the whole-milk group the average S-lipase activity values were higher than in the group fed Laktosan in a larger part of the test period. Some differences were statistically significant. It is assumed that lipase activity in blood is a reflection of the external secretion of pancreas changing according to the structure of food taken in. The activity of S-lipase was determined by the method according to Cherry and Crandall (Seligson, 1964); the average minimum value was 0.025 +/- 0.0017 u. in group A and 0.017 +/- 0.0081 u. in group B, and the average maximum value was 0.370 +/- 0.100 u. and 0.286 +/- 0.0766 u., respectively.

  14. Improved activity of immobilized horseradish peroxidase on gold nanoparticles in the presence of bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Ni, Yuyang; Li, Jun; Huang, Zhenzhen; He, Ke; Zhuang, Jiaqi; Yang, Wensheng, E-mail: wsyang@jlu.edu.cn [Jilin University, State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry (China)

    2013-11-15

    The using of macromolecular additives is known to be a simple and effective way to improve the activity of immobilized enzymes on solid support, yet the mechanism has not been well understood. Taking horseradish peroxidase (HRP) as an example, only 30 % of its catalytic activity was kept after being immobilized on the surface of 25-nm Au nanoparticles, mainly attributed to the conformational change of the heme-containing active site. The catalytic activity of HRP was significantly improved to 80 % when a certain amount of bovine serum albumin (BSA) was added at the initial stage of the immobilization. Systematic spectral investigation indicated that the addition of BSA inhibited the tertiary structure change around the active site, which was a prerequisite for improved activity of the immobilized HRP. Steady-state kinetic analyses revealed that the introduction of BSA could effectively improve the turnover rate of substrate to product in spite of slight reduced affinity to substrates, which also contributed to the improved catalytic activity.

  15. Improved activity of immobilized horseradish peroxidase on gold nanoparticles in the presence of bovine serum albumin

    Science.gov (United States)

    Ni, Yuyang; Li, Jun; Huang, Zhenzhen; He, Ke; Zhuang, Jiaqi; Yang, Wensheng

    2013-11-01

    The using of macromolecular additives is known to be a simple and effective way to improve the activity of immobilized enzymes on solid support, yet the mechanism has not been well understood. Taking horseradish peroxidase (HRP) as an example, only 30 % of its catalytic activity was kept after being immobilized on the surface of 25-nm Au nanoparticles, mainly attributed to the conformational change of the heme-containing active site. The catalytic activity of HRP was significantly improved to 80 % when a certain amount of bovine serum albumin (BSA) was added at the initial stage of the immobilization. Systematic spectral investigation indicated that the addition of BSA inhibited the tertiary structure change around the active site, which was a prerequisite for improved activity of the immobilized HRP. Steady-state kinetic analyses revealed that the introduction of BSA could effectively improve the turnover rate of substrate to product in spite of slight reduced affinity to substrates, which also contributed to the improved catalytic activity.

  16. Neomycin-phenolic conjugates: polycationic amphiphiles with broad-spectrum antibacterial activity, low hemolytic activity and weak serum protein binding.

    Science.gov (United States)

    Findlay, Brandon; Zhanel, George G; Schweizer, Frank

    2012-02-15

    Here we present a proof-of-concept study, combining two known antimicrobial agents into a hybrid structure in order to develop an emergent cationic detergent-like interaction with the bacterial membrane. Six amphiphilic conjugates were prepared by copper (I)-catalyzed 1,3-dipolar cycloaddition between a neomycin B-derived azide and three alkyne-modified phenolic disinfectants. Three conjugates displayed good activity against a variety of clinically relevant Gram positive and Gram negative bacteria, including MRSA, without the high level of hemolysis or strong binding to serum proteins commonly observed with other cationic antimicrobial peptides and detergents.

  17. Interaction mechanism between berberine and the enzyme lysozyme

    Science.gov (United States)

    Cheng, Ling-Li; Wang, Mei; Wu, Ming-Hong; Yao, Si-De; Jiao, Zheng; Wang, Shi-Long

    2012-11-01

    In the present paper, the interaction between model protein lysozyme (Lys) and antitumorigenic berberine (BBR) was investigated by spectroscopic methods, for finding an efficient and safe photosensitizer with highly active transient products using in photodynamic therapy study. The fluorescence data shows that the binding of BBR could change the environment of the tryptophan (Trp) residues of Lys, and form a new complex. Static quenching is the main fluorescence quenching mechanism between Lys and BBR, and there is one binding site in Lys for BBR and the type of binding force between them was determined to be hydrophobic interaction. Furthermore, the possible interaction mechanism between BBR and Lys under the photoexcitation was studied by laser flash photolysis method, the results demonstrated that BBR neutral radicals (BBR(-H)•) react with Trp (K = 3.4 × 109 M-1 s-1) via electron transfer to give the radical cation (Trp/NH•+) and neutral radical of Trp (TrpN•). Additionally BBR selectively oxidize the Trp residues of Lys was also observed by comparing the transient absorption spectra of their reaction products. Through thermodynamic calculation, the reaction mechanisms between 3BBR∗ and Trp or Lys were determined to be electron transfer process.

  18. Role of Serum Iron in the Activation of Lipid Peroxidation in Critical Conditions

    Directory of Open Access Journals (Sweden)

    Yu. P. Orlov

    2006-01-01

    Full Text Available Twenty-four critically ill patients due to generalized purulent peritonitis, pancreatonecrosis, thermal skin injuries, and severe poisoning by acetic acid were examined. The general regularities of the effect of high serum iron concentrations on the health status of patients, on the activity of antioxidative enzymes, and on the initiation of lipid peroxidation (LPO processes, as supported by the values of Fe2+-induced chemiluminescence, were revealed. In critically ill patients, iron metabolism occurs with the overload of a transport protein, such as transferrin, which is caused by intravascular hemolysis and hemoglobin metabolism to ionized iron. The overload of proteins responsible for iron transport leads to the tissue accumulation of free (ferrous and ferric iron that is actively involved in the processes of LPO initiation with excess synthesis of cytotoxic radicals, which in turn accounts for the severity of endotoxicosis.

  19. 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.

    1996-03-01

    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.

  20. Influence of physical and emotional activity on the metabolic profile of blood serum of race horses

    Directory of Open Access Journals (Sweden)

    T. I. Bayeva

    2016-09-01

    Full Text Available In the article data are presented on dynamics of the level of indicators of metabolic profile of blood serum of race horses of the Ukrainian riding breed in the conditions of physical and emotional loading. Clinically healthy race horses were the object of  research. Blood was taken from the jugular vein to obtain serum and for further biochemical research. For the research 12 race horses from a training group were chosen. From time to time the animals took part in competitions; they were not specially used in races and were mostly used for the training of junior riders and sportsmen of different levels. Blood was taken in conditions of relative rest after ordinary training and after emotional stress during the entertainment performances when a large number of people were present and loud music was played. In the blood serum the following biochemical indicators were defined: whole protein, urea, creatinine, uric acid, total bilirubin and its fractions, glucose, cholestererol, triacylglycerol, calcium, ferrum, lactate, pyruvate, activity of the AlAT, SGOT, GGTP, LDH, an alkaline phosphatase – which makes it possible to determine reasonably accurately the adaptation potential of a horse under various types of loading. We established that during training and psychoemotional loading of racing horses of the training group of the Ukrainian riding breed, multidirectional changes in the level of biochemical indicators of blood serum occurred, which is evidence of stress in the metabolic processes in the animals’ organisms. Concentration of a biomarker of an oxidative stress, uric acid, increased after physical loading by 8.6%, and after emotional loading by 55.1%, which demonstrates that emotional stress had the more negative effect, indicating insufficient adaptation by the horses before demonstration performances. After physical loading, reaction of transamination in the horses’ liver cells intensified, and after emotional loading its intensity

  1. Effect of smoking on activity of N-acetyl-beta-hexosaminidase in serum and urine of renal cancer patients.

    Science.gov (United States)

    Borzym-Kluczyk, Malgorzata; Radziejewska, Iwona; Zaniewska, Agnieszka; Borzym-Lewszuk, Anna; Szajda, Sławomir Dariusz; Knaś, Malgorzata; Zwierz, Krzysztof; Darewicz, Barbara

    2009-10-01

    To compare N-acetyl-beta-hexosaminidase (HEX) activity in the serum and urine of smokers as well as non-smokers with renal cancer, and healthy people. To assess hexosaminidase activity the level of p-nitrophenol released from p-nitrophenol derivatives was measured. The activity of enzyme was significantly higher in cancer group, with the highest activity in non-smokers. Cigarette smoking can inhibit, by the influence on HEX activity, catabolism of oligosaccharide chains in cancer tissues.

  2. Invertebrate lysozymes: Diversity and distribution, molecular mechanism and in vivo function

    Indian Academy of Sciences (India)

    Joris M Van Herreweghe; Chris W Michiels

    2012-06-01

    Lysozymes are antibacterial enzymes widely distributed among organisms. Within the animal kingdom, mainly three major lysozyme types occur. Chicken (c)-type lysozyme and goose (g)-type lysozyme are predominantly, but not exclusively, found in vertebrate animals, while the invertebrate (i)-type lysozyme is typical for invertebrate organisms, and hence its name. Since their discovery in 1975, numerous research articles report on the identification of i-type lysozymes in a variety of invertebrate phyla. This review describes the current knowledge on i-type lysozymes, outlining their distribution, molecular mechanism and in vivo function taking the representative from Venerupis philippinarum (formerly Tapes japonica) (Vp-ilys) as a model. In addition, invertebrate g-type and ch-type (chalaropsis) lysozymes, which have been described in molluscs and nematodes, respectively, are also briefly discussed.

  3. Concomitant Prevalence of Low Serum Diamine Oxidase Activity and Carbohydrate Malabsorption

    Directory of Open Access Journals (Sweden)

    Dietmar Enko

    2016-01-01

    Full Text Available The aim of this retrospective study was to analyze the concomitant prevalence rates for lactose malabsorption (LM, fructose malabsorption (FM, and histamine intolerance (HI in patients with so far unexplained gastrointestinal (GI symptoms. A total of 439 outpatients, who presented unclear abdominal discomfort, underwent lactose (50 g and fructose (25 g hydrogen (H2 breath tests. Additionally, serum diamine oxidase (DAO measurements were performed. Individuals with low serum DAO activity (<10 U/mL, GI symptoms, and response to histamine-free diet were diagnosed with HI. Of all 439 patients, 341 (77.7% were found with 7 various GI conditions. In total, 94 (21.4%, 31 (7.1%, and 100 (22.8% individuals presented LM, FM, or HI only, whereas 116 (26.4% patients showed an overlap of GI entities investigated here. Interestingly, 89 out of 241 (36.9% individuals with carbohydrate malabsorption were also diagnosed with HI (LM + HI: 52 [11.8%], FM + HI: 23 [5.2%], and LM + FM + HI 14 [3.2%] individuals. In conclusion different combinations of LM, FM, and HI are present in individuals with unclear abdominal discomfort/pain. In clinical practice we suggest testing for LM, FM, and additional HI in the diagnostic work-up of these patients. Depending on these various diagnoses possible, patients should get an individualized dietary advice.

  4. ADENOSINE DEAMINASE ACTIVITY AND SERUM C-REACTIVE PROTEIN AS PROGNOSTIC MARKERS OF CHAGAS DISEASE SEVERITY

    Science.gov (United States)

    BRAVO-TOBAR, Iván Darío; NELLO-PÉREZ, Carlota; FERNÁNDEZ, Alí; MOGOLLÓN, Nora; PÉREZ, Mary Carmen; VERDE, Juan; CONCEPCIÓN, Juan Luis; RODRIGUEZ-BONFANTE, Claudina; BONFANTE-CABARCAS, Rafael

    2015-01-01

    SUMMARY Chagas disease is a public health problem worldwide. The availability of diagnostic tools to predict the development of chronic Chagas cardiomyopathy is crucial to reduce morbidity and mortality. Here we analyze the prognostic value of adenosine deaminase serum activity (ADA) and C-reactive protein serum levels (CRP) in chagasic individuals. One hundred and ten individuals, 28 healthy and 82 chagasic patients were divided according to disease severity in phase I (n = 35), II (n = 29), and III (n = 18). A complete medical history, 12-lead electrocardiogram, chest X-ray, and M-mode echocardiogram were performed on each individual. Diagnosis of Chagas disease was confirmed by ELISA and MABA using recombinant antigens; ADA was determined spectrophotometrically and CRP by ELISA. The results have shown that CRP and ADA increased linearly in relation to disease phase, CRP being significantly higher in phase III and ADA at all phases. Also, CRP and ADA were positively correlated with echocardiographic parameters of cardiac remodeling and with electrocardiographic abnormalities, and negatively with ejection fraction. CRP and ADA were higher in patients with cardiothoracic index ≥ 50%, while ADA was higher in patients with ventricular repolarization disturbances. Finally, CRP was positively correlated with ADA. In conclusion, ADA and CRP are prognostic markers of cardiac dysfunction and remodeling in Chagas disease. PMID:26603224

  5. ADENOSINE DEAMINASE ACTIVITY AND SERUM C-REACTIVE PROTEIN AS PROGNOSTIC MARKERS OF CHAGAS DISEASE SEVERITY

    Directory of Open Access Journals (Sweden)

    Iván Darío BRAVO-TOBAR

    2015-10-01

    Full Text Available SUMMARY Chagas disease is a public health problem worldwide. The availability of diagnostic tools to predict the development of chronic Chagas cardiomyopathy is crucial to reduce morbidity and mortality. Here we analyze the prognostic value of adenosine deaminase serum activity (ADA and C-reactive protein serum levels (CRP in chagasic individuals. One hundred and ten individuals, 28 healthy and 82 chagasic patients were divided according to disease severity in phase I (n = 35, II (n = 29, and III (n = 18. A complete medical history, 12-lead electrocardiogram, chest X-ray, and M-mode echocardiogram were performed on each individual. Diagnosis of Chagas disease was confirmed by ELISA and MABA using recombinant antigens; ADA was determined spectrophotometrically and CRP by ELISA. The results have shown that CRP and ADA increased linearly in relation to disease phase, CRP being significantly higher in phase III and ADA at all phases. Also, CRP and ADA were positively correlated with echocardiographic parameters of cardiac remodeling and with electrocardiographic abnormalities, and negatively with ejection fraction. CRP and ADA were higher in patients with cardiothoracic index ≥ 50%, while ADA was higher in patients with ventricular repolarization disturbances. Finally, CRP was positively correlated with ADA. In conclusion, ADA and CRP are prognostic markers of cardiac dysfunction and remodeling in Chagas disease.

  6. The effect of cold on serum thyroid hormones and hepatic 5 prime mono-deiodinase activity

    Energy Technology Data Exchange (ETDEWEB)

    Hesslink, R.L. Jr.; Quesada, M.; D' Alesandro, M.; Homer, L.D.; Reed, J.L.; Christopherson, R.; Young, B.A. (Naval Medical Research Inst., Bethesda, MD (United States) Univ. of Alberta, Edmonton (Canada))

    1991-03-11

    Cold exposed swine have an increases serum concentration of triiodothyronine (T{sub 3}) and increased T{sub 3} production rate. It is thought that hepatic thyroxine (T{sub 4}) deiodination (5DI) contributes to circulating T{sub 3} concentrations. The authors investigated the effects of cold exposure (14 days) on energy intake, serum free T{sub 3} (FT{sub 3}) and free T{sub 4} (FT{sub 4}) levels; and 5DI in 5-month boars. Hepatic 5DI activity was determined by measuring the {sup 125}I generated from trace amounts of {sup 125}I T{sub 4}. FT{sub 3} and FT{sub 4} were assayed by RIA. Swine were housed in either 20C (control; n = 5) or 4C (cold; n = 7) chambers and given food ad libitum. Cold exposure increased energy intake by 42%. The increase (93%) in hepatic 5DI V{sub max} after cold exposure parallels the increase in whole animal T{sub 3} production and may account for FT{sub 3} values found after cold exposure.

  7. Serum and Plasma Cholinesterase Activity in the Cape Griffon Vulture (Gyps coprotheres).

    Science.gov (United States)

    Naidoo, Vinny; Wolter, Kerri

    2016-04-28

    Vulture (Accipitridae) poisonings are a concern in South Africa, with hundreds of birds dying annually. Although some of these poisonings are accidental, there has been an increase in the number of intentional baiting of poached rhinoceros (Rhinocerotidae) and elephant (Elephantidae) carcasses to kill vultures that alert officials to poaching sites by circling overhead. The primary chemicals implicated are the organophosphorous and carbamate compounds. Although most poisoning events can be identified by dead vultures surrounding the scavenged carcass, weak birds are occasionally found and brought to rehabilitation centers for treatment. The treating veterinarian needs to make an informed decision on the cause of illness or poisoning prior to treatment. We established the reference interval for serum and plasma cholinesterase activity in the Cape Griffon Vulture ( Gyps coprotheres ) as 591.58-1,528.26 U/L, providing a clinical assay for determining potential exposure to cholinesterase-depressing pesticides. Both manual and automated samplers were used with the butyrylthiocholine method. Species reference intervals for both serum and plasma cholinesterase showed good correlation and manual and automated measurements yielded similar results.

  8. Electroanalysis of pM-levels of urokinase plasminogen activator in serum by phosphorothioated RNA aptamer.

    Science.gov (United States)

    Jarczewska, Marta; Kékedy-Nagy, László; Nielsen, Jesper S; Campos, Rui; Kjems, Jørgen; Malinowska, Elżbieta; Ferapontova, Elena E

    2015-06-01

    Protein biomarkers of cancer allow a dramatic improvement in cancer diagnostics as compared to the traditional histological characterisation of tumours by enabling a non-invasive analysis of cancer development and treatment. Here, an electrochemical label-free assay for urokinase plasminogen activator (uPA), a universal biomarker of several cancers, has been developed based on the recently selected uPA-specific fluorinated RNA aptamer, tethered to a gold electrode via a phosphorothioated dA tag, and soluble redox indicators. The binding properties of the uPA-aptamer couple and interference from the non-specific adsorption of bovine serum albumin (BSA) were modulated by the electrode surface charge. A nM uPA electroanalysis at positively charged surfaces, complicated by the competitive adsorption of BSA, was tuned to the pM uPA analysis at negative surface charges of the electrode, being improved in the presence of negatively charged BSA. The aptamer affinity for uPA displayed via the binding/dissociation constant relationship correspondingly increased, ca. three orders of magnitude, from 0.441 to 367. Under optimal conditions, the aptasensor allowed 10(-12)-10(-9) M uPA analysis, also in serum, being practically useful for clinical applications. The proposed strategy for optimization of the electrochemical protein sensing is of particular importance for the assessment and optimization of in vivo protein ligand binding by surface-tethered aptamers.

  9. Platelet aggregation and serum adenosine deaminase (ADA) activity in pregnancy associated with diabetes, hypertension and HIV.

    Science.gov (United States)

    Leal, Claudio A M; Leal, Daniela B R; Adefegha, Stephen A; Morsch, Vera M; da Silva, José E P; Rezer, João F P; Schrekker, Clarissa M L; Abdalla, Faida H; Schetinger, Maria R C

    2016-07-01

    Platelet aggregation and adenosine deaminase (ADA) activity were evaluated in pregnant women living with some disease conditions including hypertension, diabetes mellitus and human immunodeficiency virus infection. The subject population is consisted of 15 non-pregnant healthy women [control group (CG)], 15 women with normal pregnancy (NP), 7 women with hypertensive pregnancy (HP), 10 women with gestational diabetes mellitus (GDM) and 12 women with human immunodeficiency virus-infected pregnancy (HIP) groups. The aggregation of platelets was checked using an optical aggregometer, and serum ADA activity was determined using the colorimetric method. After the addition of 5 µM of agonist adenosine diphosphate, the percentage of platelet aggregation was significantly (p < 0·05) increased in NP, HP, GDM and HIP groups when compared with the CG, while the addition of 10 µM of the same agonist caused significant (p < 0·05) elevations in HP, GDM and HIP groups when compared with CG. Furthermore, ADA activity was significantly (p < 0·05) enhanced in NP, HP, GDM and HIP groups when compared with CG. In this study, the increased platelet aggregation and ADA activity in pregnancy and pregnancy-associated diseases suggest that platelet aggregation and ADA activity could serve as peripheral markers for the development of effective therapy in the maintenance of homeostasis and some inflammatory process in these pathophysiological conditions. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  10. Adenosine deaminase activity in serum and lymphocytes of rats infected with Sporothrix schenckii.

    Science.gov (United States)

    Castro, Verônica S P; Pimentel, Victor C; Da Silva, Aleksandro S; Thomé, Gustavo R; Wolkmer, Patrícia; Castro, Jorge L C; Costa, Márcio M; da Silva, Cássia B; Oliveira, Daniele C; Alves, Sydney H; Schetinger, Maria R C; Lopes, Sonia T A; Mazzanti, Cinthia M

    2012-07-01

    Sporotrichosis is a fungal infection of subcutaneous or chronic evolution, inflammatory lesions characterized by their pyogranulomatous aspect, caused by the dimorphic fungus Sporothrix schenckii. Adenosine deaminase (ADA) is a "key" enzyme in the purine metabolism, promoting the deamination of adenosine, an important anti-inflammatory molecule. The increase in ADA activity has been demonstrated in several inflammatory conditions; however, there are no data in the literature associated with this fungal infection. The objective of this study was to evaluate the activity of serum ADA (S-ADA) and lymphocytes (L-ADA) of rats infected with S. schenckii. We used seventy-eight rats divided into two groups. In the first experiment, rats were infected subcutaneously and in the second experiment, infected intraperitoneally. Blood samples for hematologic evaluation and activities of S-ADA and L-ADA were performed at days 15, 30, and 40 post-infection (PI) to assess disease progression. In the second experiment, it was observed an acute decrease in activity of S-ADA and L-ADA (P schenckii alters the activities of S-ADA in experimentally infected rats, demonstrating the involvement of this enzyme in the pathogenesis of sporotrichosis.

  11. Pollutant-induced modulation in conformation and β-lactamase activity of human serum albumin.

    Directory of Open Access Journals (Sweden)

    Ejaz Ahmad

    Full Text Available Structural changes in human serum albumin (HSA induced by the pollutants 1-naphthol, 2-naphthol and 8-quinolinol were analyzed by circular dichroism, fluorescence spectroscopy and dynamic light scattering. The alteration in protein conformational stability was determined by helical content induction (from 55 to 75% upon protein-pollutant interactions. Domain plasticity is responsible for the temperature-mediated unfolding of HSA. These findings were compared to HSA-hydrolase activity. We found that though HSA is a monomeric protein, it shows heterotropic allostericity for β-lactamase activity in the presence of pollutants, which act as K- and V-type non-essential activators. Pollutants cause conformational changes and catalytic modifications of the protein (increase in β-lactamase activity from 100 to 200%. HSA-pollutant interactions mediate other protein-ligand interactions, such as HSA-nitrocefin. Therefore, this protein can exist in different conformations with different catalytic properties depending on activator binding. This is the first report to demonstrate the catalytic allostericity of HSA through a mechanistic approach. We also show a correlation with non-microbial drug resistance as HSA is capable of self-hydrolysis of β-lactam drugs, which is further potentiated by pollutants due to conformational changes in HSA.

  12. Impact of antibacterial drugs on human serum paraoxonase-1 (hPON1) activity:an in vitro study

    Institute of Scientific and Technical Information of China (English)

    Hakan Syt; Elif Duygu Kaya; kr Beydemir

    2014-01-01

    Objective:To investigate the in vitro effects of the antibacterial drugs, meropenem trihydrate, piperacillin sodium, and cefoperazone sodium, on the activity of human serum paraoxonase Methods: hPON1 was purified from human serum using simple chromatographic methods, including DEAE-Sephadex anion exchange and Sephadex G-200 gel filtration chromatography. Results:The three antibacterial drugs decreased in vitro hPON1 activity. Inhibition mechanisms meropenem trihydrate was noncompetitive while piperacillin sodium and cefoperazone sodium were competitive. Conclusions:Our results showed that antibacterial drugs significantly inhibit hPON1 activity, both in vitro, with rank order meropenem trihydrate piperacillin sodium cefoperazone sodium in vitro.

  13. Effect of 50% human serum on the killing activity of micafungin against eight Candida species using time-kill methodology.

    Science.gov (United States)

    Földi, Richárd; Szilágyi, Judit; Kardos, Gábor; Berényi, Réka; Kovács, Renátó; Majoros, László

    2012-08-01

    Micafungin activity was determined against 24 wild-type clinical isolates and 5 American Type Culture Collection strains belonging to 8 Candida species in RPMI-1640 with and without 50% serum using broth microdilution and time-kill methodology. MIC values increased from 4- to 128-folds in 50% serum for all Candida species. Micafungin was not fungicidal against C. albicans, C. tropicalis, and against 2 of 3 C. metapsilosis at ≥0.25, 1, and 1 μg/mL, respectively, after 48 h with 50% serum, showing good fungistatic activity. Fungicidal activity at ≥2, 4, and 32 μg/mL was noticed against C. glabrata, C. inconspicua, and C. krusei isolates, respectively. Micafungin at 8-32 μg/mL showed fungistatic activity against C. parapsilosis and C. orthopsilosis. Serum decreased the in vitro activity of micafungin. With serum binding of echinocandins taken into account, safely fungistatic or fungicidal concentrations seem to require elevated doses against some Candida species, including C. parapsilosis, C. orthopsilosis, and C. krusei.

  14. The effect of pyridoxal-5-phosphate on serum alanine aminotransferase activity in dogs suffering from canine babesiosis

    Directory of Open Access Journals (Sweden)

    E.C. Myburgh

    2009-09-01

    Full Text Available Accurate measurements of serum aminotransferase (ALT activity in dogs relies on the endogenous pro-enzyme pyridoxal 5-phosphate (P5P. The purpose of this study was to determine whether the exclusion of P5P from the analytical method causes an underestimation of serum ALT activity in dogs suffering from babesiosis and in those manifesting evidence of hepatocellular damage, and to determine if anorexia causes sufficient P5P depletion to affect in vitro serum ALT activity. One-hundred-and-twenty healthy control dogs and 105 Babesia-infected dogs were included in the study. Two methods for ALT measurement were used: Method 1 included P5P, and Method 2 excluded P5P from the reaction mixture. Higher serum ALT activity was measured with Method 1 in the Babesia-infected dogs (P < 0.001, as well as in 14 dogs with suspected hepatocellular damage (P = 0.03. Duration of anorexia had no effect, irrespective of the method used. Although inclusion of P5P to the reaction mixture consistently resulted in higher measured serum ALT activity, the differences were too small to have led to incorrect diagnoses in the Babesia-infected dogs suspected of liver disease.

  15. The interaction between physical activity and fasting on the serum lipid profile during Ramadan.

    Science.gov (United States)

    Haghdoost, A A; Poorranjbar, M

    2009-09-01

    The serum lipid profiles in Muslims change during the fasting month of Ramadan, but it is not clear whether this change is due to changes in their physical activities. In this study, we compared the patterns of the lipid profile changes in those who engaged in regular physical activity with those who did not. In a randomised trial, we assigned 93 students who took a physical education course into two groups--those who had regular physical activity after Ramadan and those who had physical activity during Ramadan. Venous blood (5 ml) was taken just before, at the end, and 40 days after Ramadan, and the fasting glucose sugar and lipid profile were measured. Fasting with physical activity decreased body weight by 1.2 kg (p-value is 0.03). Fasting blood sugar also decreased by 7 mg/dL during Ramadan, but this drop was observed in both groups. Triglyceride decreased in both groups during Ramadan, but cholesterol levels dropped considerably during and after Ramadan for those who concurrently engaged in physical activity and fasted (-12.24 and -8.4 mg/dL, respectively). The patterns of changes in the high-density lipoprotein (HDL), low-density lipoprotein (LDL) and HDL/LDL values were more or less comparable in both groups (p is greater than 0.5). Usually, people are less physically active during Ramadan, but our findings show that physical activity alone cannot explain the variations in the lipid profile. Other factors, such as changes in the diet and sleeping hours, may have more important roles.

  16. 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.

  17. Dilatometric, refractometric and viscometric study of lysozyme-cation interaction.

    Science.gov (United States)

    Abad, C; Trueba, M; Campos, A; Figueruelo, J E

    1981-11-01

    The interaction between hen egg-white lysozyme and Cu(II) or Co(II) cations has been studied by dilatometry, equilibrium dialysis-differential refractometry and viscometry at different metal cation concentrations. Delta V isotherms in copper and cobalt solutions have been obtained from dilatometry. Preferential adsorption parameters and specific viscosity have been determined from refractometric and viscosimetric measurements. It has been observed that this interaction produces structural alterations in lysozyme. The magnitude of these conformational changes depends on the metal ion and protein concentration. The results obtained using the three techniques are in good agreement.

  18. Increased serum ß2-microglobulin is associated with clinical and immunological markers of disease activity in systemic lupus erythematosus patients

    DEFF Research Database (Denmark)

    Hermansen, M-L F; Hummelshøj, L; Lundsgaard, Dorte

    2012-01-01

    The objective of this study was to explore the relationship between serum levels of ß2-microglobulin (ß2MG), which some studies suggest reflect disease activity in systemic lupus erythematosus (SLE), and various clinical and immunological markers of disease activity in SLE. Twenty-six SLE patients...

  19. Serum inflammatory mediators correlate with disease activity in electrical status epilepticus in sleep (ESES) syndrome.

    Science.gov (United States)

    van den Munckhof, Bart; de Vries, Evelien E; Braun, Kees P J; Boss, H Myrthe; Willemsen, Michèl A; van Royen-Kerkhof, Annet; de Jager, Wilco; Jansen, Floor E

    2016-02-01

    We aimed to study serum cytokine levels in 11 electrical status epilepticus in sleep (ESES) patients and 20 healthy control children. Patients showed significantly higher levels of interleukin (IL)-1α, IL-6, IL-10, chemokine (C-C motif) ligand (CCL)2 and chemokine (C-X-C motif) ligand (CXCL)8/IL-8 than controls, while macrophage migration inhibitory factor (MIF) and CCL3 were significantly lower. Follow-up analyses in five patients revealed a significant decrease of IL-6 levels after immunomodulating treatment. IL-6 changes were accompanied by clear improvement of electroencephalography (EEG) patterns and neuropsychological evaluation. We hypothesize that IL-6 correlates with disease activity and immunomodulating treatment efficacy.

  20. EFFECT OF AQUATIC ENVIRONMENT PH ON THE LEVEL OF ECTOPARASITE INFESTATION, PROTEIN AND LYSOZYME CONTENT IN SOME CYPRINID SPECIES (CYPRINIDAE)

    OpenAIRE

    L. Kurovskaya; G. Stril’ko

    2016-01-01

    Purpose. To study the effect of рН values of the aquatic environment on the level of ectoparasite infestation, protein and lysozyme content in organs and serum of some cyprinid species in experimental conditions. Methodology. The objects of the study were yearlings of Cyprinus carpio, Carassius auratus gibelio and Pseudorasbora parva caught in ponds of fish farm “Nyvka” (Kiev region) in spring. Fish were kept in experimental conditions at neutral pH water (6.8-7.2) and a temperature of 17...

  1. Serum YKL-40 Levels and Chitotriosidase Activity in Patients with Beta-Thalassemia Major

    Directory of Open Access Journals (Sweden)

    Maria Musumeci

    2014-01-01

    Full Text Available Background. YKL-40 association with human disease has been the object of many years of investigation. β-thalassemia patients are affected by hepatic siderosis, which determines a fibrotic process and tissue remodelling. Chitotriosidase has been found to be increased in thalassemic patients returning to normal in patients submitted to bone marrow transplantation. YKL-40 is associated with macrophage activation in liver and in other tissues. The aim of the study was to analyse the level of serum YKL-40 and plasma chitotriosidase activity of patients with beta-thalassemia to assess whether their expression correlates with liver disease and degree of liver siderosis. Methods. Expression of YKL-40 and chitotriosidase as a marker of inflammation in 69 thalassemic patients were evaluated. We sought to investigate whether these two chitinases could be considered as a significant biomarker to evaluate therapy effectiveness. Results. Surprisingly we found normal value of YKL-40. We, also, analysed chitotriosidase activity in the same patients that was slightly increased as a consequence of macrophage activation. Conclusions. These data would suggest a good treatment for these patients.

  2. Elevated serum level of IL-27 and VEGF in patients with ankylosing spondylitis and associate with disease activity.

    Science.gov (United States)

    Lin, Tian-Tian; Lu, Jing; Qi, Chen-Yue; Yuan, Lin; Li, Xiao-Lin; Xia, Li-Ping; Shen, Hui

    2015-05-01

    Interleukin (IL)-27 is an IL-12 family cytokine and exerts a critical role in immune regulation in the context of infection, autoimmunity, and angiogenesis. In this study, we aimed to investigate the possible pathophysiological role of IL-27 and vascular endothelial growth factor (VEGF) in ankylosing spondylitis (AS). One hundred and forty AS patients and 90 healthy controls were included in the current study. The levels of IL-27 and VEGF in serum and synovial fluid (SF) samples were measured by enzyme-linked immunosorbent assay. Erythrocyte sedimentation rate, C-reactive protein, and human leukocyte antigen (HLA)-B27 were measured by standard laboratory techniques. Disease activity in AS was scored with Bath Ankylosing Spondylitis Disease Activity Index (BASDAI). Hip involvement, peripheral arthritis, and eye involvement were also recorded. The serum levels of IL-27 were remarkably higher in AS patients than healthy groups and significantly correlated with serum levels of VEGF. Furthermore, the serum levels of IL-27 were correlated with BASDAI independent of other markers of inflammation. Elevated serum levels of IL-27 and VEGF were detected in AS patients with peripheral arthritis and HLA-B27 positive. The SF levels of IL-27 and VEGF were significantly higher than serum levels in AS patients with peripheral arthritis. By contrast, levels of IL-27 and VEGF were not increased in AS patients with hip involvement and eye involvement. IL-27 may regulate the immunological or inflammatory process of AS.

  3. Kinesiology Tape does not Affect Serum Creatine Kinase Level and Quadriceps Activity during Recovery from Delayed-Onset Muscle Soreness

    Directory of Open Access Journals (Sweden)

    Naoko Aminaka

    2017-01-01

    Full Text Available Background: Delayed-onset muscle soreness (DOMS causes muscle damage and edema that can hinder performance and increase risks for secondary injuries. Kinesiology Tape (KT may be an effective modality for aiding in recovery, however, no study has investigated the effects of KT on the physiological biomarkers such as serum creatine kinase (CK level, concurrently with measures of performance and function, during recovery from DOMS. Objective: Investigate the effects of KT on serum CK level, electromyographic (EMG activity of the quadriceps muscles, and performances of countermovement jump (CMJ and triple single-leg hop for distance (HopD during recovery from DOMS. Method: Fifty-eight healthy college-age participants were randomly assigned to KT (n=15, placebo (n=19, and control (n=24 groups. Serum CK level and quadriceps EMG activity and performance during CMJ and HopD were collected at baseline, immediately after repetitive eccentric quadriceps exercise, 48 hours, and 72 hours post-exercise. The EMG recording of rectus femoris, vastus medialis, and vastus lateralis during the CMJ and HopD were normalized to the baseline maximum voluntary isometric contraction. Results: A significant main effect of time on the serum CK level, EMG activity, and performance (p0.05. Conclusion: Taping interventions did not improve the serum CK level or muscle activity and performance during recovery from DOMS. Kinesiology tape may not be the first choice of method for enhancing recovery from DOMS in otherwise healthy individuals.

  4. Serum tau protein as a marker of disease activity in enterohemorrhagic Escherichia coli O111-induced hemolytic uremic syndrome.

    Science.gov (United States)

    Kuroda, Mondo; Shimizu, Masaki; Inoue, Natsumi; Ikeno, Iku; Nakagawa, Hiroyasu; Yokoi, Ayano; Niida, Yo; Konishi, Michio; Kaneda, Hisashi; Igarashi, Noboru; Yamahana, Junya; Taneichi, Hiromichi; Kanegane, Hirokazu; Ito, Mika; Saito, Shigeru; Furuichi, Kengo; Wada, Takashi; Nakagawa, Masaru; Yokoyama, Hitoshi; Yachie, Akihiro

    2015-01-01

    Tau protein levels in cerebrospinal fluid (CSF) and serum are elevated in patients with various central nervous system diseases. We investigated whether serum tau protein levels are useful for predicting and assessing disease activity of acute encephalopathy (AE) in enterohemorrhagic Escherichia coli (EHEC) O111-induced hemolytic uremic syndrome (HUS; EHEC encephalopathy). Serum samples were obtained from 14 patients with EHEC O111/HUS, 20 patients with non-EHEC-related AE, and 20 age- and sex-matched healthy controls. CSF samples were obtained from 2 patients with EHEC encephalopathy and 20 patients with non-EHEC-related AE. Tau protein levels and levels of several proinflammatory cytokines were quantified by enzyme-linked immunosorbent assays. Results were compared with the clinical features of EHEC encephalopathy, including magnetic resonance image (MRI) findings. Serum tau levels in patients with EHEC encephalopathy were significantly elevated compared with those in patients with EHEC O111/HUS without encephalopathy, patients with non-EHEC-related AE, and healthy controls. The ratio of CSF tau levels to serum tau levels was >1.0 in all patients with non-EHEC-related AE but encephalopathy. Serum tau protein levels increased rapidly and markedly in patients with severe EHEC 0111/HUS and encephalopathy when HUS occurred, but were not elevated in mild patients, even in the HUS phase. Furthermore, changes in serum tau protein levels in patients with EHEC encephalopathy were consistent with abnormalities on brain MRI and were positively correlated with proinflammatory cytokine levels. Our results indicate that serum tau protein might be useful to predict and assess disease activity of EHEC encephalopathy. Copyright © 2015 Elsevier Ltd. All rights reserved.

  5. Serum antioxidant enzymes activities and oxidative stress levels in patients with acute ischemic stroke: influence on neurological status and outcome.

    Science.gov (United States)

    Milanlioglu, Aysel; Aslan, Mehmet; Ozkol, Halil; Çilingir, Vedat; Nuri Aydın, Mehmet; Karadas, Sevdegül

    2016-03-01

    Oxidative stress is well believed to play a role in the pathogenesis of acute ischemic stroke. Reports on antioxidant enzyme activities in patients with stroke are conflicting. Therefore, the aim of this study was to investigate serum antioxidant enzyme activities and oxidative stress levels in patients with acute ischemic stroke within 1st, 5th, and 21st day after stroke onset and also the relationship between these results and the clinical status of patients. The current study comprised 45 patients with acute ischemic stroke and 30 healthy controls. Serum malondialdehyde (MDA) levels, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase activities were measured spectrophotometrically. Serum MDA levels were significantly higher in acute ischemic stroke patients within 24 h after stroke onset than controls (p levels did not change (p > 0.05). Serum SOD activity was significantly lower in 21st day compared to SOD activity of controls (p levels, GSH-Px, and CAT activities did not change significantly. Our study demonstrated that acute ischemic stroke patients have increased oxidative stress and decreased antioxidant enzymes activities. These findings indicated that an imbalance of oxidant and antioxidant status might play a role in the pathogenesis of acute ischemic stroke.

  6. Activity of alpha-fucosidase and beta-glucuronidase in serum and urine of patients administered parenteral nutrition.

    Science.gov (United States)

    Raczkowska, Katarzyna; Szajda, Slawomir Dariusz; Raczkowski, Krzysztof; Zasadowska, Wioletta; Chojnowska, Sylwia; Kepka, Alina; Zalewska-Szajda, Beata; Waszkiewicz, Napoleon; Knaś, Malgorzata; Snarska, Jadwiga; Zwierz, Krzysztof; Ladny, Jerzy Robert

    2013-01-01

    In hospital patients suffering from adverse clinical and biochemical symptoms of malnutrition, it is often necessary to employ parenteral nutrition to avoid the body's tissue becoming broken down by being metabolised. Thus, the patient's welfare and survival can be supported throughout any periods of medical crisis. Two of the enzymes responsible for metabolising glycoconjugates are alpha-fucosidase (FUC) and beta-glucuronidase (GLU), present in lysosomes. They release fucose or glucuronic acid from the non-reducing end of oligosaccharide chains. To determine the effect of parenteral nutrition administered to ill patients, on glycoconjugate metabolism, by measuring serum and urinary activities of FUC and GLU. Material and methods. Blood samples and the daily urine collection were taken from 23 patients' who had been undergoing parenteral nutrition for either 5 or 10 days, as well as from a baseline sample. Enzyme activities in serum and urine were determined by the method of Zwierz et al. Serum FUC activities were significantly lower after 10 days compared to 5, (p< 0.0172), whereas GLU activities were significantly lower after both 5 and 10 days, (p< 0.0007 and p< 0.0208 respectively), compared to levels before starting parenteral nutrition. GLU activities were however higher after 10 days than those after 5 days, (p< 0.0023). In urine, FUC activities were significantly decreased after 10 days compared to 5 days after starting parenteral nutrition, (p< 0.0245). Urine GLU activities were unaffected by parenteral nutrition nor was any effect seen on FUC or GLU activities when calculated per 1mg creatinine. Serum FUC and GLU activities can be used for assessing the effect of parenteral nutrition on glycoconjugate metabolism. The significant decreases of serum GLU activity observed after 5 and 10 days, may serve to indicate that the components of parental nutrition are appropriate and that the body has become suitably adapted to this form of nutrition.

  7. Antigen-specific helper activity in serum of mice primed with sheep red cells I. Definition of the test system and comparison with other systems

    NARCIS (Netherlands)

    Dijk, H. van; Rademaker, P.M.; Slotboom, A.; Willers, J.M.

    1980-01-01

    An adoptive transfer system is described to measure serum helper activity in the primary antibody response to sheep red blood cells (SRBC). Mice injected with a high dose of cyclophosphamide and reconstituted with rabbit anti-thymocyte serum-treated spleen cells were used as recipients. Serum obtain

  8. Comparison of Antibodies with Amylase Activity from Cerebrospinal Fluid and Serum of Patients with Multiple Sclerosis.

    Directory of Open Access Journals (Sweden)

    Vasilii B Doronin

    Full Text Available We have recently shown that IgGs from serum and cerebrospinal fluid (CSF of MS patients are active in hydrolysis of DNA and myelin basic protein. According to literature data, anti-DNA and anti-MBP abzymes may promote important neuropathologic mechanisms in this chronic inflammatory disorder and in MS pathogenesis development. At the same time, the involvement of antibodies with amylase activity in the pathogenesis of any autoimmune disease has not yet been identified. Electrophoretically and immunologically homogeneous IgGs were obtained by a sequential affinity chromatography of the CSF proteins on protein G-Sepharose and FPLC gel filtration. We are able to present the first unpredictable evidence showing that IgGs from CSF possess amylase activity and efficiently hydrolyze maltoheptaose; their average specific Ab activity is ~30-fold higher than that of antibodies from sera of the same MS patients. Specific average RA (SAA for IgGs from healthy volunteers was approximately ~1000 lower than that for MS patients. In addition, it was shown that a relative SAA of total proteins of CSF (including Abs ~15-fold lower than that for purified IgGs, while the relative SAA of the total sera protein is higher than that of sera IgGs by a factor of 1033. This result speaks in favor of the fact that amylolytic activity of CSF proteins is mainly caused by the activity of amylase abzymes. One cannot exclude, that amylase abzymes of CSF can play a, as yet unknown, role in the pathogenesis of MS. Some possible reasons of these findings are discussed.

  9. Drosophila pico and its mammalian ortholog lamellipodin activate serum response factor and promote cell proliferation.

    Science.gov (United States)

    Lyulcheva, Ekaterina; Taylor, Eleanor; Michael, Magdalene; Vehlow, Anne; Tan, Shengjiang; Fletcher, Adam; Krause, Matthias; Bennett, Daimark

    2008-11-01

    MIG-10/RIAM/lamellipodin (MRL) proteins link activated Ras-GTPases with actin regulatory Ena/VASP proteins to induce local changes in cytoskeletal dynamics and cell motility. MRL proteins alter monomeric (G):filamentous (F) actin ratios, but the impact of these changes had not been fully appreciated. We report here that the Drosophila MRL ortholog, pico, is required for tissue and organismal growth. Reduction in pico levels resulted in reduced cell division rates, growth retardation, increased G:F actin ratios and lethality. Conversely, pico overexpression reduced G:F actin ratios and promoted tissue overgrowth in an epidermal growth factor (EGF) receptor (EGFR)-dependent manner. Consistently, in HeLa cells, lamellipodin was required for EGF-induced proliferation. We show that pico and lamellipodin share the ability to activate serum response factor (SRF), a transcription factor that responds to reduced G:F-actin ratios via its co-factor Mal. Genetics data indicate that mal/SRF levels are important for pico-mediated tissue growth. We propose that MRL proteins link EGFR activation to mitogenic SRF signaling via changes in actin dynamics.

  10. Drosophila Pico and Its Mammalian Ortholog Lamellipodin Activate Serum Response Factor and Promote Cell Proliferation

    Science.gov (United States)

    Lyulcheva, Ekaterina; Taylor, Eleanor; Michael, Magdalene; Vehlow, Anne; Tan, Shengjiang; Fletcher, Adam; Krause, Matthias; Bennett, Daimark

    2008-01-01

    Summary MIG-10/RIAM/lamellipodin (MRL) proteins link activated Ras-GTPases with actin regulatory Ena/VASP proteins to induce local changes in cytoskeletal dynamics and cell motility. MRL proteins alter monomeric (G):filamentous (F) actin ratios, but the impact of these changes had not been fully appreciated. We report here that the Drosophila MRL ortholog, pico, is required for tissue and organismal growth. Reduction in pico levels resulted in reduced cell division rates, growth retardation, increased G:F actin ratios and lethality. Conversely, pico overexpression reduced G:F actin ratios and promoted tissue overgrowth in an epidermal growth factor (EGF) receptor (EGFR)-dependent manner. Consistently, in HeLa cells, lamellipodin was required for EGF-induced proliferation. We show that pico and lamellipodin share the ability to activate serum response factor (SRF), a transcription factor that responds to reduced G:F-actin ratios via its co-factor Mal. Genetics data indicate that mal/SRF levels are important for pico-mediated tissue growth. We propose that MRL proteins link EGFR activation to mitogenic SRF signaling via changes in actin dynamics. PMID:19000833

  11. A Rapid and Sensitive Method to Measure the Functional Activity of Shiga Toxins in Human Serum

    Directory of Open Access Journals (Sweden)

    Valentina Arfilli

    2015-11-01

    Full Text Available Shiga toxins (Stx have a definite role in the development of hemolytic uremic syndrome in children with hemorrhagic colitis caused by pathogenic Stx-producing Escherichia coli (STEC strains. The dramatic effects of these toxins on the microvasculature of different organs, particularly of the kidney, are well known, whereas there is no consensus on the mechanism by which Stx reach the endothelia of target organs and/or indirectly injure these body sites. We hereby describe a quick (4 h, radioactive, Raji cell-based method designed for the detection of Stx in human sera. The assay monitors the translation impairment induced by these powerful inhibitors of protein synthesis, which are identified properly by neutralizing their activity with specific monoclonal antibodies. By this method, we detected for the first time the functional activity of Stx in sera of STEC-infected patients during hemorrhagic colitis. Recent research has pointed to a dynamic process of Stx-induced renal intoxication in which concurrent and interactive steps are involved. Our rapid and specific method could be useful for studying the kinetics of Stx during the natural course of STEC infection and the interplay between Stx activity in serum and Stx presence in different blood fractions (neutrophils, monocytes, platelets, leukocyte-platelet aggregates, microvesicles, lipoproteins.

  12. A Rapid and Sensitive Method to Measure the Functional Activity of Shiga Toxins in Human Serum

    Science.gov (United States)

    Arfilli, Valentina; Carnicelli, Domenica; Ardissino, Gianluigi; Torresani, Erminio; Scavia, Gaia; Brigotti, Maurizio

    2015-01-01

    Shiga toxins (Stx) have a definite role in the development of hemolytic uremic syndrome in children with hemorrhagic colitis caused by pathogenic Stx-producing Escherichia coli (STEC) strains. The dramatic effects of these toxins on the microvasculature of different organs, particularly of the kidney, are well known, whereas there is no consensus on the mechanism by which Stx reach the endothelia of target organs and/or indirectly injure these body sites. We hereby describe a quick (4 h), radioactive, Raji cell-based method designed for the detection of Stx in human sera. The assay monitors the translation impairment induced by these powerful inhibitors of protein synthesis, which are identified properly by neutralizing their activity with specific monoclonal antibodies. By this method, we detected for the first time the functional activity of Stx in sera of STEC-infected patients during hemorrhagic colitis. Recent research has pointed to a dynamic process of Stx-induced renal intoxication in which concurrent and interactive steps are involved. Our rapid and specific method could be useful for studying the kinetics of Stx during the natural course of STEC infection and the interplay between Stx activity in serum and Stx presence in different blood fractions (neutrophils, monocytes, platelets, leukocyte-platelet aggregates, microvesicles, lipoproteins). PMID:26556372

  13. The effect of the binge drinking session on the activity of salivary, serum and urinary beta-hexosaminidase: preliminary data.

    Science.gov (United States)

    Waszkiewicz, Napoleon; Szajda, Slawomir Dariusz; Jankowska, Anna; Kepka, Alina; Dobryniewski, Jacek; Szulc, Agata; Zwierz, Krzysztof

    2008-01-01

    Our report is the first to show that an acute ingestion (6 h) of a relatively large, yet tolerable dose of alcohol (120-160 g), significantly increases activity of total serum beta-hexosaminidase (total beta-HEX), beta-HEX A and beta-HEX B isoenzymes, as well as salivary total beta-HEX and urinary beta-HEX A, in eight infrequent binge drinkers. An increase in the activity of serum and urinary total HEX is mainly due to its secretory isoenzyme beta-HEX A.

  14. CORRELATION BETWEEN SERUM PARAOXONASE ACTIVITY AND HIGH DENSITY LIPOPROTEIN LEVELS IN TYPE2 DIABETES PREDISPOSING TO ATEROSCLEROSIS

    Directory of Open Access Journals (Sweden)

    SK.Deepthi

    2014-11-01

    Full Text Available Human serum paraoxonase is physically associated with HDL and has been implicated in the detoxification of organophosphates and possibly in the prevention of LDL lipid peroxidation and therefore retards atherosclerosis. HDL levels are inversely related to the risk of developing atherosclerosis. We investigated the serum activity and concentration of paraoxonase and HDL levels in 104 subjects (42 diabetic patients without complications, 42 controls, 20 diabetic patients with complications.. Paraoxonase activity was found to be lower in diabetic patients than in controls. Similarly there was reduction in HDL levels in cases suggesting a positive correlation between HDL and paraoxonase levels.

  15. Self-assembly of lysozyme on the surfaces of gold nanoparticles

    Institute of Scientific and Technical Information of China (English)

    Ming Hui Xiang; Xiao Xu; Na Li; Ke An Li

    2011-01-01

    The interaction of lysozyme (Lys) and gold nanoparticles was investigated via UV-vis absorption and resonance light-scattering method. There are some changes of the plasmon absorption and resonance light-scattering of gold nanoparticles that were observed via the addition of Lys. The normalized plasmon absorption and resonance light-scattering intensity with gold nanoparticles were both linear with 1-20 nmol/L Lys. A simple model about the component of the gold nanoparticles and Lys complex was established and the calculated result was fitted well in their concentration ratio. Furthermore, the activity analysis of Lys showed that the interaction was weak and nondestructive.

  16. Relationship between serum HBV RNA levels and intrahepatic viral as well as histologic activity markers in entecavir-treated patients.

    Science.gov (United States)

    Wang, Jing; Yu, Yiqi; Li, Guojun; Shen, Chuan; Meng, Zhefeng; Zheng, Jianming; Jia, Yanhong; Chen, Shaolong; Zhang, Xiao; Zhu, MengQi; Zheng, Jiangjiang; Song, Zhangzhang; Wu, Jing; Shao, Lingyun; Qian, Peiyu; Mao, Xiaona; Wang, Xuanyi; Huang, Yuxian; Zhao, Caiyan; Zhang, Jiming; Qiu, Chao; Zhang, Wenhong

    2017-09-01

    In diagnostics, serum hepatitis B virus (HBV)-RNA levels are valuable when the HBV-DNA load in circulation is effectively suppressed by nucleos(t)ide analogue (NUC) therapy. This study aimed to determine the intrahepatic viral replication activity reflected in serum HBV-RNA and whether HBV-RNA contributes to liver histological changes in NUC-treated patients. A cross-sectional set of serum and liver biopsy samples was obtained from entecavir-treated patients with undetectable levels of serum HBV-DNA. The correlations between HBV-RNA concentration in serum and levels of peripheral and intrahepatic viral replicative forms and histological scores were analyzed. Quasispecies of serum HBV-RNA and intrahepatic viral replicative forms were examined by deep sequencing. HBV-RNA-positive hepatocytes were visualized by in situ hybridization. Serum HBV-RNA was detected in 35 of 47 patients (74.47%, 2.33-4.80 log10 copies/mL). These levels correlated not only with the intrahepatic HBV-RNA level and the ratio of intrahepatic HBV-RNA to covalently closed circular DNA (cccDNA), but also with the histological scores for grading and staging. From the view of quasispecies, serum HBV-RNA was more genetically homogenous with contemporaneously sampled intrahepatic HBV-RNA relative to cccDNA pool and dynamically changed over time in consecutive samples. In situ histology study revealed that HBV-RNA-positive hepatocytes were clustered in foci, sporadically distributed across the lobules, and co-localized with hepatitis B surface antigen. Serum HBV-RNA levels reflect intrahepatic viral transcriptional activity and are associated with liver histopathology in patients receiving NUC therapy. Our study sheds light on the nature of HBV-RNA in the pathogenesis of chronic HBV infection and has implications for the management of chronic hepatitis B during NUC therapy. Serum HBV-RNA levels are indicative of the intrahepatic transcriptional activity of covalently closed circular DNA and are

  17. Refolding of Denatured/Reduced Lysozyme Using Weak-Cation Exchange Chromatography

    Institute of Scientific and Technical Information of China (English)

    Yan WANG; Bo Lin GONG; Xin Du GENG

    2003-01-01

    Oxidative refolding of the denatured/reduced lysozyme was investigated by using weak-cation exchange chromatography (WCX). The stationary phase of WCX binds to the reduced lysozyme and prevented it from forming intermolecular aggregates. At the same time urea and ammonium sulfate were added to the mobile phase to increase the elution strength for lysozyme. Ammonium sulfate can more stabilize the native protein than a common eluting agent, sodium chloride. Refolding of lysozyme by using this WCX is successfully. It was simply carried out to obtain a completely and correctly refolding of the denatured lysozyme at high concentration of 20.0 mg/mL.

  18. Investigation on the binding activities of citalopram with human and bovine serum albumins

    Energy Technology Data Exchange (ETDEWEB)

    Lin, Jingjing; Liu, Yan, E-mail: liuyan@fjirsm.ac.cn; Chen, Mingmao; Huang, Huayin; Song, Ling, E-mail: songling@fjirsm.ac.cn

    2014-02-15

    The binding interactions of citalopram (CIT), an efficient antidepressant, with human serum albumin (HSA) and bovine serum albumin (BSA) were investigated by a series of spectroscopic methods including fluorescence, UV–vis absorption, circular dichroism (CD) and {sup 1}H nuclear magnetic resonance ({sup 1}H NMR). The fluorescence quenching and UV–vis absorption studies reveal that CIT could form complexes with both HSA and BSA. The CIT–BSA complex exhibits higher binding affinity than CIT–HSA complex. The thermodynamic study further suggests that the interactions between CIT and SAs are mainly driven by hydrophobic forces and hydrogen bonds. The {sup 1}H NMR analysis indicates that the participation of different functional groups of CIT is unequal in the complexation of CIT–HSA and CIT–BSA. Site marker competitive experiments show that the interactions between CIT and SAs primarily locate at sub-domain II A (site I). The effects of CIT on the conformation of SAs are further analyzed via synchronous fluorescence, three-dimensional fluorescence and CD spectra techniques. The results prove that the presence of CIT decreases the α-helical content of both SAs and induces the slight unfolding of the polypeptides of protein. Additionally, the conformational change of BSA induced by CIT is larger than that of HSA. -- Highlights: • The difference of binding activity between CIT–BSA and CIT–HSA is first reported. • Use spectroscopic, thermodynamic, and NMR methods. • CIT exhibits higher binding affinity to BSA than to HSA. • The binding forces between CIT and SA have been investigated. • The complexation of CIT–SA induces the conformational change of SA.

  19. The relationship between metabolic presbycusis and serum paraoxonase/arylesterase activity.

    Science.gov (United States)

    Keleş, Erol; Kapusuz, Zeliha; Gürsu, Mehmet Ferit; Karlıdag, Turgut; Kaygusuz, Irfan; Bulmuş, Funda Gülcü; Yalcın, Sinasi

    2014-01-01

    To determine the presence of a relationship between metabolic presbycusis and serum paraoxonase/arylesterase activity. A total of 30 patients who had been admitted to the Ear, Nose, and Throat (ENT) Clinic of Fırat University Medical Faculty and diagnosed as metabolic presbycusis were included in the study. The control group was composed of 30 healthy volunteers. Pure tone audiometry and impedencemeter were performed on all subjects included in the study at the audiometry laboratory of the ENT clinic. The presence of a regular hearing curve, a symmetrical sensorineural hearing loss more than 25 dB with preserved speech discrimination were accepted as criteria for metabolic presbycusis. Blood samples were drawn from the patients prior to the hearing tests. The sera were separated for measurements of total cholesterol, triglyceride, high-density lipoprotein, very low-density lipoprotein, low-density lipoprotein, human serum paraoxonase and arylesterase levels, respectively. No statistically significant difference was found between the patient and the control groups in terms of age and gender. Paraoxonase, arylesterase and paraoxonase/arylesterase, high-density lipoprotein levels were found to decrease in the study group and the difference was found to be statistically significant compared to the control group (P presbycusis. Furthermore, the results of this study make us think that there could be a relationship between metabolic presbycusis and cardiovascular diseases. In this case, metabolic presbycusis may be a determining parameter in the early diagnosis of cardiovascular diseases. We consider that this study may be the pioneer for further studies conducted with larger patient numbers.

  20. Characterization and antioxidant activity of bovine serum albumin and sulforaphane complex in different solvent systems

    Energy Technology Data Exchange (ETDEWEB)

    Dong, Xueyan; Zhou, Rui; Jing, Hao, E-mail: h200521@cau.edu.cn

    2014-02-15

    Modes and influencing factors of bovine serum albumin (BSA) and sulforaphane (SFN) interaction will help us understand the interaction mechanisms and functional changes of bioactive small molecule and biomacromolecule. This study investigated interaction mechanisms of BSA and SFN and associated antioxidant activity in three solvent systems of deionized water (dH{sub 2}O), dimethyl sulfoxide (DMSO) and ethanol (EtOH), using Fourier transform infrared spectroscopy (FT-IR), fluorescence spectroscopy, synchronous fluorescence spectroscopy, DPPH and ABTS radical scavenging assays. The results revealed that SFN had ability to quench BSA's fluorescence in static modes, and to interact with BSA at both tyrosine (Tyr) and tryptophan (Trp) residues, while the Trp residues were highly sensitive, which was demonstrated by fluorescence at 340 nm. Hydrophobic forces, hydrogen bonds and van der Waals interactions were all involved in BSA and SFN interaction, which were not significantly changed by three solvents. The binding constant values and binding site numbers were in a descending order of dH{sub 2}O>DMSO>EtOH. The values of free energy change were in a descending order of dH{sub 2}O>DMSO>EtOH, which indicated that the binding forces were in a descending order of dH{sub 2}O>DMSO>EtOH. There was no significant difference in antioxidant activity between SFN and BSA–SFN. Moreover, three solvents had not significant influence on antioxidant activity of SFN and BSA–SFN. -- Highlights: • We report interaction mechanisms of BSA and sulforaphane in three solvent systems. • We report antioxidant activity of BSA–sulforaphane complex in three solvent systems. • Decreasing the solvent polarity will decrease the binding of BSA and sulforaphane. • Three solvents had not influence on antioxidant activity of BSA–sulforaphane.

  1. Pneumococcal intracellular killing is abolished by polysaccharide despite serum complement activity.

    OpenAIRE

    Schweinle, J. E.

    1986-01-01

    Normal human serum absorbed at 0 degrees C with pneumococcal serotype 1, 12, or 25 lost the ability to support polymorphonuclear leukocyte intracellular killing of some pneumococcal serotypes even if immunoglobulin was provided. The absorbed serum contained no organisms but had residual polysaccharide when measured by counterimmunoelectrophoresis against type-specific antisera. The influence of pneumococcal polysaccharide (PPS) on serum support of intracellular polymorphonuclear leukocyte kil...

  2. Lysozyme Resistance in Streptococcus suis Is Highly Variable and Multifactorial

    NARCIS (Netherlands)

    Wichgers, P.J.; Weeghel, van C.; Rebel, J.M.J.; Smits, M.A.; Putten, van J.P.M.; Smith, H.E.

    2012-01-01

    Background Streptococcus suis is an important infectious agent for pigs and occasionally for humans. The host innate immune system plays a key role in preventing and eliminating S. suis infections. One important constituent of the innate immune system is the protein lysozyme, which is present in a v

  3. 21 CFR 862.1490 - Lysozyme (muramidase) test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Lysozyme (muramidase) test system. 862.1490 Section 862.1490 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Chemistry Test...

  4. ASSOCIATION OF LYSOZYME TO PHOSPHOLIPID SURFACES AND VESICLE FUSION

    NARCIS (Netherlands)

    ARNOLD, K; HOEKSTRA, D; OHKI, S

    1992-01-01

    Lysozyme-induced fusion of phosphatidylserine (PS) vesicles was studied as a function of pH. Fusion, monitored by lipid-mixing, was measured by following the dilution of pyrene-labelled phosphatidylcholine, incorporated in PS vesicles, into unlabelled bilayers. It is demonstrated that

  5. Antimicrobial chitosan-lysozyme (CL) films and coatings for enhancing microbial safety of mozzarella cheese.

    Science.gov (United States)

    Duan, J; Park, S-I; Daeschel, M A; Zhao, Y

    2007-11-01

    This study investigated the antimicrobial activities of chitosan-lysozyme (CL) composite films and coatings against tested microorganisms inoculated onto the surface of Mozzarella cheese. CL film-forming solutions (FFS) with a pH of 4.4 to 4.5 were prepared by incorporating 0% or 60% lysozyme (per dry weight of chitosan) into chitosan FFS with or without a pH adjustment to 5.2. Sliced cheese was subjected to 3 CL package applications: film, lamination on a multilayer coextruded film, and coating. Cheese was inoculated with Listeria monocytogenes, Escherichia coli, or Pseudomonas fluorescens at 10(4) CFU/g, or with mold and yeast at 10(2) CFU/g. Inoculated cheese was individually vacuum packaged and stored at 10 degrees C for sampling at 1, 7, and 14 d for bacteria, and at 10, 20, and 30 d for fungi. Inoculated bacteria survived but failed to multiply in untreated cheese during storage. Treated cheese received 0.43- to 1.25-, 0.40- to 1.40-, and 0.32- to 1.35-log reductions in E. coli, P. fluorescens, and L. monocytogenes, respectively. Incorporation of 60% lysozyme in chitosan FFS showed greater antimicrobial effect than chitosan alone on P. fluorescens and L. monocytogenes. The pH adjustment only affected the antimicrobial activity on L. monocytogenes, with lower pH (unadjusted) showing greater antimicrobial effect than pH 5.2. Mold and yeast increased to 10(5) CFU/g in untreated cheese after 30 d storage. Growth of mold was completely inhibited in cheese packaged with CL films, while 0.24- to 1.90- and 0.06- to 0.50-log reductions in mold populations were observed in cheese packaged with CL-laminated films and coatings, respectively. All CL packaging applications resulted in 0.01- to 0.64-log reduction in yeast populations.

  6. Two goose-type lysozymes in Mytilus galloprovincialis: possible function diversification and adaptive evolution.

    Directory of Open Access Journals (Sweden)

    Qing Wang

    Full Text Available Two goose-type lysozymes (designated as MGgLYZ1 and MGgLYZ2 were identified from the mussel Mytilus galloprovincialis. MGgLYZ1 mRNA was widely expressed in the examined tissues and responded sensitively to bacterial challenge in hemocytes, while MGgLYZ2 mRNA was predominately expressed and performed its functions in hepatopancreas. However, immunolocalization analysis showed that both these lysozymes were expressed in all examined tissues with the exception of adductor muscle. Recombinant MGgLYZ1 and MGgLYZ2 could inhibit the growth of several Gram-positive and Gram-negative bacteria, and they both showed the highest activity against Pseudomonas putida with the minimum inhibitory concentration (MIC of 0.95-1.91 µM and 1.20-2.40 µM, respectively. Protein sequences analysis revealed that MGgLYZ2 had lower isoelectric point and less protease cutting sites than MGgLYZ1. Recombinant MGgLYZ2 exhibited relative high activity at acidic pH of 4-5, while MGgLYZ1 have an optimum pH of 6. These results indicated MGgLYZ2 adapted to acidic environment and perhaps play an important role in digestion. Genomic structure analysis suggested that both MGgLYZ1 and MGgLYZ2 genes are composed of six exons with same length and five introns, indicating these genes were conserved and might originate from gene duplication during the evolution. Selection pressure analysis showed that MGgLYZ1 was under nearly neutral selection while MGgLYZ2 evolved under positive selection pressure with three positively selected amino acid residues (Y(102, L(200 and S(202 detected in the mature peptide. All these findings suggested MGgLYZ2 perhaps served as a digestive lysozyme under positive selection pressure during the evolution while MGgLYZ1 was mainly involved in innate immune responses.

  7. Cloning, purification and comparative characterization of two digestive lysozymes from Musca domestica larvae

    Directory of Open Access Journals (Sweden)

    F.C. Cançado

    2008-11-01

    Full Text Available cDNA coding for two digestive lysozymes (MdL1 and MdL2 of the Musca domestica housefly was cloned and sequenced. MdL2 is a novel minor lysozyme, whereas MdL1 is the major lysozyme thus far purified from M. domestica midgut. MdL1 and MdL2 were expressed as recombinant proteins in Pichia pastoris, purified and characterized. The lytic activities of MdL1 and MdL2 upon Micrococcus lysodeikticus have an acidic pH optimum (4.8 at low ionic strength (μ = 0.02, which shifts towards an even more acidic value, pH 3.8, at a high ionic strength (μ = 0.2. However, the pH optimum of their activities upon 4-methylumbelliferyl N-acetylchitotrioside (4.9 is not affected by ionic strength. These results suggest that the acidic pH optimum is an intrinsic property of MdL1 and MdL2, whereas pH optimum shifts are an effect of the ionic strength on the negatively charged bacterial wall. MdL2 affinity for bacterial cell wall is lower than that of MdL1. Differences in isoelectric point (pI indicate that MdL2 (pI = 6.7 is less positively charged than MdL1 (pI = 7.7 at their pH optima, which suggests that electrostatic interactions might be involved in substrate binding. In agreement with that finding, MdL1 and MdL2 affinities for bacterial cell wall decrease as ionic strength increases.

  8. Effects of Oxidized Tallow on the Rabbit Serum Lipids and Antioxidant Activity of the In-vitro Lipids.

    Science.gov (United States)

    Zeb, Alam; Rahman, Waheed Ur

    2012-09-01

    This paper describes the effects of thermally oxidized tallow on the serum lipids profile and radical scavenging activity (RSA) of the lipids extracted from the different tissues of the rabbits. Tallow was thermally oxidized at 130℃ for 9, 18, 27, 36 and 45 h respectively. Thermally oxidized tallow was fed to the local strain of Himalayan rabbits for one week. Results show that oxidation increases the formation of hydroperoxides and decrease the level of radical scavenging activity of the tallow. The rabbit serum lipids profile showed a dose dependent increase in triglyceride, total cholesterol and LDL-cholesterol. However, no statistically significant increase was observed in the HDL-cholesterol with an increase of oxidation time. Serum glucose and rabbits body weight decrease significantly (p tallow is harmful and therefore an alternative way of cooking should be used.

  9. Chaperonin-Inspired pH Protection by Mesoporous Silica SBA-15 on Myoglobin and Lysozyme.

    Science.gov (United States)

    Lynch, Michele M; Liu, Jichuan; Nigra, Michael; Coppens, Marc-Olivier

    2016-09-20

    While enzymes are valuable tools in many fields of biotechnology, they are fragile and must be protected against denaturing conditions such as unfavorable solution pH. Within living organisms, chaperonins help enzymes fold into their native shape and protect them from damage. Inspired by this natural solution, mesoporous silica SBA-15 with different pore diameters is synthesized as a support material for immobilizing and protecting enzymes. In separate experiments, the model enzymes myoglobin and lysozyme are physically adsorbed to SBA-15 and exposed to a range of buffered pH conditions. The immobilized enzymes' biocatalytic activities are quantified and compared to the activities of nonimmobilized enzymes in the same solution conditions. It has been observed that myoglobin immobilized on SBA-15 is protected from acidic denaturation from pH 3.6 to 5.1, exhibiting relative activity of up to 350%. Immobilized lysozyme is protected from unfavorable conditions from pH 6.6 to 7.6, with relative activity of up to 200%. These results indicate that the protective effects conferred to enzymes immobilized by physical adsorption to SBA-15 are driven by the enzymes' electrostatic attraction to the material's surface. The pore diameter of SBA-15 affects the quality of protection given to immobilized enzymes, but the contribution of this effect at different pH values remains unclear.

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

    Directory of Open Access Journals (Sweden)

    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.

  11. Activity of lysosomal exoglycosidases in serum and synovial fluid in patients with chronic Lyme and rheumatoid arthritis.

    Science.gov (United States)

    Pancewicz, Slawomir; Popko, Janusz; Rutkowski, Ryszard; Knaś, Malgorzata; Grygorczuk, Sambor; Guszczyn, Tomasz; Bruczko, Marta; Szajda, Slawomir; Zajkowska, Joanna; Kondrusik, Maciej; Sierakowski, Stanislaw; Zwierz, Krzysztof

    2009-01-01

    Lysosomal exoglycosidases participate in the destruction of the articular cartilage by cleaving glycoside bonds in glycoproteins and proteoglycans. The aim of the study was to determine the activity of exoglycosidases: hexosaminidase, beta-glucuronidase, beta-galactosidase, alpha-mannosidase and alpha-fucosidase in serum and synovial fluid of patients with Lyme and rheumatoid arthritis. The study group consisted of 10 patients with chronic Lyme arthritis (age 18 - 74 y), 13 with rheumatoid arthritis (age 32 - 70 y) and 10 with juvenile idiopathic arthritis (age 8 - 17 y). The control group consisted of 9 healthy volunteers (age 24 - 62 y). The activity of the exoglycosidases was determined with the p-nitrophenyl derivatives of sugars as substrates. A significant increase of the activity of all the exoglycosidases in serum and in synovial fluid of the patients with different forms of arthritis was found. The ratio of synovial fluid/serum activity of exoglycosidases was above 2.0 in LA but not in JIA and RA patients. As the main source of exoglycosidases in the joint is the synovial membrane, this result supports the appropriateness of therapeutic synovectomy in chronic Lyme arthritis with knee effusion. The serum activity of hexosaminidase may be used in monitoring the course of Lyme arthritis and the efficiency of treatment.

  12. Interaction and sonodynamic damage activity of acridine red (AD-R) to bovine serum albumin (BSA)

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Dandan; Xie, Jinhui; Wu, Qiong; Fan, Ping; Wang, Jun, E-mail: wangjun888tg@126.com

    2015-04-15

    The sonodynamic therapy (SDT) has become an attractive antitumor treatment method in recent years, but the selection of sonosensitizer, mechanism of damage biomolecule and kind of reactive oxygen species (ROS) generated during sonodynamic process have not been investigated in detail. In this paper, the acridine red (AD-R), as a sonosensitizer, combining with ultrasonic irradiation to damage bovine serum albumin (BSA) was investigated. At first, the interaction of AD-R to BSA molecules in aqueous solution was studied by fluorescence spectroscopy. As judged from the experimental results, the quenching mechanism of BSA fluorescence belongs to a static process. Synchronous fluorescence spectra demonstrate that the binding and damage sites to BSA molecules are mainly on the tryptophan residues. The generation and kind of generated ROS were also estimated by the method of oxidation and extraction photometry. This paper may offer some valuable references for the study of the sonodynamic activity and application of AD-R in SDT for tumor treatment. - Highlights: ●Acridine red (AD-R) is used to study interaction with BSA. ●Spectroscopy is used to study sonodynamic damage activity of AD-R to BSA. ●Generation of ROS caused by AD-R under ultrasonic irradiation was determined.

  13. Measurement of serum paraoxonase-1 activity in the evaluation of liver function

    Institute of Scientific and Technical Information of China (English)

    Jordi Camps; Judit Marsillach; Jorge Joven

    2009-01-01

    Paraoxonase-1 (PON1) is an esterase and lactonase synthesized by the liver and found in the circulation associated with high-density lipoproteins. The physiological function of PON1 seems to be to degrade specific oxidized cholesteryl esters and oxidized phospholipids in lipoproteins and cell membranes. PON1 is, therefore, an antioxidant enzyme. Alterations in circulating PON1 levels have been reported in a variety of diseases involving oxidative stress including chronic liver diseases. Measurement of serum PON1 activity has been proposed as a potential test for the evaluation of liver function. However, this measurement is still restricted to research and has not been extensively applied in routine clinical chemistry laboratories. The reason for this restriction is due to the problem that the substrate commonly used for PON1 measurement,paraoxon, is toxic and unstable. The recent development of new assays with non-toxic substrates makes this proposal closer to a practical development. The present editorial summarizes PON1 biochemistry and function,its involvement with chronic liver impairment, and some aspects related to the measurement of PON1 activity in circulation.

  14. The influence of starvation and water deprivation upon certain serum enzyme activity in chicks in the first week after hatching

    Directory of Open Access Journals (Sweden)

    Markovska-Gosic Bozidarka

    2009-11-01

    Full Text Available In the past decade the determination of enzyme activity in blood serum, organs and tissues gained an innocuous meaning in the veterinary medicine, especially in the early stages of diagnosis of different metabolism abnormalities. However, the experience gained from the human medicine, without a detail analyses could not be applied on domestic animals and poultry, because of the fact that the enzyme activity in the cells of different animal tissues and organs differs from those of the humans. This explains the different diagnostic meaning of the enzyme activity in animals and humans. Up to date, an extensive research of the enzyme status is done, in order to gain information for their importance in the biochemical processes in the organism, disturbances in the cell membrane transport, disease diagnosis and nutritional disorders.The influence of different stress factors and conditions upon enzyme activity both in serum and tissues is still one of the major topics in many researches. In the first week after hatch important physiological changes occur in chick organism. In the same time, in the first 24-48 hours, they are exposed to the influence of different stressors such as transportation to the hatchery along with feed and water deprivation, what gives the assumption that changes do occur in the serum enzyme activity in the first days after hatch. As literature data on the influence of food and water deprivation upon serum enzyme activity are poor and often very contradictory, this paper will try to contribute to the extensive research on the topic and clear some misunderstandings. For that purpose serum activities of the following enzymes were analysed: alanine aminotransferase, aspartat aminotarnsferase, gamma glutamile transferase, actate dehydrogenase and alkaline phosphatase.

  15. The investigation for the relationship among serum leptin, erythrocyte membrane Ca2+-ATPase activity and hypertensive disorder complicating pregnancy

    Institute of Scientific and Technical Information of China (English)

    Chunfang Li; Wenli Gou; Xuelian Chen; Shuping Zhang

    2007-01-01

    Objective: To study the significance of Leptin and the activity of erythrocyte membrane Ca2+-ATPase (EMCA) in the development of hypertensive disorder complicating pregnancy. Methods: Radioimmunoassay was used to test the level of serum Leptin,and the activity of EMCA was determined chemically in 38 pregnant women with hypertensive disorder complicating pregnancy and 36 normotensive pregnant women. Results: The level of serum Leptin in hypertensive disorder complicating pregnancy(gestational hypertension: 13.76 ± 3.46 ng/ml; preeclampsia:15.76 ± 5.47 ng/ml; eclampsia: 18.32 ± 6.38 ng/ml)was significantly higher than that in normotensive pregnant women (11.33 ± 2.93 ng/ml), respectively. The average EMCA activity of patients with hypertensive disorder complicating pregnancy (gestational hypertension: 1.65 ± 0.24 μmol· pi/mg·h; preeclampsia: 1.37 ± 0.19 μmol·pi/mg·h; eclampsia:1.12 ± 0.14 μ mol·pi/mg·h) was significantly lower than that of normotensive pregnant women(1.83 ±0.38 μ mol·pi/mg·h),respectively. There was a negative correlation between the level of serum Leptin and the activity of RMCA in hypertensive disorder complicating pregnancy (r = -0.63). Conclusion: Inhibition of EMCA activity of erythrocyte in hypertensive disorder complicating pregnancy may increase cytoplasmic free calcium, which contributes to the development of hypertensive disorder complicating pregnancy. The negative correlation between the level of serum Leptin and the activity of EMCA, also suggested that serum Leptin and the activity of EMCA may play a role in the development of hypertensive disorder complicating pregnancy.

  16. Serum hexosaminidase and ß-glucuronidase activities in infants: effects of age and sex

    Directory of Open Access Journals (Sweden)

    Mabe P.

    2003-01-01

    Full Text Available We investigated the effect of age and sex on the serum activity of hexosaminidase (HEX and ß-glucuronidase (BGLU in 275 normal term infants aged 12 h to 12 months. Up to six weeks of life, HEX was significantly higher in boys (P<=0.023. During the age period of 1-26 weeks, BGLU was also higher in boys, but differences were significant only at 2-6 and 7-15 weeks (P<=0.016. The developmental pattern of HEX and BGLU was sex dependent. HEX activity increased in both sexes from 4-7 days of life, reaching a maximum of 1.4-fold the birth value at 2-6 weeks of age in boys (P<0.001 and a maximum of 1.6-fold at 7-15 weeks in girls (P<0.001. HEX activity gradually decreased thereafter, reaching significantly lower levels at 27-53 weeks than during the first three days of life in boys (P = 0.002 and the same level of this age interval in girls. BGLU increased in both sexes from 4-7 days of age, showing a maximum increase at 7-15 weeks (3.3-fold in boys and 2.9-fold in girls, both P<0.001. Then BGLU decreased in boys to a value similar to that observed at 4-7 days of age. In girls, BGLU remained elevated until the end of the first year of life. These results indicate a variation of HEX and BGLU activities during the first year of life and a sex influence on their developmental pattern. This observation should be considered in the diagnosis of GM2 gangliosidosis and mucopolysaccharidosis type VII.

  17. Serum thymic factor, FTS, attenuates cisplatin nephrotoxicity by suppressing cisplatin-induced ERK activation.

    Science.gov (United States)

    Kohda, Yuka; Kawai, Yoshiko; Iwamoto, Noriaki; Matsunaga, Yoshiko; Aiga, Hiromi; Awaya, Akira; Gemba, Munekazu

    2005-11-01

    Serum thymic factor (FTS), a thymic peptide hormone, has been reported to attenuate the bleomycin-induced pulmonary injury and also experimental pancreatitis and diabetes. In the present study, we investigated the effect of FTS on cis-diamminedichloroplatinum II (cisplatin)-induced nephrotoxicity. We have already demonstrated that cephaloridine, a nephrotoxic antibiotic, leads to extracellular signal-regulated protein kinase (ERK) activation in the rat kidney, which probably contributes to cephaloridine-induced renal dysfunction. The aim of this study was to examine the effect of cisplatin on ERK activation in the rat kidney and also the effect of FTS on cisplatin-induced nephrotoxicity in rats. In vitro treatment of LLC-PK1 cells with FTS significantly ameliorated cisplatin-induced cell injury. Treatment of rats with intravenous cisplatin for 3 days markedly induced renal dysfunction and increased platinum contents in the kidney cortex. An increase in pERK was detected in the nuclear fraction prepared from the rat kidney cortex from days 1 to 3 after injection of cisplatin. FTS suppressed cisplatin-induced renal dysfunction and ERK activation in the kidney. FTS did not influence any Pt contents in the kidney after cisplatin administration. FTS has been shown to enhance the in vivo expression of heat shock protein (HSP) 70 in the kidney cortex. The beneficial role of FTS against cisplatin nephrotoxicity may be mediated in part by HSP70, as suggested by its up-regulation in the kidney cortex treated with FTS alone. Our results suggest that FTS participates in protection from cisplatin-induced nephrotoxicity by suppressing ERK activation caused by cisplatin.

  18. Effects of Continuous High Temperature on Digestive Enzyme and Lysozyme Activities of Young GIFT Oreochromis niloticus%持续高温对吉富罗非鱼幼鱼消化酶和溶菌酶活力的影响

    Institute of Scientific and Technical Information of China (English)

    刘加慧; 杨洪帅; 王辉

    2013-01-01

    通过人工控温,对吉富罗非鱼(GIFT Oreochromis niloticus)幼鱼实施34℃持续高温处理,研究持续高温对吉富罗非鱼幼鱼消化酶和溶菌酶活力的影响。结果表明:持续高温对吉富罗非鱼幼鱼消化酶活力的影响有统计学意义(P0.05);随着时间的推进,脂肪酶、淀粉酶活力增加,差异有统计学意义(P0.05)。表明持续高温能一定程度地提高罗非鱼的消化能力和免疫功能。%The young GIFT Oreochromis niloticus were put in water under 34℃ and the effects of continuous high temperature on digestive enzyme and lysozyme activities of young GIFT O. niloticus was studied. The results showed that effects of continuous high temperature on digestive enzyme activity of young GIFT O. niloticus had statistical significance (P0.05). With time, the activities of lipase and amylase were significantly increased, the difference had statistical significance (P 0.05). This experiment shows that continuous high temperature can ,to a certain extent, improve the digestive ability and immune function of tilapia, and provide theoretical basis for feed ratio of tilapia under continuous high temperature and prevention and treatment of diseases.

  19. Influence of Bovine Serum Albumin on the Antibacterial Activity of Endodontic Irrigants against Enterococcus Faecalis

    Science.gov (United States)

    Khedmat, Sedigheh; Aligholi, Marziyeh; Sadeghi, Samaneh

    2009-01-01

    INTRODUCTION: It has been demonstrated that organic content of the root canals can influence the antimicrobial capability of chemical irrigants. The aim of this study was to evaluate the effect of bovine serum albumin (BSA), as an organic material, on the antimicrobial activity of several intracanal irrigants. MATERIALS AND METHODS: Bactericidal activity of Ethylenediaminetetraacetic acid (EDTA) 17%, citric acid 10%, Sodium hypochlorite (NaOCl) 5.25%, Chlorhexidine 0.2% (CHX), Smear Clear and Cetrimide 0.5% were tested by means of dilution-neutralization method. Contact times were 10 and 30 seconds, 5, 10, 30, 60 minutes and 24 hours. First 950 λ of the medicament was mixed with 50 λ of the bacterial suspension in an Eppendorf test tube. The suspensions were thoroughly mixed. Sterile water served as negative controls. After each contact time, 100 λ of samples was transferred to the Eppendorf test tubes which contained neutralizers. After 5 minutes, 50 λ of serial dilutions were cultured on brain heart infusion agar and incubated in aerobic conditions. Then colonies were counted and reported as cfu/mL. In half of the samples, medicaments were suspended in BSA 0.5% 30 minutes before examination to assess its possible inhibitory effect on the antibacterial activity. RESULTS: NaOCl 5.25%, Cetrimide 0.5% and Smear Clear showed bactericidal activity within seconds after the incubation. BSA had no inhibitory effect on bactericidal activity of these three medicaments. CHX took 5 and10 minutes to kill all bacterial cells in the absence and presence of BSA, respectively. Citric acid and EDTA showed the least antibacterial activity. CONCLUSION: In this study, NaOCl 5.25%, Cetrimide 0.5% and Smear Clear were significantly more effective against E. faecalis than EDTA 17% and citric acid 10% in the presence and absence of BSA. Also, in the presence of BSA, bactericidal activity of CHX 0.2% against E. faecalis was significantly more than EDTA after 10 and 30 minutes of

  20. 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.

  1. Serum HMGB1 Serves as a Novel Laboratory Indicator Reflecting Disease Activity and Treatment Response in Ankylosing Spondylitis Patients

    Science.gov (United States)

    Miao, Ye; Huang, Yishu; Sun, Mengchen; Zhu, Yingzi; Zheng, Fang

    2016-01-01

    Objective. High mobility group box 1 (HMGB1) is a late inflammatory factor participating in the pathogenesis of various autoimmune and inflammatory diseases. In the current study, we analyzed the association between serum levels of HMGB1 and clinical features of AS patients before and during treatment. Methods. Serum HMGB1 was detected in 147 AS patients and 61 healthy controls using ELISA. We evaluated the association between HMGB1 and extra-articular manifestations as well as disease severity indices. Among these AS patients, 41 patients received close follow-up at 1, 3, and 6 months after treatment. This group comprised 25 patients treated with anti-TNF-α biologics and 16 patients receiving oral NSAIDs plus sulfasalazine. Results. The serum HMGB1 of AS patients was significantly higher than in healthy controls and positively correlated with BASDAI, BASFI, ASDAS-ESR, ASDAS-CRP, ESR, and CRP, but not with HLA-B27, anterior uveitis, and recurrent diarrhea. There was no significant difference between patients with radiographic damage of hip joints and those without. We observed that serum HMGB1 paralleled disease activity after treatment. Conclusion. Serum level of HMGB1 is higher in AS patients, and to some extent, HMGB1 can reflect the activity of AS and be used as a laboratory indicator to reflect the therapeutic response.

  2. Serum HMGB1 Serves as a Novel Laboratory Indicator Reflecting Disease Activity and Treatment Response in Ankylosing Spondylitis Patients

    Directory of Open Access Journals (Sweden)

    Chenqiong Wang

    2016-01-01

    Full Text Available Objective. High mobility group box 1 (HMGB1 is a late inflammatory factor participating in the pathogenesis of various autoimmune and inflammatory diseases. In the current study, we analyzed the association between serum levels of HMGB1 and clinical features of AS patients before and during treatment. Methods. Serum HMGB1 was detected in 147 AS patients and 61 healthy controls using ELISA. We evaluated the association between HMGB1 and extra-articular manifestations as well as disease severity indices. Among these AS patients, 41 patients received close follow-up at 1, 3, and 6 months after treatment. This group comprised 25 patients treated with anti-TNF-α biologics and 16 patients receiving oral NSAIDs plus sulfasalazine. Results. The serum HMGB1 of AS patients was significantly higher than in healthy controls and positively correlated with BASDAI, BASFI, ASDAS-ESR, ASDAS-CRP, ESR, and CRP, but not with HLA-B27, anterior uveitis, and recurrent diarrhea. There was no significant difference between patients with radiographic damage of hip joints and those without. We observed that serum HMGB1 paralleled disease activity after treatment. Conclusion. Serum level of HMGB1 is higher in AS patients, and to some extent, HMGB1 can reflect the activity of AS and be used as a laboratory indicator to reflect the therapeutic response.

  3. Serum amylase activity and renal amylase activity clearance in patients with severely impaired renal function and in patients treated with renal allotransplantation.

    Science.gov (United States)

    Pedersen, E B; Brock, A; Kornerup, H J

    1976-03-01

    Serum amylase activity was measured in 29 nondialysed patients with severe renal failure, in 24 uraemic patients treated with chronic haemodialysis, and in 29 patients treated with renal allotransplantation. Simultaneous measurement of renal amylase activity clearance (CAm) and creatinine clearance (CCr) was performed in 25 patients with severe renal failure and in 19 transplanted patients. Serum amylase activity was elevated in all three groups. CAm was significantly correlated to CCr both in the group with severe renal failure and in the transplanted group. Unlike in the group of transplanted patients, the ratio CAm/CCr was significantly increased in patients with severe impaired renal function. It is concluded that the elevation of serum amylase activity in patients with impaired renal function is primarily due to decreased glomerular filtration rate. The value of CAm/CCr for diagnosing acute pancreatitis is doubtful in patients with severe renal disease.

  4. Serum soluble Talin-1 levels are elevated in patients with multiple sclerosis, reflecting its disease activity.

    Science.gov (United States)

    Muto, Mayumi; Mori, Masahiro; Liu, Jia; Uzawa, Akiyuki; Uchida, Tomohiko; Masuda, Hiroki; Ohtani, Ryohei; Sugimoto, Kazuo; Kuwabara, Satoshi

    2017-04-15

    Previously, we identified anti-Talin-1 antibodies in the serum of MS. In this case, we measured the serum soluble Talin-1 (sTalin-1) levels by enzyme-linked immunosorbent assay. The serum sTalin-1 levels were significantly higher in 40 patients with MS than in 43 normal controls and in the acute phase of disease than in the remission phase. Interestingly, serum sTalin-1 levels were associated with a sustained increase in disability after MS attack but not with serum anti-Talin-1 antibody levels. sTalin-1 may be a biomarker for the acute phase of MS and may be used for the short-term prognosis of MS. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Species differences in avian serum B esterases revealed by chromatofocusing and possible relationships of esterase activity to pesticide toxicity.

    Science.gov (United States)

    Thompson, H M; Mackness, M I; Walker, C H; Hardy, A R

    1991-04-15

    Serum cholinesterase (BChE) and carboxylesterase (CbE) activities were investigated in ten species of birds. Multiple forms of serum BChE and CbE were also separated by chromatofocusing. Higher CbE activity and a wider range of CbE and BChE forms were present in the sera of omnivorous/herbivorous birds than carnivores. Omnivores/herbivores studied were the starling, house sparrow, tree sparrow, pigeon, partridge and magpie. Serum CbE activities of these species ranged from 0.46 to 2.93 mumol/min/mL with 2-6 forms separated by chromatofocusing. 0-6 forms of BChE were separated by the same method. The serum CbE activities of the little owl, tawny owl, barn owl and razorbill ranged from 0.19 to 0.58 mumoles/min/mL with 0-2 forms separated by chromatofocusing. No ChE forms were present within the pH gradient. These results may be significant in contributing to the understanding of the selective toxicity of organophosphorus and carbamate pesticides.

  6. Relation between serum xenobiotic induced receptor activities and sperm DNA damage and sperm apoptotic markers in European and Inuit populations

    DEFF Research Database (Denmark)

    Long, Manhai; Stronati, Alessanda; Bizzaro, Davide;

    2007-01-01

    -mediated luciferase reporter gene expression. Sperm DNA damage was measured using terminal deoxynucleotidyl transferase-driven dUTP nick labeling assay (TUNEL) and pro- (Fas) and anti-apoptotic (Bcl-xL) markers were determined by immune methods. Different features of xenobiotic-induced receptor activity in serum...

  7. Molecular cloning, inducible expression and antibacterial analysis of a novel i-type lysozyme (lyz-i2) in Pacific white shrimp, Litopenaeus vannamei.

    Science.gov (United States)

    Chen, Ting; Ren, Chunhua; Wang, Yanhong; Luo, Peng; Jiang, Xiao; Huang, Wen; Chen, Chang; Hu, Chaoqun

    2016-07-01

    The full-length cDNA coding for a novel invertebrate (i-type) lysozyme was identified in Pacific white shrimp (Litopenaeus vannamei). The newly obtained L. vannamei lysozyme is similar to the Penaeus monodon i-type lysozyme 2, but it is distant from the known L. vannamei c-type lysozyme and i-type lysozyme 1 in protein sequence; therefore, it was defined as L. vannamei i-type lysozyme 2 (lyz-i2). Expression of L. vannamei lyz-i2 transcripts were ubiquitously detected in all tissues we selected, with the highest abundance observed in the hemolymph. Challenge with Vibrio harveyi might elicit L. vannamei lyz-i2 mRNA expression in the hepatopancreas, intestine, muscle, gill and hemolymph. In the themolymph, specifically, the stimulatory effects of Vibrio and lipopolysaccharide (LPS) on lyz-i2 transcript levels were durable and transient, respectively; while Polyinosinic:polycytidylic acid [Poly (I:C)] treatment did not affect lyz-i2 expression. L. vannamei lyz-i2 recombinant protein was generated in an Escherichia coli system. By lysoplate and turbidimetric assays, the L. vannamei lyz-i2 recombinant protein showed a broad spectrum of antimicrobial properties with high activities against Micrococcaceae lysodeikticus and various Vibrio species and relatively low activity against E. coli. In conclusion, L. vannamei lyz-i2 might be a potent antibacterial protein with a role in innate immunity in Penaeid shrimp.

  8. [Activity of cathepsin D in the blood serum and urine of patients with cancer of the stomach, pancreas and liver].

    Science.gov (United States)

    Szajda, Sławomir Dariusz; Snarska, Jadwiga; Roszkowska-Jakimiec, Wiesława; Waszkiewicz, Napoleon; Siedlecka, Katarzyna; Zwierz, Krzysztof; Krupkowska, Agnieszka

    2006-12-01

    Cathepsin D is a protease involved in invasion of the cancer and metastasis formation. The purpose of the study was to evaluate a prognostic value of cathepsin D activity in blood serum and urine of patients with cancer of the stomach, pancreas and liver. The study was carried out on the samples of blood serum and urine obtained from patients with cancer of the stomach, pancreas and liver treated surgically at the First Department of General Surgery and Endocrinology of the Medical University of Białystok. The control group consisted of healthy individuals. Activity of cathepsin D was determined in serum and urine by the Folin-Ciocaltau method with the cupric modification and was expressed in nmol Tyr/ml/6h. Specific activity of cathepsin D was determined in the urine, and was expressed in nmol Tyr/mg of protein/6h. Protein concentration in serum was assessed with Lowry et al. method and results were expressed in mg/ml. A significant increase in activity of cathepsin D in serum (p = 0.0169) and urine (p = 0.0008) and an enhanced specific activity in the urine (p = 0.0085) was found in patients with cancer of the pancreas as compared with the controls. A significantly increased activity of cathepsin was revealed in serum (p = 0.0233) of patients with cancer of the stomach. No significant differences of cathepsin D activity were found in urine of the patients with cancer of the stomach when compared to the controls. Additionally, an upward tendency (almost two-fold increase) of cathepsin D activity was shown in blood serum and an increase in the activity and in specific activity was observed in urine of both patients with cancer of the liver in comparison with the healthy individuals. There were no significant differences in the activity of cathepsin D in serum of the patients with cancer of the pancreas and stomach (p = 0.4156). A statistically significantly higher activity (p = 0.0004) and specific (0.0048) cathepsin D activity was demonstrated in urine of the

  9. Programmed cell death 5 correlates with disease activity and interleukin-17 in serum and synovial fluid of rheumatoid arthritis patients

    Institute of Scientific and Technical Information of China (English)

    WANG Jun-feng; GUAN Zhen-peng; ZHANG Shao-long; PEI Zheng; CHEN Ying-yu; PAN Huan

    2013-01-01

    Background Programmed cell death 5 (PDCD5) is a novel apoptotic regulatory gene that promotes apoptosis in various tumor cells.Studies have shown that PDCD5 accelerates the apoptosis of synoviocytes in vitro,implying a potential role in rheumatoid arthritis (RA) pathogenesis.This study examined the expression of PDCD5 in serum and synovial fluid of RA patients,its effect on the expression of inflammatory cytokine,interleukin-17 (IL-17),and the assessment of disease activity in RA.Methods PDCD5 and IL-17 levels in serum and synovial fluid from 18 patients with RA and 22 patients with osteoarthritis (OA) were detected using enzyme-linked immunosorbent assay (ELISA).Concentrations of serum PDCD5 in 40 healthy people were also detected as controls.As disease activity indices,C-reactive protein (CRP),erythrocyte sedimentation rate (ESR),rheumatoid factor (RF),and X-ray grading scale were also evaluated.Results Serum and synovial fluid PDCD5 levels in RA patients were significantly higher than those in OA and healthy controls.Serum PDCD5 level was inversely correlated to CRP and ESR,and was significantly higher in the RF negative group than in the positive group.PDCD5 level was also negatively correlated with IL-17 levels both in serum and synovial fluid of RA patients.However,differences in synovial fluid PDCD5 level from RA patients at different Larsen stages were not detectable.Conclusions PDCD5 affects RA pathogenesis.Insufficient apoptosis of fibroblast-like synoviocytes and inflammatory cells in RA could increase the expression of PDCD5 protein.As PDCD5 levels correlated negatively with disease activity indices and IL-17 level,PDCD5 could become a target in the diagnosis and treatment of RA.

  10. Effects of chronic renal failure rat serum on histone acetyltransferase p300 and activation of activating transcription factor 4 of arterial smooth muscle cells cultured in vitro

    Institute of Scientific and Technical Information of China (English)

    张耀全

    2014-01-01

    Objective To investigate the effects of the rat serum with chronic renal failure(CRF)on ubiquitin-proteasome pathway,histone acetyltransferase p300 and activation of activating transcription factor 4(ATF4)of rat arterial vascular smooth muscle cells(VSMCs)cultured in vitro,and explore the possible mechanism.Methods Objective To establish the rat model of

  11. Serum γ-glutamyltransferase, alanine aminotransferase, and aspartate aminotransferase activity in Iranian healthy blood donor men

    Institute of Scientific and Technical Information of China (English)

    Hossein Khedmat; Nasrin Zarei; Farahnaz Fallahian; Hassan Abolghasemi; Bashir Hajibeigi; Zohre Attarchi; Farshid Alaeddini; Mohammad Taghi Holisaz; Masoumeh Pourali; Shahin Sharifi

    2007-01-01

    AIM: To determine serum γ-glutamyltransferase (GGT), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activity, and to assess their correlation with demographic and clinical findings in healthy blood donors.METHODS: This cross-sectional study was performed in 934 male blood donors, aged 18 to 68 years, who consecutively attended Tehran blood transfusion service in 2006. All participants were seronegative for HBV or HCV infections, non alcohol users, and all underwent a standard interview and anthropometric tests. Clinical and biochemical parameters including AST, ALT, and GGT activities were determined. Patients taking drugs known to cause hepatic fat deposition were excluded. For AST, ALT, and GGT variables, we used 33.33 and 66.66 percentiles, so that each of them was divided into three tertiles.RESULTS: Mean AST, ALT, and GGT activities were 25.26 ± 12.58 U/L (normal range 5-35 U/L), 33.13 ± 22.98 (normal range 5-35 U/L), and 25.11 ± 18.32 (normal range 6-37 U/L), respectively. By univariate analyses, there were significant associations between increasing AST, ALT, or GGT tertiles and age, body weight, body mass index, and waist and hip circumferences (P < 0.05). By multiple linear regression analyses, ALT was found to be positively correlated with dyslipidemia (B = 6.988, P = 0.038), whereas ALT and AST were negatively correlated with age. AST, ALT, and GGT levels had positive correlation with family history of liver disease (B = 15.763, P < 0.001), (B = 32.345, P < 0.001), (B =24.415, P < 0.001), respectively.CONCLUSION: Although we did not determine the cutoffs of the upper normal limits for AST, ALT, and GGT levels, we would suggest screening asymptomatic patients with dyslipidemia and also subjects with a family history of liver disease.

  12. Conductivity and Viscosity Measurements for Binary Lysozyme Chloride Aqueous Solution and Ternary Lysozyme-Salt-Water Solution

    CERN Document Server

    Buzatu, D; Buzatu, F D

    2004-01-01

    We use the conductimetric method, adequate to electrolytes, to determine the lysozyme charge in lys-water and ternary lys-salt-water systems. We measured also the viscosities for the above binary and ternary systems in the same conditions at pH$=4.5$ and T$=298$ K, measurements that allow us to see any effect of viscosity on cations mobilities and implicitly on the lysozyme charge. The method is illustrated for the lysozyme chloride aqueous solution system at 25$^o$ C, using the data reported here for pH$=4.5$ at 0.15, 0.6, 0.8, 1., 1.5, 2., 2.5, 3., 3.5 mM (mg/mL) lysozyme chloride concentrations. The method was also applied to ternary lys-salt-water systems in the same conditions at pH$=4.5$ and T$=25^o$ C. Ternary conductivities are reported for a mean concentration 0.6 mM of lysozyme chloride in all systems and a mean concentration 0.01, 0.025, 0.05, 0.1, 0.175, 0.2, 0.5, 0.7, 0.9, 1.2, 1.3 and 1.4 M for NaCl; 0.005, 0.01, 0.05, 0.1, 0.175, 0.2, 0.5, 0.7, 0.9, 1.2, 1.3, 1.4 and 1.5 M for KCl; 0.005, 0.01,...

  13. Benefits of Omega-3 Fatty Acids Supplementation on Serum Paraoxonase 1 Activity and Lipids Ratios in Polycystic Ovary Syndrome

    Directory of Open Access Journals (Sweden)

    Elahe Mohammadi

    2012-12-01

    Full Text Available Background: Polycystic ovary syndrome (PCOS is a common endocrine disorder associated with increased risk of cardiovascular disease. The purpose of this study was to investigate the ef-fects of omega-3 fatty acids on serum paraoxonase 1 activity and lipids ratios in polycystic ovary syndrome.Methods: This double-blind randomized controlled clinical trial was conducted on 64 PCOS pa-tients with 20-35 years old. Thirty two of the subjects had taken 4 g/day omega -3 fatty acids and 32 patients were given placebo for 8 weeks. Fasting blood samples, anthropometric measure-ments and dietary intakes were collected at the beginning and the end of the study. Serum total cholesterol, triglyceride, and HDL-C were measured using the enzymatic methods. LDL-C con-centration was calculated by the Friedewald formula and arylesterase activity of serum PON1 was measured. Data were analyzed using SPSS software.Results: Omega-3 fatty acids significantly decreased TC/HDL-C and LDL-C/HDL-C ratios (P = 0.009 for both and significantly increased serum PON1 activity (P = 0.048 compared with placebo. Changes in TG/HDL-C ratio were not statistically significant in omega-3 fatty acids group at the end of the study in comparison to placebo group. Reduction in TC/HDL-C, LDL-C/HDL-C and TG/HDL-C ratios and increase in serum PON1 activity were also significant in omega-3 fatty acids group at the end of the study compared with baseline values (P <0.001, P < 0.001, P = 0.004, and P = 0.001, respectively.Conclusion: Omega-3 fatty acids may decrease the risk for cardiovascular disease through the improvement in paraxonase-1 activity and reduction in some lipids ratio in PCOS women.

  14. Peroxidase-coupled method for kinetic colorimetry of total creatine kinase activity in serum.

    Science.gov (United States)

    Wimmer, M C; Artiss, J D; Zak, B

    1985-10-01

    We describe a peroxidase-coupled method involving a colorimetric indicator reaction for determining the total activity of creatine kinase (EC 2.7.3.2) in serum. The kinetically favorable reverse reaction is exploited to generate adenosine 5'-triphosphate, which is used in the glycerol kinase-catalyzed phosphorylation of glycerol. The glycerol 3-phosphate so generated is oxidized in the presence of alpha-glycerophosphate oxidase to produce hydrogen peroxide, which is reduced in the presence of peroxidase with the simultaneous oxidation and coupling of 4-aminoantipyrene and 2-hydroxy-3,5-dichlorobenzenesulfonate to produce an intensely colored red chromogen. Results of the proposed method (y) correlate well with those of the Boehringer-Mannheim "CK-NAC UV" method as applied to the Hitachi 705 chemistry analyzer (y = 1.025 chi - 18.1, r = 0.9985, n = 100, range = 19-4531 U/L). The sensitivity of the method, based on molar absorptivities, is nearly fourfold that of procedures involving the reduction of NADP+.

  15. Serum bactericidal activity from intravenous ciprofloxacin and azlocillin given alone and in combination to healthy subjects.

    Science.gov (United States)

    Orlando, P L; Barriere, S L; Hindler, J A; Frost, R W

    1990-01-01

    Ciprofloxacin plus azlocillin have been shown to exhibit in vitro synergy versus a variety of organisms, including Pseudomonas aeruginosa. This study examined this interaction in vivo, testing serum bactericidal activity (SBA) in six healthy male subjects after intravenous administration of ciprofloxacin 4 mg/kg (C), azlocillin 60 mg/kg (A), and the two simultaneously (C/A). Eight different organisms were tested: four isolates of P. aeruginosa with varying susceptibilities to C and A, and one isolate each of Escherichia coli (EC), Staphylococcus aureus (SA) Serratia marcescens (SM), and Klebsiella pneumoniae (KP), all of which were susceptible to both drugs. Blood samples were collected at the end of 30-min infusions and at 4 and 8 hr. Reciprocal titers were plotted versus time and area under the bactericidal titer curve (AUBC) calculated to assess antibacterial interactions. Results indicated that P. aeruginosa-1 (PA-1), EC, and KP were synergistically killed by C/A. AUBC for PA-1 were C = 36, A = 11, C/A = 144, p less than 0.05. AUBC for EC were C = 1059, A = 180, C/A = 1504, p = 0.05. AUBC for KP were C = 327, A = 97, C/A = 584, p = 005. Additive effects were demonstrated versus all of the other organisms except Serratia marcescens, where an indifferent effect was observed. Ciprofloxacin plus azlocillin may be a useful combination of the treatment of selected Gram-negative bacillary infections.

  16. [Regularities of formation of chlorophyll-human serum albumin functionally active complexes in the aqueous medium].

    Science.gov (United States)

    Semichaevskiĭ, V D

    1975-01-01

    In the system with constant content of the chlorophyll a and increasing amounts of human serum albumin, dependence of pigment incorporation into the complex upon interaction of its aqueous associates with protein solutions was studied by applying the gel filtration on Sephadex G-75 and by measuring light scattering and rate of sensitized photoreduction of the methyl red by ascorbic-acid. The curves were obtained after extraction of the chlorophyll by acetone from dry pigment-protein films formed after desiccation of the aqueous systems. Sigmoid character of the above dependences, their linearization in Hill's coordinates and the value of cooperativity coefficient close to 2 testifies in favour of the cooperative character of the complex formation, two pigment molecules reacting with a single protein molecule. Measurement of adsorption isotherms and their treatment with use of the Brunauer-Emmett-Teller theory of polymolecular adsorption make it possible to evaluate the maximum molar ratio of the pigment to the protein in the complex (close to 2). The pigment-pigment interaction suggests that the chlorophyll molecules adsorbed on the protein are in the state of loosely packed dimers. Deaggregation of aqueus pigment associates by the protein in the course of complex formation results in a considerable increase of the protosensitizing chlorophyll activity.

  17. Estrogen receptor β regulates endometriotic cell survival through serum and glucocorticoid-regulated kinase activation.

    Science.gov (United States)

    Monsivais, Diana; Dyson, Matthew T; Yin, Ping; Navarro, Antonia; Coon, John S; Pavone, Mary Ellen; Bulun, Serdar E

    2016-05-01

    To determine the expression and biological roles of serum and glucocorticoid-regulated kinase (SGK1) in tissues and cells from patients with endometriosis and from healthy control subjects. Case-control. University research setting. Premenopausal women. Endometriotic tissues were obtained from women with ovarian endometriosis, and normal endometrial tissues were obtained from women undergoing hysterectomy for benign conditions. Expression levels of SGK1, the role of SGK1 in endometriosis pathology, and regulation of SGK1 by estrogen receptor (ER) β. Transcript and protein levels of SGK1 were significantly higher in endometriotic tissues and cells compared with normal endometrium. SGK1 mRNA and protein levels were stimulated by E2, by the ERβ-selective agonist diarylpropionitrile, and by prostaglandin E2. SGK1 was transcriptionally regulated by ERβ based on small interfering RNA knockdown and chromatin immunoprecipitation of ERβ followed by quantitative polymerase chain reaction. SGK1 knockdown led to increased cleavage of poly(ADP-ribose) polymerase, and SGK1 activation was correlated with the phosphorylation of FOXO3a, a proapoptotic factor. ERβ leads to SGK1 overexpression in endometriosis, which contributes to the survival of endometriotic lesions through inhibition of apoptosis. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  18. Study on the NO Concentration and NOS Activity in the Serum and Tissues of Ducklings with Selenium Poisoning

    Institute of Scientific and Technical Information of China (English)

    WU Rui; KANG Shi-liang; XU Shi-wen; WANG Wei; LIN Hong-jin

    2003-01-01

    One-day-old healthy ducklings (n=100) were divided into control group and experimental group randomly, and selenium poisoning of ducklings was artificially caused by feeding ration containing 8 lng kg-1 Se every day. The dynamic changes of No content and NOS activity in the serum and tissues were determined by means of the method of nitric acid reductase. Results showed that the NO content and NOS activity in serum and tissues in experimental group increased significantly (P<0.05) and they were time-dependent.It was suggested that the high level of selenium in bodies could increase the NOS activity and NO content as it destroyed the metabolism of material and energy as well as structure and function of tissues and cells. These changes of NO content and NOS activity might be involved in the metabolism of selenium poisoning.

  19. Meningococcal surface fibril (Msf) binds to activated vitronectin and inhibits the terminal complement pathway to increase serum resistance.

    Science.gov (United States)

    Griffiths, Natalie J; Hill, Darryl J; Borodina, Elena; Sessions, Richard B; Devos, Nathalie I; Feron, Christiane M; Poolman, Jan T; Virji, Mumtaz

    2011-12-01

    Complement evasion is an important survival strategy of Neisseria meningitidis (Nm) during colonization and infection. Previously, we have shown that Nm Opc binds to serum vitronectin to inhibit complement-mediated killing. In this study, we demonstrate meningococcal interactions with vitronectin via a novel adhesin, Msf (meningococcal surface fibril, previously NhhA or Hsf). As with Opc, Msf binds preferentially to activated vitronectin (aVn), engaging at its N-terminal region but the C-terminal heparin binding domain may also participate. However, unlike Opc, the latter binding is not heparin-mediated. By binding to aVn, Msf or Opc can impart serum resistance, which is further increased in coexpressers, a phenomenon dependent on serum aVn concentrations. The survival fitness of aVn-binding derivatives was evident from mixed population studies, in which msf/opc mutants were preferentially depleted. In addition, using vitronectin peptides to block Msf-aVn interactions, aVn-induced inhibition of lytic C5b-9 formation and of serum killing could be reversed. As Msf-encoding gene is ubiquitous in the meningococcal strains examined and is expressed in vivo, serum resistance via Msf may be of significance to meningococcal pathogenesis. The data imply that vitronectin binding may be an important strategy for the in vivo survival of Nm for which the bacterium has evolved redundant mechanisms.

  20. Serum bilirubin value predicts hospital admission in carbon monoxide-poisoned patients. Active player or simple bystander?

    Directory of Open Access Journals (Sweden)

    Gianfranco Cervellin

    Full Text Available OBJECTIVES: Although carbon monoxide poisoning is a major medical emergency, the armamentarium of recognized prognostic biomarkers displays unsatisfactory diagnostic performance for predicting cumulative endpoints. METHODS: We performed a retrospective and observational study to identify all patients admitted for carbon monoxide poisoning during a 2-year period. Complete demographical and clinical information, along with the laboratory data regarding arterial carboxyhemoglobin, hemoglobin, blood lactate and total serum bilirubin, was retrieved. RESULTS: The study population consisted of 38 poisoned patients (23 females and 15 males; mean age 39±21 years. Compared with discharged subjects, hospitalized patients displayed significantly higher values for blood lactate and total serum bilirubin, whereas arterial carboxyhemoglobin and hemoglobin did not differ. In a univariate analysis, hospitalization was significantly associated with blood lactate and total serum bilirubin, but not with age, sex, hemoglobin or carboxyhemoglobin. The diagnostic performance obtained after combining the blood lactate and total serum bilirubin results (area under the curve, 0.90; 95% CI, 0.81-0.99; p<0.001 was better than that obtained for either parameter alone. CONCLUSION: Although it remains unclear whether total serum bilirubin acts as an active player or a bystander, we conclude that the systematic assessment of bilirubin may, alongside lactate levels, provide useful information for clinical decision making regarding carbon monoxide poisoning.

  1. The Correlation of Serum IL-12B Expression With Disease Activity in Patients With Inflammatory Bowel Disease

    Science.gov (United States)

    Lee, Hye Won; Chung, Sook Hee; Moon, Chang Mo; Che, Xiumei; Kim, Seung Won; Park, Soo Jung; Hong, Sung Pil; Kim, Tae Il; Kim, Won Ho; Cheon, Jae Hee

    2016-01-01

    Abstract Genetic variants in IL12B, encoding the p40 subunit common in interleukin-12 (IL-12) and interleukin-23, were identified as the susceptibility loci for inflammatory bowel disease (IBD). This study aimed to identify the correlation of serum IL-12B expression with disease activity in patients with IBD and evaluate the possibility of IL-12B as a biomarker for assessing inflammatory status in IBD. A total of 102 patients with IBD, including 38, 32, and 32 patients with Crohn's disease (CD), ulcerative colitis (UC), and intestinal Behçet's disease (intestinal BD), respectively, were included. The clinical and laboratory data from the patients were collected at the time of serum IL-12B measurement. Serum IL-12B levels were measured using an enzyme-linked immunosorbent assay. The median IL-12B levels in patients with CD, UC, and intestinal BD were significantly higher than those in controls (1.87, 2.74, and 2.73 pg/mL, respectively, vs. 1.42 pg/mL, all P <0.05). IL-12B concentrations were associated with disease activity in patients with UC and intestinal BD but not in those with CD. IL-12B levels were increased with increasing disease activity in patients with UC (P <0.001). Likewise, patients with active intestinal BD had higher IL-12B levels than those without active disease (P = 0.008). IL-12B levels were correlated with the endoscopic disease activity of UC (P = 0.002) and intestinal BD (P = 0.001) but not that of CD. Serum IL-12B levels were significantly correlated with clinical and endoscopic disease activity in patients with UC and intestinal BD, suggesting its potential use as a biomarker for assessing disease activity in these patients. PMID:27281077

  2. 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.

  3. The disinfection performance and mechanisms of Ag/lysozyme nanoparticles supported with montmorillonite clay.

    Science.gov (United States)

    Jiang, Jing; Zhang, Chang; Zeng, Guang-Ming; Gong, Ji-Lai; Chang, Ying-Na; Song, Biao; Deng, Can-Hui; Liu, Hong-Yu

    2016-11-05

    The fabrication of montmorillonite (Mt) decorated with lysozyme-modified silver nanoparticles (Ag/lyz-Mt) was reported. The lysozyme (lyz) was served as both reducing and capping reagent. Coupling the bactericidal activity of the lyz with AgNPs, along with the high porous structure and large specific surface area of the Mt, prevented aggregation of AgNPs and promoted nanomaterial-bacteria interactions, resulting in a greatly enhanced bactericidal capability against both Gram positive and Gram negative bacteria. This paper systematically elucidated the bactericidal mechanisms of Ag/lyz-Mt. Direct contact between the Ag/lyz-Mt surface and the bacterial cell was essential to the disinfection. Physical disruption of bacterial membrane was considered to be one of the bactericidal mechanisms of Ag/lyz-Mt. Results revealed that Ag(+) was involved in the bactericidal activity of Ag/lyz-Mt via tests conducted using Ag(+) scavengers. A positive ROS (reactive oxygen species) scavenging test indirectly confirmed the involvement of ROS (O2(-), H2O2, and OH) in the bactericidal mechanism. Furthermore, the concentrations of individual ROS were quantified. Results showed that Ag/lyz-Mt nanomaterial could be a promising bactericide for water disinfection.

  4. Seasonal variations of 25-OH vitamin D serum levels are associated with clinical disease activity in multiple sclerosis patients.

    Science.gov (United States)

    Hartl, Christina; Obermeier, Viola; Gerdes, Lisa Ann; Brügel, Mathias; von Kries, Rüdiger; Kümpfel, Tania

    2017-04-15

    Low 25-hydroxy vitamin D (25-[OH]-D) serum concentrations have been associated with higher disease activity in multiple sclerosis (MS) patients. In a large cross-sectional study we assessed the vitamin D status in MS patients in relation to seasonality and relapse rate. 415 MS-patients (355 relapsing-remitting MS and 60 secondary-progressive, 282 female, mean age 39.1years) of whom 25-(OH)-D serum concentrations were determined at visits between 2010 and 2013 were included in the study. All clinical data including relapse at visit and expanded disability status scale were recorded in a standardized manner by an experienced neurologist. Seasonal variations of 25-(OH)-D serum concentrations were modelled by sinusoidal regression and seasonal variability in the prevalence of relapse by cubic regression. The mean 25-(OH)-D serum concentration was 24.8ng/ml (range 8.3-140ng/ml) with peak levels of 32.2ng/ml in July/August and nadir in January/February (17.2ng/ml). The lowest modelled prevalence of relapse was in September/October (28%) and the highest modelled prevalence in March/April (47%). The nadir of 25-(OH)-D serum concentrations preceded the peak in prevalence of relapses by two months. In summary, seasonal variation of 25-(OH)-D serum levels were inversely associated with clinical disease activity in MS patients. Future studies should investigate whether vitamin D supplementation in MS patients may decrease the seasonal risk for MS relapses.

  5. Spectroscopic analysis on the binding interaction of biologically active pyrimidine derivative with bovine serum albumin

    Directory of Open Access Journals (Sweden)

    Vishwas D. Suryawanshi

    2016-02-01

    Full Text Available A biologically active antibacterial reagent, 2–amino-6-hydroxy–4–(4-N, N-dimethylaminophenyl-pyrimidine-5-carbonitrile (AHDMAPPC, was synthesized. It was employed to investigate the binding interaction with the bovine serum albumin (BSA in detail using different spectroscopic methods. It exhibited antibacterial activity against Escherichia coli and Staphylococcus aureus which are common food poisoning bacteria. The experimental results showed that the fluorescence quenching of model carrier protein BSA by AHDMAPPC was due to static quenching. The site binding constants and number of binding sites (n≈1 were determined at three different temperatures based on fluorescence quenching results. The thermodynamic parameters, enthalpy change (ΔH, free energy (ΔG and entropy change (ΔS for the reaction were calculated to be 15.15 kJ/mol, –36.11 kJ/mol and 51.26 J/mol K according to van't Hoff equation, respectively. The results indicated that the reaction was an endothermic and spontaneous process, and hydrophobic interactions played a major role in the binding between drug and BSA. The distance between donor and acceptor is 2.79 nm according to Förster's theory. The alterations of the BSA secondary structure in the presence of AHDMAPPC were confirmed by UV–visible, synchronous fluorescence, circular dichroism (CD and three-dimensional fluorescence spectra. All these results indicated that AHDMAPPC can bind to BSA and be effectively transported and eliminated in the body. It can be a useful guideline for further drug design.

  6. Effective approach to greatly enhancing selective secretion and expression of three cytoplasmic enzymes in Escherichia coli through synergistic effect of EDTA and lysozyme.

    Science.gov (United States)

    Liu, Sen-Lin; Du, Kun; Chen, Wei-Zhao; Liu, Gang; Xing, Miao

    2012-09-01

    An effective approach to greatly enhancing the selective secretion and expression of recombinant cytoplasmic enzymes in Escherichia coli was successfully developed through the synergistic effect of ethylenediaminetetraacetate (EDTA) and lysozyme. The method was applied to two endoglucanases (EGs) and an amylase. The optimal culture conditions of temperature and concentration of isopropyl-β-D: -1-thiogalactopyranoside (IPTG) were 23-30 °C and 0.2 mM, respectively, under which the three enzymes could be expressed in active form. Among all the chemicals tested, EDTA was found to be most suitable for enhancing the secretion of EG-I-1A into the medium. Addition of lysozyme alone had little influence on the secretion and expression. In contrast, on the basis of the addition of 5 g EDTA/L at the induction time of 12 h, the simultaneous addition of 0.15 g lysozyme/L further significantly increased the secretion and expression of the three enzymes, demonstrating the synergistic effect of EDTA and lysozyme. The production of EG-I-1A in the culture medium by adding 5 g EDTA/L and 0.15 g lysozyme/L under the optimal culture conditions of 23 °C and 0.2 mM IPTG was over 260-fold higher than that without EDTA and lysozyme under the standard conditions of 37 °C and 1 mM IPTG. In summary, the advantage of this novel cultivation approach for secretion was that not only did it selectively enhance the secretion of the proteins of interest, but also greatly increased the expression of the three enzymes by over 80 %.

  7. Breast cancer proliferative activity: Is it the source of serum free DNA?

    African Journals Online (AJOL)

    Taha I. Hewala

    2013-04-13

    Apr 13, 2013 ... 165 Horria Avenue, El Hadara, Alexandria 21561, Egypt. Received 30 December ... serum of patients with various types of cancers including breast cancer.1 However, the .... to the QIAamp mini spin column. After washing, the ...

  8. Effects of Iron Supplementation and Activity on Serum Iron Depletion and Hemoglobin Levels in Female Athletes

    Science.gov (United States)

    Cooter, G. Rankin; Mowbray, Kathy W.

    1978-01-01

    Research revealed that a four-month basketball training program did not significantly alter serum iron, total iron binding capacity, hemoglobin, and percent saturation levels in female basketball athletes. (JD)

  9. 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.

  10. Influence of Rifampin Therapy on Serum Bactericidal Activity in the Presence of Cloxacillin and Vancomycin

    Directory of Open Access Journals (Sweden)

    Andrew MR Mackenzie

    1990-01-01

    Full Text Available In this study the effect of rifampin on serum inhibitory and serum bactericidal titres was examined. Sera were prepared from pooled human serum to contain vancomycin (10 mg/L, cloxacillin (5 mg/L or rifampin (1 mg/L, and the combinations cloxacillin/rifampin and vancomycin/rifampin. These five sera were tested by a microtitre method for serum inhibitory power and serum bactericidal titre against 11 strains of Staphylococcus aureus. A 48 h incubation period was required to detect full colony growth for subculture plates. It was found with all strains that the effect of the addition of rifampin to the other two antibiotics was to increase the serum inhibitory power, lower the serum bactericidal titre, increase the inhibitory/cidal ratio, and slow colony growth on subculture. In the clinical part of the study it was shown that only three of 38 sera (8% from patients receiving betalactam or vanomycin but not rifampin gave an inhibitory/cidal ratio greater than 8, but that nine of 10 sera (90% from patients receiving rifampin in addition to betalactam or vancomycin gave a ratio greater than 8 (P<0.001. The study verified that the effect of rifampin in serum was to increase inhibitory power and decrease bactericidal titre. The clinical significance of these results is not known and it is suggested that a high ratio of inhibitory to bactericidal titre in the presence of rifampin is to be expected, and that a low bactericidal titre under these circumstances is not necessarily an indication to modify therapy.

  11. Impact of antibacterial drugs on human serum paraoxonase-1(hPON1)activity:an in vitro study

    Institute of Scientific and Technical Information of China (English)

    Hakan; Syt; Elif; Duygu; Kaya; Skr; Beydemir

    2014-01-01

    Objective:To investigate the in vitro effects of the antihacterial drugs,mcropenem trihydrate.piperacillin sodium,and cefoperazone sodium,on the activity of human serum paraoxonase mPOND.Methods:hPQN1 was purified from human serum using simple chromatographic methods.including DEAE-Sephadex anion exchange and Sephadex G-200 gel filtration chromatography.Results:The three antihacterial drugs decreased in vitro hPON1 activity.Inhibition mechanisms meropcnem trihydrate was noncompetitive while piperacillin sodium and cefoperazone sodium were competitive.Conclusions:Our results showed that antihacterial drugs significantly inhibit hPON1 activity,both in vitro,with rank order meropenem trihydrate piperacillin sodium cefoperazone sodium in vitro.

  12. Effects of aerobic training on serum paraoxonase activity and its relationship with PON1-192 phenotypes in women

    Institute of Scientific and Technical Information of China (English)

    Gulbin Rudarli Nalcakan; S. Rana Varol; Faruk Turgay; Mesut Nalcakan; M. Zeki Ozkol; S. Oguz Karamizrak

    2016-01-01

    Background: Paraoxonase 1 (PON1) is an antioxidant enzyme that protects high-density lipoprotein (HDL) and low-density lipoprotein against oxidation. Limited studies have addressed the influenc of exercise on PON1 activity and its relationship with PON1 phenotypes. We investigated relationships between PON1-192 phenotypes, PON1 activity, aerobic exercise, and blood lipid and lipoprotein concentrations in middle-aged women. Methods: An exercise group (n=50) engaging in regular aerobic exercise and a control group (n=41) were selected from a subset of 300 Caucasian women that met the inclusion criteria. Serum PON1, salt-stimulated PON1 (SSPON1), and arylesterase (ARE) activities;cholesterol levels and ARE activities of total HDL and HDL subgroups (HDLs) (supernatants obtained by polyethylene glycol);and blood lipid and lipoprotein concentrations were determined by standardized enzymatic methods. PON1-192 QQ (low activity), QR (moderate activity), and RR (high activity) phenotype groups were define using serum SSPON1/ARE activity ratios. The R-carries (RC) phenotype group consisted of the QR and RR groups combined. Results: All lipid and lipoprotein concentrations were greater in the exercise group than in the control group. Regardless of phenotype, no significan differences were observed between the exercise and control groups in terms of serum PON1, SSPON1, or ARE activity associated with HDLs (p>0.05), whereas PON1 activities in QQ-phenotyped women in the exercise group were significant y higher than those in the control group (p Conclusion: These results showed that a regular aerobic exercise program can improve PON1 activity depending on PON1-192 phenotype, but not on lipid and lipoprotein levels, in middle-aged Turkish women.

  13. Bioassays for estrogenic activity: development and validation of estrogen receptor (ERalpha/ERbeta) and breast cancer proliferation bioassays to measure serum estrogenic activity in clinical studies.

    Science.gov (United States)

    Li, J; Lee, L; Gong, Y; Shen, P; Wong, S P; Wise, Stephen D; Yong, E L

    2009-02-01

    Standard estrogenic prodrugs such as estradiol valerate (E2V) and increasingly popular phytoestrogen formulations are commonly prescribed to improve menopausal health. These drugs are metabolized to numerous bioactive compounds, known or unknown, which may exert combinatorial estrogenic effects in vivo. The aim of this study is to develop and validate estrogen receptor (ER) alpha/ERbeta reporter gene and MCF-7 breast cancer cell proliferation bioassays to quantify serum estrogenic activities in a clinical trial setting. We measured changes in serum estrogenicity following ingestion of E2V and compared this to mass spectrometric measurements of its bioactive metabolites, estrone and 17beta-stradiol. ERalpha bioactivity of the 192 serum samples correlated well (R = 79%) with 17beta-estradiol levels, and adding estrone improved R to 0.83 (likelihood ratio test, P estrogenic activity and that these assays suggest that the Epimedium formulation tested is unlikely to exert significant estrogenic effects in humans.

  14. Laser ablation dynamics and production of thin films of lysozyme

    DEFF Research Database (Denmark)

    Schou, Jørgen; Canulescu, Stela; Matei, Andreea;

    at the Technical University of Denmark (DTU) 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......, there was a considerable ablation weight loss of lysozyme from each shot. This is the first time the ablation by fs-lasers of a protein has been recorded quantitatively. Films of lysozyme produced by fs-laser irradiation will be analysed by MALDI in order to explore if there also is a significant amount of intact...... molecules in the films for fs-laser deposition....

  15. Comparison of Deferoxamine, Activated Charcoal, and Vitamin C in Changing the Serum Level of Fe in Iron Overloaded Rats

    Directory of Open Access Journals (Sweden)

    Reza Ghafari

    2014-02-01

    Full Text Available Background: Iron is an essential mineral for normal cellular physiology but its overload can lead to cell injury. For many years, deferoxamine injection has been used as an iron chelator for treatment of iron overload. The aim of this study is to compare oral deferoxamine, activated charcoal, and vitamin C, as an absorbent factor of Fe, in changing the serum level of iron in iron overload rats. Methods: In this experimental study, all groups were administered 150 mg iron dextran orally by gavage. After eight hours, rats in the first group received oral deferoxamine while those in the second and third groups received oral activated charcoal 1 mg/kg and oral vitamin C 150 mg, respectively. Then, serum levels of iron ware measured in all rats. Results: The mean serum level of iron in rats that received oral deferoxamine was 258.11±10.49 µg/dl, whereas mean levels of iron in charcoal and vitamin C groups were 380.88±11.21 µg/dl and 401.22±13.28 µg/dl, respectively. None of the measurements were within safety limits of serum iron. Conclusion: It seems that oral deferoxamine per se may not help physicians in the management of cases presented with iron toxicity. Activated charcoal did not reduce serum iron significantly in this study and further investigations may be warranted to assess the potential clinical utility of its mixture with oral deferoxamine as an adjunct in the clinical management of iron ingestions.

  16. Environment sensitive fluorescent analogue of biologically active oxazoles differentially recognizes human serum albumin and bovine serum albumin: Photophysical and molecular modeling studies

    Science.gov (United States)

    Maiti, Jyotirmay; Biswas, Suman; Chaudhuri, Ankur; Chakraborty, Sandipan; Chakraborty, Sibani; Das, Ranjan

    2017-03-01

    An environment sensitive fluorophore, 4-(5-(4-(dimethylamino)phenyl)oxazol-2-yl)benzoic acid (DMOBA), that closely mimics biologically active 2,5-disubstituited oxazoles has been designed to probe two homologous serum proteins, human serum albumin (HSA) and bovine serum albumin (BSA) by means of photophysical and molecular modeling studies. This fluorescent analogue exhibits solvent polarity sensitive fluorescence due to an intramolecular charge transfer in the excited state. In comparison to water, the steady state emission spectra of DMOBA in BSA is characterized by a greater blue shift ( 10 nm) and smaller Stokes' shift ( 5980 cm- 1) in BSA than HSA (Stokes'shift 6600 cm- 1), indicating less polar and more hydrophobic environment of the dye in the former than the latter. The dye-protein binding interactions are remarkably stronger for BSA than HSA which is evident from higher value of the association constant for the DMOBA-BSA complex (Ka 5.2 × 106 M- 1) than the DMOBA-HSA complex (Ka 1.0 × 106 M- 1). Fӧrster resonance energy transfer studies revealed remarkably less efficient energy transfer (8%) between the donor tryptophans in BSA and the acceptor DMOBA dye than that (30%) between the single tryptophan moiety in HSA and the dye, which is consistent with a much larger distance between the donor (tryptophan)-acceptor (dye) pair in BSA (34.5 Å) than HSA (25.4 Å). Site specific competitive binding assays have confirmed on the location of the dye in Sudlow's site II of BSA and in Sudlow's site I of HSA, respectively. Molecular modeling studies have shown that the fluorescent analogue is tightly packed in the binding site of BSA due to strong steric complementarity, where, binding of DMOBA to BSA is primarily dictated by the van der Waals and hydrogen bonding interactions. In contrast, in HSA the steric complementarity is less significant and binding is primarily guided by polar interactions and van der Waals interactions appear to be less significant in the

  17. Synthesis of biological active thiosemicarbazone and characterization of the interaction with human serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Wangshu; Shi, Lei; Hui, Guangquan [College of Chemistry and Environmental Science, Henan Normal University, Xinxiang 453007 (China); Cui, Fengling, E-mail: fenglingcui@hotmail.com [College of Chemistry and Environmental Science, Henan Normal University, Xinxiang 453007 (China)

    2013-02-15

    The synthesis of a new biological active reagent, 2-((1,4-dihydroxy)-9,10-anthraquinone) aldehyde thiosemicarbazone (DHAQTS), was designed. The interaction between DHAQTS and HSA was studied by fluorescence spectroscopy in combination with molecular modeling under simulation of physiological conditions. According to the results of fluorescence measurements, the quenching mechanism was suggested to be static. The thermodynamic parameters are calculated by van't Hoff equation, which demonstrated that hydrophobic interactions are the predominant intermolecular forces stabilizing the complex. The number of binding sites (n) was calculated. Through the site marker competitive experiment, DHAQTS was confirmed to be located in site I of HSA. The binding distance r=2.83 nm between the donor HSA and acceptor DHAQTS was obtained according to Foerster's non-radiative energy transfer theory. The three-dimensional fluorescence spectral results showed the conformation and microenvironment of HSA changed in the presence of DHAQTS. The effects of common ions on the binding of DHAQTS to HSA were also evaluated. The experimental results were in agreement with the results obtained via a molecular docking study. - Highlights: Black-Right-Pointing-Pointer 2-((1,4-dihydroxy)-9,10-anthraquinone)aldehyde thiosemicarbazone (DHAQTS) was synthesized. Black-Right-Pointing-Pointer DHAQTS can quench the fluorescence of human serum albumin (HSA) by static quenching mechanism. Black-Right-Pointing-Pointer Hydrophobic interactions were the predominant intermolecular forces. Black-Right-Pointing-Pointer The competitive experiment was carried out to identify the DHAQTS binding site on HSA. Black-Right-Pointing-Pointer Three-dimensional spectra confirmed DHAQTS caused the conformational change of HSA.

  18. Effect of cholesterol on the properties of spray-dried lysozyme-loaded liposomal powders.

    Science.gov (United States)

    Charnvanich, Dusadee; Vardhanabhuti, Nontima; Kulvanich, Poj

    2010-06-01

    The influence of cholesterol (Chol) in the liposomal bilayer on the properties of inhalable protein-loaded liposomal powders prepared by spray-drying technique was investigated. Lysozyme (LSZ) was used as a model protein. Feed solution for spray drying was prepared by direct mixing of aqueous solution of LSZ with mannitol solution and empty liposome dispersions composed of hydrogenated phosphatidylcholine and Chol at various molar ratios. The spray-dried powders were characterized with respect to morphology, thermal property, and crystallinity using scanning electron microscopy, differential scanning calorimetry, and X-ray diffraction, respectively. Most formulations gave slightly aggregated, spherical particles, and percentage yields of the spray-dried powders decreased with increasing Chol content. Degree of particle aggregation depended on the powder composition. The powders spontaneously formed liposomes which efficiently entrapped LSZ after reconstitution with HEPES buffered saline (HBS) at 37 degrees C. Lysozyme entrapment efficiency and size distribution of the reconstituted liposomes were evaluated after the powders were reconstituted with HBS. Increasing Chol content resulted in a decrease in size of the reconstituted liposomes and an increase in entrapment efficiency of LSZ. These results correlated with thermal behaviors of the reconstituted liposomes. Biological activity of LSZ was not affected by the spray-drying process. It was also demonstrated that LSZ-loaded liposomal powders could be produced without the need to preload the LSZ into liposomes prior to spray-drying process.

  19. Relationship Between Equilibrium Forms of Lysozyme Crystals and Precipitant Anions

    Science.gov (United States)

    Nadarajah, Arunan

    1996-01-01

    Molecular forces, such as electrostatic, hydrophobic, van der Waals and steric forces, are known to be important in determining protein interactions. These forces are affected by the solution conditions and changing the pH, temperature or the ionic strength of the solution can sharply affect protein interactions. Several investigations of protein crystallization have shown that this process is also strongly dependent on solution conditions. As the ionic strength of the solution is increased, the initially soluble protein may either crystallize or form an amorphous precipitate at high ionic strengths. Studies done on the model protein hen egg white lysozyme have shown that different crystal forms can be easily and reproducibly obtained, depending primarily on the anion used to desolubilize the protein. In this study we employ pyranine to probe the effect of various anions on the water structure. Additionally, lysozyme crystallization was carried out at these conditions and the crystal form was determined by X-ray crystallography. The goal of the study was to understand the physico-chemical basis for the effect of changing the anion concentration on the equilibrium form of lysozyme crystals. It will also verify the hypothesis that the anions, by altering the bulk water structure in the crystallizing solutions, alter the surface energy of the between the crystal faces and the solution and, consequently, the equilibrium form of the crystals.

  20. Comparative insight into surfactants mediated amyloidogenesis of lysozyme.

    Science.gov (United States)

    Chaturvedi, Sumit K; Khan, Javed M; Siddiqi, Mohammad K; Alam, Parvez; Khan, Rizwan H

    2016-02-01

    Electrostatic and hydrophobic interactions have an important role in the protein aggregation. In this study, we have investigated the effect of charge and hydrophobicity of oppositely charged surfactants i.e., anionic (AOT and SDS) and cationic (CTAB and DTAB) on hen egg white lysozyme at pH 9.0 and 13.0, respectively. We have employed various methods such as turbidity measurements, Rayleigh light scattering, ThT, Congo red and ANS dye binding assays, far-UV CD, atomic force microscopy, transmission electron and fluorescence microscopy. At lower molar ratio, both anionic and cationic surfactants promote amyloid fibril formation in lysozyme at pH 9.0 and 13.0, respectively. The aggregation was proportionally increased with respect to protein concentration and hydrophobicity of surfactant. The morphology of aggregates at both the pH was fibrillar in structure, as visualized by dye binding and microscopic imaging techniques. Initially, the interaction between surfactants and lysozyme was electrostatic and then hydrophobic as investigated by ITC. This study demonstrates the crucial role of charge and hydrophobicity during amyloid fibril formation.

  1. Aggregation of silica nanoparticles directed by adsorption of lysozyme.

    Science.gov (United States)

    Bharti, Bhuvnesh; Meissner, Jens; Findenegg, Gerhard H

    2011-08-16

    The interaction of the globular protein lysozyme with silica nanoparticles of diameter 20 nm was studied in a pH range between the isoelectric points (IEPs) of silica and the protein (pH 3-11). The adsorption affinity and capacity of lysozyme on the silica particles is increasing progressively with pH, and the adsorbed protein induces bridging aggregation of the silica particles. Structural properties of the aggregates were studied as a function of pH at a fixed protein-to-silica concentration ratio which corresponds to a surface concentration of protein well below a complete monolayer in the complete-binding regime at pH > 6. Sedimentation studies indicate the presence of compact aggregates at pH 4-6 and a loose flocculated network at pH 7-9, followed by a sharp decrease of aggregate size near the IEP of lysozyme. The structure of the bridged silica aggregates was studied by cryo-transmission electron microscopy (cryo-TEM) and small-angle X-ray scattering. The structure factor S(q) derived from the scattering profiles displays characteristic features of particles interacting by a short-range attractive potential and can be represented by the square-well Percus-Yevick potential model, with a potential depth not exceeding 3k(B)T.

  2. Induction of phase 2 enzymes by serum oxidized polyamines through activation of Nrf2: effect of the polyamine metabolite acrolein.

    Science.gov (United States)

    Kwak, Mi-Kyoung; Kensler, Thomas W; Casero, Robert A

    2003-06-06

    The naturally occurring polycationic polyamines including putrescine, spermidine, and spermine play an important role in cell growth, differentiation, and gene expression. However, circulating polyamines are potential substrates for several oxidizing enzymes including copper-containing serum amine oxidase. These enzymes are capable of oxidizing serum polyamines to several toxic metabolites including aldehydes and H(2)O(2). In this study, we investigated the effects of polyamines as inducers of phase 2 enzymes and other genes that promote cell survival in a cell culture system in the presence of bovine serum. Spermidine and spermine (50 microM) increased NAD(P)H quinone oxidoreductase (NQO1) activity up to 3-fold in murine keratinocyte PE cells. Transcript levels for glutathione S-transferase (GST) A1, GST M1, NQO1, gamma-glutamylcysteine ligase regulatory subunit, and UDP-glucuronyltransferase 1A6 were significantly increased by spermidine and this effect was mediated through the antioxidant response element (ARE). The ARE from the mouse GST A1 promoter was activated about 9-fold by spermine and 5-fold by spermidine treatment, but could be inhibited by the amine oxidase inhibitor, aminoguanidine, suggesting that acrolein or hydrogen peroxide generated from polyamines by serum amine oxidase may be mediators for phase 2 enzyme induction. Elevations of ARE-luciferase expression and NQO1 enzyme activity by spermidine were not affected by catalase, while both were completely repressed by aldehyde dehydrogenase treatment. Direct addition of acrolein to PE cells induced multiple phase 2 genes and elevated nuclear levels of Nrf2, a transcription factor that binds to the ARE. Expression of mutant Nrf2 repressed the activation of the ARE-luciferase reporter by polyamines and acrolein. These results indicate that spermidine and spermine increase the expression of phase 2 genes in cells grown in culture through activation of the Nrf2-ARE pathway by generating the sulfhydryl

  3. Evaluating Soluble EMMPRIN as a Marker of Disease Activity in Multiple Sclerosis: Studies of Serum and Cerebrospinal Fluid

    Science.gov (United States)

    Kaushik, Deepak K.; Yong, Heather Y. F.; Hahn, Jennifer N.; Silva, Claudia; Casha, Steven; Hurlbert, R. John; Jacques, Francois H.; Lisak, Robert; Khan, Omar; Ionete, Carolina; Larochelle, Catherine; Prat, Alex; Bar-Or, Amit; Yong, V. Wee

    2016-01-01

    Extracellular matrix metalloproteinase inducer (EMMPRIN, CD147) is an inducer of matrix metalloproteinases and has roles in leukocyte activation and migration. We reported previously that in MS and its animal model, experimental autoimmune encephalomyelitis, cell surface-associated EMMPRIN was significantly elevated in leukocytes around inflammatory perivascular cuffs in the CNS. In this study we report that activated T-cells can secrete soluble form of EMMPRIN (sEMMPRIN) upon activation. As sEMMPRIN is also present in biological fluids, we determined whether sEMMPRIN is altered in the CSF and sera of MS subjects. Sera from individuals without neurological conditions served as controls, while CSFs collected from subjects undergoing discectomy, and without evidence of CNS pathology, were used as a comparator group. We found that serum levels of sEMMPRIN from clinically stable MS patients or other inflammatory conditions did not differ from control subjects. Paired serum and CSF samples demonstrated poor correlation of sEMMPRIN. Interestingly, sEMMPRIN levels were approximately 60% higher in CSFs compared to sera. sEMMPRIN CSF levels were significantly higher in secondary progressive compared to primary progressive subjects. Thus we conclude that measurement of sEMMPRIN in serum is not informative for disease activity in MS. The differential expression of sEMMPRIN in the CSF of primary and secondary progressive MS invites hypotheses of the still undefined roles of EMMPRIN in the CNS. PMID:27727297

  4. Effect of human serum albumin upon the permeabilizing activity of sticholysin II, a pore forming toxin from Stichodactyla heliantus.

    Science.gov (United States)

    Celedón, Gloria; González, Gustavo; Gulppi, Felipe; Pazos, Fabiola; Lanio, María E; Alvarez, Carlos; Calderón, Cristian; Montecinos, Rodrigo; Lissi, Eduardo

    2013-12-01

    Sticholysin II (St II) is a haemolytic toxin isolated from the sea anemone Stichodactyla helianthus. The high haemolytic activity of this toxin is strongly dependent on the red cell status and the macromolecule conformation. In the present communication we evaluate the effect of human serum albumin on St II haemolytic activity and its capacity to form pores in the bilayer of synthetic liposomes. St II retains its pore forming capacity in the presence of large concentrations (up to 500 μM) of human serum albumin. This effect is observed both in its capacity to produce red blood cells haemolysis and to generate functional pores in liposomes. In particular, the capacity of the toxin to lyse red blood cells increases in the presence of human serum albumin (HSA). Regarding the rate of the pore forming process, it is moderately decreased in liposomes and in red blood cells, in spite of an almost total coverage of the interface by albumin. All the data obtained in red cells and model membranes show that St II remains lytically active even in the presence of high HSA concentrations. This stubbornness can explain why the toxin is able to exert its haemolytic activity on membranes immersed in complex plasma matrixes such as those present in living organisms.

  5. Glycoprotein biosynthesis in calf kidney. Glycoprotein sialyltransferase activities towards serum glycoproteins and calf Tamm-Horsfall glycoprotein.

    Science.gov (United States)

    van Dijk, W; Lasthuis, A M; van den Eijnden, D H

    1979-04-18

    CMP-AcNeu:glycoprotein sialyltransltransltransltransltransferase of calf kidney cortex was characterized using serum glycoproteins and Tamm-Horsfall glycoprotein, obtained from calf urine, as acceptors. Native calf Tamm-Horsfall glycoprotein showed the best acceptor properties, followed by desialylated calf fetuin and desialylated human alpha 1-acid glycoprotein exhibiting V values of, respectively, 114, 63 and 41 nmol/h per g wet wt. of kidney cortex and Km values of 0.12, 0.16 and 0.26 mM glycoprotein acceptor. Desialylated ovine submaxillary mucine appeared to be a very poor acceptor. Tamm-Horsfall glycoprotein sialyltransferase could be distinguished from serum glycoprotein sialyltransferase by competition studies. In addition the two glycoprotein sialyltransferase activities showed different distributions over the three regions of the calf kidney: the ratios of the Tamm-Horsfall to serum glycoprotein sialyltransferase activities decreased from 3.3 in the cortex to 0.8 and 0.4 in the medulla and the papilla, respectively. It was concluded that in calf kidney at least two different sialyltransferases exist. The high cortical Tamm-Horsfall glycoprotein sialyltransferases activity corresponds markedly to the origin of the urinary Tamm-Horsfall glycoprotein, namely the distal part of the kidney tubule. Inactivation of glycoprotein sialyltransferase activity by preincubation at various temperatures and during storage at 0 degree C, could be reduced by the addition of CMP-AcNeu. The possible relevance towards the in vivo sialylation of this finding is discussed.

  6. [Effect of low-intensity 900 MHz frequency electromagnetic radiation on rat liver and blood serum enzyme activities].

    Science.gov (United States)

    Nersesova, L S; Petrosian, M S; Gazariants, M G; Mkrtchian, Z S; Meliksetian, G O; Pogosian, L G; Akopian, Zh I

    2014-01-01

    The comparative analysis of the rat liver and blood serum creatine kinase, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and purine nucleoside phosphorylase post-radiation activity levels after a total two-hour long single and fractional exposure of the animals to low-intensity 900 MHz frequency electromagnetic field showed that the most sensitive enzymes to the both schedules of radiation are the liver creatine kinase, as well as the blood serum creatine kinase and alkaline phosphatase. According to the comparative analysis of the dynamics of changes in the activity level of the liver and blood serum creatine kinase, alanine aminotransferase, aspartate aminotransferase and purine nucleoside phosphorylase, both single and fractional radiation schedules do not affect the permeability of a hepatocyte cell membrane, but rather cause changes in their energetic metabolism. The correlation analysis of the post-radiation activity level changes of the investigated enzymes did not reveal a clear relationship between them. The dynamics of post-radiation changes in the activity of investigated enzyme levels following a single and short-term fractional schedules of radiation did not differ essentially.

  7. The assessment of carotid intima media thickness and serum Paraoxonase-1 activity in Helicobacter pylori positive subjects

    Directory of Open Access Journals (Sweden)

    Akbas Halide S

    2010-08-01

    Full Text Available Abstract Background The role of inflammation in the pathogenesis and progression of atherosclerosis has been increasingly discussed. Although the seroepidemiological studies have suggested a relationship between Helicobacter pylori (H. pylori infection and atherosclerosis; the issue is still controversial. It is well known that abnormal lipid profil is related to atherosclerosis and the measurement of carotid-intima media thickness (CIMT is one of the surrogate marker of atherosclerosis. The serum concentration of high-density lipoprotein (HDL-C has been known to have an inverse correlation with the development of atherosclerosis. Paraoxonase-1 (PON1 is a major anti-atherosclerotic component of HDL-C. PON1 activity is related to lipid peroxidation and prospective cardiovascular risk. The aim of this study was to investigate CIMT and serum PON1 activities along with lipid parameters in H. pylori positive and negative subjects. Methods Thirty H. pylori positive subjects and thirty-one negative subjects were enrolled. H. pylori infection was diagnosed by the presence of positivity of stool H. pylori antigen test or Carbon 14 labeled urea breath test. Serum PON1 activity was measured spectrophotometrically. Traditional cardiovascular risk factors were investigated and laboratory analysis included measurement of serum triglycerides (TG, total cholesterol (TC, high-density lipoprotein (HDL-C and low-density lipoprotein cholesterol (LDL-C. We assessed CIMT by high-resolution ultrasound of both common carotid arteries. Results We found that the mean and maximum values of right and overall CIMT in H. pylori positive subjects were significantly thicker than those of H. pylori negative subjects. There was no significant differences in serum HDL-C, LDL-C, TC levels and TC/HDL-C ratios between two groups. Serum TG levels of H. pylori positive subjects were significantly higher than those of H. pylori negative subjects (p = 0.014. We found that PON1

  8. Molecular cloning and characterization of c-type lysozyme gene in orange-spotted grouper, Epinephelus coioides.

    Science.gov (United States)

    Wei, Shina; Huang, Youhua; Cai, Jia; Huang, Xiaohong; Fu, Jing; Qin, Qiwei

    2012-08-01

    Lysozymes are key proteins of the host innate immune system against pathogen infection. In this study, a c-type lysozyme gene (Ec-lysC) was cloned and characterized from orange-spotted grouper, Epinephelus coioides. The full-length Ec-lysC cDNA is composed of 533 bp and encodes a polypeptide of 144-residue protein with 94% identity to lysC of Kelp grouper, Epinephelus bruneus. The genomic DNA of Ec-lysC consists of 4 exons and 3 introns, with a total length of 1897 bp. Amino acid sequence alignment showed that Ec-lysC possessed conserved catalytic residues (Glu50 and Asp67) and "GSTDYGIFQINS" motif. RT-PCR results showed that Ec-lysC transcript was most abundant in head kidney and less in muscle. The expression of Ec-lysC was differentially up-regulated in head kidney after stimulation with lipopolysaccharide (LPS), Vibrio alginolyticus and Singapore grouper iridovirus (SGIV). Subcellular localization analysis revealed that Ec-lysC was distributed predominantly in the cytoplasm. The recombinant Ec-lysC (rEc-lysC) had lytic activities against Gram-positive bacteria Micrococcus lysodeikticus, Staphylococcus aureus, Streptococcus iniae and Gram-negative bacteria V. alginolyticus. The lysozyme acted on M. lysodeikticus cell walls as shown by scanning electron microscopy (SEM). Furthermore, overexpression of Ec-lysC in grouper cells delayed the occurrence of CPE induced by SGIV and inhibited the viral gene transcription significantly. Taken together, Ec-lysC might play an important role in grouper innate immune responses to invasion of bacterial and viral pathogens. C-type lysozyme gene from E. coioides (Ec-lysC) was identified and characterized.

  9. Effect of ethanol-water mixture on the structure and dynamics of lysozyme: A fluorescence correlation spectroscopy study

    Science.gov (United States)

    Chattoraj, Shyamtanu; Mandal, Amit Kumar; Bhattacharyya, Kankan

    2014-03-01

    Effect of ethanol-water mixture on the hydrodynamic radius (rH) and conformational dynamics of lysozyme has been studied by circular dichroism, emission spectra, and fluorescence correlation spectroscopy. For this purpose, the protein lysozyme is covalently labeled near the active site with a fluorescent probe, alexa 488. The ethanol molecules are sequestered near the hydrophobic tryptophan residues as indicated by the blue shift of the emission maximum of tryptophan. It is observed that both size (rH) and time constant of conformational relaxation (τR) of lysozyme oscillate with increase in ethanol concentration. The rH of the protein fluctuates from 19 Å in the native state, to a minimum of 13 Å, and a maximum of 29 Å. It is proposed that the oscillating behavior arises from competition between mutual interaction among protein, ethanol, and water. The fluorescence intensity fluctuates because of quenching of the fluorescence of the probe (alexa) by the free amino group of certain residues (e.g., tryptophan). Rate of inter-conversion (folding dynamics) between the open (fluorescent) and closed (non-fluorescent) form has been determined and is found to exhibit similar oscillation with variation in ethanol content.

  10. Synergistic actions of complement and lysozyme in clearance of Escherichia coli from amphioxus Branchiostoma belcheri

    Institute of Scientific and Technical Information of China (English)

    Guangfeng Wang; Shicui Zhang; Zhimeng Zhuang; Zhiping Wang

    2009-01-01

    Amphioxus appears to lack free circulating blood cells. How it clears invading pathogens from its body remains unknown to date. We demonstrate here that amphioxus Branchiostoma belcheri is capable of efficiently eliminating the invading bacterium Escherichia coil from its humorai fluid, and the complement and lysozyme are both involved in the elimination of the invading pathogen. Both the com-plement and lysozyme act in concert against the invading bacterium, but the complement appears to play a more dominant role than the lysozyme.

  11. A vote for robustness: Monitoring serum enzyme activity by thin-layer chromatography of dabsylated bradykinin products.

    Science.gov (United States)

    Bayer, Malte; König, Simone

    2017-09-05

    High-end analytical methods provide excellent data but may lack the robustness required in large analytical studies. In particular complex chemical matrices may cause difficulties and increase the need for extensive sample preparation. For screening of patients we thus developed a low-tech assay to monitor bradykinin degradation by serum proteases. The bradykinin concentration mirrors the activity of angiotensin-converting enzyme (ACE). Dabsylated bradykinin (DBK) and its labeled fragments DBK1-8 and DBK1-5 were visualized by thin-layer chromatography using only 3μL of serum. Lower DBK1-5 levels indicated reduced ACE activity due to medication (ACE-inhibitors) or disease. Provided that purified DBK is available, the assay protocol itself is very simple and does not require any expensive high-end equipment. Copyright © 2017 Elsevier B.V. All rights reserved.