Sample records for serum proteins including

  1. Protein Crystal Serum Albumin

    NASA Technical Reports Server (NTRS)

    1998-01-01

    As the most abundant protein in the circulatory system albumin contributes 80% to colloid osmotic blood pressure. Albumin is also chiefly responsible for the maintenance of blood pH. It is located in every tissue and bodily secretion, with extracellular protein comprising 60% of total albumin. Perhaps the most outstanding property of albumin is its ability to bind reversibly to an incredible variety of ligands. It is widely accepted in the pharmaceutical industry that the overall distribution, metabolism, and efficiency of many drugs are rendered ineffective because of their unusually high affinity for this abundant protein. An understanding of the chemistry of the various classes of pharmaceutical interactions with albumin can suggest new approaches to drug therapy and design. Principal Investigator: Dan Carter/New Century Pharmaceuticals

  2. A dietary pattern including nopal, chia seed, soy protein, and oat reduces serum triglycerides and glucose intolerance in patients with metabolic syndrome.

    PubMed

    Guevara-Cruz, Martha; Tovar, Armando R; Aguilar-Salinas, Carlos A; Medina-Vera, Isabel; Gil-Zenteno, Lidia; Hernández-Viveros, Isaac; López-Romero, Patricia; Ordaz-Nava, Guillermo; Canizales-Quinteros, Samuel; Guillen Pineda, Luz E; Torres, Nimbe

    2012-01-01

    Metabolic syndrome (MetS) is a health problem throughout the world and is associated with cardiovascular disease and diabetes. Thus, the purpose of the present work was to evaluate the effects of a dietary pattern (DP; soy protein, nopal, chia seed, and oat) on the biochemical variables of MetS, the AUC for glucose and insulin, glucose intolerance (GI), the relationship of the presence of certain polymorphisms related to MetS, and the response to the DP. In this randomized trial, the participants consumed their habitual diet but reduced by 500 kcal for 2 wk. They were then assigned to the placebo (P; n = 35) or DP (n = 32) group and consumed the reduced energy diet plus the P or DP beverage (235 kcal) minus the energy provided by these for 2 mo. All participants had decreases in body weight (BW), BMI, and waist circumference during the 2-mo treatment (P < 0.0001); however, only the DP group had decreases in serum TG, C-reactive protein (CRP), and AUC for insulin and GI after a glucose tolerance test. Interestingly, participants in the DP group with MetS and the ABCA1 R230C variant had a greater decrease in BW and an increase in serum adiponectin concentration after 2 mo of dietary treatment than those with the ABCA1 R230R variant. The results from this study suggest that lifestyle interventions involving specific DP for the treatment of MetS could be more effective if local foods and genetic variations of the population are considered. PMID:22090467

  3. Serum Protein Profile Alterations in Hemodialysis Patients

    SciTech Connect

    Murphy, G A; Davies, R W; Choi, M W; Perkins, J; Turteltaub, K W; McCutchen-Maloney, S L; Langlois, R G; Curzi, M P; Trebes, J E; Fitch, J P; Dalmasso, E A; Colston, B W; Ying, Y; Chromy, B A

    2003-11-18

    Background: Serum protein profiling patterns can reflect the pathological state of a patient and therefore may be useful for clinical diagnostics. Here, we present results from a pilot study of proteomic expression patterns in hemodialysis patients designed to evaluate the range of serum proteomic alterations in this population. Methods: Surface-Enhanced Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (SELDI-TOFMS) was used to analyze serum obtained from patients on periodic hemodialysis treatment and healthy controls. Serum samples from patients and controls were first fractionated into six eluants on a strong anion exchange column, followed by application to four array chemistries representing cation exchange, anion exchange, metal affinity and hydrophobic surfaces. A total of 144 SELDI-TOF-MS spectra were obtained from each serum sample. Results: The overall profiles of the patient and control samples were consistent and reproducible. However, 30 well-defined protein differences were observed; 15 proteins were elevated and 15 were decreased in patients compared to controls. Serum from one patient exhibited novel protein peaks suggesting possible additional changes due to a secondary disease process. Conclusion: SELDI-TOF-MS demonstrated dramatic serum protein profile differences between patients and controls. Similarity in protein profiles among dialysis patients suggests that patient physiological responses to end-stage renal disease and/or dialysis therapy have a major effect on serum protein profiles.

  4. Determination of serum and tissue levels of phenazines including clofazimine

    Microsoft Academic Search

    Robert O'Connor; J. F. O'Sullivan; Richard O'Kennedy

    1996-01-01

    A rapid and sensitive HPLC method is described for the analysis of synthetic phenazines, including clofazimine, from a variety of biological samples. Phenazines were extracted from serum, tissue and fat using a mixture of dichloromethane and sodium hydroxide. The drugs were then quantified on a reversed-phase C18 column using a mobile phase consisting of 594 ml of water, 400 ml

  5. Quantitating Metabolites in Protein Precipitated Serum Using NMR Spectroscopy

    PubMed Central

    2015-01-01

    Quantitative NMR-based metabolite profiling is challenged by the deleterious effects of abundant proteins in the intact blood plasma/serum, which underscores the need for alternative approaches. Protein removal by ultrafiltration using low molecular weight cutoff filters thus represents an important step. However, protein precipitation, an alternative and simple approach for protein removal, lacks detailed quantitative assessment for use in NMR based metabolomics. In this study, we have comprehensively evaluated the performance of protein precipitation using methanol, acetonitrile, perchloric acid, and trichloroacetic acid and ultrafiltration approaches using 1D and 2D NMR, based on the identification and absolute quantitation of 44 human blood metabolites, including a few identified for the first time in the NMR spectra of human serum. We also investigated the use of a “smart isotope tag,” 15N-cholamine for further resolution enhancement, which resulted in the detection of a number of additional metabolites. 1H NMR of both protein precipitated and ultrafiltered serum detected all 44 metabolites with comparable reproducibility (average CV, 3.7% for precipitation; 3.6% for filtration). However, nearly half of the quantified metabolites in ultrafiltered serum exhibited 10–74% lower concentrations; specifically, tryptophan, benzoate, and 2-oxoisocaproate showed much lower concentrations compared to protein precipitated serum. These results indicate that protein precipitation using methanol offers a reliable approach for routine NMR-based metabolomics of human blood serum/plasma and should be considered as an alternative to ultrafiltration. Importantly, protein precipitation, which is commonly used by mass spectrometry (MS), promises avenues for direct comparison and correlation of metabolite data obtained from the two analytical platforms to exploit their combined strength in the metabolomics of blood. PMID:24796490

  6. Comparison of biotin binding protein of pregnant rat serum with rat serum albumin

    Microsoft Academic Search

    P. B. Seshagiri; P. R. Adiga

    1989-01-01

    The purified biotin binding protein of pregnant rat serum was shown to be immunologically similar to rat serum albumin as\\u000a assessed by a sensitive radioimmunoassay. In radioimmunoassay for rat biotin binding protein, the binding of [125I] rat biotin binding protein to anti-chicken egg yolk biotin binding protein antibodies was displaced by both rat serum (10–100\\u000a nl) and purified rat serum

  7. Arsenic trioxide binding to serum proteins.

    PubMed

    Shooshtary, Sara; Behtash, Sima; Nafisi, Shohreh

    2015-07-01

    Arsenic trioxide (ATO) also known as Trisenox, is an anticancer chemotherapeutic drug which has been used in treating diagnosed and relapsed patients with acute promyelocytic leukemia (APL). Serum albumin is the most abundant of the proteins in blood plasma and is the major transporter for delivering several drugs in vivo. The current study was designed to evaluate the potential ability of human and bovine serum albumin for delivering arsenic trioxide. Therefore, interaction of arsenic trioxide with HSA and BSA was investigated in aqueous solution at physiological conditions using a constant protein concentration and various drug contents. FTIR and UV-Vis spectroscopic methods were used to analyze arsenic trioxide and protein binding modes, the binding constants and the effect of drug complexation on HSA and BSA stability and conformation. Results of this study showed that drug complexation altered protein conformation by major reduction of ?-helix and increase of turn structure which is indicative of a partial protein destabilization. Structural analysis revealed that arsenic trioxide bind HSA and BSA with overall binding constants of KATO-HSA=1.07 (±0.01)×10(4)M(-1) and KATO-BSA=1.27(±0.02)×10(4)M(-1). It could be concluded that serum albumins can be considered as good carriers for delivering arsenic trioxide to target tissue. PMID:25863441

  8. Electrophoretic serum protein fractions in dwarf and normal layer hens

    E-print Network

    Paris-Sud XI, Université de

    , belonging to 7 sire families, were compared for serum protein profiles, using a Polyacrylamide gel, dwarfism, serum protein, egg production. Résumé Fractions électrophorétiques des protéines du sérum chezNote Electrophoretic serum protein fractions in dwarf and normal layer hens and their correlations

  9. Serum protein concentrations in Plasmodium falciparum malaria.

    PubMed

    Graninger, W; Thalhammer, F; Hollenstein, U; Zotter, G M; Kremsner, P G

    1992-12-01

    In patients with uncomplicated Plasmodium falciparum infection cytokine-mediated serum protein levels of C-reactive protein (CRP), coeruloplasmin (COE), beta 2-microglobulin (B2M), alpha 1-acid glycoprotein (AAG), alpha 1-antitrypsin (AAT), haptoglobin (HPT), prealbumin (PRE), retinol binding protein (RBP), albumin (ALB) and transferrin (TRF) were measured in an endemic area of the Amazonian rain forest. Semi-immune (SI) and nonimmune (NI) patients were investigated. In both patient groups the serum concentrations of CRP, COE and B2M were elevated on admission. In addition AAG and AAT concentrations were increased in NI patients compared to control subjects. Significantly lower serum concentrations of HPT, PRE, RBP, ALB and TRF were seen in both patient groups during the acute phase of the disease, and were more pronounced in NI patients. After a 28-day follow-up, AAT and B2M were normal in SI patients but HPT, AAT and B2M were still significantly altered in NI patients. PMID:1283805

  10. Sertoli cells secrete both testis-specific and serum proteins.

    PubMed Central

    Wright, W W; Musto, N A; Mather, J P; Bardin, C W

    1981-01-01

    The secretions of the Sertoli cell were examined with two polyvalent antisera--one prepared against proteins in rat serum and the other against testis-specific proteins in rete testis fluid. These antisera detected 12 serum and 9 testis-specific proteins in rete testis fluid. To determine the origin of these proteins, primary cultures enriched in Sertoli cells were incubated with [35S]methionine, and the radiolabeled proteins in the medium were immunoprecipitated. Gel electrophoresis of the two immunoprecipitates resolved eight serum and nine testis-specific proteins. These two sets of proteins were specifically bound to their respective antiserum and were immunologically distinct. Medium from Sertoli cell cultures contained 10 times more of the testis-specific proteins than did cultures enriched for testicular myoid or interstitial cells. The concentration of the serum proteins in Sertoli cell medium was 5 and 10 times greater, respectively, than in myoid or interstitial cell preparations. The proteins from Sertoli cells were next characterized on two-dimensional gels. Seven of the proteins recognized by antiserum against serum proteins had identical molecular weights and isoelectric points as serum proteins. Three of these proteins were ceruloplasmin, transferrin, and glycoprotein 2. In addition to the proteins immunoprecipitated by the two antisera, more than 60 other proteins were detected on two-dimensional gels of the total secretory proteins. We conclude that the Sertoli cell secretes many proteins, some of which are specific to the testis and others of which are similar to serum proteins. Images PMID:6950398

  11. Serum protein and zinc levels in patients with thoracic empyema

    Microsoft Academic Search

    M. Erkan Balkan; Hilal Özgüne?

    1996-01-01

    The element Zn is the metal component or activator of many important enzymes. The tissue concentrations and activities of\\u000a Zn metalloenzymes direct the rate of protein and nucleic acid syntheses, thereby influencing tissue growth and reperative\\u000a processes. Most of the serum Zn is normally bound to circulating proteins. Low serum Zn concentrations might result from depletion\\u000a of Zn-binding proteins. Serum

  12. Multiplexed microfluidic quantification of proteins in serum

    NASA Astrophysics Data System (ADS)

    Rajan, Nitin; Rajauria, Sukumar; Cleland, Andrew

    2015-03-01

    Rapid and low cost immunoassays targeting proteins in blood or other bodily fluids are highly sought after for point-of-care devices and early screening of patients. Immunoturbidimetric assays utilize latex particles functionalized with antibodies, with particle aggregation in the presence of the analyte detected by a change in absorbance. Using a high throughput micro-fluidic particle analyzer based solely on electrical signals (resistive pulse sensing), we are able to accurately quantify the degree of aggregation by analyzing the changes in the particle size distribution. Thus we study the aggregation of streptavidin (SAv) coated beads in the presence of biotinylated bovine serum albumin as a proof-of-principle assay and extract the binding capacity of the SAv beads from the dose-response curve. We also use our aggregation measurement platform to characterize a commercial C-reactive protein (CRP) immunoturbidimetric assay (hsCRP, Diazyme Inc.). We obtain a linear calibration curve as well as a better limit of detection of CRP than that obtained by absorbance measurements. By using different bead sizes functionalized with different antibodies, multiplexed analyte detection is also possible. We demonstrate this by combining the commercial anti-CRP functionalized beads (0.4 microns) with biotin coated beads (1.0 microns), and carry out the simultaneous detection of SAv and CRP in a single sample.

  13. Interpretation of Serum Calcium in Patients with Abnormal Serum Proteins

    Microsoft Academic Search

    R. B. Payne; A. J. Little; R. B. Williams; J. R. Milner

    1973-01-01

    Two hundred consecutive specimens received in this laboratory for “liver function tests” showed a wide range of abnormal protein concentrations. Calcium concentration correlated closely with albumin (r = 0·867) but less closely with total protein (r = 0·682). A simple formula for adjusting calcium concentration was derived from the regression equation of calcium on albumin. Adjusted calcium = calcium -

  14. Discovery and fine mapping of serum protein loci through transethnic meta-analysis

    Microsoft Academic Search

    Nora Franceschini; Frank J A van Rooij; Bram P Prins; Mary F Feitosa; Mahir Karakas; John H Eckfeldt; Aaron R Folsom; Jeffrey Kopp; Ahmad Vaez; Jeanette S Andrews; Jens Baumert; Vesna Boraska; Linda Broer; Caroline Hayward; Julius S Ngwa; Yukinori Okada; Ozren Polasek; Harm-Jan Westra; Ying A Wang; Fabiola Del Greco M; Nicole L Glazer; Karen Kapur; Ido P Kema; Lorna M Lopez; Arne Schillert; Albert V Smith; Cheryl A Winkler; Lina Zgaga; Stefania Bandinelli; Sven Bergmann; Mladen Boban; Murielle Bochud; Y D Chen; Gail Davies; Abbas Dehghan; Jingzhong Ding; Angela Doering; J Peter Durda; Luigi Ferrucci; Oscar H Franco; Lude Franke; Grog Gunjaca; Albert Hofman; Fang-Chi Hsu; Ivana Kolcic; Aldi Kraja; Michiaki Kubo; Karl J Lackner; Lenore Launer; Laura R Loehr; Guo Li; Christa Meisinger; Yusuke Nakamura; Christine Schwienbacher; John M Starr; Atsushi Takahashi; Vesela Torlak; Andr'e G Uitterlinden; Veronique Vitart; Melanie Waldenberger; Philipp S Wild; Mirna Kirin; Tanja Zeller; Tatijana Zemunik; Qunyuan Zhang; Andreas Ziegler; Stefan Blankenberg; Eric Boerwinkle; Ingrid B Borecki; Harry Campbell; Ian J Deary; Timothy M Frayling; Christian Gieger; Tamara B Harris; Andrew A Hicks; Wolfgang Koenig; Christopher J O' Donnell; Caroline S Fox; Peter P Pramstaller; Bruce M Psaty; Alex P Reiner; Jerome I Rotter; Igor Rudan; Harold Snieder; Toshihiro Tanaka; Cornelia M van Duijn; Peter Vollenweider; Gerard Waeber; James F Wilson; Jacqueline C M Witteman; Bruce H R Wolffenbuttel; Alan F Wright; Qingyu Wu; Yongmei Liu; Nancy S Jenny; Kari E North; Janine F Felix; Behrooz Z Alizadeh; L Adrienne Cupples; John R B Perry; Andrew P Morris

    2012-01-01

    Many disorders are associated with altered serum protein concentrations, including malnutrition, cancer, and cardiovascular, kidney, and inflammatory diseases. Although these protein concentrations are highly heritable, relatively little is known about their underlying genetic determinants. Through transethnic meta-analysis of European-ancestry and Japanese genome-wide association studies, we identified six loci at genome-wide significance (p 5 × 10(-8)) for serum albumin (HPN-SCN1B, GCKR-FNDC4, SERPINF2-WDR81,

  15. Fractionation of Serum Proteins by Zone Electrophoresis in Glass Powder

    Microsoft Academic Search

    C. J. Bradish; Nan V. Smart

    1954-01-01

    IN connexion with recent electrophoretic studies of ox serum1, it became necessary to establish a reliable method for the fractionation of serum proteins which would allow numerous samples to be separated into distinct fractions of sufficient volume and concentration for subsequent physico-chemical and biological characterization. Zone electrophoresis2 appeared to be the method of choice, and the apparatus here described is

  16. Serum protein pattern in ewe with pregnancy toxemia

    Microsoft Academic Search

    Gul Fatma Yarim; Gulay Ciftci

    2009-01-01

    Pregnancy toxemia is a metabolic disease of pregnant ewes which causes significant economic losses in sheep industry. The\\u000a pathophysiology and metabolic changes of this disorder remain poorly understood. We conducted this study to describe the serum\\u000a protein pattern associated with the pregnancy toxemia in ewes. In this study, the electrophoretic pattern of serum proteins\\u000a of 15 ewes with naturally occuring

  17. Spectroscopic imaging of serum proteins using quantum cascade lasers.

    PubMed

    Mukherjee, Anadi; Bylund, Quentin; Prasanna, Manu; Margalit, Yotam; Tihan, Tarik

    2013-03-01

    First measurements of biomedical imaging using quantum cascade lasers (QCL) are presented. We report spectroscopic imaging of serum proteins using QCLs as an example for monitoring surface biocontamination. We found that dry smears of human serum can be spectroscopically imaged, identified, and quantified with high sensitivity and specificity. The core parts of the imaging platform consist of optically multiplexing three QCLs and an uncooled microbolometer camera. We show imaging of human serum proteins at 6.1, 9.25, and 9.5 ?m QCLs with high sensitivity and specificity. The sensitivity limit of 3???g/cm² of the human serum spot was measured at an S/N=3.The specificity of human serum detection was measured at 99% probability at a threshold of 77???g/cm². We anticipate our imaging technique to be a starting point for more sophisticated biomolecular diagnostic applications. PMID:23515866

  18. Interaction of Serum Proteins with Surface of Hemodialysis Fiber Membranes

    NASA Astrophysics Data System (ADS)

    Afrin, Rehana; Shirako, Yuji; Kishimoto, Kikuo; Ikai, Atsushi

    2012-08-01

    The poly(vinyl pyrrolidone)-covered hydrophilic surface of hollow-fiber membranes (fiber membrane, hereafter) for hemodialysis was mechanically probed using modified tips on an atomic force microscope (AFM) with covalent crosslinkers and several types of serum protein. The retraction part of many of the force extension (F-E) curves obtained with AFM tips coated with serum albumin had a long and smooth extension up to 200-300 nm indicating forced elongation of poly(vinyl pyrrolidone) chains. When fibrinogen-coated tips were used, long extension F-E curves up to 500 nm with multiple peaks were obtained in addition to smooth curves most likely reflecting the unfolding of fibrinogen molecules. The results indicated that individual polymer chains had a significant affinity toward serum proteins. The adhesion frequency of tips coated with serum proteins was lower on the poly(vinyl pyrrolidone) surface than on the uncoated hydrophobic polysulfone surface.

  19. Detection of human serum proteins using Raman and SERS spectroscopy

    NASA Astrophysics Data System (ADS)

    Ruan, Qiuyong; Liao, Fadian; Lin, Juqiang; Liu, Nenrong; Lin, Jinyong; Zeng, Yongyi; Li, Ling; Huang, Zufang; Chen, Rong

    2014-09-01

    The use of normal Raman (NR) spectroscopy and surface enhanced Raman scattering (SERS) spectroscopy to analyze the biochemical information of human serum proteins and hence distinguish between normal and primary hepatic carcinoma (PHC) serum samples was investigated. The serum samples were obtained from patients who were clinically diagnosed with PHC (n=20) and healthy volunteers (n=20). All spectra were collected in the spectral range of 400-1800 cm-1 and analyzed through the multivariate statistical methods of principal component analysis (PCA). The results showed that both NR and SERS combined with PCA had good performance in distinguishing the human serum proteins between PHC patients and healthy volunteers with high sensitivity and specificity of 100%. And we can get more detail information of component and conformation of human serum proteins by considering NR and SERS spectrum. Our results support the concept again that serum protein Raman and SERS spectroscopy combined with PCA analysis both can become noninvasive and rapid diagnostic tools to detect the primary hepatic carcinoma.

  20. INFLUENCE OF X-RAY IRRADIATION UPON SERUM PROTEINS

    Microsoft Academic Search

    Shibata

    1960-01-01

    Changes of serum proteins of x irradiatsd rabbits were used, total ; protein and albumin decreased as the total dose increased. In the early period, ; alpha globuin increased and gamma globulin decreased, wbile in the later ; period, alpha globulin decreased and gamma globulin increased. More marked ; changes were observed after liver irradiation than after irradiation of the

  1. Erythropoietin binding protein from mammalian serum

    DOEpatents

    Clemons, G.K.

    1997-04-29

    Purified mammalian erythropoietin binding-protein is disclosed, and its isolation, identification, characterization, purification, and immunoassay are described. The erythropoietin binding protein can be used for regulation of erythropoiesis by regulating levels and half-life of erythropoietin. A diagnostic kit for determination of level of erythropoietin binding protein is also described. 11 figs.

  2. Erythropoietin binding protein from mammalian serum

    DOEpatents

    Clemons, Gisela K. (Berkeley, CA)

    1997-01-01

    Purified mammalian erythropoietin binding-protein is disclosed, and its isolation, identification, characterization, purification, and immunoassay are described. The erythropoietin binding protein can be used for regulation of erythropoiesis by regulating levels and half-life of erythropoietin. A diagnostic kit for determination of level of erythropoietin binding protein is also described.

  3. Magnetic nanoparticles-serum proteins bioconjugates for binding of irinotecan.

    PubMed

    Tamyurek, Ecem; Maltas, Esra; Bas, Salih Zeki; Ozmen, Mustafa; Yildiz, Salih

    2015-02-01

    The binding of irinotecan to serum proteins (hemoglobin, globulin and human serum albumin) was studied on the surface of epoxide modified superparamagnetic iron oxide nanoparticles (GPTS-SPIONs), which were synthesized by the coprecipitation of ferrous and ferric salts with NH4OH and then modified with [3-(2,3-epoxypropoxy)propyl] trimethoxy silane (GPTS) to obtain functional epoxide groups on the SPIONs' surface. Results were compared to find an alternative as drug carries system. Data showed that binding amount of human serum albumin (HSA), globulin (Glb) and hemoglobin (Hb) found to be as 44, 21.2 and 32.6 ?g per 20 mg of GPTS modified SPIONs, respectively. The thermal behavior of the serum protein-Ir interaction on GPTS-SPIONs was also studied by using thermo gravimetric analysis (TGA) technique and then the kinetic parameters for the thermal decomposition were determined using Horowitz-Metzger method. PMID:25445689

  4. Distribution of Serum Total Protein in Elderly Chinese

    PubMed Central

    Tian, Chang-Rong; Qian, Li; Shen, Xiao-Zhu; Li, Jia-Jing; Wen, Jiang-Tao

    2014-01-01

    The serum total protein levels of the elderly possibly decrease gradually with aging. However, serum total protein levels are not suitable as a uniform reference standard for the elderly at different ages and genders. Thus, we investigated the total serum protein distribution in different gender and age groups of 11,453 elderly individuals aged ?60 years and without liver or renal disease from Lianyungang, Jiangsu, China. The total protein levels (TPL) of these individuals exhibited normal distribution (Z?=?1.206, P?=?0.109), whereas the reference range (95% CI) was 54.1 g/L to 82.3 g/L. TPL was higher in females than in males for those aged between 60 and 75 years, whereas no significant difference was observed for those aged between 80 and 95 years. TPL was negatively correlated with age in males (r?=??0.1342, P<0.05), females (r?=??0.304, P<0.05), and the total group (r?=??0.2136, P<0.05). TPL also decreased with aging and showed a faster rate in women than in men. These results indicated that an appropriate range of serum total protein based on age and gender differences should be used for clinical applications. PMID:24967900

  5. Distribution of serum total protein in elderly Chinese.

    PubMed

    Tian, Chang-Rong; Qian, Li; Shen, Xiao-Zhu; Li, Jia-Jing; Wen, Jiang-Tao

    2014-01-01

    The serum total protein levels of the elderly possibly decrease gradually with aging. However, serum total protein levels are not suitable as a uniform reference standard for the elderly at different ages and genders. Thus, we investigated the total serum protein distribution in different gender and age groups of 11,453 elderly individuals aged ?60 years and without liver or renal disease from Lianyungang, Jiangsu, China. The total protein levels (TPL) of these individuals exhibited normal distribution (Z?=?1.206, P?=?0.109), whereas the reference range (95% CI) was 54.1 g/L to 82.3 g/L. TPL was higher in females than in males for those aged between 60 and 75 years, whereas no significant difference was observed for those aged between 80 and 95 years. TPL was negatively correlated with age in males (r?=?-0.1342, P<0.05), females (r?=?-0.304, P<0.05), and the total group (r?=?-0.2136, P<0.05). TPL also decreased with aging and showed a faster rate in women than in men. These results indicated that an appropriate range of serum total protein based on age and gender differences should be used for clinical applications. PMID:24967900

  6. Detection of serum proteins in the electrophoretic patterns of total proteins of mycoplasma cells.

    PubMed Central

    Yaguzhinskaya, O. E.

    1976-01-01

    The contamination of mycoplasma cell preparations by serum proteins originating from culture medium was studied. A. laidlawii and M. arthritidis cells were grown in the presence of [14C]-aminoacids, and the cells were washed with 0-9% NaC1 by threefold centrifugation. Total proteins of the washed cells were analysed by SDS gel electrophoresis. Coomassie-stained electrophoretic patterns were compared with autoradiographs of the same gels. The stained electrophoretic pattern of washed A. laidlawii grown without serum was identical with autoradiographs of the same cells grown without or with serum. That of washed A. laidlawii grown with serum differed from the corresponding autoradiography by the presence of extra protein bands I, II, III, and IV with molecular weights of over 160,000, 80,000-87,000, 55,000 and 25,000, respectively. The same extra bands were found in stained electrophoretic patterns of washed: (a) A. laidlawii cells grown without serum and mixed with serum in the stationary phase, (b) M. arthritidis cells, as compared with their autoradiographs, (c) serum precipitate. The bands III and IV may be due to the heavy and light chains of gamma-globulin, the band II might belong to transferrin or to some component of complement. Acidification of serum to pH 5 brought about 100-fold rise of amount of serum precipitate, the number of bands in the electrophoretic pattern of the precipitate being also increased. Stained electrophoretic patterns of cells purified by twofold centrifugation in step sucrose density gradient (1-20-1-27 g./cm.3 for A. laidlawii, and 1-15-1-25 for M. arthritidis) contained no extra bands and matched completely with their autoradiographs. It was concluded that contamination of washed mycoplasma cells by serum proteins is mainly due to co-precipitation of aggregated serum proteins together with cells during centrifugation rather than to adsorption of serum proteins on the cell surface. Images Plate 1 PMID:10333

  7. Homology of beta Lactoglobulin, Serum Retinol-Binding Protein, and Protein HC

    Microsoft Academic Search

    Syed Pervaiz; Keith Brew

    1985-01-01

    The milk protein beta -lactoglobulin has been extensively studied but its function has not been identified. A clue regarding the function of a protein can be obtained by discovering a genetic relationship with a protein of known function through comparisons of amino acid sequence. Such comparisons revealed that beta -lactoglobulin is similar to human serum retinol-binding protein and to another

  8. QSAR modeling of ?-lactam binding to human serum proteins

    Microsoft Academic Search

    L. Mark Hall; Lowell H. Hall; Lemont B. Kier

    2003-01-01

    The binding of beta-lactams to human serum proteins was modeled with topological descriptors of molecular structure. Experimental data was the concentration of protein-bound drug expressed as a percent of the total plasma concentration (percent fraction bound, PFB) for 87 penicillins and for 115 ß-lactams. The electrotopological state indices (E-State) and the molecular connectivity chi indices were found to be the

  9. SERUM PROTEIN PROFILES IN THE SUCKLING AND NON SUCKLING PIGLET

    E-print Network

    Paris-Sud XI, Université de

    SERUM PROTEIN PROFILES IN THE SUCKLING AND NON SUCKLING PIGLET: THE IMPORTANCE OF COLOSTRUM J PORCELET NOUVEAU-NE : IMPOR- TANCE DU COLOSTRUM. ― Le profil des protéines sériques a été étudié, qui ont reçu le colostrum, un profil de type adulte est observé dans les deux heures qui suivent la

  10. Changes in serum protein profiles of chickens with tibial dyschondroplasia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Differences in serum protein profiles were analyzed to identify biomarkers associated with a poultry leg problem named tibial dyschondroplasia (TD) that can cause lameness. We used a bead-based affinity matrix containing a combinatorial library of hexapeptides (ProteoMinerTM) to deplete high abundan...

  11. Serum Immune-Related Proteins are Differentially Expressed during Hibernation in the American Black Bear

    PubMed Central

    Chow, Brian A.; Donahue, Seth W.; Vaughan, Michael R.; McConkey, Brendan; Vijayan, Mathilakath M.

    2013-01-01

    Hibernation is an adaptation to conserve energy in the face of extreme environmental conditions and low food availability that has risen in several animal phyla. This phenomenon is characterized by reduced metabolic rate (?25% of the active basal metabolic rate in hibernating bears) and energy demand, while other physiological adjustments are far from clear. The profiling of the serum proteome of the American black bear (Ursus americanus) may reveal specific proteins that are differentially modulated by hibernation, and provide insight into the remarkable physiological adaptations that characterize ursid hibernation. In this study, we used differential gel electrophoresis (DIGE) analysis, liquid chromatography coupled to tandem mass spectrometry, and subsequent MASCOT analysis of the mass spectra to identify candidate proteins that are differentially expressed during hibernation in captive black bears. Seventy serum proteins were identified as changing by ±1.5 fold or more, out of which 34 proteins increased expression during hibernation. The majority of identified proteins are involved in immune system processes. These included ?2-macroglobulin, complement components C1s and C4, immunoglobulin ? and J chains, clusterin, haptoglobin, C4b binding protein, kininogen 1, ?2-HS-glycoprotein, and apoplipoproteins A-I and A-IV. Differential expression of a subset of these proteins identified by proteomic analysis was also confirmed by immunodetection. We propose that the observed serum protein changes contribute to the maintenance of the hibernation phenotype and health, including increased capacities for bone maintenance and wound healing during hibernation in bears. PMID:23825529

  12. Serum protein electrophoresis as a prognostic marker of chronic liver disease in dogs

    Microsoft Academic Search

    E Sevelius; M Andersson

    1995-01-01

    In a previous study in dogs with chronic liver disease it was found that a combination of clinical features and laboratory findings was useful in differentiating liver diseases but could not be used to evaluate a dog's prognosis. When an electrophoretic analysis of the serum proteins was included, marked decreases in the concentrations of albumin and the alpha-globulins alpha-1-antitrypsin and

  13. Elevation of serum riboflavin carrier protein in breast cancer.

    PubMed

    Rao, P N; Levine, E; Myers, M O; Prakash, V; Watson, J; Stolier, A; Kopicko, J J; Kissinger, P; Raj, S G; Raj, M H

    1999-11-01

    Presently available tumor markers have had a limited clinical impact. Riboflavin carrier protein (RCP) is an estrogen inducible protein that occupies a key position in riboflavin metabolism. Because other vitamin carrier proteins (VCP) have been shown to be overexpressed in patients with malignant disease, we evaluated serum RCP levels in patients with adenocarcinoma of the breast. In this prospective blinded study, patients with breast cancer, benign breast disease, and healthy controls were analyzed for RCP levels. Using a highly sensitive RIA, we observed that serum RCP levels were significantly elevated in women with breast cancer (n = 52) as compared with control subjects [n = 50; 6.06 +/- 7.27 ng/ml versus 0.70 +/- 0.19 ng/ml (mean +/- SD), respectively; P < 0.0001]. A serum RCP level of > or = 1.0 ng/ml was highly predictive of the presence of breast cancer, detecting 88% of tumors in stages I-II and 100% of tumors in stages III-IV. Overall, this RCP assay has a sensitivity of 92.3%, a specificity of 88%, a positive predictive value of 88.9%, and a negative predictive value of 91.7%. These results show increased serum levels of RCP in breast adenocarcinoma patients and suggest that RCP levels may be useful as a new marker for breast cancer. The positive predictive value in early-stage breast cancer suggests that the RCP assay may be a useful adjunct to present screening technology. PMID:10566553

  14. Serum proteins of wild turkey vultures (Cathartes aura).

    PubMed

    Apanius, V; Temple, S A; Bale, M

    1983-01-01

    We separated and identified the major serum proteins of turkey vultures (Cathartes aura): albumin, HDL, LDL, IgG and M, transferrin, alpha 2M and putatively, IgA, ceruloplasmin and fibronectin. Turkey vulture HDL and LDL had similar electrophorectic mobility and solution properties as those from gallinaceous birds. Turkey vulture IgG and M, and their subunits, had molecular weights comparable to other birds. Serum IgG and M levels in wild turkey vultures were within the range of values reported for domestic avian species. PMID:6661906

  15. The interaction of serum proteins with carbon nanotubes depend on the physicochemical properties of nanotubes.

    PubMed

    Du, Jiangfeng; Ge, Cuicui; Liu, Ying; Bai, Ru; Li, Denghua; Yang, Yanlian; Liao, Lifu; Chen, Chunying

    2011-11-01

    With more and more potential applications of carbon nanotubes (CNTs) in different fields, the risk of exposure to CNTs is increasing. The interaction between CNTs and protein in biological media can affect the way cells interact with, recognize and process the nanoparticles, and this has important implications for safety considerations. In this study, the interaction of single-walled and multiwall CNTs with various serum proteins was investigated. The adsorption kinetics of protein to CNTs was investigated and a semi-qualitative analysis was provided by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Matrix assisted laser desorption ionization/time of flight mass spectrometry (MALDI-TOF MS) was used to identify the protein species binding to CNTs and atomic force microscopy (AFM) was used to vividly demonstrate the adsorption model of protein on CNTs. All the experimental results showed that the adsorption capacity of CNTs for protein was highly dependent on the type, arrangement model, size and surface modification of CNTs. Significant quantity of proteins in serum could be quickly adsorbed by CNTs, mainly including albumin, prealbumin, transferrin, and immunoglobulin. Noncovalent functionalization of CNTs by polyethylene glycol (PEG) could decrease the protein adsorption on CNTs. These results provide crucial insights into human serum proteins binding to different kinds of CNTs, which is important for understanding the safe application of carbon nanotubes. PMID:22413351

  16. Serum C-reactive protein levels and falciparum malaria.

    PubMed

    Naik, P; Voller, A

    1984-01-01

    A microplate ELISA was developed to measure C-reactive protein (CRP) and it was used to establish the relationship between CRP levels and malaria. Highest serum CRP levels were found in African patients with high Plasmodium falciparum parastaemia. However, even African children with lower parasitaemia had higher CRP levels than others without parasitaemia. All African groups studied had CRP levels above those of a control UK group. PMID:6398534

  17. Serum proteins of Canada goose (Branta canadensis) subspecies

    USGS Publications Warehouse

    Morgan, R.P., II; Sulkin, S.T.; Henny, C.J.

    1977-01-01

    Serum proteins from nine subspecies of Canada Geese (Brunta canadensis) were analyzed through the use of column and slab acrylamide electrophoresis. Variation was minimal within a subspecies, although all the subspecies were closely related. B. c. leucopareia appeared to be the most distinct subspecies, while maxima and moffitti were the most similar. Our preliminary findings suggest that the electrophoresis techniques are sensitive enough to identify some of the subspecies; however, baseline data from breeding ranges of all subspecies are required.

  18. Detection of pancreatic cancer using serum protein profiling

    PubMed Central

    Velstra, Berit; Bonsing, Bert A; Mertens, Bart J; Burgt, Yuri E M; Huijbers, Anouck; Vasen, Hans; Mesker, Wilma E; Deelder, André M; Tollenaar, Rob A E M

    2013-01-01

    Background Currently, no suitable biomarkers for the early detection of pancreatic cancer (PC) are available. Proteins present in the serum could reflect a state of the disease. In this study, these profiles as a diagnostic marker for PC were evaluated. Methods Serum samples were obtained from PC patients (n = 50 calibration set, n = 39 validation set) and healthy volunteers (n = 110 and n = 75 respectively) according to a uniform standardized collection and processing protocol. For peptide and protein isolation, automated solid-phase extraction (SPE) with Weak Cation Exchange (WCX) magnetic beads (MB) was performed using a 96-channel liquid handling platform. Protein profiles were obtained by mass spectrometry (MS) and evaluated by linear discriminant analysis with double cross-validation. Results A discriminating profile for PC has been identified, with a sensitivity of 78% and a specificity of 89% in the calibration set with an area under the curve (AUC) of 90%. These results were validated with a sensitivity of 74% and a specificity of 91% (AUC 90%). Conclusion Serum profiles of healthy controls and PC can be discrimated between. Further research is warranted to evaluate specificity and whether this biosignature can be used for early detection in a high risk population. PMID:23458426

  19. p53 protein is absent from the serum of patients with lung cancer.

    PubMed Central

    Levesque, M. A.; D'Costa, M.; Diamandis, E. P.

    1996-01-01

    p53 protein, which accumulates intracellularly in over half of all human tumours, has also been reported to be present in the sera of patients with various malignancies, including lung cancer. Using a quantitative immunoassay, we measured p53 protein concentrations in 216 sera from 114 lung cancer patients of whom 75 provided matched lung tumour tissues, which were also assayed for p53 protein. p53 protein levels above the detection limit of 0.04 ng ml-1 were detected in only two sera from lung cancer patients (0.14 ng ml-1 and 0.27 ng ml-1), but not in any of 13 sera from non-malignant lung disease patients or in 100 sera from normal non-diseased individuals. The presence of these apparent traces of serum p53 protein concentrations could not be related either to the p53 protein expression status of the primary lung tumours or to the tumour stage, grade or histological type. By pretreating these two sera with anti-p53 antibody linked to solid phase, and by the addition of mouse serum to neutralise possible heterophilic antibodies, the signals arising from these sera were shown to be non-specific and possibly caused by heterophilic antibodies. We conclude that our data do not support previous reports of p53 protein in the sera of lung cancer patients. Since immunoassays are subject to numerous sources of interference in serum, including heterophilic antibodies, we suggest that the results of p53 protein analysis of serum specimens should be interpreted with caution. PMID:8912541

  20. Interactions of apomorphine with serum and tissue proteins

    SciTech Connect

    Smith, R.V.; Velagapudi, R.B.; McLean, A.M.; Wilcox, R.E.

    1985-05-01

    Physical and covalent interactions of apomorphine with serum and tissue proteins could influence the drug's disposition and pharmacological activities in mammals. Ultrafiltration, equilibrium dialysis, and ultraviolet spectrophotometric methods have been used to study the reversible binding of apomorphine to bovine, human, rat, and swine plasma proteins. The degree of binding was generally greater than 90%, but variations were noted in some instances on the basis of drug concentrations and pH over the range of 6.8-7.8. Incubation of (8,9-/sup 3/H2)apomorphine with bovine serum albumin led to retention of radioactivity and a stoichiometrically controlled released of tritium which arose from the reaction of an electrophilic drug oxidation product and protein, producing drug-protein conjugates. In vitro experiments with mouse striatal brain preparations indicated parallel covalent binding reactions. In vivo experiments in mice indicated accumulation of radioactivity in brain regions and other tissues following daily injections of (8,9-/sup 3/H2)apomorphine for 14 days. The physical and covalent interactions of apomorphine with mammalian tissue proteins could be the cause of longer disposition half-lives in mammals than those previously reported. The covalent interactions, in particular, may be important in elucidating the mechanism of apomorphine-induced behavioral effects in mice.

  1. Development of Gel-Filter Method for High Enrichment of Low-Molecular Weight Proteins from Serum

    PubMed Central

    Chen, Lingsheng; Zhai, Linhui; Li, Yanchang; Li, Ning; Zhang, Chengpu; Ping, Lingyan; Chang, Lei; Wu, Junzhu; Li, Xiangping; Shi, Deshun; Xu, Ping

    2015-01-01

    The human serum proteome has been extensively screened for biomarkers. However, the large dynamic range of protein concentrations in serum and the presence of highly abundant and large molecular weight proteins, make identification and detection changes in the amount of low-molecular weight proteins (LMW, molecular weight ? 30kDa) difficult. Here, we developed a gel-filter method including four layers of different concentration of tricine SDS-PAGE-based gels to block high-molecular weight proteins and enrich LMW proteins. By utilizing this method, we identified 1,576 proteins (n = 2) from 10 ?L serum. Among them, 559 (n = 2) proteins belonged to LMW proteins. Furthermore, this gel-filter method could identify 67.4% and 39.8% more LMW proteins than that in representative methods of glycine SDS-PAGE and optimized-DS, respectively. By utilizing SILAC-AQUA approach with labeled recombinant protein as internal standard, the recovery rate for GST spiked in serum during the treatment of gel-filter, optimized-DS, and ProteoMiner was 33.1 ± 0.01%, 18.7 ± 0.01% and 9.6 ± 0.03%, respectively. These results demonstrate that the gel-filter method offers a rapid, highly reproducible and efficient approach for screening biomarkers from serum through proteomic analyses. PMID:25723528

  2. Oxidative modification of serum proteins in multiple sclerosis.

    PubMed

    Sadowska-Bartosz, Izabela; Adamczyk-Sowa, Monika; Galiniak, Sabina; Mucha, Sebastian; Pierzchala, Krystyna; Bartosz, Grzegorz

    2013-11-01

    Multiple sclerosis (MS) has been demonstrated to involve oxidative stress and augmented glycoxidation. In this study, several markers of protein oxidative damage and glycoxidation have been compared in 14 relapsing remittent in MS (RRMS) patients without immunomodifying treatment, 10 patients in clinical relapse, and clinically stable patient groups treated with interferon ? 1a (18) , ? 1b (19) and glatiramer acetate (GA; 6) in relation to healthy subjects (12). The glycophore content was increased in RRSM patients without treatment and in patients treated with GA. The level of advanced protein oxidation products (AOPP) was increased in RRSM patients without treatment and in patients with clinical relapse. The level of protein carbonyls was elevated in RRSM patients without treatment and in patients treated with interferon ? 1b. The levels of dityrosine level and N'-formylkynureine were elevated in RRSM patients without treatment while serum protein thiol groups were decreased in RRSM patients in clinical relapse as well as RRMS patients treated with interferon ? 1a. Several markers of protein modification showed correlation with the C-reactive protein level and white blood cell count, suggesting that oxidative protein modifications are linked to the inflammatory processes in MS. Results of this study confirm the occurrence of protein oxidative and glycoxidative damage in MS and show that spectrophotometric and fluorimetric markers of this damage, especially the AOPP level, may be useful in monitoring oxidative stress in the course of therapy of MS. PMID:24036284

  3. Serum proteins in magnesium-deficient rat Y. RAYSSIGUIER P. LARVOR Y. AUGUSTI J. DURLACH

    E-print Network

    Paris-Sud XI, Université de

    Serum proteins in magnesium-deficient rat Y. RAYSSIGUIER P. LARVOR Y. AUGUSTI J. DURLACH (1 Cochin, Paris Summary. Hypoproteinemia appears early in the magnesium-deficient rat with a drop in serum. In the magnesium-deficient rat, we have shown serum protein modifications related to immunologic and allergic

  4. Elevated Serum Leptin Level Is Associated with Body Mass Index But Not with Serum C-reactive Protein and Erythrocyte Sedimentation Rate Values in Hemodialysis Patients

    PubMed Central

    Dervisevic, Amela; Subo, Anela; Avdagic, Nesina; Zaciragic, Asija; Babic, Nermina; Fajkic, Almir; Lepara, Orhan; Hadzovic-Dzuvo, Almira

    2015-01-01

    Objectives: Aim of the present study was to investigate serum concentration of leptin and its association with values of body mass index (BMI), C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR) in hemodialysis (HD) patients. Methods: This cross-sectional study included 60 HD patients (34 male, 26 female) and 30 age- and sex-matched (4 males, 26 females) apparently healthy subjects. Serum leptin concentration was determined by an enzyme-linked immunosorbent assay (ELISA). Serum CRP concentration was measured by means of particle-enhanced immunonephelometry. ESR value was determined by Western Green method. BMI was calculated as weight (kg) divided by height squared (m2). Results: Results have shown that median serum leptin concentration (30.65 ng/mL; 12.48-86.40 ng/mL) was statistically significantly higher in HD patients compared to median serum leptin concentration (15.75 ng/mL; 9.15-30.65 ng/mL) in the control group of healthy subjects (p<0.05). Likewise, median serum CRP concentration (5.5 mg/L; 1.93-8.9 mg/L) and median ESR value (57.5 mm/h; 40.5-77.0 mm/h) were significantly higher in HD patients compared to median serum CRP concentration (0.8 mg/L; 0.38-1.43 mg/L) (p<0.001) and median ESR value (10.0 mm/h; 6.5-14.0 mm/h) (p<0.001) determined in the control group. Statistically significant positive correlation was found between BMI values and serum leptin concentration in HD patients (rho=0.434; p<0.001). Positive, although not significant, correlation was observed between serum CRP and leptin levels in HD patients (rho=0.171; p>0.05). Negative correlation between ESR values and serum leptin concentrations in HD patients was determined but it was not statistically significant (rho= -0.029; p>0.05). Conclusions: Increased serum concentration of leptin as pro-inflammatory cytokine as well as elevated serum values of CRP and ESR indicate presence of systemic micro inflammation in HD patients. Results of the present study point to possible use of serum leptin concentration as an indicator of nutritional status in HD patients based on observed significant positive correlation between serum leptin concentrations and BMI values. However, absence of significant association between serum leptin and CRP levels as well as between serum leptin concentrations and ESR values in HD patients requires further investigation and clarification. PMID:26005385

  5. Inflammation and dietary protein intake exert competing effects on serum albumin and creatinine in hemodialysis patients

    Microsoft Academic Search

    George A Kaysen; Glenn M Chertow; Rohini Adhikarla; Belinda Young; Claudio Ronco; Nathan W Levin

    2001-01-01

    Inflammation and dietary protein intake exert competing effects on serum albumin and creatinine in hemodialysis patients.BackgroundCross-sectional studies have shown an inverse correlation between serum C-reactive protein (CRP) and serum albumin concentration in hemodialysis patients. The net effects of inflammation and dietary protein intake on nutritional markers over time are unknown.MethodsTo explore the effects of CRP and normalized protein catabolic rate

  6. Embryo culture in teratological surveillance and serum proteins in development. Progress report, 1979-1980

    SciTech Connect

    Klein, N.W.

    1980-07-01

    Research progress for the period 1979-1980 is reported. The feasibility of using rat embryo cultures to test the teratogenic activity of serum was studied. The mechanisms regulating the synthesis of serum proteins were investigated. (ACR)

  7. Improved medium and culture conditions for clonal growth with minimal serum protein and for enhanced serum-free survival of Swiss 3T3 cells

    Microsoft Academic Search

    Gary D. Shipley; Richard G. Ham

    1981-01-01

    Summary  Improved culture conditions have been developed that will support clonal growth of Swiss mouse embryo 3T3 cells at concentrations\\u000a of serum protein as low as 125?g\\/ml. Survival of the cells under completely protein-free conditions also is enhanced greatly.\\u000a The improvements that made these results possible include: (a) use of medium MCDB 402, which was developed specifically for\\u000a Swiss 3T3 cells

  8. Isotachophoretic Preconcentration of Cardiac Proteins from Human Serum

    NASA Astrophysics Data System (ADS)

    Dutta, Prashanta; Hossan, Mohammad; Jubery, Talukder; Bottenus, Danny; Ivory, Cornelius

    2011-11-01

    Cationic isotachophoresis (ITP) is used to concentrate and detect a cardiac biomarker, cardiac troponin I (cTnI), spiked into depleted human serum in a cascade poly(methyl methacrylate) (PMMA) microfluidic channel. PMMA microchannel was formed using solvent imprinting and temperature-assisted bonding. A 100x reduction in cross-sectional area is implemented by gradually decreasing the channel width and height to increase the sensitivity of ITP. The ITP was performed in peak mode with potassium ion as the leading electrolyte and hydronium ions as the terminating electrolyte. The cross-sectional area reductions in combination with ITP allowed visualization of lower concentrations of fluorescently labeled cTnI. Experimental results show that ITP can detect cTnI at initial concentrations as low as 46 ng/mL in the presence of human serum proteins, and ITP can obtain cTnI concentrations factors as high as 10,000. This demonstrates the detection of cTnI in depleted human serum at clinically relevant concentrations without the use of antibodies and relying solely on ITP in a cascade microchip.

  9. Identification of Protein Carbonyls in serum of the fetal and neonatal pig.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Oxidation of serum proteins leads to non-reversible carbonyl formation which alters their function and has long been associated with stress-related disease processes. The primary objective of this study was to identify oxidized serum protein biomarkers of metabolic stress in baby pigs. Protein carb...

  10. Sensitive detection of C-reactive protein in serum by immunoprecipitation-microchip capillary gel electrophoresis.

    PubMed

    Herwig, Ela; Marchetti-Deschmann, Martina; Wenz, Christian; Rüfer, Andreas; Redl, Heinz; Bahrami, Soheyl; Allmaier, Günter

    2015-06-01

    Sepsis represents a significant cause of mortality in intensive care units. Early diagnosis of sepsis is essential to increase the survival rate of patients. Among others, C-reactive protein (CRP) is commonly used as a sepsis marker. In this work we introduce immune precipitation combined with microchip capillary gel electrophoresis (IP-MCGE) for the detection and quantification of CRP in serum samples. First high-abundance proteins (HSA, IgG) are removed from serum samples using affinity spin cartridges, and then the remaining proteins are labeled with a fluorescence dye and incubated with an anti-CRP antibody, and the antigen/antibody complex is precipitated with protein G-coated magnetic beads. After precipitation the complex is eluted from the beads and loaded onto the MCGE system. CRP could be reliably detected and quantified, with a detection limit of 25ng/?l in serum samples and 126pg/?l in matrix-free samples. The overall sensitivity (LOQ=75ng/?l, R(2)=0.9668) of the method is lower than that of some specially developed methods (e.g., immune radiometric assay) but is comparable to those of clinically accepted ELISA methods. The straightforward sample preparation (not prone to mistakes), reduced sample and reagent volumes (including the antibodies), and high throughput (10 samples/3h) are advantages and therefore IP-MCGE bears potential for point-of-care diagnosis. PMID:25778394

  11. Serum antibody immunoreactivity to equine zona protein after SpayVac vaccination.

    PubMed

    Mask, Tracy A; Schoenecker, Kathryn A; Kane, Albert J; Ransom, Jason I; Bruemmer, Jason E

    2015-07-15

    Immunocontraception with porcine ZP (pZP) can be an effective means of fertility control in feral horses. Previous studies suggest that antibodies produced after pZP vaccination may both inhibit fertilization and cause follicular dysgenesis. Zonastat-H, PZP-22, and SpayVac are three pZP vaccines proposed for use in horses. Although all these vaccines contain the pZP antigen, variations in antigen preparation and vaccine formulation lead to differences in antigenic properties among them. Likewise, despite numerous efficacy and safety studies of Zonastat-H and PZP-22, the contraceptive mechanisms of SpayVac remain unclear. The preparation of pZP for SpayVac is thought to include more nonzona proteins, making it less pure than the other two vaccines. This may result in increased antigenicity of the vaccine. We therefore investigated the immunoreactivity of serum antibodies from SpayVac-vaccinated mares to equine zona protein. Western blot analyses revealed an immunoreactivity of these antibodies to protein isolated from mature equine oocytes, ZP, follicular tissues, and ovarian tissues. Immunohistochemical analyses were used to locate the binding of serum antibodies to the ZP of immature oocytes in ovarian stromal tissue. We also found serum antibodies from SpayVac-treated mares to be predominantly specific for zona protein 3. Collectively, our results suggest a model where serum antibodies produced in response to SpayVac vaccination are immunoreactive to equine zona protein in vitro. Our study lends insight into the contraceptive mechanisms underlying the infertility observed after SpayVac vaccination. PMID:25922172

  12. Serum tau protein as a marker of disease activity in enterohemorrhagic Escherichia coli O111-induced hemolytic uremic syndrome.

    PubMed

    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 <1.0 in 2 patients with EHEC 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. PMID:25895963

  13. Bayesian hierarchical reconstruction of protein profiles including a digestion model

    E-print Network

    Paris-Sud XI, Université de

    Bayesian hierarchical reconstruction of protein profiles including a digestion model Pierre to recover the protein biomarkers content in a robust way. We will focus on the digestion step since and each branch to a molecular processing such as digestion, ionisation and LC-MS separation

  14. Association of serum protein levels with egg productivity in Taiwan red-feathered country chickens

    Microsoft Academic Search

    M. L. Liou; S. Y. Huang; Y. C. Liu; J. H. Lin; C. K. Chuang; W. C. Lee

    2007-01-01

    This study investigated the relationship between serum protein levels and egg productivity in Taiwan red-feathered country chickens (TRFCC). The total egg numbers of TRFCC (n=157) were recorded from 25 to 48 weeks of age. Serum samples were collected at 24 and 35 weeks of age, then classified by total egg number into four groups according to mean±1S.D. Serum X protein

  15. DEVELOPMENTAL CHANGES IN SERUM PROTEINS, LIPIDS AND CHOLESTEROL DURING THE COURSE

    E-print Network

    Paris-Sud XI, Université de

    the #12;quantitative changes in serum proteins, lipids and cholestrol during the course of force feeding changes in serum proteins, lipids and cholestrol during the course of force feeding in geese. The effect by paper electrophoresis suggested by DiTThtER and ANrro (1965). The examination of the cholestrol was done

  16. Serum Protein MALDI Profiling to Distinguish Upper Aerodigestive Tract Cancer Patients From Control Subjects

    Microsoft Academic Search

    David Sidransky; Rafael Irizarry; Joseph A. Califano; Xianbin Li; Hening Ren; Nicole Benoit; Li Mao

    2003-01-01

    Background: There are no reliable blood markers for the early detection and monitoring of aerodigestive tract tumors. Recent studies have suggested that serum protein patterns may be able to distinguish cancer patients from control subjects. Methods: We used matrix-assisted laser desorption and ionization (MALDI) mass spectroscopy to obtain serum protein patterns from patients with head and neck cancer (n 99)

  17. Influence of protein intake associated with coconut or salmon oil on serum,

    E-print Network

    Paris-Sud XI, Université de

    Influence of protein intake associated with coconut or salmon oil on serum, VLDL, LDL and HDL of protein depletion associated with salmon (rich in w3 PUFA) or co- conut oil (poor in EFA) on various serum% casein + 5% salmon oil), SAd (2% casein + 5% salmon oil), COC (20% casein + 5% coconut oil), COd (2

  18. Serum Free Light Chains: An Alternative to the Urine Bence Jones Proteins Screening Test for Monoclonal Gammopathies

    Microsoft Academic Search

    Peter G. Hill; Julia M. Forsyth; Baldeep Rai; Stewart Mayne

    Background: Retrospective analyses have established the role of quantitative serum free light chains (FLCs) in the diagnosis of monoclonal light chain disorders. The aims of this study were to assess (a) whether the addition of serum FLCs to serum protein electrophoresis (SPEP) identified additional patients with monoclonal gammopathies; (b) whether serum FLC measurements could replace urinalysis for Bence Jones protein

  19. Serum Free Light Chains: An Alternative Test to Urine Bence Jones Proteins When Screening for Monoclonal Gammopathies

    Microsoft Academic Search

    Peter G. Hill; Julia M. Forsyth; Baldeep Rai; Stewart Mayne

    Background: Retrospective analyses have established the role of quantitative serum free light chains (FLCs) in the diagnosis of monoclonal light chain disorders. The aims of this study were to assess (a) whether the addition of serum FLCs to serum protein electrophoresis (SPEP) identified additional patients with monoclonal gammopathies; (b) whether serum FLC measurements could replace urinalysis for Bence Jones protein

  20. THE INFLUENCE OF PROLONGED, INTENSIVE PLASMAPHERESIS UPON THE ABILITY OF THE ORGANISM TO REGENERATE SERUM PROTEIN

    PubMed Central

    Melnick, Daniel; Cowgill, George R.

    1937-01-01

    1. When the dog is subjected to quantitative plasmapheresis and fed appropriate "synthetic" artificial rations, it is possible to evaluate the ability of the organism to regenerate serum protein from both exogenous and endogenous sources. Approximately 44 per cent of the protein, casein, fed in excess of the minimal amount needed normally to meet the general nitrogen requirements, is utilized for the formation of new serum protein. Under our experimental conditions the dog can regenerate each week solely from endogenous sources approximately 0.6 gm. of this blood protein per kilo of optimal body weight. This is equivalent to about 21 per cent of the total amount of serum protein normally present in the plasma. 2. When the dog is fed an adequate protein diet and is subjected to a régime of prolonged intensive plasmapheresis (period of 16 consecutive weeks), no impairment in the ability of the organism to regenerate serum protein from either exogenous or endogenous sources occurs. Under our conditions of experimentation the dog appears to be able to form each week an amount of the blood protein approximately equal to that normally present in the plasma. Because of this remarkable ability of the normal organism to regenerate rapidly serum protein over a relatively long period of time, it seems that loss of protein alone in conditions of Bright's disease cannot be the etiologic agent responsible for the persistence of the hypoproteinemia. An additional factor, the "specific" ability of such individuals to regenerate serum protein, must be taken into consideration. PMID:19870679

  1. Temperature dependence of serum protein adsorption in PEGylated PNIPAm microgels.

    PubMed

    Trongsatitkul, Tatiya; Budhlall, Bridgette M

    2013-03-01

    The effect of PEGylation on the thermal response and protein adsorption resistance of crosslinked PNIPAm microgels was investigated. It was found that the presence of PEG, its molecular weight (M(n) 300 and 1100 g/mol) and its concentration (10, 20, and 30 wt.%) each significantly influenced both the value and breadth of the volume phase transition temperature (VPTT) and the adsorption of bovine serum albumin (BSA) on the surface of the microgels. Specifically, as the degree of PEGylation increased, the value and breadth of the VPTT increased, and the adsorption of BSA decreased significantly. The critical concentration that minimizes protein adsorption on PNIPAm-co-PEGMa microgels was found to be 20 wt.% of PEGMa. This critical concentration was confirmed qualitatively using laser scanning confocal microscopy (LSCM). Evidence for the effect of the molecular weight of PEG on the structure of PNIPAm-co-PEGMa microgels was provided by thermal analysis using differential scanning calorimetry. The VPTT study revealed significant differences in the composition of the microgels when PEGMa samples with two different molecular weights were used as comonomers with PNIPAm. It was determined that the molecular weight and concentration of PEGMa controls the structure of the microgels, which in turn influences their temperature response and protein adsorption resistance properties of the microgels. Our work establishes specific design concepts for controlling the molecular architecture of the hydrogels in order to tune their temperature response and biocompatibility for use in a variety of biomedical applications such as, cell encapsulation, drug delivery and tissue engineering applications. PMID:23201744

  2. Electrochemical detection of glycan and protein epitopes of glycoproteins in serum.

    PubMed

    Shah, Alok K; Hill, Michelle M; Shiddiky, Muhammad J A; Trau, Matt

    2014-11-21

    Aberrant protein glycosylation is associated with a range of pathological conditions including cancer and possesses diagnostic importance. Translation of glycoprotein biomarkers will be facilitated by the development of a rapid and sensitive analytical platform that simultaneously interrogates both the glycan and protein epitopes of glycoproteins in body fluids such as serum or saliva. To this end, we developed an electrochemical biosensor based on the immobilization of a lectin on the gold electrode surface to recognize/capture a target glycan epitope conjugated to glycoproteins, followed by detection of the protein epitope using a target protein-specific antibody. Electrochemical signals are generated by label-free voltammetric or impedimetric interrogation of a ferro/ferricyanide redox couple (e.g. [Fe(CN)6](3-/4-)) on the sensing surface, where the change in voltammetric current or interfacial electron transfer resistance was measured. The detection system was demonstrated using the model glycoprotein chicken ovalbumin with Sambucus nigra agglutinin type I (SNA lectin), and exhibits femtomolar sensitivity in the background of diluted human serum. The results obtained in this proof-of-concept study demonstrate the possibility of using electrochemical detection for developing cheap point-of-care diagnostics with high specificity and sensitivity for blood glycoprotein biomarkers. PMID:25267970

  3. Overcoming inactivation of the lung surfactant by serum proteins: a potential role for fluorocarbons?

    PubMed

    Krafft, Marie Pierre

    2015-08-14

    In many pulmonary conditions serum proteins interfere with the normal adsorption of components of the lung surfactant to the surface of the alveoli, resulting in lung surfactant inactivation, with potentially serious untoward consequences. Here, we review the strategies that have recently been designed in order to counteract the biophysical mechanisms of inactivation of the surfactant. One approach includes protein analogues or peptides that mimic the native proteins responsible for innate resistance to inactivation. Another perspective uses water-soluble additives, such as electrolytes and hydrophilic polymers that are prone to enhance adsorption of phospholipids. An alternative, more recent approach consists of using fluorocarbons, that is, highly hydrophobic inert compounds that were investigated for partial liquid ventilation, that modify interfacial properties and can act as carriers of exogenous lung surfactant. The latter approach that allows fluidisation of phospholipid monolayers while maintaining capacity to reach near-zero surface tension definitely warrants further investigation. PMID:26110877

  4. Rate of elimination of parenterally administered serum proteins in relation to their species origin

    Microsoft Academic Search

    P. N. Kosyakov; Z. I. Rovnova

    1962-01-01

    One ml each of the serum proteins of chicken (or duck), dog, cat, human, pig, horse, sheep and bull were introduced simultaneously into the blood of 12 rabbits. The fate of the proteins administered was investigated by the precipitin method with the aid of immune specific sera against the proteins named. The investigation showed that avian proteins disappear from rabbit

  5. Correlation between some hematological parameters, acute phase proteins and serum immunoglobulins in experimental caprine besnoitiosis.

    PubMed

    Hosseini, A; Namazi, F; Oryan, A; Nazifi, S

    2015-06-01

    The present study was undertaken to investigate correlation between some hematological parameters, acute phase proteins and immunoglobulins in the experimentally infected goats with Besnoitia caprae from the time of infection till 360 days post infection (DPI). Six male goats, approximately 12-16 months old, were inoculated subcutaneously with approximately 1.3 × 10(8) bradyzoites of B. caprae and blood samples were collected at weekly intervals from the jugular vein of the goats. Total leukocyte count and differential leukocyte counts were determined. Acute phase proteins (APPs) including serum amyloid A (SAA), haptoglobin (Hp), fibrinogen and ceruloplasmin were undertaken at weekly intervals. We evaluated an enzyme-linked immunosorbent assay (ELISA) (using a somatic antigen of bradyzoite) to detect anti-B. caprae antibodies in caprine sera. Cysts were present in the skin biopsies of the distal parts of the leg of the infected goats from 28 DPI. From 30 to 360 DPI, results showed that the APPs concentrations including SAA, Hp, fibrinogen and ceruloplasmin were enhanced in the serum of infected goats. However, there were some variation in hematological parameters; the differences were not significant with those of the normal values. Some variations were seen in the levels of specific antibodies against this parasite and they had correlation with some hematological parameters and acute-phase proteins. PMID:26063991

  6. Functionalization of graphene oxide generates a unique interface for selective serum protein interactions.

    PubMed

    Tan, Xiaofang; Feng, Liangzhu; Zhang, Jing; Yang, Kai; Zhang, Shuai; Liu, Zhuang; Peng, Rui

    2013-02-01

    Potential toxicity and risk of inducing allergy and inflammation have always been a great concern of using nanomaterials in biomedicine. In this work, we investigate the serum behaviors of graphene oxide (GO) and how such behaviors are affected by its surface modification such as PEGylation. The results show that, when incubated with human sera, unfunctionalized GO adsorbs a significant amount of serum proteins and strongly induces complement C3 cleavage (part of the complement activation cascade), generating C3a/C3a(des-Arg), an anaphylatoxin involved in local inflammatory responses, whereas PEGylated nano-GO (nGO-PEG) exhibits dramatic reductions in both protein binding in general and complement C3 activation. Moreover, we uncover that PEGylation on GO nanosheets apparently generates an interesting nanointerface, evidenced by the acquired certain selectivity and increased binding capacities of nGO-PEG toward a few serum proteins. Further mass spectrometry analysis identifies six nGO-PEG binding proteins, four of which are immune-related factors, including C3a/C3a(des-Arg). A series of Western blot analysis demonstrate that nGO-PEG binds up to 2-fold amount of C3a/C3a(des-Arg) than unfunctionalized GO, and can efficiently decrease the level of C3a/C3a(des-Arg) in treated sera, preventing the normal interaction of C3a with its receptor. In a proof-of-concept experiment, we demonstrate that nGO-PEG may serve to help eliminate the C3a/C3a(des-Arg) induced by other nanomaterials such as as-made GO, indicating a new strategy to modulate the immune responses evoked by one nanomaterial through the addition of another type of nanomaterial. Our results highlight the great importance of nanobio interface in regulating the biological effects of nanomaterials. PMID:23360681

  7. The disposition of serum proteins as drug-carriers in mice bearing Sarcoma 180.

    PubMed

    Tanaka, T; Kaneo, Y; Shiramoto, S; Iguchi, S

    1993-12-01

    The tumor distribution and the disposition of serum proteins, such as albumin, fetuin, transferrin, and IgG, were investigated in mice bearing Sarcoma 180. Serum proteins labeled with fluoresceinisothiocyanate (FITC) were administered to the mice. The FITC-labeled proteins acylated with glutaric anhydride were also administered to the mice in order to investigate the effect of chemical modification. The plasma concentration of each glutarylated serum protein was significantly lower, about 15 to 46-fold, in comparison to that of the non-acylated protein at 24 h after administration. The tissue distributions of the glutarylated serum proteins were also decreased compared to those of the non-acylated proteins. Especially, the hepatic distribution of albumin and IgG was significantly reduced with glutarylation. The urinary excretion of albumin and transferrin, and fecal excretion of IgG, were significantly increased with glutarylation. The serum proteins were accumulated effectively in the tumor tissue in mice bearing Sarcoma 180. It was found that the tumor distributions were not impaired by the glutarylation, except involving fetuin. It was suggested, therefore, that the glutarylated serum proteins were valuable for relative tumor-selectivity and might be utilized in a macromolecular carrier system for antitumor drugs. PMID:8130778

  8. Proteomic Analysis of Bovine Pregnancy-specific Serum Proteins by 2D Fluorescence Difference Gel Electrophoresis

    PubMed Central

    Lee, Jae Eun; Lee, Jae Young; Kim, Hong Rye; Shin, Hyun Young; Lin, Tao; Jin, Dong Il

    2015-01-01

    Two dimensional-fluorescence difference gel electrophoresis (2D DIGE) is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. The purpose of this study was to investigate bovine pregnancy-specific proteins in the proteome between bovine pregnant and non-pregnant serum using DIGE technique. Serums of 2 pregnant Holstein dairy cattle at day 21 after artificial insemination and those of 2 non-pregnant were used in this study. The pre-electrophoretic labeling of pregnant and non-pregnant serum proteins were mixed with Cy3 and Cy5 fluorescent dyes, respectively, and an internal standard was labeled with Cy2. Labeled proteins with Cy2, Cy3, and Cy5 were separated together in a single gel, and then were detected by fluorescence image analyzer. The 2D DIGE method using fluorescence CyDye DIGE flour had higher sensitivity than conventional 2D gel electrophoresis, and showed reproducible results. Approximately 1,500 protein spots were detected by 2D DIGE. Several proteins showed a more than 1.5-fold up and down regulation between non-pregnant and pregnant serum proteins. The differentially expressed proteins were identified by MALDI-TOF mass spectrometer. A total 16 protein spots were detected to regulate differentially in the pregnant serum, among which 7 spots were up-regulated proteins such as conglutinin precursor, modified bovine fibrinogen and IgG1, and 6 spots were down-regulated proteins such as hemoglobin, complement component 3, bovine fibrinogen and IgG2a three spots were not identified. The identified proteins demonstrate that early pregnant bovine serum may have several pregnancy-specific proteins, and these could be a valuable information for the development of pregnancy-diagnostic markers in early pregnancy bovine serum. PMID:25925056

  9. Use of plant protein hydrolysates for varicella virus production in Serum-Free Medium

    Microsoft Academic Search

    Bok-Hwan Chun; Yong Kwon Lee; Won-Gi Bang; Namhyun Chung

    2005-01-01

    Serum-free media containing no animal-derived components were assessed for their efficacy to produce live attenuated varicella virus. Serum-free medium containing an ultrafiltrate of soy protein acid hydrolysate and lipid resulted in a viral production yield comparable to media containing fetal bovine serum, indicating that varicella virus can be produced without the risk of contamination associated with the use of bovine

  10. Effects of Adherence, Activation and Distinct Serum Proteins on the In Vitro Human Monocyte Maturation Process

    Microsoft Academic Search

    Yukio Akiyama; Richard Griffith; Paul Miller; G. W. Stevenson; Stacy Lund; D. J. Kanapa; Henry C. Stevenson

    Elutriator-punfied human monocytes were cultured in a serum-free (SF) medium, and various serum proteins and functional activating agents were assessed for their effects on the in vitro maturation of human monocytes to macrophages. Following 3 days of suspension culture in Teflon Iabware, 60% of the monocytes were easily recovered. When varying concentrations of human AB serum (HuAB) were employed, human

  11. Inheritance patterns of enzymes and serum proteins of mallard-black duck hybrids

    USGS Publications Warehouse

    Morgan, R.P., II; Meritt, D.W.; Block, S.B.; Cole, M.

    1984-01-01

    From 1974 to 1976, a breeding program was used to produce hybrids of black ducks and mallards for the evaluation of inheritance patterns of serum proteins and serum, liver and muscle enzymes. In addition to the crosses designed to produce hybrids, a series of matings in 1975 and 1976 were designed to evaluate inheritance patterns of a hybrid with either a black duck or mallard. At the F1 level, hybrids were easily distinguished using serum proteins. However, once a hybrid was crossed back to either a mallard or black duck, only 12-23% of the progeny were distinguishable from black ducks or mallards using serum proteins and 23-39% using esterases. Muscle, serum and liver enzymes were similar between the two species.

  12. Inheritance patterns of enzymes and serum proteins of mallard-black duck hybrids

    USGS Publications Warehouse

    Morgan, R.P., II; Meritt, D.W.; Block, S.B.; Cole, M.A.; Sulkin, S.T.; Lee, F.B.; Henny, C.J.

    1984-01-01

    From 1974 to 1976, a breeding program was used to produce hybrids of black ducks and mallards for the evaluation of inheritance patterns of serum proteins and serum, liver and muscle enzymes. In addition to the crosses designed to produce hybrids, a series of matings in 1975 and 1976 were designed to evaluate inheritance patterns of a hybrid with either a black duck or mallard. At the F1 level, hybrids were easily distinguished using serum proteins. However, once a hybrid was crossed back to either a mallard or black duck, only 12?23% of the progeny were distinguishable from black ducks or mallards using serum proteins and 23?39% using esterases. Muscle, serum and liver enzymes were similar between the two species.

  13. Serum Levels of the Adipokine Adipocyte Fatty Acid–binding Protein Are Increased in Preeclampsia

    Microsoft Academic Search

    Mathias Fasshauer; Jeannette Seeger; Theresa Waldeyer; Susanne Schrey; Thomas Ebert; Jürgen Kratzsch; Ulrike Lössner; Matthias Blüher; Michael Stumvoll; Renaldo Faber; Holger Stepan

    2008-01-01

    BackgroundPreeclampsia (PE) is a serious complication of pregnancy which is associated with an increased future metabolic and cardiovascular risk for both mother and newborn. Recently, adipocyte fatty acid–binding protein (AFABP) was introduced as a novel adipokine, serum levels of which independently correlate with the development of the metabolic syndrome and cardiovascular disease in humans. In this study, we investigated serum

  14. Serum levels of adipocyte fatty acid binding protein are increased in gestational diabetes mellitus

    Microsoft Academic Search

    Susan Kralisch; Holger Stepan; Jürgen Kratzsch; Michael Verlohren; Hans-Joachim Verlohren; Kathrin Drynda; U. Lossner; M. Bluher; Michael Stumvoll; Mathias Fasshauer

    2008-01-01

    Objective: Adipocyte fatty acid binding protein (AFABP) was recently introduced as a novel adipokine, serum levels of which independently correlate with the development of the metabolic syndrome and cardiovascular disease in humans. In the current study, we investigated serum concentrations of AFABP in patients with gestational diabetes mellitus (GDM) as compared with healthy pregnant controls matched for gestational age and

  15. Serum C-Reactive Protein Levels in Patients with Cervical Neoplasia

    Microsoft Academic Search

    Felix Wong; Mano Arumanayagam; R. Swaminathan; Allan Chang

    1989-01-01

    Serum C-reactive protein (CRP) levels were determined by enzyme immunoassay in 56 patients with invasive carcinoma of the cervix, 34 patients with cervical dysplasia and 24 women with no evidence of cervical pathology. The logarithmic CRP values were significantly higher in patients with invasive carcinoma when compared with the controls (p < 0.05). Measurement of serum CRP in cancer patients

  16. Acute phase response of serum amyloid A protein and C reactive protein to the common cold and influenza

    Microsoft Academic Search

    J T Whicher; R E Chambers; J Higginson; L Nashef; P G Higgins

    1985-01-01

    C reactive protein (CRP) and serum amyloid A protein (SAA) are sensitive and rapid acute phase reactants, and their measurement for monitoring inflammatory disease and assessing the prognosis in secondary amyloidosis is gaining widespread acceptance. The changes in these proteins in eight subjects suffering from natural colds, 15 subjects with experimentally induced colds (rhinoviruses E1, 3, 9, 14, or 31),

  17. Comparison of composition, sensory, and volatile components of thirty-four percent whey protein and milk serum protein concentrates1

    Microsoft Academic Search

    J. Evans; J. Zulewska; M. Newbold; M. A. Drake; D. M. Barbano

    2009-01-01

    The objectives of this study were to identify and compare the composition, flavor, and volatile compo- nents of serum protein concentrate (SPC) and whey protein concentrate (WPC) containing about 34% protein made from the same milk to each other and to commercial 34% WPC from 6 different factories. The SPC and WPC were manufactured in triplicate with each pair of

  18. Serum protein identification and quantification of the corona of 5, 15 and 80 nm gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Schäffler, Martin; Semmler-Behnke, Manuela; Sarioglu, Hakan; Takenaka, Shinji; Wenk, Alexander; Schleh, Carsten; Hauck, Stefanie M.; Johnston, Blair D.; Kreyling, Wolfgang G.

    2013-07-01

    When nanoparticles (NP) enter the body they come into contact with body fluids containing proteins which can adsorb to their surface. These proteins may influence the NP interactions with the biological vicinity, eventually determining their biological fate inside the body. Adsorption of the most abundantly binding proteins was studied after an in vitro 24 hr incubation of monodisperse, negatively charged 5, 15 and 80 nm gold spheres (AuNP) in mouse serum by a two-step analysis: proteomic protein identification and quantitative protein biochemistry. The adsorbed proteins were separated from non-adsorbed proteins by centrifugation and gel electrophoresis and identified using a MALDI-TOF-MS-Proteomics-Analyzer. Quantitative analysis of proteins in gel bands by protein densitometry, required the focus on predominantly binding serum proteins. Numerous proteins adsorbed to the AuNP depending on their size, e.g. apolipoproteins or complement C3. The qualitative and quantitative amount of adsorbed proteins differed between 5, 15 and 80 nm AuNP. Band intensities of adsorbed proteins decreased with increasing AuNP sizes based not only on their mass but also on their surface area. Summarizing, the AuNP surface is covered with serum proteins containing transport and immune related proteins among others. Hence, protein binding depends on the size, surface area and curvature of the AuNP.

  19. Differences in the serum binding determinants of isradipine and darodipine--consequences for serum protein binding in various diseases.

    PubMed Central

    Pinquier, J L; Urien, S; Chaumet-Riffaud, P; Tillement, J P

    1989-01-01

    1. Serum protein binding of isradipine and darodipine, and serum concentrations of alpha 1-acid glycoprotein (AAG), albumin (HSA) and non-esterified fatty acids (NEFA) were measured in three groups of patients, I: healthy subjects (n = 20); II: patients with inflammatory disorders (n = 15) and III: patients with hepatic insufficiency (n = 17). 2. AAG was increased significantly in group II patients (P less than 0.001) and decreased in group III patients (P less than 0.001); HSA was decreased significantly in group II and group III patients (P less than 0.001). 3. The free percentage of isradipine was decreased significantly in group II patients (P less than 0.05) and increased in group III patients (P less than 0.05) and multivariate analysis showed that these variations were inversely related to changes in AAG concentration. 4. The free percentage of darodipine was increased significantly in group II and III patients (P less than 0.05) due to a decrease in HSA concentration, as shown by multivariate analysis. 5. The changes in free serum percentages of isradipine and darodipine were inversely related to concomitant changes in the concentration of the serum protein for which they showed the highest affinity, AAG for isradipine and HSA for darodipine, respectively. 6. The unexplained variability in the binding data was greater when AAG was the major determinant of binding (isradipine). PMID:2531607

  20. Effect of water salinity on total protein and electrophoretic pattern of serum proteins of grass carp, Ctenopharyngodon idella.

    PubMed

    Peyghan, Rahim; Khadjeh, Gholam Hosain; Enayati, Ala

    2014-01-01

    In this study the effects of water salinity on serum total protein and its components in grass carp were investigated. The aim of this study was to determine the effect of salinity tolerance of fish on total serum protein level and its components as an indicator of liver and kidney activity. One hundred and twenty grass carp were divided into four groups, randomly. The first three groups were reared in concentration of 4, 8 and 12 g L(-1) of salt solution, respectively, and the fourth group was reared in freshwater and served as control. After 3 weeks, blood samples were collected and after harvesting the blood serum, serum total protein and protein components were measured with Biuret and electrophoresis methods, respectively. Results showed that mean value of serum total protein in the control and three salinities groups were 2.75, 3.28, 2.90 and 3.13 g dL(-1), respectively. Five fractions of serum protein were electrophoretically observed as: albumin (Alb), alpha-1 globulin (?1-glu), alpha-2 globulin (?2-glu), beta globulin (?-glu) and gamma globulin (?-glu). There were not any significant differences between the average mean of serum total protein of experimental and control groups (p > 0.05). However, Alb, ?1-glu and ?-glu levels in the experimental groups were significantly higher than in the control group (p < 0.05). The average of ?2-glu and ?-glu revealed no significant difference between the experimental groups (p > 0.05). In conclusion, our results showed that increasing water salinity could have a significant effect on Alb, ?1-glu and ?-glu levels but not on total serum protein in grass carp. PMID:25568723

  1. Correlation Between Hypertension, C-Reactive Protein and Serum Uric Acid With Psychological Well-being

    PubMed Central

    Maleki, Ali; Samandari, Saeid; Almeida, Osvaldo; Jafarian Kerman, Scott Reza; Abdolvand, Mahdi; Aliyari, Farshid; Foroughi, Saeid

    2014-01-01

    Background: Multiple population-based human studies have established a strong association between increasing levels of serum C-reactive protein, uric acid and subsequent development of hypertension. Objectives: We aimed to investigate the association between mental well-being with presence of hypertension, hyperuricemia and hs-CRP levels. ?? Patients and Methods: This was a cross sectional study of 801 individuals aged 35-85 years old in Broujerd, Iran, included by randomized cluster sampling. General Health Questionnaire (GHQ-12) for assessing mental health/distress level, MONICA standard questions for evaluating hypertension history, serum hs-CRP and Serum Uric Acid (SUA) were evaluated Data were analyzed by appropriate statistical test such as chi-square, T-test and correlation. Results: One hundred eighty five patients (23.1%) had high distress/minor psychiatric disorders. SUA had significant association with hypertension (r = 0.64, P = 0.034). No significant relation was observed between hs-CRP and hypertension. The correlation between GHQ and hs-CRP was not significant but a weak and negative correlation was found between GHQ and SUA SUA (P = 0.012, r = -0.089). Conclusions: The weak and strong correlation among these parameters indicate that mental wellbeing relays on physical wellness and interact with each other; therefore, controlling hypertension along with uric acid control may effect mental health of any kind of patients. PMID:25237581

  2. Correlation between Serum Level of Monocyte Chemoattractant Protein-1 and Postoperative Recurrence of Spinal Tuberculosis in the Chinese Han Population

    PubMed Central

    He, Dan; Zhang, Xiaolu; Gao, Qile; Huang, Rongfu; Deng, Zhansheng; Guo, Chaofeng; Guo, Qiang; Huang, Jia; Zhang, Hongqi

    2015-01-01

    Objective To correlate serum level of monocyte chemoattractant protein-1 (MCP-1) with postoperative recurrence of spinal tuberculosis in the Chinese Han population. Methods Patients of Han nationality with newly diagnosed spinal tuberculosis were consecutively included in this study. At different time points postoperatively, serum level of MCP-1 was determined using an enzyme linked immunosorbent assay. Recurrence of spinal tuberculosis after surgery and during the follow-up period was recorded. The correlation between serum MCP-1 level and recurrence of spinal tuberculosis was analyzed. Results A total of 169 patients with spinal tuberculosis were included in the study and followed up for an average of2.2±1.3 years (range, 1–5 years). Of these patients, 11 had postoperative recurrence of spinal tuberculosis. The patients’ serum level of MCP-1 increased significantly after postoperative recurrence of spinal tuberculosis. Once the symptoms of recurrence were cured, the serum level of MCP-1 decreased significantly and it did not differ from patients without disease recurrence. Conclusion Postoperative recurrence of spinal tuberculosis is likely to increase the serum level of MCP-1. PMID:25962150

  3. A serum-free medium for the culture of insect cells and production of recombinant proteins

    Microsoft Academic Search

    W. F. Hink

    1991-01-01

    Summary  A low protein aqueous lipid supplement (Ex-Cyte VLE), in combination with pluronic polyol, is an effective replacement for\\u000a fetal bovine serum for insect Sf-9 cells. Serum-free medium with lipid supplement and pluronic (SFM-LP) supported higher cell\\u000a viability and maximum cell populations than serum-supplemented medium. No adaptation procedures are required when switching\\u000a cells from serum-containing medium to SFM-LP, and growth rates

  4. Quantitative profiling of serum samples using TMT protein labelling, fractionation and LC–MS\\/MS

    Microsoft Academic Search

    John Sinclair; John F. Timms

    2011-01-01

    Blood-borne biomarkers are urgently required for the early detection, accurate diagnosis and prognosis of disease. Additionally, improved methods of profiling serum and plasma proteins for biomarker discovery efforts are needed. Herein, we report a quantitative method based on amino-group labelling of serum proteins (rather than peptides) with isobaric tandem mass tags (TMT) and incorporating immune-based depletion, gel-based and strong anion

  5. Cardiac isoform of ?-2 macroglobin, a novel serum protein, may induce cardiac hypertrophy in rats

    Microsoft Academic Search

    Chellam Rajamanickam; Sadayappan Sakthivel; Gopal Jegadeesh Babu; Friedrich Lottspeich; Bernhard Kadenbach

    2001-01-01

    Earlier studies from this laboratory have identified a novel high molecular weight (182 kDa) serum protein suggested to be\\u000a involved in the development of cardiac hypertrophy. In the present case the role of this novel serum protein in the development\\u000a of pressure-induced cardiac hypertrophy and the molecular events associated with it in experimental rats has been investigated.\\u000a Multiple injections of

  6. A study of serum proteins from horses infected with equine infectious anemia virus

    E-print Network

    Folks, Thomas Murill

    1972-01-01

    of MASTER OF SCZENCE Flay 1972 Major Subject: Veterinary Microbiolo'y A STUDY OF SERUM PROTEINS FROM HORSES INFECTED NITH EQUINE ZNFECTIOUS ANEMIA VIRUS A Thesis THOMAS MURZLL FOLKS Approved as to style and content by: o. (Chairman of Committee...) (Head of' Department) (Member) (Member) (Member) (Member) (Member) May 1972 ABS RAUT . ', Study of Serum Proteins from Horses Infected with Equine Infectious Anemia Virus. (May 1972) Thomas Murill Folks, B. A. , Univer- sity oz" Texas at Austin...

  7. Serum levels of club (Clara) cell secretory protein predict cancer mortality in adults

    PubMed Central

    Guerra, Stefano; Vasquez, Monica M.; Spangenberg, Amber; Halonen, Marilyn; Martinez, Fernando D.

    2014-01-01

    Background Club (formerly Clara) cell secretory protein (CC16) is produced mainly by bronchiolar club cells and has been shown to have protective effects against airway inflammation and oxidative stress from cigarette smoking and related carcinogens. The goal of this study was to determine whether serum CC16 levels predict all-cause and cancer-specific mortality in adults. Methods We used data from the population-based TESAOD study, a prospective cohort study of respiratory health initiated in Tucson, AZ in 1972. At baseline, participants completed standardized respiratory questionnaires and lung function tests. Serum CC16 was measured in cryopreserved serum samples. A review of vital status of participants as of January 1st, 2011 was completed through contact with next of kin, collection of death certificates, and linkage with the National Death Index. Findings A total of 1086 participants who were 21 to 70 years old at enrollment were included. Of these, 653 (60%) died by 2011 and cause of death was ascertained for 649 (99%). In Cox proportional hazards models adjusted for sex, age, education, body mass index categories, smoking and pack-years, and baseline levels of lung function, serum CC16 levels at the baseline survey were inversely associated with mortality risk over the study follow-up. Mortality risk increased by 16% for each standard deviation (SD) decrease in CC16 (Hazard Ratio (HR), 95% CI: 1.16, 1.06 – 1.26; p = 0.0007). When data on cause-specific mortality were analyzed, each SD decrease in serum CC16 was associated with >40% increased risk of dying of cancer (adjusted HR=1.41, 1.19 – 1.67; p < 0.0001). Among smokers, the corresponding adjusted HRs for mortality by lung cancer were 1.52 (1.14 – 2.03; p = 0.004). Interpretation Serum CC16 levels predict mortality risk in the general adult population. The excess risk associated with lower CC16 is largely explained by cancer, particularly lung cancer. PMID:24461757

  8. Comparison of functional properties of 34% and 80% whey protein and milk serum protein concentrates.

    PubMed

    Luck, P J; Vardhanabhuti, B; Yong, Y H; Laundon, T; Barbano, D M; Foegeding, E A

    2013-09-01

    This study compared the functional properties of serum protein concentrate (SPC) with whey protein concentrate (WPC) made from the same milk and with commercial WPC. The experimental SPC and WPC were produced at 34% or 80% protein from the same lot of milk. Protein contents of WPC and SPC were comparable; however, fat content was much lower in SPC compared with WPC and commercial WPC. The effect of drying methods (freeze vs. spray drying) was studied for 34% WPC and SPC. Few differences due to drying method were found in turbidity and gelation; however, drying method made a large difference in foam formation for WPC but not SPC. Between pH 3 and 7, SPC was found to have lower turbidity than WPC; however, protein solubility was similar between SPC and WPC. Foaming and gelation properties of SPC were better than those of WPC. Differences in functional properties may be explained by differences in composition and extent of denaturation or aggregation. PMID:23871371

  9. Serum protein electrophoresis by using high-resolution agarose gel in clinically healthy and Aspergillus species-infected falcons.

    PubMed

    Kummrow, Maya; Silvanose, Christudas; Di Somma, Antonio; Bailey, Thomas A; Vorbrüggen, Susanne

    2012-12-01

    Serum protein electrophoresis has gained importance in avian medicine during the past decade. Interpretation of electrophoretic patterns should be based on species-specific reference intervals and the electrophoresis gel system. In this study, serum protein electrophoresis by using high-resolution agarose gels was performed on blood samples collected from 105 falcons, including peregrine falcons (Falco peregrinus), gyrfalcons (Falco rusticolus), saker falcons (Falco cherrug), red-naped shaheens (Falco pelegrinoides babylonicus), and hybrid falcons, that were submitted to the Dubai Falcon Hospital (Dubai, United Arab Emirates) between 2003 and 2006. Reference values were established in clinically healthy birds and compared with values from falcons infected with Aspergillus species (n = 32). Falcons with confirmed aspergillosis showed significantly lower prealbumin values, which is a novel finding. Prealbumin has been documented in many avian species, but further investigation is required to illuminate the diagnostic significance of this negative acute-phase protein. PMID:23409432

  10. Serum Golgi protein 73 levels and liver pathological grading in cases of chronic hepatitis B

    PubMed Central

    XU, ZHENGJU; PAN, XINGNAN; WEI, KAIPENG; DING, HONGBING; WEI, MEIJUAN; YANG, HUANWEN; LIU, QIAN

    2015-01-01

    The present study was designed to assess the correlation between serum Golgi protein 73 (GP73) and liver pathological grading and staging in patients with chronic hepatitis B (CHB). Two hundred and fifty-three patients with chronic hepatitis B virus (HBV) infections were enrolled in the present study. All patients received a serum GP73 test, and 91 CHB patients underwent liver biopsy. GP73 expression in liver tissue was assessed by immunohistochemical analysis. The results indicated that serum GP73 levels were positively correlated with disease progression in patients with chronic HBV infection (r=0.677). There was no significant difference in serum GP73 levels between hepatitis B e antigen-positive and ?negative patients (P>0.05). There were also no significant differences in serum GP73 levels among specimens with varying HBV DNA contents (P>0.05). Serum GP73 levels were positively correlated with increased liver pathological grading (r=0.737) and staging (r=0.692), and immunohistochemical analysis indicated that GP73 protein expression increased concurrently with liver pathological grading and staging. In conclusion, serum GP73 was found to be correlated with liver pathological grading and staging in patients with CHB, and may be an effective indicator for the evaluation of disease progression. However, serum GP73 levels were not associated with HBV replication. PMID:25524053

  11. Cockroach Larval-specific Protein, a Tyrosine-rich Serum Protein* (Received for publication, June9, 1983)

    E-print Network

    Kunkel, Joseph G.

    Cockroach Larval-specific Protein, a Tyrosine-rich Serum Protein* (Received for publication, June9 in the hemolymph of cockroaches shortly be- fore molting, but is rapidly cleared from the hemo- lymph during themolting cycle of cockroaches has been studied in syn- chronouslymoltingcultures (1-3). Among

  12. Relationship between Acute Phase Proteins and Serum Fatty Acid Composition in Morbidly Obese Patients

    PubMed Central

    Fernandes, Ricardo; Beserra, Bruna Teles Soares; Cunha, Raphael Salles Granato; Hillesheim, Elaine; Camargo, Carolina de Quadros; Pequito, Danielle Cristina Tonello; de Castro, Isabela Coelho; Fernandes, Luiz Cláudio; Nunes, Everson Araújo; Trindade, Erasmo Benício Santos de Moraes

    2013-01-01

    Background. Obesity is considered a low-grade inflammatory state and has been associated with increased acute phase proteins as well as changes in serum fatty acids. Few studies have assessed associations between acute phase proteins and serum fatty acids in morbidly obese patients. Objective. To investigate the relationship between acute phase proteins (C-Reactive Protein, Orosomucoid, and Albumin) and serum fatty acids in morbidly obese patients. Methods. Twenty-two morbidly obese patients were enrolled in this study. Biochemical and clinical data were obtained before bariatric surgery, and fatty acids measured in preoperative serum. Results. Orosomucoid was negatively correlated with lauric acid (P = 0.027) and eicosapentaenoic acid (EPA) (P = 0.037) and positively with arachidonic acid (AA) (P = 0.035), AA/EPA ratio (P = 0.005), and n-6/n-3 polyunsaturated fatty acids ratio (P = 0.035). C-Reactive Protein (CRP) was negatively correlated with lauric acid (P = 0.048), and both CRP and CRP/Albumin ratio were negatively correlated with margaric acid (P = 0.010, P = 0.008, resp.). Albumin was positively correlated with EPA (P = 0.027) and margaric acid (P = 0.008). Other correlations were not statistically significant. Conclusion. Our findings suggest that serum fatty acids are linked to acute phase proteins in morbidly obese patients. PMID:24167354

  13. A panel of regulated proteins in serum from patients with cervical intraepithelial neoplasia and cervical cancer.

    PubMed

    Boichenko, Alexander P; Govorukhina, Natalia; Klip, Harry G; van der Zee, A G J; Güzel, Co?kun; Luider, Theo M; Bischoff, Rainer

    2014-11-01

    We developed a discovery-validation mass-spectrometry-based pipeline to identify a set of proteins that are regulated in serum of patients with cervical intraepithelial neoplasia (CIN) and squamous cell cervical cancer using iTRAQ, label-free shotgun, and targeted mass-spectrometric quantification. In the discovery stage we used a "pooling" strategy for the comparative analysis of immunodepleted serum and revealed 15 up- and 26 down-regulated proteins in patients with early- (CES) and late-stage (CLS) cervical cancer. The analysis of nondepleted serum samples from patients with CIN, CES, an CLS and healthy controls showed significant changes in abundance of alpha-1-acid glycoprotein 1, alpha-1-antitrypsin, serotransferrin, haptoglobin, alpha-2-HS-glycoprotein, and vitamin D-binding protein. We validated our findings using a fast UHPLC/MRM method in an independent set of serum samples from patients with cervical cancer or CIN and healthy controls as well as serum samples from patients with ovarian cancer (more than 400 samples in total). The panel of six proteins showed 67% sensitivity and 88% specificity for discrimination of patients with CIN from healthy controls, a stage of the disease where current protein-based biomarkers, for example, squamous cell carcinoma antigen (SCCA), fail to show any discrimination. Additionally, combining the six-protein panel with SCCA improves the discrimination of patients with CES and CLS from healthy controls. PMID:25232869

  14. Serum Vascular Adhesion Protein-1 Predicts 10-Year Cardiovascular and Cancer Mortality in Individuals With Type 2 Diabetes

    PubMed Central

    Li, Hung-Yuan; Jiang, Yi-Der; Chang, Tien-Jyun; Wei, Jung-Nan; Lin, Mao-Shin; Lin, Cheng-Hsin; Chiang, Fu-Tien; Shih, Shyang-Rong; Hung, Chi Sheng; Hua, Cyue-Huei; Smith, David J.; Vanio, Jani; Chuang, Lee-Ming

    2011-01-01

    OBJECTIVE Vascular adhesion protein-1 (VAP-1) participates in inflammation and catalyzes the breakdown of amines to produce aldehyde, hydrogen peroxide, and ammonia. Serum VAP-1 correlates positively with both acute hyperglycemia and diabetes. We conducted a cohort study to evaluate whether serum VAP-1 predicts 10-year survival in type 2 diabetic patients. RESEARCH DESIGN AND METHODS Between July 1996 and June 2003, we enrolled 661 type 2 diabetic subjects at National Taiwan University Hospital. Serum VAP-1 in the samples obtained at enrollment was measured by time-resolved immunofluorometric assay. The vital status of all subjects was ascertained by linking their data with computerized death certificates in Taiwan. RESULTS The medium follow-up period was 10.4 years. Subjects with serum VAP-1 in the highest tertile had a hazard ratio (HR) of 2.19 (95% CI 1.17–4.11) for all-cause mortality adjusted for age, sex, smoking, history of cardiovascular disease, obesity, hypertension, hemoglobin A1c, diabetes duration, total cholesterol, use of statins, abnormal ankle-brachial index, estimated glomerular filtration rate (eGFR), and proteinuria. The adjusted HRs for logarithmically transformed serum VAP-1 were 5.83 (95% CI 1.17–28.97) for cardiovascular mortality, 6.32 (95% CI 1.25–32.00) for mortality from cardiovascular and diabetic causes, and 17.24 (95% CI 4.57–65.07) for cancer mortality. There were four variables, including age, serum VAP-1, proteinuria, and eGFR, which could enhance mortality prediction significantly. CONCLUSIONS Serum VAP-1 can predict 10-year all-cause mortality, cardiovascular mortality, and cancer mortality independently in type 2 diabetic subjects. Serum VAP-1 is a novel biomarker that improves risk prediction over and above established risk factors. PMID:21282368

  15. Effect of nutrient supplementation on serum homocysteine, iron and proteins in psychogeriatric patients.

    PubMed

    Bartels, Piet C M; Schoorl, Marianne; Peetoom, Jan J

    2003-01-01

    In a longitudinal follow-up study the effect of pharmaceutical supplementation of nutrients (folate, vitamin B12, B6, B1, C, iron and proteins) was established in 25 psychogeriatric patients (subject group). A reference group of 30 apparently healthy elderly subjects was used for comparison and statistical evaluation. At the time of hospitalization percentages concerning the incidence of decreased serum concentrations reflecting an inappropriate nutrient state in the subject group amounted to 28% for vitamin B12, 20% for folate, 36% for iron, 12% for transferrin and 56% for albumin concentrations. Increased plasma concentrations of homocysteine combined with decreased folate concentrations were found in 16% of the psychogeriatric patients. If compared with the initial results at admission, after three weeks of nutrient supplementation the vitamin B12 and folate serum concentrations were increased. Results for serum iron concentrations remained below the reference range interval in 5 of the 25 subjects reflecting iron deficiency. Initially decreased serum transferrin concentrations did not return to the reference range. Serum albumin levels still further decreased after admission to the hospital, resulting after three weeks in albumin concentrations below the reference range for 68% of the subjects. It is concluded that supplementation of folate and vitamin B12 lowered homocysteine plasma concentrations successfully. Supplementation of protein nutrients is not appropriate in order to restore disturbances of protein metabolism. Persisting low concentrations of proteins in serum are indicative of irreversible decreased synthesis. PMID:12593473

  16. Effect of lithium on hepatic and serum elemental status under different dietary protein regimens.

    PubMed

    Tandon, A; Nagpaul, J P; Bandhu, H; Singh, N; Dhawan, D K

    1999-04-01

    Lithium carbonate at the dose level of 1.1 g/kg was administered in diet to normal (18% protein), low-protein- (LP; 8%) and high-protein (HP; 30% diet)-fed rats for a period of 1 mo. The LP diet resulted in a significant decrease in the hepatic levels of zinc, iron, copper, manganese, calcium, and serum levels of calcium and sodium. The HP diet caused a marked decrease in copper and calcium levels in liver, but an increase in potassium levels in serum was observed. Lithium treatment to normal rats led to a significant reduction in the hepatic contents of zinc, copper, potassium, calcium, and serum contents of potassium and sodium, whereas an elevation in serum contents of calcium was noticed. Administration of lithium to protein-deficient rats increased the hepatic concentration of manganese and serum concentration of calcium and the levels almost reached the normal limits. On the other hand, there was a marked depression in potassium contents in the serum of LP- as well as HP-fed rats following lithium treatment when compared to LP and HP groups, respectively. PMID:10208656

  17. A Serum Protein Profile Predictive of the Resistance to Neoadjuvant Chemotherapy in Advanced Breast Cancers*

    PubMed Central

    Hyung, Seok-Won; Lee, Min Young; Yu, Jong-Han; Shin, Byunghee; Jung, Hee-Jung; Park, Jong-Moon; Han, Wonshik; Lee, Kyung-Min; Moon, Hyeong-Gon; Zhang, Hui; Aebersold, Ruedi; Hwang, Daehee; Lee, Sang-Won; Yu, Myeong-Hee; Noh, Dong-Young

    2011-01-01

    Prediction of the responses to neoadjuvant chemotherapy (NACT) can improve the treatment of patients with advanced breast cancer. Genes and proteins predictive of chemoresistance have been extensively studied in breast cancer tissues. However, noninvasive serum biomarkers capable of such prediction have been rarely exploited. Here, we performed profiling of N-glycosylated proteins in serum from fifteen advanced breast cancer patients (ten patients sensitive to and five patients resistant to NACT) to discover serum biomarkers of chemoresistance using a label-free liquid chromatography-tandem MS method. By performing a series of statistical analyses of the proteomic data, we selected thirteen biomarker candidates and tested their differential serum levels by Western blotting in 13 independent samples (eight patients sensitive to and five patients resistant to NACT). Among the candidates, we then selected the final set of six potential serum biomarkers (AHSG, APOB, C3, C9, CP, and ORM1) whose differential expression was confirmed in the independent samples. Finally, we demonstrated that a multivariate classification model using the six proteins could predict responses to NACT and further predict relapse-free survival of patients. In summary, global N-glycoproteome profile in serum revealed a protein pattern predictive of the responses to NACT, which can be further validated in large clinical studies. PMID:21799047

  18. Detection of   and   Light Chain Monoclonal Proteins in Human Serum: Automated Immunoassay versus Immunofixation Electrophoresis

    Microsoft Academic Search

    Troy D. Jaskowski; Christine M. Litwin; Harry R. Hill

    2006-01-01

    Recently, turbidimetric immunoassays for detecting and quantifying and free light chains (FLC) have become available and are promoted as being more sensitive than immunofixation electrophoresis (IFE) in detecting FLC monoclonal proteins. In this study, we assessed the ability of these turbidimetric assays to detect serum monoclonal proteins involving both free and heavy-chain-bound and light chains compared to standard immunofixation electrophoresis.

  19. Reproducibility of SELDI-TOF protein patterns in serum: comparing datasets from different experiments

    Microsoft Academic Search

    Keith A. Baggerly; Jeffrey S. Morris; Kevin R. Coombes

    2004-01-01

    Motivation: There has been much interest in using patterns derived from surface-enhanced laser desorption and ioniza- tion (SELDI) protein mass spectra from serum to differentiate samples from patients both with and without disease. Such patterns have been used without identification of the underly- ing proteins responsible. However, there are questions as to the stability of this procedure over multiple experiments.

  20. Glutathione-Coated Luminescent Gold Nanoparticles: A Surface Ligand for Minimizing Serum Protein Adsorption

    PubMed Central

    2015-01-01

    Ultrasmall glutathione-coated luminescent gold nanoparticles (GS-AuNPs) are known for their high resistance to serum protein adsorption. Our studies show that these NPs can serve as surface ligands to significantly enhance the physiological stability and lower the serum protein adsorption of superparamagnetic iron oxide nanoparticles (SPIONs), in addition to rendering the NPs the luminescence property. After the incorporation of GS-AuNPs onto the surface of SPIONs to form the hybrid nanoparticles (HBNPs), these SPIONs’ protein adsorption was about 10-fold lower than those of the pure glutathione-coated SPIONs suggesting that GS-AuNPs are capable of providing a stealth effect against serum proteins. PMID:25029478

  1. Effect of bone morphogenetic protein-4 on cardiac differentiation from mouse embryonic stem cells in serum-free and low-serum media

    Microsoft Academic Search

    Masoumeh Fakhr Taha; Mojtaba Rezazadeh Valojerdi

    2008-01-01

    In spite of previous reports, the precise role of bone morphogenetic proteins (BMPs) on cardiomyocyte differentiation, especially in the absence or presence of minimum amount of serum in culture medium is still unclear. So, the aim of the present study was to investigate the effect of BMP-4 on mouse embryonic stem cells (ESCs)-derived cardiomyocyte differentiation in serum-free and low-serum media.

  2. Serum zinc and copper level in children with protein energy malnutrition.

    PubMed

    Gautam, B; Deb, K; Banerjee, M; Ali, M S; Akhter, S; Shahidullah, S M; Hoque, M R

    2008-07-01

    This case control study was carried out in the department of Biochemistry, Mymensingh Medical College in co-operation with the Pediatric wards of Mymensingh Medical College Hospital and Ganashasthya Nagar Hospital, Dhaka during the period from July 2005 to June 2006. The aim of the study was to explore the status of serum zinc and copper level in Bangladeshi children with Protein Energy Malnutrition (PEM) as a means to monitor the possibility of management of these children as each of these mineral deficiencies produce typical deficiency syndromes. A total of 68 children aging from five months to five years were included in this study. Subjects were divided into two groups-Group I (Control; n=20)-children with normal growth, weight for age between 3rd and 97th centile curve, Centers for Disease Control (CDC) growth chart, USA, 2000 and group II-(children with PEM; n=48)-children with retarded growth, weight for age below 3rd centile of CDC growth chart, USA, 2000. Group II was again divided into three subgroups according to Wellcome classification of PEM and clinical features. These were Group IIA: Marasmus (n=19), Group IIB: Kwashiorkor (n=14) and Group IIC: Marasmic Kwashiorkor (n=15). Serum zinc and copper levels were determined by Atomic Absorption Spectrophotometric method. Statistical analysis was done by using Statistical Package for the Social Sciences (SPSS) window package. Among the different groups of children mean+/-SD (Standard Deviation) of serum zinc in PEM (59.85+/-11.18 microg/dl), Marasmus (66.73+/-8.23 microg/dl), Kwashiorkor (49.69+/-10.35 microg/dl) and Marasmic Kwashiorkor (60.63+/-8.04 microg/dl) were all significantly lower (p<0.001) than in control group (106.16+/-13.36 microg/dl). Similarly mean+/-SD of serum copper in PEM (82.73+/-16.35 microg/dl), Marasmus (93.72+/-9.77 microg/dl), Kwashiorkor (63.75+/-13.12 microg/dl) and Marasmic Kwashiorkor (86.52+/-8.68 microg/dl) were all also significantly lower (p<0.001) than in control group (135.88+/-11.88 microg/dl). It is evident from the study that serum zinc and copper level significantly decrease in children with PEM. PMID:18946444

  3. Association between serum levels of C-reactive protein and personality traits in women

    Microsoft Academic Search

    Susanne Henningsson; Fariba Baghaei; Roland Rosmond; Göran Holm; Mikael Landén; Henrik Anckarsäter; Agneta Ekman

    2008-01-01

    BACKGROUND: While low-grade inflammation has consistently been observed in subjects with depression, studies on the possible relationship between inflammation and other aspects of brain function are as yet sparse. In this study, we aimed to investigate the possible association between serum levels of the inflammation marker C-reactive protein (CRP) and personality traits. METHODS: In this study, serum levels of high-sensitivity

  4. Effects of psychological stress on serum immunoglobulin, complement and acute phase protein concentrations in normal volunteers

    Microsoft Academic Search

    Michael Maes; Dirk Hendriks; An Van Gastel; Paul Demedts; Annick Wauters; Hugo Neels; Aleksandar Janca; Simon Scharpé

    1997-01-01

    The aim of this study was to examine the effects of academic examination stress on serum immunoglobulins (Igs), i.e. IgA, IgG, IgM, complement factors, i.e. C3c and C4, and acute phase proteins, i.e. ?1-acid glycoprotein (?1-S), haptoglobin (Hp), and ?2-macroglobulin (?2-M). Thirty-seven university students participated in this study. Serum was sampled a few weeks before and after as well as

  5. Comparison of Serum Protein-Bound Carbohydrate and Glycoprotein Patterns of Man, Monkey and Rat

    PubMed Central

    Cockerell, G. L.; Beisel, W. R.

    1973-01-01

    Baseline concentrations of serum protein-bound carbohydrates and glycoprotein patterns of man, rhesus monkey and albino rat were compared. In all cases patterns of monkey resembled man more closely than did rat. Based on these data it is concluded that the rhesus monkey can serve as a relatively more accurate model than the albino rat to study alterations in concentrations of serum glycoproteins that occur during various infectious and inflammatory processes in man. ImagesFig. 2Fig. 3 PMID:4632122

  6. Effects of soy protein and saponins on serum, tissue and feces steroids in rat.

    PubMed

    Sautier, C; Doucet, C; Flament, C; Lemonnier, D

    1979-11-01

    Four groups of rats were fed, for 45 days, one of the following semipurified diets containing sucrose 55% (w/w) and (a) casein 25%, (b) casein 24%, saponins (from Saponaria officinalis) 1%, (c) isolated soy protein 25%, (d) soy protein 24%, saponins 1%. The soy protein diet, compared to the casein one, produced an increase in the fecal excretion of neutral sterols on the 29th and 42nd days, without any modification in the liver, aorta and serum cholesterol concentrations. The effect of soy protein cannot be attributed to its saponin content but other substances associated to soy protein may interfere. With the casein diet, added saponins increased the fecal excretion of neutral sterols and bile acids and decreased liver and aorta cholesterol levels. Serum cholesterol was found unchanged. The effects of saponins were suppressed or greatly reduced with the soy protein diet. These results could be explained by binding of the sterols in insoluble forms. PMID:574769

  7. Probing thyroglobulin in undiluted human serum based on pattern recognition and competitive adsorption of proteins

    NASA Astrophysics Data System (ADS)

    Wang, Ran; Huang, Shuai; Li, Jing; Chae, Junseok

    2014-10-01

    Thyroglobulin (Tg) is a sensitive indicator of persistent or recurrent differentiated thyroid cancer of follicular cell origin. Detection of Tg in human serum is challenging as bio-receptors, such as anti-Tg, used in immunoassay have relatively weak binding affinity. We engineer sensing surfaces using the competitive adsorption of proteins, termed the Vroman Effect. Coupled with Surface Plasmon Resonance, the "cross-responsive" interactions of Tg on the engineered surfaces produce uniquely distinguishable multiple signature patterns, which are discriminated using Linear Discriminant Analysis. Tg-spiked samples, down to 2 ng/ml Tg in undiluted human serum, are sensitively and selectively discriminated from the control (undiluted human serum).

  8. Preoperative elevation of serum C-reactive protein as an independent prognostic indicator for gastric cancer

    Microsoft Academic Search

    Tadahiro Nozoe; Tomohiro Iguchi; Eisuke Adachi; Akito Matsukuma; Takahiro Ezaki

    2011-01-01

    Purpose  Preoperative elevation of serum C-reactive protein (CRP) is a prognostic indicator for some malignant tumors. We investigated\\u000a the clinicopathologic importance of an elevated preoperative serum CRP value in gastric carcinoma patients.\\u000a \\u000a \\u000a \\u000a \\u000a Methods  We studied the relationship between the preoperative serum CRP value and clinicopathologic characteristics in 204 patients\\u000a who underwent curative resection of gastric carcinoma.\\u000a \\u000a \\u000a \\u000a \\u000a Results  The patients with preoperative CRP elevation

  9. Absence of serum growth hormone binding protein in patients with growth hormone receptor deficiency (Laron dwarfism)

    SciTech Connect

    Daughaday, W.H.; Trivedi, B.

    1987-07-01

    It has recently been recognized that human serum contains a protein that specifically binds human growth hormone (hGH). This protein has the same restricted specificity for hGH as the membrane-bound GH receptor. To determine whether the GH-binding protein is a derivative of, or otherwise related to, the GH receptor, the authors have examined the serum of three patients with Laron-type dwarfism, a condition in which GH refractoriness has been attributed to a defect in the GH receptor. The binding of /sup 125/I-labeled hGH incubated with serum has been measured after gel filtration of the serum through an Ultrogel AcA 44 minicolumn. Results are expressed as percent of specifically bound /sup 125/I-hGH and as specific binding relative to that of a reference serum after correction is made for endogenous GH. The mean +/- SEM of specific binding of sera from eight normal adults (26-46 years of age) was 21.6 +/- 0.45%, and the relative specific binding was 101.1 +/- 8.6%. Sera from 11 normal children had lower specific binding of 12.5 +/- 1.95% and relative specific binding of 56.6 +/- 9.1%. Sera from three children with Laron-type dwarfism lacked any demonstrable GH binding, whereas sera from 10 other children with other types of nonpituitary short stature had normal relative specific binding. They suggest that the serum GH-binding protein is a soluble derivative of the GH receptor. Measurement of the serum GH-binding protein may permit recognition of other abnormalities of the GH receptor.

  10. The vitamin E-binding protein afamin increases in maternal serum during pregnancy

    PubMed Central

    Hubalek, Michael; Buchner, Hannes; Mörtl, Manfred G.; Schlembach, Dietmar; Huppertz, Berthold; Firulovic, Branka; Köhler, Wolfgang; Hafner, Erich; Dieplinger, Benjamin; Wildt, Ludwig; Dieplinger, Hans

    2014-01-01

    Background Afamin is a liver-derived plasma glycoprotein with vitamin E-binding properties and a putative function in fertility. This study evaluated serum afamin concentrations during and postpartum to uncomplicated pregnancies and investigated a potential association between afamin concentrations and pregnancy outcome. Methods Afamin serum concentrations were measured in women with uncomplicated pregnancies in a retrospective cohort (n = 466) at different gestational ages and a prospective observational study (n = 76) in the first, second and third trimester. Furthermore, afamin was determined in the first trimester in a cross-sectional pilot study including women with preeclampsia (PE), pregnancy-induced hypertension (PIH) and women without pregnancy complications (n = 13 each). Finally, expression of afamin was investigated in human placental tissue by RT-PCR and immunohistochemistry. Results Afamin concentrations increased linearly almost two-fold during pregnancy in both retrospective and prospective studies in women without pregnancy complications with median afamin serum concentrations of 61.9 mg/l, 79.6 mg/l, and 98.6 mg/l in the first, second, and third trimester, respectively. After delivery, median afamin concentrations decreased to baseline values of 54.6 mg/l. In the pilot study with pregnancy complications, women with PE displayed significantly higher median afamin concentrations than did women with uncomplicated pregnancy (70.0 mg/l vs. 55.4 mg/l, P = 0.007). Expression analyses revealed no placental afamin expression at either mRNA or protein level in uncomplicated pregnancy. Conclusion A linear increase in the maternally expressed glycoprotein afamin during pregnancy may serve as basic reference for subsequent investigations of afamin in pregnancy-related disorders. PMID:24768783

  11. Quantitative profiling of serum samples using TMT protein labelling, fractionation and LC-MS/MS.

    PubMed

    Sinclair, John; Timms, John F

    2011-08-01

    Blood-borne biomarkers are urgently required for the early detection, accurate diagnosis and prognosis of disease. Additionally, improved methods of profiling serum and plasma proteins for biomarker discovery efforts are needed. Herein, we report a quantitative method based on amino-group labelling of serum proteins (rather than peptides) with isobaric tandem mass tags (TMT) and incorporating immune-based depletion, gel-based and strong anion exchange separation of proteins prior to differential endoproteinase treatment and liquid chromatography tandem mass spectrometry. We report a generally higher level of quantitative coverage of the serum proteome compared to other peptide-based isobaric tagging approaches and show the potential of the method by applying it to a set of unique samples that pre-date the diagnosis of pancreatic cancer. PMID:21397697

  12. Gross protein influence upon blood plasma and serum self organization processes in patients with coronary heart disease

    NASA Astrophysics Data System (ADS)

    Malinova, Lidia I.; Sergeeva, Uliya V.; Simonenko, Georgy V.; Denisova, Tatiana P.; Tuchin, Valery V.

    2007-05-01

    Blood plasma pattern formation is a process sensitive to environment and carrier properties, and plasma biochemical content. 96 patients with coronary heart disease (CHD) were involved in the study. Control group include 12 practically health persons (PHP). Platelets poor plasma and serum were used to study functional morphology. Plasma and serum samples of equal volume were placed on degreased glass carrier with after going wedge dehydration. The result of wedge dehydration is a formation of a special structure called facia. To the samples of compare albumin solution was added. Morphology of prepared facies was studied by means of light microscopy ("Lomo Biolam P2-1") with 10 times magnification. All received facies were of the same principle structure with central, intermediate and edge zones. Zone index was increasing in samples with albumin adding. Special structures obligatory to atherosclerosis, vessels stiffness increase and hypoxia were found in facies of plasma and serum of patients with CHD. Quantity of these structures correlated to protein concentration (p = 0.021). Samples' drying period was also increasing in samples of compare, and differed significantly in patients with CHD and PHP. In our study gross proteins concentration increase modified plasma and serum morphology. Albumin solution can be proposed as a probe to elucidate differences of facies of patients with CHD and PHP.

  13. Isoforms of procarboxypeptidase B, (pancreas-specific protein, PASP) in human serum, pancreatic tissue and juice.

    PubMed

    Fernstad, R; Kylander, C; Tsai, L; Tydén, G; Pousette, A

    1993-01-01

    Human Pancreas-Specific Protein (PASP) has been described as a useful serum marker for pancreatic graft rejection and acute pancreatitis. By molecular cloning PASP has recently been identified as procarboxypeptidase B (PCPB). By use of SDS-gel electrophoresis and Western blots, PASP isoforms (proteins interacting with PASP-antiserum) have now been explored in serum and pancreatic tissue and juice. In serum from healthy volunteers, six different isoforms could be visualised. Their MW varied as follows: > 100 K, about 100 K, 45 K, 35-40 K (two bands) about 30 K and 16 K. The 45 K band, corresponding to PASP purified from pancreatic tissue, was always the major band. In pancreatic cytosol and pancreatic juice, the major band corresponded to PASP (45 K) but weak bands were also seen at MW of 40, 30 and 16 K. In serum from 15 patients with acute pancreatitis, PASP was highly increased. Bands corresponding to PASP/PCPB as well as carboxypeptidase B (CPB) were seen simultaneously. During recovery PASP in serum was normalised and the CPB band disappeared. During 12 episodes of pancreatic graft rejection, PASP in serum was also increased but no changes of isoforms in serum were detected. However during these episodes isoforms in pancreatic juice changed dramatically. The band corresponding to PASP disappeared or became much weaker and only bands with lower MW were seen. We have thus observed changes in PASP/PCB serum isoforms during episodes of acute pancreatitis but not at graft rejection episodes although the total increase of PASP in serum was of the same magnitude.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8322019

  14. Relationship of the serum protein-bound iodine to rates of gain in beef cattle

    E-print Network

    Stokes, David Kershaw

    1956-01-01

    RELATIONSHIP OP THE SERUM PROTEIN-BOUND IODINE TO RATES OF GAIN IN BEEP CATTLE A Dissertation b7 DAVID KERSHAW STOKES, JR. Approved as to style and content by: January 1956 L IB R ARY R ft M COLLEGE OF TEXAS RELATIONSHIP OP THE SERUM... PROTEIN-BOUND IODINE TO RATES OF GAIN IN BEEP CATTLE A Dissertation RE DAVID KERSHAW STOKES, JR. Subiaitted to the Graduate School of the Agricultural and Mechanical College of Texas in partial fulfillment of the requirements for the degree...

  15. Signaling by retinol and its serum binding protein.

    PubMed

    Noy, Noa

    2015-02-01

    Vitamin A, retinol, circulates in blood bound to retinol-binding protein (RBP) which, in turn, associates with transthyretin (TTR) to form a retinol-RBP-TTR ternary complex. At some tissues, retinol-bound (holo-) RBP is recognized by a membrane protein termed STRA6, which transports retinol from extracellular RBP into cells and, concomitantly, activates a JAK2/STAT3/5 signaling cascade that culminates in induction of STAT target genes. STRA6-mediated retinol transport and cell signaling are critically inter-dependent, and they both require the presence of cellular retinol-binding protein 1 (CRBP1), an intracellular retinol acceptor, as well as a retinol-metabolizing enzyme such as lecithin:retinol acyltransferase (LRAT). STRA6 thus functions as a "cytokine signaling transporter" which couples vitamin A homeostasis and metabolism to cell signaling, thereby regulating gene transcription. Recent studies provided molecular level insights into the mode of action of this unique protein. PMID:25481334

  16. Synthesis of serum proteins by the posthaematopoietic feline yolk sac

    Microsoft Academic Search

    K. Tiedemann; W. W. Minuth

    1980-01-01

    The feline yolk sac persists even after the end of its haematopoietic phase with prominent ER-cisternae in the endoderm suggesting biosynthetic capacity. Therefore, yolk sac explants from the 54th day and 57th day were incubated with [3H]-l-leucine in order to study its protein biosynthesis. Newly synthesized proteins were discovered in sliced SDS-polyacrylamide gels by the use of scintillation technique and

  17. Correlation between serum levels of C-reactive protein and infant pneumonia: A meta-analysis

    PubMed Central

    XIAO, XIAO; XUE, LONG; SHENG, HUI-LIN; XIAO, ZHI-HUI

    2015-01-01

    Experimental data imply that serum C-reactive protein (CRP) is a marker of general systemic inflammation, and inflammation may have a role in the development of pneumonia. The aim of the present study was to investigate the associations of serum CRP levels in infant patients with pneumonia by meta-analysis. The Science Citation Index, Cochrane Library, PubMed, Embase, CINAHL, Current Contents and two Chinese (CMB and CNKI) databases were searched. Studies were pooled and standard mean difference (SMD) and the corresponding 95% confidence interval (CI) were calculated. Subgroup analyses and publication bias detection were also conducted. The statistical analysis was conducted using Stata software version 12.0. Serum CRP levels were analyzed in 10 clinical case-control studies (652 infants with pneumonia and 845 healthy controls); significant differences in serum CRP levels were observed between infants with pneumonia and the healthy controls (SMD=4.41, 95% CI: 3.34–5.47, P<0.001). Ethnicity-stratified subgroup analysis detected that high levels of serum CRP may be the main risk factor for infant pneumonia in Asian, African and Caucasian populations (all P<0.05). Serum CRP levels were statistically higher in infants with pneumonia than in healthy infants, and thus serum levels of CRP may have independent diagnostic value for pneumonia in children. PMID:26136982

  18. Serum from human burn victims impairs myogenesis and protein synthesis in primary myoblasts

    PubMed Central

    Corrick, Katie L.; Stec, Michael J.; Merritt, Edward K.; Windham, Samuel T.; Thomas, Steven J.; Cross, James M.; Bamman, Marcas M.

    2015-01-01

    The pathophysiological response to a severe burn injury involves a robust increase in circulating inflammatory/endocrine factors and a hypermetabolic state, both of which contribute to prolonged skeletal muscle atrophy. In order to characterize the role of circulating factors in muscle atrophy following a burn injury, human skeletal muscle satellite cells were grown in culture and differentiated to myoblasts/myotubes in media containing serum from burn patients or healthy, age, and sex-matched controls. While incubation in burn serum did not affect NF?B signaling, cells incubated in burn serum displayed a transient increase in STAT3 phosphorlyation (Tyr705) after 48 h of treatment with burn serum (? + 70%; P < 0.01), with these levels returning to normal by 96 h. Muscle cells differentiated in burn serum displayed reduced myogenic fusion signaling (phospho-STAT6 (Tyr641), ??75%; ADAM12, ?-20%; both P < 0.01), and reduced levels of myogenin (??75%; P < 0.05). Concomitantly, myotubes differentiated in burn serum demonstrated impaired myogenesis (assessed by number of nuclei/myotube). Incubation in burn serum for 96 h did not increase proteolytic signaling (assessed via caspase-3 and ubiquitin levels), but reduced anabolic signaling [p-p70S6k (Ser421/Thr424), ?30%; p-rpS6 (Ser240/244), ?-50%] and impaired protein synthesis (?24%) (P < 0.05). This resulted in a loss of total protein content (?18%) and reduced cell size (?33%) (P < 0.05). Overall, incubation of human muscle cells in serum from burn patients results in impaired myogenesis and reduced myotube size, indicating that circulating factors may play a significant role in muscle loss and impaired muscle recovery following burn injury. PMID:26136691

  19. Serum from human burn victims impairs myogenesis and protein synthesis in primary myoblasts.

    PubMed

    Corrick, Katie L; Stec, Michael J; Merritt, Edward K; Windham, Samuel T; Thomas, Steven J; Cross, James M; Bamman, Marcas M

    2015-01-01

    The pathophysiological response to a severe burn injury involves a robust increase in circulating inflammatory/endocrine factors and a hypermetabolic state, both of which contribute to prolonged skeletal muscle atrophy. In order to characterize the role of circulating factors in muscle atrophy following a burn injury, human skeletal muscle satellite cells were grown in culture and differentiated to myoblasts/myotubes in media containing serum from burn patients or healthy, age, and sex-matched controls. While incubation in burn serum did not affect NF?B signaling, cells incubated in burn serum displayed a transient increase in STAT3 phosphorlyation (Tyr705) after 48 h of treatment with burn serum (? + 70%; P < 0.01), with these levels returning to normal by 96 h. Muscle cells differentiated in burn serum displayed reduced myogenic fusion signaling (phospho-STAT6 (Tyr641), ?-75%; ADAM12, ?-20%; both P < 0.01), and reduced levels of myogenin (?-75%; P < 0.05). Concomitantly, myotubes differentiated in burn serum demonstrated impaired myogenesis (assessed by number of nuclei/myotube). Incubation in burn serum for 96 h did not increase proteolytic signaling (assessed via caspase-3 and ubiquitin levels), but reduced anabolic signaling [p-p70S6k (Ser421/Thr424), -30%; p-rpS6 (Ser240/244), ?-50%] and impaired protein synthesis (-24%) (P < 0.05). This resulted in a loss of total protein content (-18%) and reduced cell size (-33%) (P < 0.05). Overall, incubation of human muscle cells in serum from burn patients results in impaired myogenesis and reduced myotube size, indicating that circulating factors may play a significant role in muscle loss and impaired muscle recovery following burn injury. PMID:26136691

  20. Identification and characterization of lipopolysaccharide binding protein (LBP) as an estrogen receptor ? specific serum biomarker

    PubMed Central

    Chisamore, Michael J; Hong, Kwok-lam Karen; Cheng, Chun; Alves, Stephen E; Rohrer, Susan P; Wilkinson, Hilary A

    2012-01-01

    Estrogen Receptor a (ER?) and Estrogen Receptor ? (ER?) are steroid nuclear receptors that transduce estrogen signaling to control diverse physiological processes linked to reproduction, bone remodeling, behavior, immune response and endocrine-related diseases. In order to differentiate between ER? and ER? mediated effects in vivo, ER subtype selective biomarkers are essential. We utilized ER? knockout (AERKO) and ER? knockout (BERKO) mouse liver RNA and genome wide profiling to identify novel ER? selective serum biomarker candidates. Results from the gene array experiments were validated using real-time RT-PCR and subsequent ELISA's to demonstrate changes in serum proteins. Here we present data that Lipopolysacharide Binding Protein (LBP) is a novel liver-derived ER? selective biomarker that can be measured in serum. PMID:22299632

  1. Identification and characterization of lipopolysaccharide binding protein (LBP) as an estrogen receptor ? specific serum biomarker.

    PubMed

    Chisamore, Michael J; Hong, Kwok-Lam Karen; Cheng, Chun; Alves, Stephen E; Rohrer, Susan P; Wilkinson, Hilary A

    2012-03-01

    Estrogen Receptor ? (ER?) and Estrogen Receptor ? (ER?) are steroid nuclear receptors that transduce estrogen signaling to control diverse physiological processes linked to reproduction, bone remodeling, behavior, immune response and endocrine-related diseases. In order to differentiate between ER? and ER? mediated effects in vivo, ER subtype selective biomarkers are essential. We utilized ER? knockout (AERKO) and ER? knockout (BERKO) mouse liver RNA and genome wide profiling to identify novel ER? selective serum biomarker candidates. Results from the gene array experiments were validated using real-time RT-PCR and subsequent ELISA's to demonstrate changes in serum proteins. Here we present data that Lipopolysacharide Binding Protein (LBP) is a novel liver-derived ER? selective biomarker that can be measured in serum. PMID:22299632

  2. Serum proteins prevent aggregation of Fe2O3 and ZnO nanoparticles

    PubMed Central

    Wells, Mark A.; Abid, Aamir; Kennedy, Ian M.; Barakat, Abdul I.

    2014-01-01

    Aggregation of metal oxide nanoparticles in aqueous media complicates interpretation of in vitro studies of nanoparticle–cell interactions. We used dynamic light scattering to investigate the aggregation dynamics of iron oxide and zinc oxide nanoparticles. Our results show that iron oxide particles aggregate more readily than zinc oxide particles. Pretreatment with serum stabilises iron oxide and zinc oxide nanoparticles against aggregation. Serum-treated iron oxide is stable only in pure water, while zinc oxide is stable in water or cell culture media. These findings, combined with zeta potential measurements and quantification of proteins adsorbed on particle surface, suggest that serum stabilisation of iron oxide particles occurs primarily through protein adsorption and resulting net surface charge. Zinc oxide stabilisation, however, also involves steric hindrance of particle aggregation. Fluid shear at levels used in flow experiments breaks up iron oxide particle aggregates. These results enhance our understanding of nanoparticle aggregation and its consequences for research on the biological effects of nanomaterials. PMID:22149273

  3. Measurement of serum PSP\\/ reg -protein concentration in various diseases with a newly developed enzyme-linked immunosorbent assay

    Microsoft Academic Search

    Yoshitake Satomura; Norio Sawabu; Ikurou Mouri; Osamu Yamakawa; Hiroyuki Watanabe; Yoshiharu Motoo; Takashi Okai; Takako Ito; Kazue Kaneda; Hiroshi Okamoto

    1995-01-01

    An enzyme-linked immunosorbent assay, based on two monoclonal antibodies (Hreg1-1 and Hreg101-1) specific for pancreatic stone protein (PSP)\\/reg-protein, was developed to determine the concentration of this protein in serum from individuals with various diseases. The\\u000a serum concentration of PSP\\/reg-protein was significantly higher in patients with various pancreatic diseases than in normal controls, and was also significantly\\u000a higher in patients with

  4. Annexin A1 protein regulates the expression of PMVEC cytoskeletal proteins in CBDL rat serum-induced pulmonary microvascular remodeling

    PubMed Central

    2013-01-01

    Background Hepatopulmonary syndrome (HPS) is characterized by advanced liver disease, hypoxemia and intrapulmonary vascular dilatation (IPVD). The pathogenesis of HPS is not completely understood. Recent findings have established the role of proliferation and phenotype differentiation of pulmonary microvascular endothelial cells (PMVECs) in IPVD of HPS; the change in cytoskeletal proteins and their molecular mechanism play an essential role in the proliferation, phenotype modulation and differentiation of PMVECs. However, little is known about the relevance of cytoskeletal protein expression and its molecular mechanism in IPVD of HPS. In addition, ANX A1 protein has been identified as a key regulator in some important signaling pathways, which influences cytoskeletal remodeling in many diseases, such as lung cancer, liver cancer, etc. Methods PMVECs were cultured from the normal rats and then divided into three groups(Ad-ANXA1-transfected group, a non-transfected group, and an adenovirus empty vector group) and incubated by nomal rat serum or hepatopulmonary syndrome rat serum respectively. mRNA level was evaluated by real time reverse transcription polymerase chain reaction, and protein expression was detected by western blot. Cell proliferation was determined by the MTT and thymidine incorporation assay. Results In this study, we found that the serum from a common bile duct ligation(CBDL) Rat model decreased the expression levels of the ANX A1 mRNA and protein by at least two-fold in human PMVECs. We also found the expression of cytoskeletal proteins (Destrin, a1-actin, and a1-tubulin) in PMVECs significantly increased. After stimulating ANX A1 over-expression in PMVECs by adenovirus-mediated ANX A1 (Ad-ANXA1) transfection, we found the expression levels of cytoskeletal proteins were significantly suppressed in PMVECs at all time points. Additionally, we report here that serum from a CBDL Rat model increases the proliferation of PMVECs by nearly two-fold and that over-expression of Ad-ANXA1 significantly inhibits HPS-rat-serum-induced PMVEC proliferation (p <0.05). These findings suggest that the ANX A1 down-regulation of PMVEC proliferation in the presence of HPS-rat-serum may be the major cause of aberrant dysregulation of cytoskeletal proteins (Destrin, a1-actin, and a1-tubulin) and may, therefore, play a fundamental role in the proliferation and phenotype differentiation of PMVECs in the PVD of HPS. Conclusion Finally, the fact that transfection with Ad-ANXA1 results in inhibition of the aberrant dysregulation of cytoskeletal proteins and proliferation of PMVECs suggests a potential therapeutic effect on PVD of HPS. PMID:23587191

  5. The effects of temperature on thyroid hormone binding to serum proteins in sea turtles 

    E-print Network

    Haynes, Shane Patrick

    1990-01-01

    THE EFFECTS OF TEMPERATURE ON THYROID HORMONE BINDING TO SERUN PROTEINS IN SEA TURTLES A Thesis by SHANE PATRICK HAYNES Submitted to the Office of Graduate Studies of Texas A&M University in partial fulfillment of the requirements... for the degree of MASTER OF SCIENCE August 1990 Major Subject: Zoology THE EFFECTS OF TEMPERATURE ON THYROID HORMONE BINDING TO SERUM PROTEINS IN SEA TURTLES A Thesis by SHANE PATRICK HAYNES Approved as to style and content by: Duncan S. Mac...

  6. Serum surfactant protein D is increased in acute and chronic inflammation in mice

    Microsoft Academic Search

    Masaki Fujita; John M. Shannon; Hiroshi Ouchi; Dennis R. Voelker; Yoichi Nakanishi; Robert J. Mason

    2005-01-01

    Surfactant protein A (SP-A) and surfactant protein D (SP-D) are important components of innate immunity that can modify the inflammatory response. However, alterations and regulation of SP-A and SP-D in acute and chronic inflammation are not well defined. In addition, serum SP-D may serve as a biomarker of lung inflammation.We determined the expression of SP-A and SP-D in murine models.

  7. Effect of Citrus reticulata on Serum Protein Fractions of Mice After Schistosoma mansoni Infection

    Microsoft Academic Search

    Nagy-Saba El-Rigal; Mona H. Hetta

    2006-01-01

    The effect of the ethanolic extract of Citrus reticulata roots or the oleo-resin extract from Myrrh of Commiphora molmol tree (Mirazid) were studied as new antischistosomal drugs. The total protein and the different serum proteins namely albumin, prealbumin, beta-lipoproteins, macroglobulins, alpha-1-acid glycoprotein, alpha-1-antitrypsin, cholinesterase, ceruloplasmin, haemopexin, haptoglobin, transferrin and lipoprotein were measured in both infected and treated mice. Also, liver

  8. Hyperglycemia and non-enzymatic glycation of serum and tissue proteins in chickens

    Microsoft Academic Search

    H. Klandorf; S. B. Holt; J. A. McGowan; Y. Pinchasov; D. Deyette; R. A. Peterson

    1995-01-01

    The objectives of these studies were to determine whether elevated plasma glucose concentrations in broiler breeder chickens (200–250 mg\\/dl) can result in the non-enzymatic attachment of glucose to serum proteins (fructosamine) and eventual cross-linking of tissue proteins (basement membrane thickness), and to investigate the effects of a factor that may influence this cross-linking process. In response to feeding the satiety

  9. Serum electrolyte and protein modification during different workload in jumper horse

    Microsoft Academic Search

    G. Piccione; C. Giannetto; A. Assenza; F. Fazio; G. Caola

    2007-01-01

    Five clinically healthy Italian saddle horses were used to assess serum electrolyte and protein modification during different\\u000a workloads. Blood samples were collected from each horse at rest, immediately after each exercise and at 30 and 60 min after\\u000a the end of exercise. Our results confirm that exercise has variable effects, depending on work intensity, on some electrolytes,\\u000a total protein and haematocrit.

  10. Synthetic serum substitute (SSS): A globulin-enriched protein supplement for human embryo culture

    Microsoft Academic Search

    Paul S. Weathersbee; Thomas B. Pool; Teri Ord

    1995-01-01

    Objective: The purpose of the present study was to evaluate whether an IVF protein supplement prepared from human serum albumin (HSA) and human globulins would retain performance characteristics equivalent to those reported for the commercial plasma expanders, Plasmatein (Alpha Therapeutics, Los Angeles, California) and Plasmanate (Cutter Biological, Miles Inc., Elkhart, Indiana). Methods: Pronuclear-stage human embryos were randomly divided and cultured

  11. Serum protein changes in immune and nonimmune pigeons infected with various strains of Trichomonas gallinae

    USGS Publications Warehouse

    Kocan, R.M.; Herman, C.M.

    1970-01-01

    Serum protein changes were studied in immune and nonimmune pigeons infected with three different strains of Trichomonas gallinae. Strain I (nonvirulent) produced no change in the relative concentration of serum components. Strains II (oral canker) and III (Jones' Barn) produced decreases in albumin and alpha globulins, and increases in beta and gamma globulins between the 7th and 20th days post infection. Birds infected with strain II began to return to normal by the 20th day, while all those infected with strain III were dead between 10 and 14 days post infection. Two serum protein patterns resulted from infection of immune birds with the Jones' Barn strain. One showed no change in relative protein concentrations and no tissue invasion by the parasite while the other was similar to that seen in nonimmune birds infected with a strain producing oral canker. These also showed evidence of tissue invasion by the parasite. It was concluded that tissue invasion was necessary to evoke a quantitative change in serum protein concentrations.

  12. Changes of Growth and of Serum Proteins in Ducklings Intoxicated with Cobalt

    Microsoft Academic Search

    S. Paulov

    1971-01-01

    Changes of growth and serum proteins were found in ducklings nourished with commercial diet enriched with cobalt. It was found that a high concentration of this element in diet involves a significant diminution of growth, an increase of ?-globulins and a decrease of albumin. It was observed too that the diminution of concentration of cobalt in diet involves after- wards

  13. Peculiarities of the Genetic Variability of Blood Serum Proteins in Some Species of the Family Anatidae

    Microsoft Academic Search

    Sigita Slav?nait?; Aniolas Sruoga; Algimantas Paulauskas; Elena Mozalien?

    2000-01-01

    We investigated blood serum of the following eight waterfowl species from the family Anatidae in polyacrylamide gel: Goosander (Mergus merganser), Smew (Mergus albellus), Eider (Somateria mollissima), Spectacled Eider (Somateria fisheri), Common Scoter (Melanitta nigra), Harlequin (Histronicus histronicus), Goldeneye (Bucephala clangula) and Long-tailed Duck (Clangula hyemalis). During our investigations we identified the systems of general protein and enzymes in polyacrylamide gel.

  14. Serum protein levels and neonatal growth rate of Nubian goat kids in Taiwan area

    Microsoft Academic Search

    J. C Chen; C. J Chang; H. C Peh; S. Y Chen

    1999-01-01

    Levels of serum total protein and ?-globulin were determined in 43 newborn goat kids of up to 5 days of age on two goat farms in Taiwan using colorimetric assay and an agarose gel electrophoresis kit, respectively. The kids were sufficiently bottle-fed with the maternal colostrum every 6h for the first day and every 12h since 2 days. The overall

  15. Iron Dextran treatment does not induce serum protein carbonyls in the newborn pig

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Oxidation of serum proteins can lead to carbonyl formation which alters their function and is often associated with stress-related diseases. Since it is recommended that all pigs reared in modern production facilities be given supplemental iron at birth to prevent anemia, and metals can catalyze th...

  16. Serum C-reactive protein as a prognostic factor in patients with epithelial ovarian cancer

    Microsoft Academic Search

    Junichi Kodama; Yasunari Miyagi; Noriko Seki; Keizo Tokumo; Mitsuo Yoshinouchi; Yuuji Kobashi; Hiroyuki Okuda; Takafumi Kudo

    1999-01-01

    Objective: It is well known that the serum level of Interleukin-6 (IL-6) correlates with the level of C-reactive protein (CRP). The purpose of this study is to determine the significance of CRP as a prognostic factor in epithelial ovarian cancer. Study Design: The present study is comprised of 120 patients with epithelial ovarian cancer from 1985 to 1992. In this

  17. High Throughput Quantitative Analysis of Serum Proteins Using Glycopeptide Capture and Liquid Chromatography

    Microsoft Academic Search

    Mass Spectrometry; Hui Zhang; Eugene C. Yi; Xiao-jun Li; Parag Mallick; Karen S. Kelly-Spratt; Christophe D. Masselon; David G. Camp II; Richard D. Smith; Christopher J. Kemp; Ruedi Aebersold

    It is expected that the composition of the serum proteome can provide valuable information about the state of the human body in health and disease and that this informa- tion can be extracted via quantitative proteomic measure- ments. Suitable proteomic techniques need to be sensi- tive, reproducible, and robust to detect potential biomarkers below the level of highly expressed proteins,

  18. Analysis of blood gases, serum fat and serum protein: a new approach to estimate survival chances of stranded Harbor seal (Phoca vitulina) pups from the German North Sea

    PubMed Central

    2014-01-01

    Background Facing numerous challenges, such as illness, storms or human disturbance, some harbor seal (Phoca vitulina) pups lose contact to their dams and are found abandoned along the North Sea coast. In Schleswig-Holstein, pups with the prospect of surviving rehabilitation are admitted to the Seal Center Friedrichskoog. Despite elaborate clinical health assessments on admission, including differential hematology, in 2010, 17% of 108 admitted pups did not survive the first 20 days. The death rate during the years 2006 and 2009 varied between 9 and 19%. To broaden the spectrum of variables which could be predictive for survival, blood gas and serum analyses were performed for 99 pups using venous blood. Variables included total CO2, pH, partial CO2, HCO3–, base excess and anion gap as well as glucose, urea nitrogen, sodium, potassium and chloride. Moreover, total serum protein and fat (triglyceride) concentrations were measured for all pups on admission. Results Repeated measurements of 12 randomly selected individuals revealed a significant (p = 0.002) positive influence of time in rehabilitation on triglyceride concentrations. This trend probably shows the improvement of the pups’ nutritional status as a consequence of the shift from milk replacer formula to fish. No such positive influence was detected for total protein concentrations though. Hematologic values, including blood gases, were not predictive for survival. Conclusions For the first time blood gas values are reported in this study for a large sample size (N = 99) of seal pups (regardless of their health status). The ranges and medians calculated from the data can serve as a stepping stone towards the establishment of reference values for neonate harbor seals. However, future investigations on the development of blood gases in harbor seals with different health conditions and ages over time are necessary to allow for a better understanding of acid–base regulation in harbor seals. PMID:24490584

  19. Altered protein expression in serum from endometrial hyperplasia and carcinoma patients

    PubMed Central

    2011-01-01

    Background Endometrial carcinoma is one of the most common gynecological malignancies in women. The diagnosis of the disease at early or premalignant stages is crucial for the patient's prognosis. To date, diagnosis and follow-up of endometrial carcinoma and hyperplasia require invasive procedures. Therefore, there is considerable demand for the identification of biomarkers to allow non-invasive detection of these conditions. Methods In this study, we performed a quantitative proteomics analysis on serum samples from simple endometrial hyperplasia, complex endometrial hyperplasia, atypical endometrial hyperplasia, and endometrial carcinoma patients, as well as healthy women. Serum samples were first depleted of high-abundance proteins, labeled with isobaric tags (iTRAQ™), and then analyzed via two-dimensional liquid chromatography and tandem mass spectrometry. Protein identification and quantitation information were acquired by comparing the mass spectrometry data against the International Protein Index Database using ProteinPilot software. Bioinformatics annotation of identified proteins was performed by searching against the PANTHER database. Results In total, 74 proteins were identified and quantified in serum samples from endometrial lesion patients and healthy women. Using a 1.6-fold change as the benchmark, 12 proteins showed significantly altered expression levels in at least one disease group compared with healthy women. Among them, 7 proteins were found, for the first time, to be differentially expressed in atypical endometrial hyperplasia. These proteins are orosomucoid 1, haptoglobin, SERPINC 1, alpha-1-antichymotrypsin, apolipoprotein A-IV, inter-alpha-trypsin inhibitor heavy chain H4, and histidine-rich glycoprotein. Conclusions The differentially expressed proteins we discovered in this study may serve as biomarkers in the diagnosis and follow-up of endometrial hyperplasia and endometrial carcinoma. PMID:21489304

  20. Mass spectrometric identification of human prostate cancer-derived proteins in serum of xenograft-bearing mice.

    PubMed

    van den Bemd, Gert-Jan C M; Krijgsveld, Jeroen; Luider, Theo M; van Rijswijk, Angelique L; Demmers, Jeroen A A; Jenster, Guido

    2006-10-01

    Lack of sensitivity and specificity of current tumor markers has intensified research efforts to find new biomarkers. The identification of potential tumor markers in human body fluids is hampered by large variability and complexity of both control and patient samples, laborious biochemical analyses, and the fact that the identified proteins are unlikely produced by the diseased cells but are due to secondary body defense mechanisms. In a new approach presented here, we eliminate these problems by performing proteomic analysis in a prostate cancer xenograft model in which human prostate cancer cells form a tumor in an immune-incompetent nude mouse. Using this concept, proteins present in mouse serum that can be identified as human will, by definition, originate from the human prostate cancer xenograft and might have potential diagnostic and prognostic value. Using one-dimensional gel electrophoresis, liquid chromatography, and mass spectrometry, we identified tumor-derived human nm23/nucleoside-diphosphate kinase (NME) in the serum of a nude mouse bearing the androgen-independent human prostate cancer xenograft PC339. NME is known to be involved in the metastatic potential of several tumor cells, including prostate cancer cells. Furthermore we identified six human enzymes involved in glycolysis (fructose-bisphosphate aldolase A, triose-phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, alpha enolase, and lactate dehydrogenases A and B) in the serum of the tumor-bearing mice. The presence of human NME and glyceraldehyde-3-phosphate dehydrogenase in the serum of PC339-bearing mice was confirmed by Western blotting. Although the putative usefulness of these proteins in predicting prognosis of prostate cancer remains to be determined, the present data illustrate that our approach is a promising tool for the focused discovery of new prostate cancer biomarkers. PMID:16714762

  1. Prognostic evaluation of preoperative serum C-reactive protein concentration in patients with epithelial ovarian cancer

    PubMed Central

    LU, YAPING; HUANG, SEN; LI, PING; CHEN, BINGYU; LIU, WEILING; CHEN, ZHIQING; YIN, FUBO

    2015-01-01

    The aim of the present study was to evaluate correlations between preoperative C-reactive protein (CRP) serum concentration and the clinical-pathological parameters of epithelial ovarian cancer (EOC), as well as cancer antigen 125 (CA125) concentration, in patients with EOC. The preoperative serum CRP concentrations of 107 patients with EOC were compared with values of 44 control patients and defined as positive if values were >8 mg/l. Correlations between CRP expression and serum CA125 concentrations, International Federation of Gynecologists and Obstetricians (FIGO) staging, lymph node metastasis, ascites and size of postoperative residual lesions were evaluated for the EOC cases. A total of 69% (74/107) of the EOC cases were CRP-positive, with a mean serum CRP concentration of 14.32 mg/l (versus 2.18 mg/l in the control patients) (P<0.001). A rank correlation analysis showed a positive correlation between serum CRP and CA125 concentrations (P<0.001). The five-year survival rate of the CRP-positive patients with EOC was significant lower than that of the patients that were CRP-negative (33.3 vs. 75.8%, P<0.001). Non-parametric and multivariate analyses showed significant correlations between CRP concentrations and FIGO staging (P=0.001), lymph node metastasis (P=0.001) and ascites (P=0.033). In contrast to previous reports, the mean CRP serum concentration of the CRP-positive patients with EOC also correlated significantly with the CA125 values, and the increases in the CRP serum concentration in the Chinese patients were lower than those described for Caucasian EOC cases. In conclusion, the CRP serum concentration may be a useful clinical marker, solely or in combination with CA125, in patients with EOC.

  2. Analysis of protein oxidation in serum of fetal and newborn piglets and the influence of iron dextran on induction of protein carbonyls.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Methods were employed to evaluate serum biomarkers associated with protein oxidative stress and damage, to determine potential sources of metabolic stress in baby pigs. Protein carbonyls in serum were converted to dinitrophenyl (DNP) derivatives with DNP-hydrazine, precipitated with TCA, extracted i...

  3. Serum-dependent expression of promyelocytic leukemia protein suppresses propagation of influenza virus

    SciTech Connect

    Iki, Shigeo [Department of Microbiology, Sapporo Medical University School of Medicine, Chuo-ku, Sapporo 060-8556 (Japan); Hokkaido Institute of Public Health, Kita-ku, Sapporo 060-0819 (Japan); Yokota, Shin-ichi [Department of Microbiology, Sapporo Medical University School of Medicine, Chuo-ku, Sapporo 060-8556 (Japan); Okabayashi, Tamaki [Department of Microbiology, Sapporo Medical University School of Medicine, Chuo-ku, Sapporo 060-8556 (Japan); Yokosawa, Noriko [Department of Microbiology, Sapporo Medical University School of Medicine, Chuo-ku, Sapporo 060-8556 (Japan); Nagata, Kyosuke [Department of Infection Biology, Graduate School of Comprehensive Human Sciences and Institute of Basic Medical Sciences, University of Tsukuba, Tsukuba 305-8575 (Japan); Fujii, Nobuhiro [Department of Microbiology, Sapporo Medical University School of Medicine, Chuo-ku, Sapporo 060-8556 (Japan)]. E-mail: fujii@sapmed.ac.jp

    2005-12-05

    The rate of propagation of influenza virus in human adenocarcinoma Caco-2 cells was found to negatively correlate with the concentration of fetal bovine serum (FBS) in the culture medium. Virus replicated more rapidly at lower FBS concentrations (0 or 2%) than at higher concentrations (10 or 20%) during an early stage of infection. Basal and interferon (IFN)-induced levels of typical IFN-inducible anti-viral proteins, such as 2',5'-oligoadenylate synthetase, dsRNA-activated protein kinase and MxA, were unaffected by variation in FBS concentrations. But promyelocytic leukemia protein (PML) was expressed in a serum-dependent manner. In particular, the 65 to 70 kDa isoform of PML was markedly upregulated following the addition of serum. In contrast, other isoforms were induced by IFN treatment, and weakly induced by FBS concentrations. Immunofluorescence microscopy indicated that PML was mainly formed nuclear bodies in Caco-2 cells at various FBS concentrations, and the levels of the PML-nuclear bodies were upregulated by FBS. Overexpression of PML isoform consisting of 560 or 633 amino acid residues by transfection of expression plasmid results in significantly delayed viral replication rate in Caco-2 cells. On the other hand, downregulation of PML expression by RNAi enhanced viral replication. These results indicate that PML isoforms which are expressed in a serum-dependent manner suppress the propagation of influenza virus at an early stage of infection.

  4. Immunoglobulin production in the European pond tortoise, Emys orbicularis, immunized with serum protein antigens

    PubMed Central

    Lykakis, J. J.

    1968-01-01

    The immunological responses of the European pond tortoise, Emys orbicularis, to BGG and sheep serum proteins indicate that in this tortoise there is no serum component corresponding with mammalian albumin and that both ?G (7S) and ?M (19S) immunoglobulins are involved when antibodies are produced. A prolonged period of ?M antibody production occurs after primary immunization. The separation of the ?M and ?G immunoglobulins was performed using starch block electrophoresis and Sephadex G-200 gel filtration. The separated immunoglobulins were characterized by immunodiffusion and by starch gel, agar gel and immunoelectrophoresis. ImagesFIG. 1-3FIG. 6FIG. 8FIG. 9FIG. 10 PMID:4173673

  5. The Relationship between Bone Turnover and Body Weight, Serum Insulin-Like Growth Factor (IGF) I, and Serum IGF-Binding Protein Levels in Patients with Anorexia Nervosa

    Microsoft Academic Search

    MARI HOTTA; IZUMI FUKUDA; KANJI SATO; NAOMI HIZUKA; TAMOTSU SHIBASAKI; KAZUE TAKANO

    2010-01-01

    Malnutrition is one of the risk factors for bone loss in patients with anorexia nervosa (AN). To clarify the effects of nutritional status on bone metabolism, we examined the relationship between serum levels of nutritional indicators (insulin-like growth factor I (IGF-I), IGF- binding protein-2 (IGFBP-2), and IGFBP-3) and markers for bone metabolism (serum osteocalcin and urinary excretion of C-terminal telopeptide

  6. Synchrony in serum antibody response to conserved proteins of Streptococcus pneumoniae in young children.

    PubMed

    Ren, Dabin; Almudevar, Anthony L; Pichichero, Michael E

    2015-01-01

    Conserved Streptococcus pneumoniae (Spn) proteins are currently under investigation as vaccine candidates. We recently identified a subset of children prone to frequent acute otitis media (AOM) that we refer to as stringently-defined otitis prone (sOP). We investigated the synchrony of serum antibody responses against 5 Spn protein vaccine antigens, PhtD, LytB, PcpA, PhtE, and PlyD1 resulting from nasopharyngeal colonization and AOM in sOP children (49 observations) and non-otitis prone (NOP) children (771 observations). Changes in serum IgG and IgM were quantitated with ELISA. IgG antibody concentrations against PhtD, PcpA, and PlyD1 rose in synchrony in sOP and NOP children; that is, the proteins appeared equally and highly immunogenic in children at age 6 to 15 months and then leveled off in their rise at 15 to 25 months. In contrast, rises in concentrations to PhtE and LytB were significantly slower and had not peaked in children even at 25 months of age, consistent with lower immunogenicity. Serum IgM responses against PhtD and PlyD1 were in synchrony in children at age 6-25 months old. PcpA did not induce a significant increase of serum IgM response in children, suggesting that primary responses to PcpA occurred prior to children attaining age 6 months old. PhtD, PcpA, and Ply elicit a synchronous natural acquisition of serum antibody in young children suggesting that a trivalent Spn protein vaccine combining PhtD, PcpA, and PlyD1 would be less likely to display antigen competition when administered as a combination vaccine in young children. PMID:25692218

  7. Serum C-reactive protein: a prognostic factor in metastatic urothelial cancer of the bladder.

    PubMed

    Eggers, Hendrik; Seidel, Christoph; Schrader, Andres Jan; Lehmann, Rieke; Wegener, Gerd; Kuczyk, Markus A; Steffens, Sandra

    2013-12-01

    Until today, there is no reliable prognostic or predictive parameter for the prognosis of patients with metastatic urothelial cancer of the bladder prior to chemotherapy. Recently, serum C-reactive protein (CRP) level has been shown to be associated with survival of patients with various malignancies including localized and metastatic renal cell carcinoma, upper urinary tract as well as penile cancer. The aim of this study was to evaluate the prognostic impact of the pretreatment CRP serum level in patients with metastatic urothelial cancer of the bladder. We retrospectively evaluated 34 patients with metastatic urothelial cancer of the bladder and information about the CRP level prior to chemotherapy. The CRP level was correlated with patient- and tumor-specific characteristics. Kaplan-Meier and log-rank analyses were employed to calculate progression-free (PFS) and overall survival (OS). Receiver operating characteristics (ROC) analysis was used to determine an optimal prognostic CRP cutoff value to predict cancer-specific death. The median PFS to first-line chemotherapy and the OS for the whole cohort were 3.3 and 24.3 months, respectively. Serum CRP in mg/l was significantly associated with patients' survival (HR 1.02, p < 0.001, univariate Cox-regression). ROC analysis identified a CRP value of 80 mg/l to be the optimal cutoff. The median PFS was 4.5 and 3.0 months (p = 0.08; Mann-Whitney test), and the calculated 1-year OS was 82.6 and 22.2 % for patients with a CRP <80 and ?80 mg/l, respectively (log-rank, p < 0.001). In contrast, neither T-stage, tumor grade, sex, age nor the body mass index was related to the CRP level or associated with overall survival. This is the first analysis revealing that the CRP value prior to systemic treatment might be of prognostic significance and could enable better risk stratification for patients with metastatic urothelial cancer of the bladder. PMID:24005810

  8. Protein and microRNA biomarkers from lavage, urine, and serum in military personnel evaluated for dyspnea

    PubMed Central

    2014-01-01

    Background We have identified candidate protein and microRNA (miRNA) biomarkers for dyspnea by studying serum, lavage fluid, and urine from military personnel who reported serious respiratory symptoms after they were deployed to Iraq or Afghanistan. Methods Forty-seven soldiers with the complaint of dyspnea who enrolled in the STudy of Active Duty Military Personnel for Environmental Dust Exposure (STAMPEDE) underwent comprehensive pulmonary evaluations at the San Antonio Military Medical Center. The evaluation included fiber-optic bronchoscopy with bronchoalveolar lavage. The clinical findings from the STAMPEDE subjects pointed to seven general underlying diagnoses or findings including airway hyperreactivity, asthma, low diffusivity of carbon monoxide, and abnormal cell counts. The largest category was undiagnosed. As an exploratory study, not a classification study, we profiled proteins or miRNAs in lavage fluid, serum, or urine in this group to look for any underlying molecular patterns that might lead to biomarkers. Proteins in lavage fluid and urine were identified by accurate mass tag (database-driven) proteomics methods while miRNAs were profiled by a hybridization assay applied to serum, urine, and lavage fluid. Results Over seventy differentially expressed proteins were reliably identified both from lavage and from urine in forty-eight dyspnea subjects compared to fifteen controls with no known lung disorder. Six of these proteins were detected both in urine and lavage. One group of subjects was distinguished from controls by expressing a characteristic group of proteins. A related group of dyspnea subjects expressed a unique group of miRNAs that included one miRNA that was differentially overexpressed in all three fluids studied. The levels of several miRNAs also showed modest but direct associations with several standard clinical measures of lung health such as forced vital capacity or gas exchange efficiency. Conclusions Candidate proteins and miRNAs associated with the general diagnosis of dyspnea have been identified in subjects with differing medical diagnoses. Since these markers can be measured in readily obtained clinical samples, further studies are possible that test the value of these findings in more formal classification or case–control studies in much larger cohorts of subjects with specific lung diseases such as asthma, emphysema, or some other well-defined lung disease. PMID:25282157

  9. Capillary tube isotachophoresis of proteins in early postmortem serum, cerebrospinal fluid and pericardial fluid.

    PubMed

    Takahama, K; Seo, Y; Tomono, S; Yukawa, N

    1989-06-01

    Serum, cerebrospinal fluid (CSF) and pericardial fluid (PDF) obtained from 22 early cadavers were subjected to capillary tube isotachophoresis. Separated proteins were grouped into three main fractions, namely, Fr. 1, Fr. 2 and Fr. 3, in order of their decreasing effective (net) mobilities. The percentages of main fractions to total content were calculated by UV absorbance integration. It was found that Fr. 1 decreased and Fr. 2 increased in all three kinds of specimens with an increase in time following death. In serum and also in CSF, the degree of these changes between 6 and 58 hours following death seemed large enough (about 30%) to be used for crude estimations of early postmortem intervals. Changes in PDF proceeded more gradually than in serum and or in CSF, with only 5-20% changes during this period. PMID:2810897

  10. Association between brain metastasis from lung cancer and the serum level of myelin basic protein

    PubMed Central

    LIU, WEI; ZHAO, JING; WEI, YUJUAN

    2015-01-01

    The aim of the present study was to determine the association between the expression of myelin basic protein in the serum and the metastasis of lung cancer to the brain. A total of 68 lung cancer patients, treated in the Department of Respiratory Medicine of the People’s Hospital of Rizhao (Rizhao, China), were divided into two groups, those with brain metastasis (32 cases) and those without brain metastasis (36 cases). The expression levels of myelin basic protein were measured for all the patients. The results indicated that the expression levels of myelin basic protein in the brain metastasis group were significantly higher when compared with those in the group without metastasis (P<0.05). However, there was no statistically significant correlation between the size of the brain metastasis and the expression levels of myelin basic protein (P>0.05). Furthermore, no statistically significant difference was found in the average level of myelin basic protein between the two subgroups of patients with brain tumor diameters of >1.5 cm and <1.5 cm (P>0.05). Therefore, the results demonstrated a statistically significant correlation between the expression of myelin basic protein in the serum and the metastasis of lung cancer to the brain. Myelin basic protein may thus prove useful in the early diagnosis of brain metastases in lung cancer patients. PMID:25667676

  11. Large-scale serum protein biomarker discovery in Duchenne muscular dystrophy

    PubMed Central

    Hathout, Yetrib; Brody, Edward; Clemens, Paula R.; Cripe, Linda; DeLisle, Robert Kirk; Furlong, Pat; Gordish-Dressman, Heather; Hache, Lauren; Henricson, Erik; Hoffman, Eric P.; Kobayashi, Yvonne Monique; Lorts, Angela; Mah, Jean K.; McDonald, Craig; Mehler, Bob; Nelson, Sally; Nikrad, Malti; Singer, Britta; Steele, Fintan; Sterling, David; Sweeney, H. Lee; Williams, Steve; Gold, Larry

    2015-01-01

    Serum biomarkers in Duchenne muscular dystrophy (DMD) may provide deeper insights into disease pathogenesis, suggest new therapeutic approaches, serve as acute read-outs of drug effects, and be useful as surrogate outcome measures to predict later clinical benefit. In this study a large-scale biomarker discovery was performed on serum samples from patients with DMD and age-matched healthy volunteers using a modified aptamer-based proteomics technology. Levels of 1,125 proteins were quantified in serum samples from two independent DMD cohorts: cohort 1 (The Parent Project Muscular Dystrophy–Cincinnati Children’s Hospital Medical Center), 42 patients with DMD and 28 age-matched normal volunteers; and cohort 2 (The Cooperative International Neuromuscular Research Group, Duchenne Natural History Study), 51 patients with DMD and 17 age-matched normal volunteers. Forty-four proteins showed significant differences that were consistent in both cohorts when comparing DMD patients and healthy volunteers at a 1% false-discovery rate, a large number of significant protein changes for such a small study. These biomarkers can be classified by known cellular processes and by age-dependent changes in protein concentration. Our findings demonstrate both the utility of this unbiased biomarker discovery approach and suggest potential new diagnostic and therapeutic avenues for ameliorating the burden of DMD and, we hope, other rare and devastating diseases. PMID:26039989

  12. Large-scale serum protein biomarker discovery in Duchenne muscular dystrophy.

    PubMed

    Hathout, Yetrib; Brody, Edward; Clemens, Paula R; Cripe, Linda; DeLisle, Robert Kirk; Furlong, Pat; Gordish-Dressman, Heather; Hache, Lauren; Henricson, Erik; Hoffman, Eric P; Kobayashi, Yvonne Monique; Lorts, Angela; Mah, Jean K; McDonald, Craig; Mehler, Bob; Nelson, Sally; Nikrad, Malti; Singer, Britta; Steele, Fintan; Sterling, David; Sweeney, H Lee; Williams, Steve; Gold, Larry

    2015-06-01

    Serum biomarkers in Duchenne muscular dystrophy (DMD) may provide deeper insights into disease pathogenesis, suggest new therapeutic approaches, serve as acute read-outs of drug effects, and be useful as surrogate outcome measures to predict later clinical benefit. In this study a large-scale biomarker discovery was performed on serum samples from patients with DMD and age-matched healthy volunteers using a modified aptamer-based proteomics technology. Levels of 1,125 proteins were quantified in serum samples from two independent DMD cohorts: cohort 1 (The Parent Project Muscular Dystrophy-Cincinnati Children's Hospital Medical Center), 42 patients with DMD and 28 age-matched normal volunteers; and cohort 2 (The Cooperative International Neuromuscular Research Group, Duchenne Natural History Study), 51 patients with DMD and 17 age-matched normal volunteers. Forty-four proteins showed significant differences that were consistent in both cohorts when comparing DMD patients and healthy volunteers at a 1% false-discovery rate, a large number of significant protein changes for such a small study. These biomarkers can be classified by known cellular processes and by age-dependent changes in protein concentration. Our findings demonstrate both the utility of this unbiased biomarker discovery approach and suggest potential new diagnostic and therapeutic avenues for ameliorating the burden of DMD and, we hope, other rare and devastating diseases. PMID:26039989

  13. MALDI-TOF-MS serum protein profiling for developing diagnostic models and identifying serum markers for discogenic low back pain

    PubMed Central

    2014-01-01

    Background The identification of the cause of chronic low back pain (CLBP) represents a great challenge to orthopedists due to the controversy over the diagnosis of discogenic low back pain (DLBP) and the existence of a number of cases of CLBP of unknown origin. This study aimed to develop diagnostic models to distinguish DLBP from other forms of CLBP and to identify serum biomarkers for DLBP. Methods Serum samples were collected from patients with DLBP, chronic lumbar disc herniation (LDH), or CLBP of unknown origin, and healthy controls (N), and randomly divided into a training set (n?=?30) and a blind test set (n?=?30). Matrix-assisted laser desorption ionization time-of-flight mass spectrometry was performed for protein profiling of these samples. After the discriminative ability of two most significantly differential peaks from each two groups was assessed using scatter plots, classification models were developed using differential peptide peaks to evaluate their diagnostic accuracy. The identity of peptides corresponding to three representative differential peaks was analyzed. Results The fewest statistically significant differential peaks were identified between DLBP and CLBP (3), followed by CLBP vs. N (5), DLBP vs. N (9), LDH vs. CLBP (20), DLBP vs. LDH (23), and LDH vs. N (43). The discriminative ability of two most significantly differential peaks was poor in classifying DLBP vs. CLBP but good in classifying DLBP vs. LDH. The accuracy of models for classification of DLBP vs. CLBP was not very high in the blind test (forecasting ability, 67.24%; sensitivity, 70%), although a higher accuracy was observed for classification of DLBP vs. LDH and LDH vs. N (forecasting abilities, ~90%; sensitivities, >90%). A further investigation of three representative differential peaks led to the identification of two peaks as peptides of complement C3, and one peak as a human fibrinogen peptide. Conclusions Our findings benefit not only the diagnosis of CLBP but also the understanding of the differences between different forms of DLBP. The ability to distinguish between different causes of CLBP and the identification of serum biomarkers may be of great value to diagnose different causes of DLBP and predict treatment efficacy. PMID:24889399

  14. Detection of Serum Protein Biomarkers for the Diagnosis and Staging of Hepatoblastoma

    PubMed Central

    Zhao, Wei; Li, Juan; Zhang, Junjie; Gao, Pengfei; Pei, Hang; Wang, Lei; Guo, Fei; Yu, Jiekai; Zheng, Shu; Wang, Jiaxiang

    2015-01-01

    The present study aimed to identify serum biomarkers for the detection of hepatoblastoma (HB). Serum samples were collected from 71 HB patients (stage I, n = 19; stage II, n = 19, stage III, n = 19; and stage IV, n = 14) and 23 age- and sex-matched healthy children. Differential expression of serum protein markers were screened using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS), and the target proteins were isolated and purified using HPLC and identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), SEQUEST, and bioinformatics analysis. Differential protein expression was confirmed by enzyme-linked immunosorbent analysis (ELISA). SELDI-TOF-MS screening identified a differentially expressed protein with an m/z of 9348 Da, which was subsequently identified as Apo A–I; its expression was significantly lower in the HB group as compared to the normal control group (1546.67 ± 757.81 vs. 3359.21 ± 999.36, respectively; p < 0.01). Although the expression level decreased with increasing disease stage, pair-wise comparison revealed significant differences in Apo A–I expression between the normal group and the HB subgroups (p < 0.01). ELISA verified the reduced expression of Apo A–I in the HB group. Taken together, these results suggest that Apo A–I may represent a serum protein biomarker of HB. Further studies will assess the value of using Apo A–I expression for HB diagnosis and staging. PMID:26053398

  15. Solid-phase immunoradiometric assay for serum amyloid A protein using magnetisable cellulose particles.

    PubMed

    De Beer, F C; Dyck, R F; Pepys, M B

    1982-10-29

    An immunoradiometric assay for human serum amyloid A protein (SAA) was developed using magnetisable cellulose particles as the solid phase. Rabbit antiserum to to SAA was raised by immunization with SAA isolated from acute-phase serum by gel filtration in formic acid. The antiserum was rendered monospecific for SAA by solid-phase immunoabsorption with normal human serum, which contains only traces of SAA, and some was coupled covalently to the cellulose particles. Immunopurified anti-SAA antibodies were isolated from the monospecific anti-SAA serum by binding to, and elution from insolubilized acute-phase serum and were radiolabelled with 125I. The assay was calibrated with an acute phase serum which contained 6000 times more SAA than normal sera with the lowest detectable level of SAA, and an arbitrary value of 6000 U/l was assigned to this standard. Sera were tested in the native, undenatured state and there was no increase in SAA immunoreactivity following alkali treatment or heating. The assay range was from 1-2000 U/l so that all SAA levels above 6 U/l could be measured on a single (1:6) dilution of serum. The intra- and interassay coefficients of variation were 11.7 and 15.0% respectively. Among 100 healthy normal subjects (50 male, 50 female) the median SAA level was 9 U/l, range less than 1-100, with 93% below 20 U/l and only 2% below the lower limit of sensitivity of the assay (1 U/l). PMID:7175191

  16. Smoking and lung cancer-induced changes in N-glycosylation of blood serum proteins

    PubMed Central

    Vasseur, Jacqueline A; Goetz, John A; Alley, William R; Novotny, Milos V

    2012-01-01

    Glycosylation is a key post-translational protein modification which appears important in malignant transformation and tumor metastasis. Abnormal glycosylation of different proteins can often be measured in the blood serum. In this study, we extend our serum-based structural investigations to samples provided by patients diagnosed with lung cancer, paying particular attention to the effects of smoking on the serum glycomic traces. Following a battery of glycomic tests, we find that several fucosylated tetra-antennary structures with varying degrees of sialylation are increased in their abundances in control samples provided by the former smokers, with further elevations in the lung cancer patients who were former smokers. Further detailed investigations demonstrated that the level of outer-arm fucosylation was elevated in the control samples of the former smokers and again in the lung cancer samples provided by the former smokers. This trend was particularly noticeable for the tri- and tetra-antennary structures. Different ratios of sialylation linkages were also observed that could be correlated with the different states of health and smoking status. Decreases in the abundance levels of isomers with two and three ?2,3-linked sialic acids and an increased abundance of an isomer with two ?2,6-linked sialic acids were noted for a fucosylated tri-sialylated tri-antennary glycan. These results demonstrate the long-term effects of smoking on glycomic profiles and that this factor needs to be considered in these and other serum-based analyses. PMID:22781126

  17. Spherical Nucleic Acid Nanoparticle Conjugates Enhance G-Quadruplex Formation and Increase Serum Protein Interactions**

    PubMed Central

    Chinen, Alyssa B.; Guan, Chenxia M.

    2014-01-01

    To understand the effect of three-dimensional oligonucleotide structure on protein corona formation, we studied the identity and quantity of human serum proteins that bind to spherical nucleic acid (SNA) nanoparticle conjugates. SNAs exhibit cellular uptake properties that are remarkably different from those of linear nucleic acids, which have been related to their interaction with certain classes of proteins. Through a proteomic analysis, this work shows that the protein binding properties of SNAs are sequence-specific and supports the conclusion that the oligonucleotide tertiary structure can significantly alter the chemical composition of the SNA protein corona. This knowledge will impact our understanding of how nucleic acid-based nanostructures, and SNAs in particular, function in complex biological milieu. PMID:25393322

  18. Liver Retinol Transporter and Receptor for Serum Retinol-binding Protein (RBP4)*

    PubMed Central

    Alapatt, Philomena; Guo, Fangjian; Komanetsky, Susan M.; Wang, Shuping; Cai, Jinjin; Sargsyan, Ashot; Rodríguez Díaz, Eduardo; Bacon, Brandon T.; Aryal, Pratik; Graham, Timothy E.

    2013-01-01

    Vitamin A (retinol) is absorbed in the small intestine, stored in liver, and secreted into circulation bound to serum retinol-binding protein (RBP4). Circulating retinol may be taken up by extrahepatic tissues or recycled back to liver multiple times before it is finally metabolized or degraded. Liver exhibits high affinity binding sites for RBP4, but specific receptors have not been identified. The only known high affinity receptor for RBP4, Stra6, is not expressed in the liver. Here we report discovery of RBP4 receptor-2 (RBPR2), a novel retinol transporter expressed primarily in liver and intestine and induced in adipose tissue of obese mice. RBPR2 is structurally related to Stra6 and highly conserved in vertebrates, including humans. Expression of RBPR2 in cultured cells confers high affinity RBP4 binding and retinol transport, and RBPR2 knockdown reduces RBP4 binding/retinol transport. RBPR2 expression is suppressed by retinol and retinoic acid and correlates inversely with liver retinol stores in vivo. We conclude that RBPR2 is a novel retinol transporter that potentially regulates retinol homeostasis in liver and other tissues. In addition, expression of RBPR2 in liver and fat suggests a possible role in mediating established metabolic actions of RBP4 in those tissues. PMID:23105095

  19. Chromatographic adsorption of serum albumin and antibody proteins in cryogels with benzyl-quaternary amine ligands.

    PubMed

    Yun, Junxian; Cheng, Xiuhong; Ye, Jialei; Shen, Shaochuan; Yang, Gensheng; Yao, Kejian; Kirsebom, Harald; Lin, Dong-Qiang; Guan, Yi-Xin; Yao, Shan-Jing

    2015-02-13

    The preparation and characterization of mixed-mode adsorbents for a typical separation purpose are of great importance in bioseparation areas. In this work, we prepared a new monolithic cryogel with a combination of ion-exchange and hydrophobic functions by employing benzyl-quaternary amine groups. The fundamental cryogel properties, protein equilibrium adsorption isotherm and chromatographic adsorption in the cryogel were measured experimentally. The results showed that, by using bovine serum album as the model protein, the dual functional cryogel has protein binding capability even in salt solution and the buffer with pH close or below the protein isoelectric point due to both the electrostatic and hydrophobic interactions. A capillary-based adsorption model was developed, which provided satisfied insights of the microstructure, axial dispersion, mass transfer as well as protein adsorption characteristics within the cryogel bed. The chromatographic isolation of bioactive proteins from rabbit blood serum was carried out by the cryogel. Immunoglobulin G antibody with a purity of 98.2% and albumin with a purity of 96.8% were obtained, indicating that the cryogel could be an interesting and promising adsorbent in bioseparation areas. PMID:25618356

  20. Formation of long-lived reactive species of blood serum proteins by the action of heat.

    PubMed

    Bruskov, Vadim I; Popova, Nelly R; Ivanov, Vladimir E; Karp, Olga E; Chernikov, Anatoly V; Gudkov, Sergey V

    2014-01-17

    It has been previously established that heat induces the formation of reactive oxygen species (ROS) in aqueous solutions. In biological systems, ROS cause oxidative damage predominantly to proteins due to their abundance and sensitivity to oxidation. Proteins oxidized by the action of X-rays represent long-lived reactive species, which trigger the secondary generation of ROS (Bruskov et al. (2012) [25]). Here we studied the possibility of formation of long-lived species of the blood serum proteins bovine serum albumin and bovine gamma-globulin in air-saturated solutions under the action of heat. It is shown that heat induces the generation of long-lived protein species, which in turn generate ROS ((1)?2, (·)O2(-), (·)O?, and H2O2). The formation of the long-lived reactive species of BSA and BGG with a half-life of about 4h induced by moderate hyperthermia was revealed using the chemiluminescence of protein solutions. It was found that long-lived reactive species of BSA and BGG cause prolonged generation of H2O2. The results obtained suggest that H2O2 produced by proteins after heating represents a messenger in signaling pathways and produces therapeutic effects in living organisms. PMID:24361896

  1. Effect of altered eating pattern on serum fructosamine: total protein ratio and plasma glucose level.

    PubMed

    Ch'ng, S L; Cheah, S H; Husain, R; Duncan, M T

    1989-05-01

    The effect of alteration of eating pattern during Ramadan on body mass index (BMI), serum fructosamine: total protein ratio (F/TP), and glucose level in 18 healthy male Asiatic Moslems were studied. The results showed a significant decrease (p less than 0.025) in F/TP at the second week of Ramadan in 11 subjects who experienced continuous decrease in BMI throughout Ramadan. The remaining 7 subjects showed no significant changes in BMI and F/TP. No evidence of hypoglycaemia was observed in the subjects during the study. Serum fructosamine: total protein ratio in subjects with altered eating pattern preferably should be interpreted along with the change in body mass index. PMID:2774480

  2. Serum Lipopolysaccharide Binding Protein Levels Predict Severity of Lung Injury and Mortality in Patients with Severe Sepsis

    Microsoft Academic Search

    Jesús Villar; Lina Pérez-Méndez; Elena Espinosa; Carlos Flores; Jesús Blanco; Arturo Muriel; Santiago Basaldúa; Mercedes Muros; Lluis Blanch; Antonio Artigas; Robert M. Kacmarek; Rory Edward Morty

    2009-01-01

    BackgroundThere is a need for biomarkers insuring identification of septic patients at high-risk for death. We performed a prospective, multicenter, observational study to investigate the time-course of lipopolysaccharide binding protein (LBP) serum levels in patients with severe sepsis and examined whether serial serum levels of LBP could be used as a marker of outcome.Methodology\\/Principal FindingsLBP serum levels at study entry,

  3. Effect of bleaching permeate from microfiltered skim milk on 80% serum protein concentrate.

    PubMed

    Campbell, Rachel E; Adams, Michael C; Drake, Maryanne; Barbano, David M

    2013-03-01

    Whey proteins that have been removed before the cheese-making process are referred to as "native" whey proteins or milk serum proteins. Because serum proteins isolated directly from milk are not exposed to the cheese-making process, they are free from functional or sensory effects arising from this process. Whey proteins used in food and beverage applications are largely derived from annatto-colored Cheddar cheese. Some of the annatto is left in the whey and this color is converted to a colorless compound by bleaching. The effect of bleaching serum proteins on flavor and functionality of spray-dried protein provides a platform to investigate the effect of bleaching free from the confounding effects of cheese manufacture. The objective of this study was to characterize and compare the sensory and functional properties of 80% milk serum protein concentrate (SPC80) produced from bleached and unbleached microfiltration (MF) permeate made from skim milk with and without added annatto color. Colored and uncolored MF permeates were bleached with benzoyl peroxide (BP) or hydrogen peroxide (HP), ultrafiltered, diafiltered, and spray-dried. The SPC80 from unbleached colored and uncolored MF permeates were manufactured as controls. All treatments were manufactured in triplicate. All SPC80 were evaluated by sensory testing, instrumental analyses, functionality, color, and proximate analysis. The HP-bleached SPC80 was higher in lipid oxidation compounds than BP-bleached or unbleached SPC80, specifically hexanal, heptanal, nonanal, decanal, and 2,3-octadienone. The HP treatments were higher in aroma intensity and cardboard and fatty flavors compared with the unbleached and BP-bleached SPC80. The SPC80 bleached with BP had lower concentrations of norbixin compared with SPC80 bleached with HP. Functionality testing demonstrated that HP treatments had more soluble protein after 10min of heating at 90°C and pH 4.6 and pH 7 compared with the no bleach and BP treatments, regardless of additional color. Foams generated from bleached SPC80 were more stable than those from unbleached SPC80, and those bleached with HP were lower in yield stress than other SPC80. Overall, HP bleaching destroyed less norbixin and caused more lipid oxidation and subsequent off-flavors than did BP bleaching. However, the heat stability of SPC80 was enhanced by HP bleaching compared with control treatments or BP bleaching. PMID:23295111

  4. Elevated Serum C-Reactive Protein Levels and Early Arterial Changes in Healthy Children

    Microsoft Academic Search

    Mikko J. Järvisalo; Aimo Harmoinen; Maarit Hakanen; Ulla Paakkunainen; Jorma Viikari; Jaakko Hartiala; Terho Lehtimäki; Olli Simell; Olli T. Raitakari

    Objective—Elevated serum concentration of C-reactive protein (CRP) predicts cardiovascular events in adults. Because atherosclerosis begins in childhood, we undertook a study to determine whether changes in brachial artery endothelial function and the thickness of the carotid intima-media complex, 2 markers of early atherosclerosis, are related to CRP levels in healthy children. Methods and Results—Brachial artery flow-mediated dilatation (FMD) and carotid

  5. Rat islet culture in serum-free medium containing silk protein sericin

    Microsoft Academic Search

    Mitsuhiro Morikawa; Toshihisa Kimura; Makoto Murakami; Kanji Katayama; Satoshi Terada; Akio Yamaguchi

    2009-01-01

    Background  The development of islet cultures is desirable for successful clinical islet transplantation. Fetal bovine serum (FBS) has\\u000a been used as a supplement in islet culture medium, but it may be an unsuitable supplement due recent animal health problems.\\u000a We have evaluated the use of the silk protein, sericin, derived from Bombyx mori as a replacement for FBS in islet culture

  6. Comparison of inhibitory effects of oxygen radicals and calf serum protein on surfactant activity

    Microsoft Academic Search

    M. M. Lee; F. H. Y. Green; S. Schürch; S. Cheng; S. G. Bjarnason; S. Leonard; W. Wallace; F. Possmayer; V. Vallyathan

    2004-01-01

    The effects of the reactive oxygen species (ROS) superoxide anion (O\\u000a2\\u000a.\\u000a–) and hydroxyl radical (OH) on the surface tension lowering properties of bovine lipid extract surfactant (BLES) were compared to the effects of calf serum protein (CSP) in a captive bubble surfactometer (CBS). O\\u000a2\\u000a.\\u000a– was generated from xanthine\\/xanthine oxidase (X\\/XO), and OH was generated

  7. Acute-Phase Proteins during Hemodialysis: Correlations with Serum Interleukin1? Levels and Different Dialysis Membranes

    Microsoft Academic Search

    Eero Honkanen; Carola Grönhagen-Riska; Anna-Maija Teppo; C. P. J. Maury; Seppo Meri

    1991-01-01

    The effects of hemodialysis (HD) on the levels of serum amyloid A (SAA), C-reactive protein (CRP) and interleukin-1? (IL-1?) were studied in 8 patients. Bicarbonate dialysate was used exclusively, and three different membranes, Cuprophan® (CU), cellulose acetate (CA), and polymethylmetachrylate (PMMA) were compared. The SAA levels increased significantly with each membrane. With CU, they rose from 4.0 ± 2.0 (mg\\/l,

  8. Serum Levels of Surfactant Protein D Are Increased in Mice with Lung Tumors1

    Microsoft Academic Search

    Feijie Zhang; William Pao; Sarah M. Umphress; Sonia B. Jakowlew; Amy M. Meyer; Lori D. Dwyer-Nield; Larry D. Nielsen; Katsuyuki Takeda; Erwin W. Gelfand; James H. Fisher; Lening Zhang; Alvin M. Malkinson; Robert J. Mason

    2003-01-01

    Most murine lung tumors are composed of differentiated epithelial cells. We have reported previously that surfactant protein (SP)-D is expressed in urethane-induced tumors. Serum levels of SP-D are increased in patients with interstitial lung disease and acute respiratory distress syndrome and in rats with acute lung injury but have not been measured in mice. In this study, we sought to

  9. Rouleaux-Forming Serum Proteins Are Involved in the Rosetting of Plasmodium falciparum-Infected Erythrocytes

    Microsoft Academic Search

    Carl Johan Treutiger; Carin Scholander; Johan Carlson; Keith P. McAdam; John G. Raynes; Lars Falksveden; Mats Wahlgren

    1999-01-01

    Treutiger, C. J., Scholander, C., Carlson, J., McAdam, K. P., Raynes, J. G., Falksveden, L., and Wahlgren, M. 1999. Rouleaux-forming serum proteins are involved in the rosetting of Plasmodium falciparum-infected erythrocytes. Experimental Parasitology93, 215–224. Excessive sequestration of Plasmodium falciparum-infected (pRBC) and uninfected erythrocytes (RBC) in the microvasculature, cytoadherence, and rosetting, have been suggested to be correlated with the development of

  10. Identification of specific protein carbonylation sites in model oxidations of human serum albumin

    Microsoft Academic Search

    Ani Temple; Ten-Yang Yen; Scott Gronert

    2006-01-01

    Human serum albumin (HSA) was subjected to oxidative stress and the locations of the resulting protein carbonyls were determined\\u000a using mass spectrometry in conjunction with a hydrazide labeling scheme. To model oxidative stress, HSA samples were subjected\\u000a to metal-catalyzed oxidation (MCO) conditions or treated with hypochlorous acid (HOCl). Oxidation led to the conversion of\\u000a lysine residues to 2-aminoadipic semi-aldehyde residues,

  11. Olmesartan reduces arterial stiffness and serum adipocyte fatty acid-binding protein in hypertensive patients

    Microsoft Academic Search

    Toru MiyoshiMasayuki; Masayuki Doi; Satoshi Hirohata; Shigeshi Kamikawa; Shinichi Usui; Hiroko Ogawa; Kosuke Sakane; Reishi Izumi; Yoshifumi Ninomiya; Shozo Kusachi

    Adipocyte fatty acid binding protein (A-FABP) has been reported to be involved in insulin resistance, lipid metabolism, and\\u000a atherosclerosis; however, little is known about the effect of medication on the change in circulating A-FABP in human subjects.\\u000a We evaluated the effects of angiotensin II type 1 receptor blocker (ARB) on arterial stiffness and its association with serum\\u000a A-FABP in patients

  12. Early Postnatal Surge of Serum Clara Cell Secretory Protein in Newborn Infants

    Microsoft Academic Search

    A. S. Loughran-Fowlds; J. W. Lin; J. Oei; J. Michalowski; R. Henry; K. Lui

    2012-01-01

    Background: Clara cell secretory protein (CCSP) is an anti-inflammatory mediator, but its role in neonatal lung adaptation and diseases is uncertain. Objective: To characterize postnatal changes in serum CCSP in relation to gestation, respiratory disease (RDS) and bronchopulmonary dysplasia (BPD) in comparison with other anti-inflammatory cytokines (IL-4, -10 and -13). Methods: Blood was collected from 76 infants (26 of 23–29

  13. Genetic Polymorphisms in Blood Serum Transferrins and Certain Milk Proteins of the Canadian Breed1

    Microsoft Academic Search

    E. M. Wilson; H. F. MacRae; J. M. Belanger

    1967-01-01

    The seruln B-globulin (transferrin), a~-casein, fi-casein, and fl-lactoglobulin types of 58 individual cows in one herd of the Canadian breed were studied by means of polyacrylamide-gel eleetrophore- sis in a discontinuous buffer system. The distributions of genotypes for each of the proteins were as follows: serum trans- ferrin--3 A 'A, 27 D\\/D, 13 A\\/'D, 4 A\\/E, and 11. D, E;

  14. Regulation of serum amyloid A protein expression during the acute-phase response.

    PubMed Central

    Jensen, L E; Whitehead, A S

    1998-01-01

    The acute-phase (AP) serum amyloid A proteins (A-SAA) are multifunctional apolipoproteins which are involved in cholesterol transport and metabolism, and in modulating numerous immunological responses during inflammation and the AP response to infection, trauma or stress. During the AP response the hepatic biosynthesis of A-SAA is up-regulated by pro-inflammatory cytokines, and circulating concentrations can increase by up to 1000-fold. Chronically elevated A-SAA concentrations are a prerequisite for the pathogenesis of secondary amyloidosis, a progressive and fatal disease characterized by the deposition in major organs of insoluble plaques composed principally of proteolytically cleaved A-SAA, and may also contribute to physiological processes that lead to atherosclerosis. There is therefore a requirement for both positive and negative control mechanisms that permit the rapid induction of A-SAA expression until it has fulfilled its host-protective function(s) and subsequently ensure that its expression can be rapidly returned to baseline. These mechanisms include modulation of promoter activity involving, for example, the inducer nuclear factor kappaB (NF-kappaB) and its inhibitor IkappaB, up-regulatory transcription factors of the nuclear factor for interleukin-6 (NF-IL6) family and transcriptional repressors such as yin and yang 1 (YY1). Post-transcriptional modulation involving changes in mRNA stability and translation efficiency permit further up- and down-regulatory control of A-SAA protein synthesis to be achieved. In the later stages of the AP response, A-SAA expression is effectively down-regulated via the increased production of cytokine antagonists such as the interleukin-1 receptor antagonist (IL-1Ra) and of soluble cytokine receptors, resulting in less signal transduction driven by pro-inflammatory cytokines. PMID:9729453

  15. Serum levels of selected liver proteins following partial hepatectomy in the female rat.

    PubMed

    Hau, J; Cervinková, Z; O'Brien, D; Stodulski, G; Simek, J

    1995-04-01

    Liver regeneration in the rat following partial hepatectomy (PH) is a frequently used model to study regulatory mechanisms in relation to cell growth and differentiation. In the present study, we analysed quantitative changes in the peripheral circulation of a number of important serum proteins following PH and laparotomy. Alpha-fetoprotein synthesis was induced in the PH rats and remained at very low levels in non-operated controls. Pregnancy-associated murine protein-1 levels and fibronectin levels were lower in the circulation of all operated animals compared with the intact controls. The serum levels of alpha-2 macroglobulin were elevated in all operated animals compared with the non-operated controls. Circulating complement factors C3c and C4 were present at significantly lower levels in PH animals than in rats following laparotomy. Small growth stimulating molecules (< 10 kDa) synthesized by different organs during the regenerative process have been described in the literature, but dialysates of the spleen and liver from non-operated rats and PH rats showed no significant impact on serum levels of the proteins in the present study. PMID:7541490

  16. Serum concentrations of cartilage oligomeric matrix protein and bone sialoprotein in hip osteoarthritis: A one year prospective study

    Microsoft Academic Search

    Thierry Conrozier; Tore Saxne; Charles Shan Sei Fan; Pierre Mathieu; Anne-Marie Tron; Dick Heinegård; Eric Vignon

    1998-01-01

    OBJECTIVETo evaluate serum concentations of cartilage oligomeric matrix protein (COMP) and bone sialoprotein (BSP) as predictors of disease progression in hip osteoarthrtitis (OA).METHODSForty eight consecutive patients, referred to hospital for symptomatic hip OA, (ACR criteria) were monitored in a one year prospective trial with radiographs and serum samples. The radiographs were graded for joint space narrowing, osteophytes, and sclerosis and

  17. Simple Stratification of Survival Using Bone Scan and Serum C-Reactive Protein in Prostate Cancer Patients with Metastases

    Microsoft Academic Search

    Jun Nakashima; Eiji Kikuchi; Akira Miyajima; Ken Nakagawa; Mototsugu Oya; Takashi Ohigashi; Masaru Murai

    2008-01-01

    Background: IL-6 has been reported to be a significant prognostic factor for prostate cancer and induces synthesis of C-reactive protein (CRP) by hepatocytes. The present study was undertaken to evaluate the clinical value of serum CRP in prostate cancer patients with metastases. Methods: The prognostic significance of serum CRP as well as tumor histology, extent of disease (EOD) on bone

  18. Structural modification of serum vitamin D3-binding protein and immunosuppression in AIDS patients.

    PubMed

    Yamamoto, N; Naraparaju, V R; Srinivasula, S M

    1995-11-01

    A serum glycoprotein, vitamin D3-binding protein (Gc protein), can be converted by beta-galactosidase of stimulated B lymphocytes and sialidase of T lymphocytes to a potent macrophage-activating factor (MAF), a protein with N-acetylgalactosamine as the remaining sugar moiety. Thus, Gc protein is a precursor for MAF. Treatment of purified Gc protein with immobilized beta-galactosidase and sialidase generates an extremely high-titered MAF (GcMAF). When peripheral blood monocytes/macrophages of 46 HIV-infected patients were treated with GcMAF (100 pg/ml), the monocytes/macrophages of all patients were efficiently activated. However, the MAF precursor activity of plasma Gc protein was low in 16 (35%) of of these patients. Loss of the MAF precursor activity appeared to be due to deglycosylation of plasma Gc protein by alpha-N-acetylgalactosaminidase found in the patient blood stream. Levels of plasma alpha-N-acetylgalactosaminidase activity in individual patients had an inverse correlation with the MAF precursor activity of their plasma Gc protein. Thus, precursor activity of Gc protein and alpha-N-acetylgalactosaminidase activity in patient blood can serve as diagnostic and prognostic indices. PMID:8573395

  19. Influence of sulfur-amino acid content variation in plant vs animal protein on serum and tissue lipids in rats

    Microsoft Academic Search

    Susan M. Potter; Constance V. Kies

    1990-01-01

    The objective of the study was to determine the effects of methionine and cysteine supplementation of soy protein isolate and casein on serum and tissue lipid levels in rats. Sixty male, weanling, Wistar-Kyoto rats were fed two sources of protein (casein or soy protein isolate) and three variations of sulfur-amino acid supplementation (none, methionine, or cysteine). At this level of

  20. THE INFLUENCE OF SERUM BINDING PROTEINS AND FEEDBACK CONTROL OF SERUM ESTRADIOL LEVELS ON THE COMPARATIVE POTENCY OF ENDOCRINE ACTIVE COMPOUNDS

    EPA Science Inventory

    THE INFLUENCE OF SERUM BINDING PROTEINS ON THE COMPARATIVE RECEPTOR BINDING POTENCY OF ENDOCRINE ACTIVE COMPOUNDS. JG Teeguarden1 and HA Barton2. 1ICF Consulting, Research Triangle Park NC; 2US EPA, ORD, NHEERL, ETD, Pharmacokinetics Branch, RTP, NC. Accurate comparison of...

  1. Loss of Metastatic and Primary Tumor Factor X Activator Capabilities by Lewis Lung Carcinoma Cells Cultured in Vitamin K-dependent Protein Deficient Serum1

    Microsoft Academic Search

    George E. Eagan; Adrian C. Wilson; John F. Gierthy; David L. Lincoln

    A highly metastatic line of Lewis lung tumor cells established in fetal bovine serum (10%) was subcultured into normal rodent (mouse or rat) serum or rodent serum made deficient in functional vitamin K-dependent proteins (barium sulfate adsorption or warfarin treatment of animals). Following injection of cells cultured in normal rodent serum into C57BL\\/ 6 mice, Factor \\\\ activator activity in

  2. Estrogen Downregulation of Albumin and a 170-kDa Serum Protein in the Turtle, Trachemys scripta

    Microsoft Academic Search

    Kyle W. Selcer; Brent D. Palmer

    1995-01-01

    We examined changes in serum protein composition after estradiol-17? treatment of ovariectomized female Trachemys scripta, with the objective of identifying proteins that are repressed by estrogen. The experimental protocol was validated by measuring serum estradiol-17? levels with a specific radioimmunoassay. Control turtle sera contained little or no estradiol-17? (mean = 25.8 pg\\/ml) while estrogen-treated turtle sera had elevated estradiol-17? levels

  3. Serum anti-lysosomal-associated membrane protein-2 antibody levels in cutaneous polyarteritis nodosa.

    PubMed

    Kawakami, Tamihiro; Ishizu, Akihiro; Arimura, Yosihiro; Soma, Yoshinao

    2013-01-01

    Lysosomal-associated membrane protein-2 (LAMP-2) is a target antigen for anti-neutrophil cytoplasmic antibodies (ANCAs), which are closely linked to a subset of primary systemic vasculitides. Cutaneous polyarteritis nodosa (CPN) is a necrotizing vasculitis of small to medium-sized arteries within the skin. We measured levels of serum anti-LAMP-2 antibody in 50 patients with CPN, 8 with microscopic polyangiitis (MPA), and 34 healthy persons. We also investigated the presence of ANCA in patients with CPN using indirect immunofluorescence (IIF), a direct ELISA and a capture ELISA specific for myeloperoxidase (MPO) and proteinase 3 (PR3). Serum anti-LAMP-2 antibody levels differed significantly between patients with CPN (0.263 U/ml) and those with MPA (0.180 U/ml) (p = 0.0102). Serum of all patients with CPN was negative for MPO-ANCA and PR3-ANCA by both direct ELISA and capture ELISA. In contrast, IIF assay revealed ANCA in 42 (84.0%) of the 50 CPN patients. Serum anti-LAMP-2 antibody levels in the perinuclear ANCA (P-ANCA) group were significantly elevated compared with the non-ANCA group (p = 0.0147). We suggest that anti-LAMP-2 antibody could play an important role in the pathogenesis of CPN in the presence of P-ANCA detected by IIF. PMID:22854803

  4. Trefoil factor family (TFF) proteins as potential serum biomarkers in patients with metastatic colorectal cancer.

    PubMed

    Vocka, M; Langer, D; Petrtyl, J; Vockova, P; Hanus, T; Kalousova, M; Zima, T; Petruzelka, L

    2015-01-01

    Trefoil factor family (TFF) is composed of three secretory proteins (TFF1, TFF2 and TFF3) that play an important role in mucosal protection of gastrointestinal tract. Their overexpression in colorectal tumors seems to be associated with more aggressive disease. We collected serum samples from 79 healthy controls and 97 patients with metastatic colorectal cancer at the time of diagnosis or at progression. Serum levels of TTF1-3, CEA and CA19-9 were measured by ELISA. Serum TFF1 and TFF3 levels were significantly higher in patients with colorectal cancer compared to healthy controls (p?serum levels of TFF3 correlated with extent of liver involvement in patient without pulmonary metastases and patients with higher TFF3 levels had significantly worse outcome (p < 0.0001). Compared to CEA and CA19-9, TFF3 had higher sensitivity and the same specificity. Our results indicate that TFF3 is an effective biomarker in patients with metastatic colorectal cancer with higher sensitivity than CEA a CA19-9. TFF3 levels strongly correlate with extension of liver disease and seem to have prognostic value. PMID:25866228

  5. Release of Glial Tissue-Specific Proteins After Acute Stroke A Comparative Analysis of Serum Concentrations of Protein S-100B and Glial Fibrillary Acidic Protein

    Microsoft Academic Search

    Manfred Herrmann; Pieter Vos; Michael T. Wunderlich; Chris H. M. M. de Bruijn; Karl J. B. Lamers

    Background and Purpose—This study was aimed at the comparative analysis of serum concentrations of glial fibrillary acidic protein (GFAP) and protein S-100B in patients with acute stroke. Methods—We investigated 32 patients with stroke symptoms consistent with cerebral ischemia in the anterior territory of vascular supply. Serial venous blood samples were taken after admission to the hospital and during the first

  6. Single-Molecule enzyme-linked immunosorbent assay detects serum proteins at subfemtomolar concentrations

    PubMed Central

    Rissin, David M.; Kan, Cheuk W.; Campbell, Todd G.; Howes, Stuart C.; Fournier, David R.; Song, Linan; Piech, Tomasz; Patel, Purvish P.; Chang, Lei; Rivnak, Andrew J.; Ferrell, Evan P.; Randall, Jeffrey D.; Provuncher, Gail K.; Walt, David R.; Duffy, David C.

    2010-01-01

    The detection of single protein molecules1,2 in blood could help identify many new diagnostic protein markers. We report an approach for detecting hundreds to thousands of individual protein molecules simultaneously that enables the detection of very low concentrations of proteins. Proteins are captured on microscopic beads and labeled with an enzyme, such that each bead has either one or zero enzyme-labeled proteins. By isolating these beads in arrays of 50-femtoliter reaction chambers, single proteins can be detected by fluorescence imaging. By singulating molecules in these arrays, ~10–20 enzymes can be detected in 100 ?L (~10?19 M). Single molecule enzyme-linked immunosorbent assays (digital ELISA) based on singulation of enzyme labels enabled the detection of clinically-relevant proteins in serum at concentrations (<10?15 M) much lower than conventional ELISA3-5. Digital ELISA detected prostate specific antigen in all tested sera from patients who had undergone radical prostatectomy, down to 14 fg/mL (0.4 fM). PMID:20495550

  7. Deglycosylation of serum vitamin D3-binding protein leads to immunosuppression in cancer patients.

    PubMed

    Yamamoto, N; Naraparaju, V R; Asbell, S O

    1996-06-15

    Serum vitamin D3-binding protein (Gc protein) can be converted by beta-galactosidase of B cells and sialidase of T cells to a potent macrophage activating factor, a protein with N-acetylgalactosamine as the remaining sugar moiety. Thus, Gc protein is the precursor of the macrophage activating factor (MAF). Treatment of Gc protein with immobilized beta-galactosidase and sialidase generates an extremely high titered MAF, Gc-MAF. When peripheral blood monocytes/macrophages of 52 patients bearing various types of cancer were incubated with 100 pg/ml of GcMAF, the monocytes/macrophages of all patients were efficiently activated. However, the MAF precursor activity of patient plasma Gc protein was found to be severely reduced in about 25% of this patient population. About 45% of the patients had moderately reduced MAF precursor activities. Loss of the precursor activity was found to be due to deglycosylation of plasma Gc protein by alpha-N-acetylgalactosaminidase detected in the patient's bloodstream. The source of the enzyme appeared to be cancerous cells. Radiation therapy decreased plasma alpha-N-acetylgalactosaminidase activity with concomitant increase of precursor activity. This implies that radiation therapy decreases the number of cancerous cells capable of secreting alpha-N-acetylgalactosaminidase. Both alpha-N-acetylgalactosaminidase activity and MAF precursor activity of Gc protein in patient bloodstream can serve as diagnostic and prognostic indices. PMID:8665521

  8. Biophysical analysis of the interaction of the serum protein human ?2GPI with bacterial lipopolysaccharide

    PubMed Central

    Gries, Anna; Prassl, Ruth; Fukuoka, Satoshi; Rössle, Manfred; Kaconis, Yani; Heinbockel, Lena; Gutsmann, Thomas; Brandenburg, Klaus

    2014-01-01

    There are several human serum proteins for which no clear role is yet known. Among these is the abundant serum protein beta2-glycoprotein-I (?2GPI), which is known to bind to negatively charged phospholipids as well as to bacterial lipopolysaccharides (LPS), and was therefore proposed to play a role in the immune response. To understand the details of these interactions, a biophysical analysis of the binding of ?2GPI to LPS and phosphatidylserine (PS) was performed. The data indicate only a moderate tendency of the protein (1) to influence the LPS-induced cytokine production in vitro, (2) to react exothermally with LPS in a non-saturable way, and (3) to change its local microenvironment upon LPS association. Additionally, we found that the protein binds more strongly to phosphatidylserine (PS) than to LPS. Furthermore, ?2GPI converts the LPS bilayer aggregates into a stronger multilamellar form, and reduces the fluidity of the hydrocarbon moiety of LPS due to a rigidification of the acyl chains. From these data it can be concluded that ?2GPI plays a role as an immune-modulating agent, but there is much less evidence for a role in immune defense against bacterial toxins such as LPS. PMID:24918058

  9. Effects of Aerobic Exercise on Serum Retinol Binding Protein4, Insulin Resistance and Blood Lipids in Obese Women

    PubMed Central

    TAGHIAN, Farzaneh; ZOLFAGHARI, Maryam; HEDAYATI, Mehdi

    2014-01-01

    Abstract Background Retinol binding protein4 (RBP4) is a type of adipokine which transports vitamin A to serum. RBP4 could be a bridge between obesity and insulin resistance. This study aimed to investigate the effects of aerobic exercises on RBP4 serum’s concentration and metabolic syndrome risk factors in obese women. Methods Twenty obese women with body max index 35.81±3.67Kg/m2, fat percentage 43.98±4.02, and waist to hip ratio 1.03±0.05 were included and were randomly assigned to experimental and control groups. The experimental group received aerobic exercises for a period of 12 weeks each three sessions on treadmill workout. The treadmill speed were based on a 60-65 and 80-85 maximal heart rate percentage and duration of 15-20 and 45-50 minutes, at the beginning and the end of exercise, respectively. Body composition, serum glucose, insulin, TG, LDL-C, HDL-C, total cholesterol, and RBP4, were measured in both groups before and after the treatment by ELISA method. Insulin resistance was measured by HOMA-IR. To compare within group differences and between group comparisons t-correlated and t-independent tests were used, respectively. Results After 12 week aerobic exercises; weight, fat percentage, WHR, and BMI in the experimental group was significantly decreased (P<0.05). RBP4, insulin, insulin resistance, TG and HDL-C had significant differences between two groups. The cholesterol level, LDL-C and glucose did not have any significant changes. Conclusion The aerobic exercises can decrease body composition, insulin resistance, TG, and RBP4, so it can be beneficial for obese women’s health, because it. PMID:26060767

  10. Effect of bone morphogenetic protein-4 on cardiac differentiation from mouse embryonic stem cells in serum-free and low-serum media.

    PubMed

    Taha, Masoumeh Fakhr; Valojerdi, Mojtaba Rezazadeh

    2008-06-23

    In spite of previous reports, the precise role of bone morphogenetic proteins (BMPs) on cardiomyocyte differentiation, especially in the absence or presence of minimum amount of serum in culture medium is still unclear. So, the aim of the present study was to investigate the effect of BMP-4 on mouse embryonic stem cells (ESCs)-derived cardiomyocyte differentiation in serum-free and low-serum media. The mouse ESCs differentiation to cardiomyocytes was induced by embryoid bodies' (EBs') development through hanging drop, suspension and plating stages. Different models of differentiation were designed according to addition of fetal bovine serum (FBS) or knockout serum replacement (KoSR) to the medium of three stages. 10 ng/ml BMP-4 was added throughout the suspension period. Up to 30 days after plating, contraction and beating frequency were monitored and evaluated daily. The growth characteristics of cardiomyocytes were assessed by cardioactive drugs, immunocytochemistry, transmission electron microscopy (TEM) and reverse transcription-polymerase chain reaction (RT-PCR). In the complete absence of serum, neither control nor BMP-4 treated groups resulted in cardiac differentiation. Addition of FBS to hanging drop stage resulted in the appearance of beating cardiac clusters in some BMP-4 treated EBs. In the best designed differentiation model in which only hanging drop and the first 24 h of plating stage was carried out at the presence of FBS, the BMP-4 treatment resulted in cardiac differentiation in EBs characterized by positive immunostaining for the applied antibodies, chronotropic response to the cardioactive drugs and cardiac-specific genes expression at different developmental stages. These cardiomyocytes showed immature myofibrils and numerous intercellular junctions. In conclusion, BMP-4 is unable to induce cardiomyocyte differentiation from mouse ESCs in serum-free models, and at least small amount of FBS in hanging drop stage is necessary. Furthermore, serum factors are not strictly necessary after the initial activation, but they do favor a better differentiation of cardiomyocytes. PMID:17714812

  11. Immunochemical and ultrastructural study of multiple myeloma with a heavy chain protein in the serum.

    PubMed Central

    Bartoloni, C; Flamini, G; Gentiloni, N; Russo, M A; Barone, C; Gambassi, G; Terranova, T

    1980-01-01

    A patient with multiple myeloma had antigenically related monoclonal Fc-gamma fragments and complete IgG-kappa molecules in the serum. The urine contained only Fc-gamma fragments in the absence of Bence-Jones protein. The two distinct M-components in the serum showed electrophoretic identity but could be separated by chromatography. The simultaneous presence of complete monoclonal IgG molecules and Fc-gamma fragments, though difficult to detect, could be a frequent occurrence in multiple myeloma, and it could be defined as 'double paraproteinaemia'. A detailed ultrastructural study was performed in this case and showed fibril bundles being released from the malignant plasma cells; such fibrils could be the supramolecular organisation of the neosynthesised heavy chain fragments. Images Fig. 10 Fig. 1 Fig. 4 Fig. 5 Fig. 6 Fig. 7 Fig. 8 Fig. 9 Fig. 11 PMID:6776150

  12. Isolation and characterization of C-reactive protein and serum amyloid P component in the rat.

    PubMed Central

    de Beer, F C; Baltz, M L; Munn, E A; Feinstein, A; Taylor, J; Bruton, C; Clamp, J R; Pepys, M B

    1982-01-01

    C-reactive protein (RP) and serum amyloid P component (SAP) have been identified for the first time in rat serum and isolated by calcium-dependent affinity chromatography. Rat CRP closely resembled human CRP in its amino acid composition, in having five subunits per molecule and in its electron microscopic appearance as a pentameric annular disc. It differed, however, from all other mammalian CRP's characterised hitherto in being a glycoprotein bearing a single complex oligosaccharide on each polypeptide subunit. Furthermore one pair of tis subunits per molecule was linked by a interchain disulphide bridges whereas in other animals the subunits of both CRP and SAP are all non-covalently associated. The serum concentration of CRP in normal healthy laboratory rats and in specific pathogen-free rats was 300-600 micrograms/ml which is much greater than has been described in any other species and exceeds even maximal acute phase levels of CRP in man. Following injections of casein or croton oil, serum CRP levels rose to a maximum of about 900 micrograms/ml. Rat CRP bound to pneumococcal C-polysaccharide (CPS( but, in marked contrast to the behaviour of CRP from man, rabbit and marine teleost fish, it did not precipitate with CPS solutions, agglutinate CPS-coated sheep erythrocytes or initiate complement activation. Rat SAP, like SAP of other species, was a glycoprotein but unlike them it was composed only of a single pentameric disc not two such discs interacting face-to-face. The normal level of SAP in rat serum was 20-50 micrograms/ml, very similar to the levels seen in man, and it did not behave as an acute phase reactant in response to casein or croton-oil injections. In this respect it resembled human SAP but differed from murine SAP which is a major acute phase reactant. Images Figure 2 Figure 3 Figure 5 Figure 6 Figure 7 Figure 8 Figure 11 PMID:7056568

  13. Metallothionein gene expression is regulated by serum factors and activators of protein kinase C.

    PubMed Central

    Imbra, R J; Karin, M

    1987-01-01

    The exact physiological role of metallothionein (MT) is not clear. It has been suggested that these low-molecular-weight, highly inducible, heavy-metal-binding proteins serve in the regulation of intracellular Zn metabolism. Among the Zn-requiring systems are several enzymes involved in DNA replication and repair. Therefore, during periods of active DNA synthesis there is likely to be an increased demand for Zn, which could be met by elevated MT synthesis. For that reason, we examined whether stimulation of cellular proliferation leads to increased expression of MT. We report here that treatment of cultured mammalian cells with serum growth factors and activators of protein kinase C, all of which are known to have growth stimulatory activity, led to induction of MT mRNA. One of the required steps in the signal transduction pathways triggered by these agents, ending in MT induction, appears to be the activation of protein kinase C. Images PMID:3600629

  14. Serum Levels of Retinol-Binding Protein 4 (RBP4) and Risk of Colon Adenoma

    PubMed Central

    Abola, Matthew V.; Thompson, Cheryl L.; Chen, Zhengyi; Chak, Amitabh; Berger, Nathan A.; Kirwan, John P.; Li, Li

    2015-01-01

    Retinol binding-protein 4 (RBP4), a recently identified adipokine and retinol transporter, has been shown to play a causative role in insulin resistance, an underpinning between obesity and colon neoplasia. Yet, the relationship between RPB4 and cancer, including colon neoplasia is largely unexplored. We carried out a cross-sectional study to determine the risk association between RBP4 and colon adenomas. We determined pre-diagnostic serum levels of RBP4 in 626 patients undergoing screening colonoscopies from January 2006 to March 2007. The cases had statistically significant higher levels of RBP4 than the controls (58.5µg/mL ± 38.2 vs. 51.9µg/mL ± 32.5, p=0.03). Multivariate logistic regression model revealed a statistically significant overall association of RBP4 with risk of colon adenoma (OR = 3.10 for each increment of 35µg/mL, CI = 1.15 – 8.66; p = 0.03). Stratified analysis by the median BMI showed that the risk association was largely limited to those with BMI < 27.8 kg/m2. Compared to those in the bottom tertile of RBP4, the ORs for the 2nd and 3rd tertiles were 1.84 (CI = 0.89–3.8) and 2.14 (CI = 1.08–4.23) respectively (p for trend = 0.03); there was little evidence for such an association among those with BMI ? 27.8 kg/m2. This is the first study to show colon adenoma risk association with high circulating levels of RBP4. Further study is merited to investigate the mechanism that underlies the RBP4-colon neoplasia link. PMID:25712946

  15. Identification of differential pattern of protein expression in canine osteoarthritis serum after anterior cruciate ligament transection: a proteomic analysis.

    PubMed

    Gharbi, M; Sanchez, C; Mazzucchelli, G; De Pauw, E; Henrotin, Y

    2013-09-01

    Osteoarthritis (OA) management remains a great challenge and there is considerable effort to understand its pathophysiology and to identify new therapeutic targets and biomarkers. Canine OA surgically induced by the transection of the anterior cruciate ligament (ACLT) is a widely used and relevant model. This study reports a proteome mapping of dog serum and an analysis of the differentially expressed proteins between before and after ACLT. In the first part of the study, 261 picked protein spots were identified from preparative 2D gels and 71 different proteins were identified among the 261 spots present on the reference map. Canine serum proteome mapping reveals the presence of proteins of interest, such as fetuin B, complement C3 and C1s and pregnancy zone protein. The comparison between serum from dogs before and after ACLT reveals the differential expression of several proteins that could play a key role in the pathogenesis of OA. A number of proteins, such as fetuin B and complement C3, were increased in dog OA serum whereas others, such as hyaluronan binding protein 2, inter-alpha-trypsin inhibitor H4 (ITIH4), complement C1s and C4 and haptoglobin were decreased. Some of these proteins could be candidate biomarkers for diagnosis, prognosis and treatment evaluation. The results of the study also reinforced the similarities between dog experimental OA and human cases of OA. PMID:23831215

  16. Serum-free culture alters the quantity and protein composition of neuroblastoma-derived extracellular vesicles.

    PubMed

    Li, Jinghuan; Lee, Yi; Johansson, Henrik J; Mäger, Imre; Vader, Pieter; Nordin, Joel Z; Wiklander, Oscar P B; Lehtiö, Janne; Wood, Matthew J A; Andaloussi, Samir El

    2015-01-01

    Extracellular vesicles (EVs) play a significant role in cell-cell communication in numerous physiological processes and pathological conditions, and offer promise as novel biomarkers and therapeutic agents for genetic diseases. Many recent studies have described different molecular mechanisms that contribute to EV biogenesis and release from cells. However, little is known about how external stimuli such as cell culture conditions can affect the quantity and content of EVs. While N2a neuroblastoma cells cultured in serum-free (OptiMEM) conditions did not result in EVs with significant biophysical or size differences compared with cells cultured in serum-containing (pre-spun) conditions, the quantity of isolated EVs was greatly increased. Moreover, the expression levels of certain vesicular proteins (e.g. small GTPases, G-protein complexes, mRNA processing proteins and splicing factors), some of which were previously reported to be involved in EV biogenesis, were found to be differentially expressed in EVs under different culture conditions. These data, therefore, contribute to the understanding of how extracellular factors and intracellular molecular pathways affect the composition and release of EVs. PMID:26022510

  17. Serum-free culture alters the quantity and protein composition of neuroblastoma-derived extracellular vesicles

    PubMed Central

    Li, Jinghuan; Lee, Yi; Johansson, Henrik J.; Mäger, Imre; Vader, Pieter; Nordin, Joel Z.; Wiklander, Oscar P. B.; Lehtiö, Janne; Wood, Matthew J. A.; Andaloussi, Samir EL

    2015-01-01

    Extracellular vesicles (EVs) play a significant role in cell–cell communication in numerous physiological processes and pathological conditions, and offer promise as novel biomarkers and therapeutic agents for genetic diseases. Many recent studies have described different molecular mechanisms that contribute to EV biogenesis and release from cells. However, little is known about how external stimuli such as cell culture conditions can affect the quantity and content of EVs. While N2a neuroblastoma cells cultured in serum-free (OptiMEM) conditions did not result in EVs with significant biophysical or size differences compared with cells cultured in serum-containing (pre-spun) conditions, the quantity of isolated EVs was greatly increased. Moreover, the expression levels of certain vesicular proteins (e.g. small GTPases, G-protein complexes, mRNA processing proteins and splicing factors), some of which were previously reported to be involved in EV biogenesis, were found to be differentially expressed in EVs under different culture conditions. These data, therefore, contribute to the understanding of how extracellular factors and intracellular molecular pathways affect the composition and release of EVs. PMID:26022510

  18. BZP, a novel serum-responsive zinc finger protein that inhibits gene transcription.

    PubMed Central

    Franklin, A J; Jetton, T L; Shelton, K D; Magnuson, M A

    1994-01-01

    We report the fortuitous isolation of cDNA clones encoding a novel zinc finger DNA-binding protein termed BZP. The protein encoded is 114 kDa and contains eight zinc finger motifs, seven of which are present in two clusters at opposite ends of the molecule. Both finger clusters bound to the 9-bp sequence AAAGGTGCA with apparent Kds of approximately 2.5 nM. Two of the finger motifs within the amino- and carboxy-terminal finger clusters share 63% amino acid identity. BZP inhibited transcription of the herpes simplex virus thymidine kinase promoter when copies of the 9-bp target motif were linked in cis, suggesting that it functions as a transcriptional repressor. BZP mRNA and immunoreactivity were detected in several established cell lines but were most abundant in hamster insulinoma (HIT) cells, the parental source of the cDNAs. In mouse tissues, BZP mRNA and immunoreactivity were identified in cells of the endocrine pancreas, anterior pituitary, and central nervous system. Interestingly, in HIT cells proliferating in culture, BZP immunoreactivity was predominately nuclear in location, whereas it was usually located in the cytoplasm in most neural and neuroendocrine tissues. Serum deprivation of HIT cells caused BZP immunoreactivity to become predominantly cytoplasmic in location and attenuated its inhibitory effect on transcription, thereby suggesting that the both the subcellular location and the function of this protein are modulated by factors in serum. Images PMID:7935395

  19. Comparison of serum amyloid A protein and C-reactive protein concentrations in cancer and non-malignant disease.

    PubMed Central

    Raynes, J G; Cooper, E H

    1983-01-01

    Serum amyloid A (SAA) concentrations correlate well with C-reactive protein (CRP) concentrations. However, SAA is sometimes raised in disease when CRP is normal. This appears to occur more often in certain diseases such as rheumatoid arthritis, primary biliary cirrhosis and chronic active hepatitis. SAA concentrations did not distinguish between cancer with and without metastases as previously indicated, although mean concentrations were higher in more advanced tumours. Despite the higher sensitivity of SAA over CRP in the inflammatory response, SAA has little advantage over CRP in the assessment of malignant disease. PMID:6863571

  20. Riboflavin carrier protein: a serum and tissue marker for breast carcinoma.

    PubMed

    Karande, A A; Sridhar, L; Gopinath, K S; Adiga, P R

    2001-09-20

    We have earlier shown that the estrogen-modulated riboflavin carrier protein (RCP) first isolated from the chicken egg is evolutionarily conserved in mammals and is elaborated by lactating mammary gland as demonstrated with rat mammary epithelial cells in culture and confirmed by isolation of the vitamin carrier from bovine milk. In view of several earlier reports that many milk proteins as well as other estrogen-inducible proteins are up-regulated and secreted into circulation in animal models and in women with neoplastic breast disease, we analyzed serum RCP levels in a double-blind study using a specific radioimmunoassay in pre- and post-menopausal women with clinically diagnosed breast cancer at early and advanced stages of the disease and compared these levels with those in normal age-matched control volunteers. Our data reveal that the serum RCP levels in cycling breast cancer patients are 3- to 4-fold higher (p < 0.01) than those in their normal counterparts. This difference in circulatory RCP levels between cancer patients and their age-matched normal counterparts is further magnified to 9- to 11-fold (p < 0.005) at the post-menopausal stage. In addition, there seems to be a good correlation between rising RCP levels and disease progression, since significantly higher RCP concentrations (p < 0.005) are encountered in patients with advanced metastasizing breast cancer versus those with early disease. Using specific monoclonal antibodies, RCP could be localized immunohistochemically in the cytoplasm of invading neoplastic cells of lobular and ductal carcinomas of the breast, indicating that the malignant cells are probably the source of the elevated serum RCP levels in breast cancer. These findings suggest that measurement of circulatory RCP and the immunohistochemical staining pattern of RCP in biopsy specimens could be exploited as an additional marker in diagnosis/prognosis of breast cancer in women. PMID:11494224

  1. Serum Vitamin D, Vitamin D Binding Protein, and Risk of Colorectal Cancer

    PubMed Central

    Anic, Gabriella M.; Weinstein, Stephanie J.; Mondul, Alison M.; Männistö, Satu; Albanes, Demetrius

    2014-01-01

    Background We previously reported a positive association between serum 25-hydroxyvitamin D (25(OH)D) and colorectal cancer risk. To further elucidate this association, we examined the molar ratio of 25(OH)D to vitamin D binding protein (DBP), the primary 25(OH)D transport protein, and whether DBP modified the association between 25(OH)D and colorectal cancer risk. Methods In a nested case-control study within the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study, controls were 1?1 matched to 416 colorectal cancer cases based on age and date of blood collection. Logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CI) for quartiles of 25(OH)D, DBP, and the molar ratio of 25(OH)D:DBP, a proxy for free, unbound circulating 25(OH)D. Results Comparing highest to lowest quartiles, DBP was not associated with colorectal cancer risk (OR?=?0.91; 95% CI: 0.58, 1.42, p for trend ?=?0.58); however, a positive risk association was observed for the molar ratio of 25(OH)D:DBP (OR?=?1.44; 95% CI: 0.92, 2.26, p for trend ?=?0.04). In stratified analyses, the positive association between 25(OH)D and colorectal cancer was stronger among men with DBP levels above the median (OR?=?1.89; 95% CI: 1.07, 3.36, p for trend ?=?0.01) than below the median (OR?=?1.20; 95% CI: 0.68, 2.12, p for trend ?=?0.87), although the interaction was not statistically significant (p for interaction ?=?0.24). Conclusion Circulating DBP may influence the association between 25(OH)D and colorectal cancer in male smokers, with the suggestion of a stronger positive association in men with higher DBP concentrations. This finding should be examined in other populations, especially those that include women and non-smokers. PMID:25036524

  2. Speciation of trace elements in proteins in human and bovine serum by size exclusion chromatography and inductively coupled plasma-mass spectrometry with a magnetic sector mass spectrometer.

    PubMed

    Wang, J; Houk, R S; Dreessen, D; Wiederin, D R

    1999-10-01

    Proteins are separated by size exclusion chromatography while atomic ions from the inorganic elements are detected on-line by inductively coupled plasma-mass spectrometry. A double focusing mass analyzer provides very high sensitivity, low background, and sufficient spectral resolution to separate the atomic ions of interest from most polyatomic ions at the same nominal m/z value. The chromatograms show the distribution of the elements of interest between protein-bound and free fractions and provide the approximate molecular weights of those protein fractions that contain the elements monitored. The distribution of various elements, including V, Mo, Fe, Co, Mn, and lanthanides, in human or bovine serum samples are shown. Alkali metals and Tl are present primarily as free metal ions and are not bound to proteins. Inorganic elements spiked into the serum samples can be followed into various proteins. EDTA does not remove Fe, Pb, Sn, or Th from the proteins but does extract Mn from some proteins. Procedures for determining the effects of breaking disulfide linkages on the metal binding characteristics of proteins are also described. PMID:10550683

  3. Differential expression profiling of serum proteins and metabolites for biomarker discovery

    NASA Astrophysics Data System (ADS)

    Roy, Sushmita Mimi; Anderle, Markus; Lin, Hua; Becker, Christopher H.

    2004-11-01

    A liquid chromatography-mass spectrometry (LC-MS) proteomics and metabolomics platform is presented for quantitative differential expression analysis. Proteome profiles obtained from 1.5 [mu]L of human serum show ~5000 de-isotoped and quantifiable molecular ions. Approximately 1500 metabolites are observed from 100 [mu]L of serum. Quantification is based on reproducible sample preparation and linear signal intensity as a function of concentration. The platform is validated using human serum, but is generally applicable to all biological fluids and tissues. The median coefficient of variation (CV) for ~5000 proteomic and ~1500 metabolomic molecular ions is approximately 25%. For the case of C-reactive protein, results agree with quantification by immunoassay. The independent contributions of two sources of variance, namely sample preparation and LC-MS analysis, are respectively quantified as 20.4 and 15.1% for the proteome, and 19.5 and 13.5% for the metabolome, for median CV values. Furthermore, biological diversity for ~20 healthy individuals is estimated by measuring the variance of ~6500 proteomic and metabolomic molecular ions in sera for each sample; the median CV is 22.3% for the proteome and 16.7% for the metabolome. Finally, quantitative differential expression profiling is applied to a clinical study comparing healthy individuals and rheumatoid arthritis (RA) patients.

  4. Serum-stable quantum dot--protein hybrid nanocapsules for optical bio-imaging

    NASA Astrophysics Data System (ADS)

    Lee, Jeong Yu; Nam, Dong Heon; Oh, Mi Hwa; Kim, Youngsun; Choi, Hyung Seok; Jeon, Duk Young; Beum Park, Chan; Nam, Yoon Sung

    2014-05-01

    We introduce shell cross-linked protein/quantum dot (QD) hybrid nanocapsules as a serum-stable systemic delivery nanocarrier for tumor-targeted in vivo bio-imaging applications. Highly luminescent, heavy-metal-free Cu0.3InS2/ZnS (CIS/ZnS) core-shell QDs are synthesized and mixed with amine-reactive six-armed poly(ethylene glycol) (PEG) in dichloromethane. Emulsification in an aqueous solution containing human serum albumin (HSA) results in shell cross-linked nanocapsules incorporating CIS/ZnS QDs, exhibiting high luminescence and excellent dispersion stability in a serum-containing medium. Folic acid is introduced as a tumor-targeting ligand. The feasibility of tumor-targeted in vivo bio-imaging is demonstrated by measuring the fluorescence intensity of several major organs and tumor tissue after an intravenous tail vein injection of the nanocapsules into nude mice. The cytotoxicity of the QD-loaded HSA-PEG nanocapsules is also examined in several types of cells. Our results show that the cellular uptake of the QDs is critical for cytotoxicity. Moreover, a significantly lower level of cell death is observed in the CIS/ZnS QDs compared to nanocapsules loaded with cadmium-based QDs. This study suggests that the systemic tumor targeting of heavy-metal-free QDs using shell cross-linked HSA-PEG hybrid nanocapsules is a promising route for in vivo tumor diagnosis with reduced non-specific toxicity.

  5. Serum levels of insulin-like growth factor-I, IGF-binding protein 1 and 3, and insulin and endometrial cancer risk

    Microsoft Academic Search

    E Weiderpass; K Brismar; R Bellocco; H Vainio; R Kaaks

    2003-01-01

    Insulin-like growth factor-I (IGF-I) and IGF-binding protein-1 and 3 (IGFPB-1, IGFPB-3) are expressed in normal and neoplastic endometrium. Their role and the role of insulin in the aetiology of endometrial cancer, is unclear. We performed a population-based case-control study in Sweden, including 288 endometrial cancer patients and 392 control women and analysed total serum IGF-I, IGFBP-1, IGFBP-3, insulin and BMI

  6. Serum Albumin Prevents Protein Aggregation and Amyloid Formation and Retains Chaperone-like Activity in the Presence of Physiological Ligands

    PubMed Central

    Finn, Thomas E.; Nunez, Andrea C.; Sunde, Margaret; Easterbrook-Smith, Simon B.

    2012-01-01

    Although serum albumin has an established function as a transport protein, evidence is emerging that serum albumin may also have a role as a molecular chaperone. Using established techniques to characterize chaperone interactions, this study demonstrates that bovine serum albumin: 1) preferentially binds stressed over unstressed client proteins; 2) forms stable, soluble, high molecular weight complexes with stressed client proteins; 3) reduces the aggregation of client proteins when it is present at physiological levels; and 4) inhibits amyloid formation by both WT and L55P transthyretin. Although the antiaggregatory effect of serum albumin is maintained in the presence of physiological levels of Ca2+ and Cu2+, the presence of free fatty acids significantly alters this activity: stabilizing serum albumin at normal levels but diminishing chaperone-like activity at high concentrations. Moreover, here it is shown that depletion of albumin from human plasma leads to a significant increase in aggregation under physiologically relevant heat and shear stresses. This study demonstrates that serum albumin possesses chaperone-like properties and that this activity is maintained under a number of physiologically relevant conditions. PMID:22549788

  7. Integration of binding peptide selection and multifunctional particles as tool-box for capture of soluble proteins in serum.

    PubMed

    Cusano, Angela Maria; Causa, Filippo; Moglie, Raffaella Della; Falco, Nunzia; Scognamiglio, Pasqualina Liana; Aliberti, Anna; Vecchione, Raffaele; Battista, Edmondo; Marasco, Daniela; Savarese, Marika; Raucci, Umberto; Rega, Nadia; Netti, Paolo Antonio

    2014-10-01

    In this paper, we report on a general approach for the detection of a specific tumoural biomarker directly in serum. Such detection is made possible using a protein-binding peptide selected through an improved phage display technique and then conjugated to engineered microparticles (MPs). Protein biomarkers represent an unlimited source of information for non-invasive diagnostic and prognostic tests; MP-based assays are becoming largely used in manipulation of soluble biomarkers, but their direct use in serum is hampered by the complex biomolecular environment. Our technique overcomes the current limitations as it produces a selective MP--engineered with an antifouling layer--that 'captures' the relevant protein staying impervious to the background. Our system succeeds in fishing-out the human tumour necrosis factor alpha directly in serum with a high selectivity degree. Our method could have great impact in soluble protein manipulation and detection for a wide variety of diagnostic applications. PMID:25100324

  8. Structural insights into the binding of uranyl with human serum protein apotransferrin structure and spectra of protein-uranyl interactions.

    PubMed

    Benavides-Garcia, Maria G; Balasubramanian, Krishnan

    2009-09-01

    Ab initio quantum mechanical computational studies for the structure and IR spectra of the uranyl complex with human serum apotransferrin (TF) protein are carried out to model uranyl intake into the human cell through endocytosis and formation of a coordination complex with the protein binding sites. The computed IR spectra and structure of the uranyl-protein complex facilitate interpretation of the observed spectra and confirm the primary binding sites of the transferrin protein with the uranyl ion. Our computed equilibrium geometry and the IR spectra of the uranyl-TF complex reveal that uranyl ion is bound to two tyrosines, one aspartate group, and one carbonate ion. Our IR spectra indicate that histidine is not involved in binding to uranyl with transferrin protein. Our computations reveal a short, strong hydrogen bond, which could play an important role in the stabilization and formation of the uranyl-TF complex. Computed Laplacian charge plots indicate high chemical reactivity on this complex as both an electrophile and a nucleophile, facilitating binding to different receptors and thus entry into a number of target organs and the blood-brain barrier. The Mulliken charge density plots and the three-dimensional charge density plots suggest a donor-acceptor mechanism in the complex formation. PMID:19678663

  9. Correlation between Ocular Demodex Infestation and Serum Immunoreactivity to Bacillus Proteins in Patients with Facial Rosacea

    PubMed Central

    Li, Jianjing; O'Reilly, Niamh; Sheha, Hosam; Katz, Raananah; Raju, Vadrevu K.; Kavanagh, Kevin; Tseng, Scheffer C. G.

    2010-01-01

    Purpose To investigate correlation between ocular Demodex infestation and serum. Design A prospective study to correlate clinical findings with laboratory data. Participants We consecutively enrolled 59 patients: 34 men and 25 women with a mean age of 60.4±17.6 years (range, 17–93). Methods Demodex counting was performed based on lash sampling. Serum immunoreactivity to two 62-kDa and 83-kDa proteins derived from B oleronius was determined by Western blot analysis. Facial rosacea, lid margin, and ocular surface inflammation were documented by photography and graded in a masked fashion. Main Outcome Measures Statistical significance based on correlative analyses of clinical and laboratory data. Results These 59 patients were age matched, but not gender matched, regarding serum immunoreactivity, ocular Demodex infestation, or facial rosacea. There was a significant correlation between serum immunoreactivity and facial rosacea (P = 0.009), lid margin inflammation (P = 0.040), and ocular Demodex infestation (P = 0.048), but not inferior bulbar conjunctival inflammation (P = 0.573). The Demodex count was significantly higher in patients with positive facial rosacea (6.6±9.0 vs. 1.9±2.2; P = 0.014). There was a significant correlation of facial rosacea with lid margin inflammation (P = 0.016), but not with inferior bulbar conjunctival inflammation (P = 0.728). Ocular Demodex infestation was less prevalent in patients with aqueous tear-deficiency dry eye than those without (7/38 vs. 12/21; P = 0.002). Conclusions The strong correlation provides a better understanding of comorbidity between Demodex mites and their symbiotic B oleronius in facial rosacea and blepharitis. Treatments directed to both warrant future investigation. PMID:20079929

  10. Differentially Expressed RNA from Public Microarray Data Identifies Serum Protein Biomarkers for Cross-Organ Transplant Rejection and Other Conditions

    PubMed Central

    Chen, Rong; Sigdel, Tara K.; Li, Li; Kambham, Neeraja; Dudley, Joel T.; Hsieh, Szu-chuan; Klassen, R. Bryan; Chen, Amery; Caohuu, Tuyen; Morgan, Alexander A.; Valantine, Hannah A.; Khush, Kiran K.; Sarwal, Minnie M.; Butte, Atul J.

    2010-01-01

    Serum proteins are routinely used to diagnose diseases, but are hard to find due to low sensitivity in screening the serum proteome. Public repositories of microarray data, such as the Gene Expression Omnibus (GEO), contain RNA expression profiles for more than 16,000 biological conditions, covering more than 30% of United States mortality. We hypothesized that genes coding for serum- and urine-detectable proteins, and showing differential expression of RNA in disease-damaged tissues would make ideal diagnostic protein biomarkers for those diseases. We showed that predicted protein biomarkers are significantly enriched for known diagnostic protein biomarkers in 22 diseases, with enrichment significantly higher in diseases for which at least three datasets are available. We then used this strategy to search for new biomarkers indicating acute rejection (AR) across different types of transplanted solid organs. We integrated three biopsy-based microarray studies of AR from pediatric renal, adult renal and adult cardiac transplantation and identified 45 genes upregulated in all three. From this set, we chose 10 proteins for serum ELISA assays in 39 renal transplant patients, and discovered three that were significantly higher in AR. Interestingly, all three proteins were also significantly higher during AR in the 63 cardiac transplant recipients studied. Our best marker, serum PECAM1, identified renal AR with 89% sensitivity and 75% specificity, and also showed increased expression in AR by immunohistochemistry in renal, hepatic and cardiac transplant biopsies. Our results demonstrate that integrating gene expression microarray measurements from disease samples and even publicly-available data sets can be a powerful, fast, and cost-effective strategy for the discovery of new diagnostic serum protein biomarkers. PMID:20885780

  11. Protein phosphorylation patterns in serum from schizophrenia patients and healthy controls.

    PubMed

    Jaros, Julian A J; Martins-de-Souza, Daniel; Rahmoune, Hassan; Rothermundt, Matthias; Leweke, F Markus; Guest, Paul C; Bahn, Sabine

    2012-12-01

    Most proteomic studies to date have attempted to identify changes in protein levels without considering the effects of post-translational modifications (PTM). However, characteristic changes of PTM such as phosphorylation could be biologically informative, as these can give insights into disease and drug mechanisms of action at the functional level. With this in mind, we have conducted a comparative proteomic and phosphoproteomic analysis of blood sera from 20 antipsychotic-naïve schizophrenia patients and 20 matched healthy controls. We used immobilised metal ion affinity chromatography (IMAC) for enrichment of phosphoproteins combined with label-free liquid chromatography-mass spectrometry (LC-MS(E)) for identification and measurement of protein and phosphoprotein levels. The LC-MS(E) analysis of both IMAC-fractions resulted in identification of 35 proteins with altered levels in schizophrenia. Analysis of the enriched fraction resulted in identification of 72 phosphoproteins with altered phosphorylation patterns. Of these, 59 showed changes in phosphorylation only, with no overall change in protein levels. This study provided evidence that schizophrenia patients feature serum abnormalities in phosphorylation of proteins involved in acute phase response and coagulation pathways. Further studies of such phosphorylation-specific changes could lead to a better understanding of the molecular aetiology of schizophrenia, and provide a means of biomarker identification for clinical studies. This article is part of a Special Issue entitled: Integrated omics. PMID:22641159

  12. The Anion Gap and Routine Serum Protein Measurements in Monoclonal Gammopathies

    PubMed Central

    Joseph, Rosy E.; Gaughan, William J.; McBride, Laura; Bilotti, Elizabeth; McNeill, Ann; Schmidt, Linda; Schillen, Danielle; Siegel, David S.

    2011-01-01

    Summary Background and objectives An abnormal anion gap and an increased total protein and globulin are clues to the diagnosis of monoclonal gammopathy. We explored the utility of these markers in IgG, IgA, IgM, and free light chain monoclonal gammopathies. Design, Setting, Participants, & Measurements The anion gap, Na+ – (Cl– + HCO3–), corrected for hypoalbuminemia, was calculated in patients with monoclonal gammopathies. Exclusion criteria were serum calcium >10.5 mg/dl and/or creatinine >2 mg/dl. Results Among 287 patients, 242 remained after applying exclusion criteria (109 IgG, 64 IgA, 21 IgM, and 48 light chain); 36% of 242 patients required correction for hypoalbuminemia. The anion gap was decreased (<10) in 22% of IgG and increased (>15) in 31% of IgA monoclonal gammopathies. IgM did not affect the gap. In light chain gammopathies, the anion gap showed no consistent trend (15% increased, 17% decreased). Mean clonal IgG, IgA, and IgM concentrations were 10-fold higher than mean clonal free light chain concentrations in the respective monoclonal gammopathies (P < 0.001). These paraprotein level disparities were reflected in significantly increased mean serum total protein and globulin concentrations in IgG, IgA, and IgM versus free light chain monoclonal gammopathies, where mean total protein and globulin levels were within normal limits (P < 0.001). Conclusions The anion gap was significantly altered in IgG and IgA monoclonal gammopathies, but it was not a sensitive tool for suspecting the diagnosis. In light chain monoclonal gammopathies, the anion gap, total protein, and globulin did not provide reliable diagnostic clues. PMID:22157711

  13. A Multiplexed Device Based on Tunable Nanoshearing for Specific Detection of Multiple Protein Biomarkers in Serum

    PubMed Central

    Vaidyanathan, Ramanathan; van Leeuwen, Lara Michelle; Rauf, Sakandar; Shiddiky, Muhammad J. A.; Trau, Matt

    2015-01-01

    Microfluidic flow based multiplexed devices have gained significant promise in detecting biomarkers in complex biological samples. However, to fully exploit their use in bioanalysis, issues such as (i) low sensitivity and (ii) high levels of nonspecific adsorption of non-target species have to be overcome. Herein, we describe a new multiplexed device for the sensitive detection of multiple protein biomarkers in serum by using an alternating current (ac) electrohydrodynamics (ac-EHD) induced surface shear forces based phenomenon referred to as nanoshearing. The tunable nature (via manipulation of ac field) of these nanoshearing forces can alter the capture performance of the device (e.g., improved fluid transport enhances number of sensor-target collisions). This can also selectively displace weakly (nonspecifically) bound molecules from the electrode surface (i.e., fluid shear forces can be tuned to shear away nonspecific species present in biological samples). Using this approach, we achieved sensitive (100?fg mL?1) naked eye detection of multiple protein targets spiked in human serum and a 1000-fold enhancement in comparison to hydrodynamic flow based devices for biomarker detection. We believe that this approach could potentially represent a clinical diagnostic tool that can be integrated into resource-limited settings for sensitive detection of target biomarkers using naked eye. PMID:25978807

  14. A Serum Protein-Based Algorithm for the Detection of Alzheimer's Disease

    PubMed Central

    O'Bryant, Sid E.; Xiao, Guanghua; Barber, Robert; Reisch, Joan; Doody, Rachelle; Fairchild, Thomas; Adams, Perrie; Waring, Steven; Diaz-Arrastia, Ramon

    2011-01-01

    Background Alzheimer's disease (AD) is the most common form of age-related dementia and one of the most serious health problems in the industrialized world. Biomarker approaches to diagnostics would be more time and cost effective and may also be useful for identifying endophenotypes within AD patient populations. Methods We analyzed serum protein-based multiplex biomarker data from 197 patients diagnosed with AD and 203 controls from a longitudinal study of Alzheimer's disease being conducted by the Texas Alzheimer's Research Consortium to develop an algorithm that separates AD from controls. The total sample was randomized equally into training and test sets and random forest methods were applied to the training set to create a biomarker risk score. Findings The biomarker risk score had a sensitivity and specificity of 0.80 and 0.91, respectively and an AUC of 0.91 in detecting AD. When age, gender, education, and APOE status were added to the algorithm, the sensitivity, specificity, and AUC were 0.94, 0.84, and 0.95, respectively. Interpretation These initial data suggest that serum protein-based biomarkers can be combined with clinical information to accurately classify AD. Of note, a disproportionate number of inflammatory and vascular markers were weighted most heavily in analyses. Additionally, these markers consistently distinguished cases from controls in SAM, logistic regression and Wilcoxon analyses, suggesting the existence of an inflammatory-related endophenotype of AD that may provide targeted therapeutic opportunities for this subset of patients. PMID:20837851

  15. Short-term space flight on nitrogenous compounds, lipoproteins, and serum proteins

    NASA Technical Reports Server (NTRS)

    Leach, C. S.; Lane, H. W.; Krauhs, J. M.

    1994-01-01

    Biochemical variables in blood were measured in venous blood samples from 38 to 72 Space Shuttle astronauts before and immediately after flights of 2 to 11 days. Mean pre- and postflight values were compared using the paired t-test or the Wilcoxon signed-rank test. The largest change in serum enzymes was a 21% increase (P = .0014) in gamma-glutamyl-transpeptidase, which may have been related to stress. The median value of apolipoprotein (apo) A-I decreased from 152 to 127 mg/dL (P < .0001), but the change in apo B (77 to 73 mg/dL) was not statistically significant, and the mean apo A-I/apo B ratio remained well above 1.5. A decrease in dietary fat and cholesterol intake during shuttle missions may have been a cause of the change in apo A-I. Twelve of the 16 nonenzyme serum proteins measured were significantly elevated (P < .05), possibly because of hemoconcentration and increased protein catabolism. The 56% increase in haptoglobin may be related to release of suppressed erythropoiesis at landing.

  16. A multiplexed device based on tunable nanoshearing for specific detection of multiple protein biomarkers in serum.

    PubMed

    Vaidyanathan, Ramanathan; van Leeuwen, Lara Michelle; Rauf, Sakandar; Shiddiky, Muhammad J A; Trau, Matt

    2015-01-01

    Microfluidic flow based multiplexed devices have gained significant promise in detecting biomarkers in complex biological samples. However, to fully exploit their use in bioanalysis, issues such as (i) low sensitivity and (ii) high levels of nonspecific adsorption of non-target species have to be overcome. Herein, we describe a new multiplexed device for the sensitive detection of multiple protein biomarkers in serum by using an alternating current (ac) electrohydrodynamics (ac-EHD) induced surface shear forces based phenomenon referred to as nanoshearing. The tunable nature (via manipulation of ac field) of these nanoshearing forces can alter the capture performance of the device (e.g., improved fluid transport enhances number of sensor-target collisions). This can also selectively displace weakly (nonspecifically) bound molecules from the electrode surface (i.e., fluid shear forces can be tuned to shear away nonspecific species present in biological samples). Using this approach, we achieved sensitive (100?fg mL(-1)) naked eye detection of multiple protein targets spiked in human serum and a 1000-fold enhancement in comparison to hydrodynamic flow based devices for biomarker detection. We believe that this approach could potentially represent a clinical diagnostic tool that can be integrated into resource-limited settings for sensitive detection of target biomarkers using naked eye. PMID:25978807

  17. A solvent system for delipidation of plasma or serum without protein precipitation

    Microsoft Academic Search

    Bill E. Cham; Brian R. Knowlee

    A technique has been developed which attains ih 30 minutes complete removal of triglyceride, cholesterol, phos- pholipid, and unesterified fatty acids from plasma without protein denaturation. Plasma is agitated at room temperature with a mixture of butanol and di-isopropyl ether in a 40:60 (v\\/v) ratio. The plasma proteins, including the apolipo- proteins, remain in solution in the aqueous phase, while

  18. Use of serum retinol-binding protein for prediction of vitamin A deficiency: effects of HIV1 infection, protein malnutrition, and the acute phase response1-3

    Microsoft Academic Search

    Jared M Baeten; Barbra A Richardson; Daniel D Bankson; Mark H Wener; Joan K Kreiss; Ludo Lavreys; Kishorchandra Mandaliya; Job J Bwayo; R Scott McClelland

    Background: Serum retinol is the most commonly used indicator of vitamin A status. Retinol is transported in a 1-to-1 complex with retinol-binding protein (RBP). RBP is easy and inexpensive to measure, and studies have shown a high correlation between concentrations of RBP and concentrations of retinol. The perfor- mance of RBP in the context of infection or protein malnutrition, however,

  19. Impact of Serum Proteins on MRI Contrast Agents: Cellular Binding and T2 relaxation.

    PubMed

    Hill, Alexandra; Payne, Christine K

    2014-01-01

    Superparamagnetic iron oxide nanoparticles (SPIONs) used as MRI contrast agents or for theranostic applications encounter a complex mixture of extracellular proteins that adsorb on the SPION surface forming a protein corona. Our goal was to understand how cellular binding and T2 relaxation times are affected by this protein corona. Our studies focused on carboxymethyl dextran-modified SPIONs, chosen for their similarity to Resovist SPIONs used to detect liver lesions. Using a combination of fluorescence microscopy and flow cytometry, we find that the cellular binding of SPIONs to both macrophages and epithelial cells is significantly inhibited by serum proteins. To determine if this decreased binding is due to the iron oxide core or the carboxymethyl dextran surface coating, we functionalized polystyrene nanoparticles with a similar carboxymethyl dextran coating. We find a comparable decrease in cellular binding for the carboxymethyl dextran-polystyrene nanoparticles indicating that the carbohydrate surface modification is the key factor in SPION-cell interactions. NMR measurements showed that T2 relaxation times are not affected by corona formation. These results indicate that SPIONs have a decreased binding to cells under physiological conditions, possibly limiting their use in theranostic applications. We expect these results will be useful in the design of SPIONs for future diagnostic and therapeutic applications. PMID:25485101

  20. A time-resolved immunoassay to measure serum antibodies to the rotavirus VP6 capsid protein

    PubMed Central

    Kavanagh, Owen; Zeng, Xi-Lei; Ramani, Sasirekha; Mukhopadhya, Indrani; Crawford, Sue E.; Kang, Gagandeep; Estes, Mary K.

    2013-01-01

    The rotavirus (RV) inner capsid protein VP6 is widely used to evaluate immune response during natural infection and in vaccine studies. Recombinant VP6 from the most prevalent circulating rotavirus strains in each subgroup (SG) identified in a birth cohort of children in southern India [SGII (G1P[8]) and SGI (G10P[11])] were produced. The purified proteins were used to measure VP6-specific antibodies in a Dissociation-Enhanced Lanthanide Fluorometric Immunoassay (DELFIA). The ability of the assay to detect a ?2 fold rise in IgG level in a panel of serum samples from a longitudinal study was compared to a gold standard virus-capture ELISA. A strong association was observed between the assays (p < 0.001; chi-squared test) with assay performances remaining similar when the samples were subdivided as having a fold change increase in VP6 antibody levels (a) within 90 days of RV RNA detection in stool or (b) if no RV RNA was detected within that time period. This study demonstrates the suitability of using recombinant proteins to measure anti-RV immune responses and serves as a “proof of principle” to examine the antibody responses generated to other recombinant RV proteins and thereby possibly identify a correlate of protection. PMID:23183143

  1. Prognostic value of serum alpha-fetoprotein in fulminant hepatic failure including patients treated by charcoal haemoperfusion

    Microsoft Academic Search

    I M Murray-Lyon; A H Orr; B Gazzard; J Kohn; R Williams

    1976-01-01

    Serum alpha-fetoprotein (AFP) levels have been measured sequentially by a radio-immunoassay method in 64 patients with fulminant hepatic failure. In 15 of the 64 patients (23%) AFP levels were raised but in only two did they exceed 500 ng\\/ml. Of the 23 survivors 11 (48%) had raised AFP levels compared with four of the 41 (9-8%) fatal cases (P less

  2. In vitro serum protein-binding characteristics of bis-(2-ethylhexyl) phthalate and its principal metabolite, mono-(2-ethylhexyl) phthalate

    SciTech Connect

    Griffiths, W.C.; Camara, P.D.; Saritelli, A.; Gentile, J.

    1988-04-01

    The metabolism and toxicity of the ubiquitous plasticizer, bis-(2-ethylhexyl) phthalate (DEHP), and its principal metabolite, mono-(2-ethylhexyl) phthalate (MEHP), have been extensively investigated. In an attempt to understand their disposition in man, the authors studied the in vitro serum protein-binding characteristics of these compounds, using ultracentrifugation and agarose gel electrophoresis. The association of DEHP and lipoproteins was shown to be highly dependent upon, and proportional to, the lipid concentration of the serum. It appears that more than half of the serum DEHP is bound to proteins with density greater than 1.21 g/mL when the concentration of cholesterol is below 300 mg/dL or the cholesterol and triglyceride total concentration is less than 600 mg/dL. As the cholesterol and triglyceride concentrations increase, the percent DEHP bound to VLDL, IDL, and LDL increases. MEHP is bound principally to nonlipoprotein constituents in the serum, and this binding distribution is unaffected by lipid concentration. The percent binding of DEHP and MEHP to individual proteins was also found to be unaffected by their concentrations in serum. These data indicate that the protein-binding characteristics of these compounds, in vitro, is somewhat more complex than previously reported.

  3. Role of Serum, a Biological Cue, in Adherence of Enterococcus faecalis to Extracellular Matrix Proteins, Collagen, Fibrinogen, and Fibronectin

    PubMed Central

    Nallapareddy, Sreedhar R.; Murray, Barbara E.

    2009-01-01

    Background Most previous reports have found that Enterococcus faecalis isolates do not show significant adherence to fibronectin and fibrinogen Methods The influence of various conditions on E. faecalis adherence to ECM proteins was evaluated using a radiolabeled adherence assay. Results Among conditions studied, growth in 40% serum, a biological cue with potential clinical relevance, elicited adherence of all 46 E. faecalis strains tested to fibronectin and fibrinogen, but not to elastin; the adherence levels were independent of strain source and was eliminated by treating cells with trypsin. As previously reported, serum also elicited collagen adherence. While prolonged exposure to serum during growth was needed for enhancement of adherence to fibrinogen, brief exposure (<5 min) to serum had an immediate, although partial, effect on adherence to fibronectin and collagen. Pre-treatment of bacteria with chloramphenicol did not decrease the enhanced fibronectin and collagen adherence, indicating that protein synthesis is not required for the latter effect. Conclusion Together, these data suggest that serum components may serve i) as host environmental stimuli to induce production of extracellular matrix protein binding adhesin(s), as previously seen with collagen adherence, and also ii) as activators of adherence, perhaps by forming bridges between ECM proteins and adhesins. PMID:18462135

  4. Proteomics analysis of serum protein profiling in pancreatic cancer patients by DIGE: up-regulation of mannose-binding lectin 2 and myosin light chain kinase 2

    Microsoft Academic Search

    Yefei Rong; Dayong Jin; Chenrui Hou; Jianwen Hu; Wenchuan Wu; Xiaolin Ni; Dansong Wang; Wenhui Lou

    2010-01-01

    BACKGROUND: Pancreatic cancer has significant morbidity and mortality worldwide. Good prognosis relies on an early diagnosis. The purpose of this study was to develop techniques for identifying cancer biomarkers in the serum of patients with pancreatic cancer. METHODS: Serum samples from five individuals with pancreatic cancer and five individuals without cancer were compared. Highly abundant serum proteins were depleted by

  5. Primary structure of duck amyloid protein A. The form deposited in tissues may be identical to its serum precursor.

    PubMed

    Ericsson, L H; Eriksen, N; Walsh, K A; Benditt, E P

    1987-06-22

    The amino acid sequence has been determined for the major protein that accumulates in amyloid fibrils in tissues of the Pekin duck. With the exception of 16 residues at the amino terminus, this 106-residue protein is homologous with human serum amyloid protein A (104-residue apoSAA), which is the putative precursor of the 76-residue protein that accumulates in human patients with amyloidosis. Duck serum is shown to contain a protein that is immunologically related and approximately equal in size (12 kDa) to the deposited form in ducks. These results indicate that proteolytic processing of the precursor is not a necessary step in the deposition of amyloid fibrils, at least in the duck. PMID:3109944

  6. Mass spectrometric identification, characterization and validation of the haptoglobin ?-chain protein as a lung cancer serum biomarker.

    PubMed

    Ayyub, Asima; Saleem, Mahjabeen; Musharraf, Syed Ghulam; Naz, Mamoona; Tariq, Asma; Hashmi, Naghma

    2015-09-01

    Lung cancer is the major contributor to overall cancer-related mortality. Biomarkers are important in early detection and prognosis, in addition to developing treatment regimes, which may improve the patient survival rates. Biomarkers may also assist in investigating the in depth metabolic pathways and in establishing a set of therapeutic agents leading to early detection of the disease. The present study was designed to identify and confirm a lung cancer protein biomarker and to correlate the differential expression of the protein to a particular histological disease type. A total of 100 lung cancer patients and 50 healthy controls were included in the present study and were categorized into the two main histological types of lung cancer; non?small cell lung cancer (NSCLC; n=88) and small cell lung cancer (SCLC; n=12). NSCLC was further subclassified into three histological types; adenocarcinoma (n=34), squamous cell carcinoma (n=48) and large cell carcinoma (n=6). The patient and control serum samples underwent sodium dodecyl sulphate polyacrylamide gel electrophoresis characterization followed by two?dimensional gel electrophoresis. Following mass spectrometry, human haptoglobin was identified with a mass of ~42?46 kDa and an isoelectric point (pI) of ~5.5?6.2. The experimental mass of the protein was found to be 45.8 kDa with a pI of 6.13. The matrix?assisted laser desorption/ionization time?of?flight/time?of?flight data exhibited spectral peaks of 1146.134, 1724.191, 1345.339 and 2210.319 m/z and Mascot search analysis identified these peaks as haptoglobin (accession no. P00738; Mascot score 87; sequence coverage 23%). This protein was significantly overexpressed in squamous cell carcinoma and adenocarcinoma, as compared with the control. The present study described differentially expressed human haptoglobin as a lung cancer serum protein biomarker, which may serve as a diagnostic and therapeutic target and set a standard criteria for the evaluation of histological types of lung cancer compared with other disease types. PMID:26005016

  7. Each row in the table corresponds to one protein group. A. Protein IDs: SGD ORFs, including all proteins that could be identified from the peptides identified.

    E-print Network

    Martin, Alain

    : SGD ORFs, including all proteins that could be identified from the peptides identified. Protein ids are ordered by number of peptides identified (most to least number of peptide identified. C. SGD Id: Mapping from first id to SGD

  8. Ret finger protein inhibits muscle differentiation by modulating serum response factor and enhancer of polycomb1

    PubMed Central

    Kee, H J; Kim, J-R; Joung, H; Choe, N; Lee, S E; Eom, G H; Kim, J C; Geyer, S H; Jijiwa, M; Kato, T; Kawai, K; Weninger, W J; Seo, S B; Nam, K-I; Jeong, M H; Takahashi, M; Kook, H

    2012-01-01

    Skeletal myogenesis is precisely regulated by multiple transcription factors. Previously, we demonstrated that enhancer of polycomb 1 (Epc1) induces skeletal muscle differentiation by potentiating serum response factor (SRF)-dependent muscle gene activation. Here, we report that an interacting partner of Epc1, ret finger protein (RFP), blocks skeletal muscle differentiation. Our findings show that RFP was highly expressed in skeletal muscles and was downregulated during myoblast differentiation. Forced expression of RFP delayed myoblast differentiation, whereas knockdown enhanced it. Epc1-induced enhancements of SRF-dependent multinucleation, transactivation of the skeletal ?-actin promoter, binding of SRF to the serum response element, and muscle-specific gene induction were blocked by RFP. RFP interfered with the physical interaction between Epc1 and SRF. Muscles from rfp knockout mice (Rfp?/?) mice were bigger than those from wild-type mice, and the expression of SRF-dependent muscle-specific genes was upregulated. Myotube formation and myoblast differentiation were enhanced in Rfp?/? mice. Taken together, our findings highlight RFP as a novel regulator of muscle differentiation that acts by modulating the expression of SRF-dependent skeletal muscle-specific genes. PMID:21637294

  9. Changes of Serum Retinol Binding Protein 4 Levels Following 8 Weeks Moderate Aerobic Exercise

    PubMed Central

    Ahmadi, Narges; Moghadasi, Mehrzad; Nuri, Reza

    2013-01-01

    Purpose The purpose of this study was to examine the effects of 8 weeks moderate intensity aerobic exercise on serum retinol binding protein 4 (RBP4) levels in female athletes. Methods Twenty female karate athletics were randomly assigned to one of the exercise group (n = 10) or control group (n = 10). The training group performed endurance training 3 days a week for 8 weeks at an intensity corresponding to 50-60% individual maximum oxygen consumption for 45 min. Results Body mass and body mass index increased (P < 0.05) after 8 weeks aerobic exercise compared to the control group. For waist to hip ratio (WHR), body fat percentage and maximal oxygen consumption there were no significant differences between the exercise group and the control group. There were virtually no changes in body fat percentage, fasting glucose, insulin, insulin resistance and RBP4 levels after 8 weeks training. Conclusion Serum RBP4 level was not affected by 8-week moderate aerobic exercise in female athletes. PMID:24427480

  10. Characteristics of retinol accumulation from serum retinol-binding protein by cultured sertoli cells

    SciTech Connect

    Shingleton, J.L.; Skinner, M.K.; Ong, D.E. (Vanderbilt Univ. School of Medicine, Nashville, TN (USA))

    1989-12-12

    The uptake of retinol was examined in cultured Sertoli cells when retinol was provided as a complex with the transport protein retinol-binding protein (RBP). Sertoil cells accumulated ({sup 3}H)retinol in a time- and temperature-dependent manner. The change in rate of retinol accumulation occurred when the cells had accumulated approximately 0.53 pmol of retinol/{mu}g of cellular DNA. Extraction and HPLC analysis of the cell-associated radioactivity yielded retinol and retinyl esters, indicating that a significant proportion of the accumulated retinol was esterified. Excess unlabeled retinol-RBP competed with ({sup 3}H)retinol-RBP for ({sup 3}H)retinol delivery to the cells, indicating that RBP delivery of retinol was a saturable and competable process. However, free ({sup 3}H)retinol associated with Sertoli cells in a noncompetable manner. The transport constant for specific retinol accumulation from RBP was 3.0 {mu}M. Neither iodinated nor reductively methylated RBP was accumulated by or tightly bound to Sertoli cells. Competition studies indicated, however, that protein recognition is important in the retinol uptake process. RBP, CRBP, and CRBP(II) competed with ({sup 3}H)retinol-RBP for ({sup 3}H)retinol accumulation, but free retinol, retinol-bovine serum albumin, and retinol-{beta}-lactoglobulin did not. These studies indicated that Sertoli cell uptake of retinol involved recognition of the retinol-RBP complex at the cell surface with subsequent internalization of retinol, but not RBP.

  11. Protective effect of serum antibodies against a 110-kilodalton protein of Actinobacillus actinomycetemcomitans following periodontal therapy.

    PubMed

    Beikler, T; Karch, H; Ehmke, B; Klaiber, B; Flemmig, T F

    1999-10-01

    Thirty-four adult patients with untreated periodontitis were randomly assigned to receive full mouth scaling alone or scaling with an adjunctive antimicrobial therapy, both followed by supportive periodontal therapy. At 24 months, specific serum immunoglobulin A (IgA), IgG and IgG subclass antibody reactivities against a 110-kDa protein of Actinobacillus actinomycetemcomitans were assessed by Western blot. In patients harboring A. actinomycetemcomitans intraorally, the IgG4 antibody reactivity against the 110-kDa protein of A. actinomycetemcomitans was associated with significantly increased survival rates of teeth and of sites not exhibiting 2 mm or more of probing attachment loss. The same trend was found for IgG3 and IgG2 antibody reactivities, but it was statistically insignificant. No association with clinical treatment outcome was observed for IgA, IgG and IgG1 antibody reactivities. The results indicated that systemic IgG4 antibody reactivity against the 110-kDa protein of A. actinomycetemcomitans may have a protective effect against periodontal disease progression in patients harboring A. actinomycetemcomitans and receiving periodontal therapy. PMID:10551154

  12. Thiolated DAB dendrimer/ZnSe nanoparticles for C-reactive protein recognition in human serum.

    PubMed

    Algarra, M; Campos, B B; Gomes, D; Alonso, B; Casado, C M; Arrebola, M M; Diez de los Rios, M J; Herrera-Gutiérrez, M E; Seller-Pérez, G; Esteves da Silva, J C G

    2012-09-15

    A nanocomposite obtained by a thiol DAB-dendrimer (generation 5), coated with fluorescent ZnSe quantum dots, was successfully synthesized for the selective recognition of C-reactive protein. The procedure presented was carried out by a novel, cheap and non-toxic bottom up synthesis. The nanocomposite showed an excitation at 180 nm, with two emission bands at 411 and 465 nm, with a full-width at half-maximum of 336 nm. The Stokes shift was influenced by the presence of coating molecules and the intensity was dependent on pH due to the presence of a charge transfer process. The transmission electron microscopy images demonstrated that the spherical nanoparticles obtained displayed a regular shape of 30 nm size. The fluorescence intensity was markedly quenched by the presence of C-reactive protein, with a dynamic Stern-Volmer constant of 0.036 M(-1). The quenching profile shows that about 51% of the ZnSe QDs are located in the external layer of the thiol dendrimer accessible to the quencher. The precision of the method obtained as relative standard deviation was 3.76% (4 mg L(-1), n=3). This water soluble fluorescent nanocomposite showed a set of favorable properties to be used as a sensor for the C-reactive protein in serum samples, at concentrations of risk levels. PMID:22967596

  13. Human serum amyloid A protein. Behaviour in aqueous and urea-containing solutions and antibody production.

    PubMed

    Strachan, A F; Shephard, E G; Bellstedt, D U; Coetzee, G A; van der Westhuyzen, D R; de Beer, F C

    1989-10-15

    Human serum amyloid A protein (apo-SAA) can be prepared by gel filtration of delipidated acute-phase high-density lipoprotein in the presence of urea. The resultant apo-SAA is soluble (greater than 90% solubility) in a wide range of buffer solutions, with all of the six major isoforms of apo-SAA being equally soluble. In urea-containing solutions the isoforms behave qualitatively differently in various urea concentrations, probably reflecting subtle primary-structure variations. The higher-pI isoforms are only completely unfolded at greater than 7 M-urea. By immunizing with apo-SAA adsorbed to acid-treated bacteria (Salmonella minnesota R595), high-titre antibodies can easily be elicited in rabbits. PMID:2597108

  14. [Detection and determination of alpha-feto-protein in human serum. Use in hepato-gastroenterology].

    PubMed

    Borel-Giraud, N; Berthet, P; Nyssen, M; Dorche, J; Girard, M; Bel, A; Vachon, A; Plauchu, A; Saint Cyr, M

    1975-01-01

    The presence of alpha-foeto-protein (A.F.P.) was sought in the serum of 520 patients, suffering from various digestive disorders, using two methods: double immuno-diffusion and counter immuno-electrophoresis on cellogel--a more sensitive technique. A.F.P. was detected on 90 of the 520 patients. A.F.P. was measured by radial immunodiffusion or by electro-immunodiffusion on cellogel. Study of the values obtained in various kinds of hepatic disorders indicates that high levels (more than 10 mg/1) are seen almost solely in association with primary carcinoma of the liver, whilst lower levels (less than 10 mg/1) are found in the majority of hepatic disorders, without having any precise significance. The interest of the clinician should therefore be directed not towards the presence or absence of A.F.P. but to its level, which alone is significant. PMID:49879

  15. Different Changes in Protein and Phosphoprotein Levels Result from Serum Starvation of High-Grade Glioma and Adenocarcinoma Cell Lines

    PubMed Central

    Levin, Victor A.; Panchabhai, Sonali C.; Shen, Li; Kornblau, Steven M.; Qiu, Yihua; Baggerly, Keith A.

    2012-01-01

    Tumor cells undergoing serum starvation in vitro partially mimic metabolically stressed cells trying to adjust to a changed environment in vivo by inducing signal transduction and gene expression so that the tumor continues to grow. Our hypothesis is that the changes in protein and phosphoprotein levels after serum starvation may reflect the adapted phenotype of the tumor, which could be targeted for therapy. We used reverse-phase protein microarrays to interrogate five high-grade glioma cell lines and seven adenocarcinoma cell lines for differences in the level of 81 proteins and 25 phosphoproteins. All cell lines were studied in the well-fed condition of growth with 10% FBS and the starved condition of 0.5% FBS. Protein expression levels were normalized to ?-actin and trichotomized as increased (+1, upper 75th quartile), decreased (?1, lowest 25th quartile), or unchanged (0, others) to focus on the patterns of the biggest (and hopefully most robust) changes in protein and phosphoprotein levels. We examined these trichotomized values to better understand Starved-Fed differences among the cell lines and thereby gain better/clearer insight into the effects of serum starvation on potential cellular responses. In general, the expression of proteins and phosphoproteins 24 h after FBS starvation increased more often in glioma lines than in adenocarcinoma lines, which appeared to have fewer increased protein scores and more decreased scores. Many of the proteins increased in gliomas were downstream targets of the PTEN-PI-3 kinase-AKT, EGFR-MAPK-Stat, and transcription activator-polyamine signaling pathways. In adenocarcinomas, the expression of proteins and phosphoproteins generally increased in apoptosis pathways, while there were minor fluctuations in the other pathways above. Contrawise, gliomas become resistant to apoptosis after 24 h of serum starvation and upregulate transcription activators and polyamines more so than adenocarciomas. PMID:19894763

  16. Diagnostic utility and limitations of glutamine synthetase and serum amyloid-associated protein immunohistochemistry in the distinction of focal nodular hyperplasia and inflammatory hepatocellular adenoma.

    PubMed

    Joseph, Nancy M; Ferrell, Linda D; Jain, Dhanpat; Torbenson, Michael S; Wu, Tsung-Teh; Yeh, Matthew M; Kakar, Sanjay

    2014-01-01

    Inflammatory hepatocellular adenoma can show overlapping histological features with focal nodular hyperplasia, including inflammation, fibrous stroma, and ductular reaction. Expression of serum amyloid-associated protein in inflammatory hepatocellular adenoma and map-like pattern of glutamine synthetase in focal nodular hyperplasia can be helpful in this distinction, but the pitfalls and limitations of these markers have not been established. Morphology and immunohistochemistry were analyzed in 54 inflammatory hepatocellular adenomas, 40 focal nodular hyperplasia, and 3 indeterminate lesions. Morphological analysis demonstrated that nodularity, fibrous stroma, dystrophic blood vessels, and ductular reaction were more common in focal nodular hyperplasia, while telangiectasia, hemorrhage, and steatosis were more common in inflammatory hepatocellular adenoma, but there was frequent overlap of morphological features. The majority of inflammatory hepatocellular adenomas demonstrated perivascular and/or patchy glutamine synthetase staining (73.6%), while the remaining cases had diffuse (7.5%), negative (3.8%), or patchy pattern of staining (15%) that showed subtle differences from the classic map-like staining pattern and was designated as pseudo map-like staining. Positive staining for serum amyloid-associated protein was seen in the majority of inflammatory hepatocellular adenomas (92.6%) and in the minority of focal nodular hyperplasia (17.5%). The glutamine synthetase staining pattern was map-like in 90% of focal nodular hyperplasia cases, with the remaining 10% of cases showing pseudo map-like staining. Three cases were labeled as indeterminate and showed focal nodular hyperplasia-like morphology but lacked map-like glutamine synthetase staining pattern; these cases demonstrated a patchy pseudo map-like glutamine synthetase pattern along with the expression of serum amyloid-associated protein. Our results highlight the diagnostic errors that can be caused by variant patterns of staining with glutamine synthetase and serum amyloid-associated protein in inflammatory hepatocellular adenoma and focal nodular hyperplasia. PMID:23807780

  17. Grafting of bovine serum albumin proteins on plasma-modified polymers for potential application in tissue engineering

    NASA Astrophysics Data System (ADS)

    Kasálková, Nikola Slepi?ková; Slepi?ka, Petr; Kolská, Zde?ka; Hoda?ová, Petra; Ku?ková, Št?pánka; Švor?ík, Václav

    2014-04-01

    In this work, an influence of bovine serum albumin proteins grafting on the surface properties of plasma-treated polyethylene and poly- l-lactic acid was studied. The interaction of the vascular smooth muscle cells with the modified polymer surface was determined. The surface properties were characterized by X-ray photoelectron spectroscopy, atomic force microscopy, nano-LC-ESI-Q-TOF mass spectrometry, electrokinetic analysis, and goniometry. One of the motivations for this work is the idea that by the interaction of the cell with substrate surface, the proteins will form an interlayer between the cell and the substrate. It was proven that when interacting with the plasma-treated high-density polyethylene and poly- l-lactic acid, the bovine serum albumin protein is grafted on the polymer surface. Since the proteins are bonded to the substrate surface, they can stimulate cell adhesion and proliferation.

  18. Soy proteins and isoflavones reduce interleukin-6 but not serum lipids in older women: a randomized controlled trial?,??

    PubMed Central

    Mangano, Kelsey M.; Hutchins-Wiese, Heather L.; Kenny, Anne M.; Walsh, Stephen J.; Abourizk, Robin H.; Bruno, Richard S.; Lipcius, Rosanne; Fall, Pamela; Kleppinger, Alison; Kenyon-Pesce, Lisa; Prestwood, Karen M.; Kerstetter, Jane E.

    2015-01-01

    Soy foods contain several components, notably, isoflavones and amino acids, that may improve cardiovascular health. We evaluated the long-term effect of soy protein and/or soy isoflavones supplementation on serum lipids and inflammatory markers using a 1-year randomized, double-blind, placebo-control, clinical trial in 131 healthy ambulatory women older than 60 years. We hypothesized that soy protein, in combination with isoflavones, would have the largest positive effect on coronary heart disease risk factors (serum lipids and inflammatory markers) compared with either intervention alone and that, within groups receiving isoflavones, equol producers would have more positive effects on coronary heart disease risk factors than nonequol producers. After a 1-month baseline period, participants were randomized into 1 of 4 intervention groups: soy protein (18 g/d) and isoflavone tablets (105 mg/d isoflavone aglycone equivalents), soy protein and placebo tablets, control protein and isoflavone tablets, or control protein and placebo tablets. T Tests were used to assess differences between equol and nonequol producers. Ninety-seven women completed the trial. Consumption of protein powder and isoflavone tablets did not differ among groups, and compliance with study powder and tablets was 79% and 90%, respectively. After 1 year, in the entire population, there were either no or little effects on serum lipids and inflammatory markers, regardless of treatment group. Equol producers, when analyzed separately, had significant improvements in total cholesterol/high-density lipoprotein and low-density lipoprotein/high-density lipoprotein ratios (?5.9%, P = .02; ?7.2%, P = .04 respectively). Soy protein and isoflavone (either alone or together) did not impact serum lipids or inflammatory markers. Therefore, they should not be considered an effective intervention to prevent cardiovascular disease because of lipid modification in healthy late postmenopausal women lacking the ability to produce equol. PMID:24267042

  19. QSAR modeling of beta-lactam binding to human serum proteins.

    PubMed

    Hall, L Mark; Hall, Lowell H; Kier, Lemont B

    2003-01-01

    The binding of beta-lactams to human serum proteins was modeled with topological descriptors of molecular structure. Experimental data was the concentration of protein-bound drug expressed as a percent of the total plasma concentration (percent fraction bound, PFB) for 87 penicillins and for 115 beta-lactams. The electrotopological state indices (E-State) and the molecular connectivity chi indices were found to be the basis of two satisfactory models. A data set of 74 penicillins from a drug design series was successfully modeled with statistics: r2 = 0.80, s = 12.1, q2 = 0.76, spress = 13.4. This model was then used to predict protein binding (PFB) for 13 commercial penicillins, resulting in a very good mean absolute error, MAE = 12.7 and correlation coefficient, q2 = 0.84. A group of 28 cephalosporins were combined with the penicillin data to create a dataset of 115 beta-lactams that was successfully modeled: r2 = 0.82, s = 12.7, q2 = 0.78, spress = 13.7. A ten-fold 10% leave-group-out (LGO) cross-validation procedure was implemented, leading to very good statistics: MAE = 10.9, spress = 14.0, q2 (or r2press) = 0.78. The models indicate a combination of general and specific structure features that are important for estimating protein binding in this class of antibiotics. For the beta-lactams, significant factors that increase binding are presence and electron accessibility of aromatic rings, halogens, methylene groups, and =N- atoms. Significant negative influence on binding comes from amine groups and carbonyl oxygen atoms. PMID:13677479

  20. Oxidative damage of bovine serum albumin and other enzyme proteins by iron-chelate complexes.

    PubMed

    Ogino, T; Okada, S

    1995-12-14

    Direct oxidative protein damage by iron-nitrilotriacetate (NTA), as well as physiological iron complexes, iron-citrate and iron-ADP was studied in the presence or absence of H2O2, using bovine serum albumin (BSA), glucose-6-phosphate dehydrogenase (G-6-PD), glutathione reductase (GSSGRase) and catalase as the target proteins. Both Fe(III)NTA+H2O2 and Fe(II)NTA+H2O2 caused marked BSA fragmentation which accompanied the decrease in the intrinsic tryptophan fluorescence and appearance of bityrosine fluorescence. However, Fe(III)citrate+H2O2 showed only slight BSA fragmentation. In the absence of H2O2, Fe(II) NTA but not Fe(III)NTA caused similar but slight BSA fragmentation, which depended on the molecular oxygen. Fe(II)citrate also showed O2-dependent BSA fragmentation to a comparable degree, however, Fe(II)ADP showed no detectable BSA damage. BSA fragmentation by Fe(II)NTA+O2 and by Fe(III)NTA+H2O2 resulted in the appearance of the new alpha-amino groups. Electron spin resonance study using 5,5-dimethyl-1-pyrroline N-oxide (DMPO) as a spin trapping reagent showed DMPO-OH spin adduct, which suggests the presence of hydroxyl radical, in Fe(III)NTA+H2O2, but not in Fe(II)NTA+O2 system. Fe(II)NTA inactivated G-6-PD and GSSGRase in a O2-dependent manner, however, G-6-PD was more susceptible to the damage. This enzyme inactivation also accompanied the protein fragmentation and was not due to simple sulfhydryl oxidation. Catalase was not significantly inactivated nor fragmented by Fe(II)NTA+O2. These findings suggest that the interaction between proteins and iron-chelate complexes is important in iron catalyzed oxidative damage, and that the structure of the chelating agent may determine the target molecules. PMID:8541312

  1. The influence of naturally occurring heterophilic anti-immunoglobulin antibodies on direct measurement of serum proteins using sandwich ELISAs

    Microsoft Academic Search

    Christian Hennig; Lothar Rink; Ursula Fagin; Wolfram J Jabs; Holger Kirchner

    2000-01-01

    Sandwich ELISAs have become a widely used method for the quantitative detection of serum proteins. However, they can be biased by a variety of interfering substances. As reported recently, we observed false-positive levels of interferon (IFN)-? and -? in up to 27% of sera from healthy blood donors using commercial ELISAs. We now demonstrate that two different groups of naturally

  2. Bone Origin of the Serum Complex of Calcium, Phosphate, Fetuin, and Matrix Gla Protein: Biochemical Evidence for

    E-print Network

    Price, Paul A.

    Bone Origin of the Serum Complex of Calcium, Phosphate, Fetuin, and Matrix Gla Protein: Biochemical-remodeling compartment (BRC), a cancellous bone compartment in which the concentrations of calcium and phosphate will cause a sharp rise in the concentrations of calcium and phosphate in the aqueous solution of the BRC

  3. THE INFLUENCE OF SERUM BINDING PROTEINS AND CLEARANCE ON THE COMPARATIVE RECEPTOR BINDING POTENCY OF ENDOCRINE ACTIVE COMPOUNDS

    EPA Science Inventory

    THE INFLUENCE OF SERUM BINDING PROTEINS AND CLEARANCE ON THE COMPARATIVE RECEPTOR BINDING POTENCY OF ENDOCRINE ACTIVE COMPOUNDS. JG Teeguarden1 and HA Barton2. 1ENVIRON International, Ruston LA; 2US EPA, ORD, NHEERL, ETD, Pharmacokinetics Branch, RTP, NC. One measure of th...

  4. A Sensitive and Specific Quantitation Method for Determination of Serum Cardiac Myosin Binding Protein-C by Electrochemiluminescence Immunoassay

    PubMed Central

    Kuster, Diederik W.D.; Barefield, David; Govindan, Suresh; Sadayappan, Sakthivel

    2013-01-01

    Biomarkers are becoming increasingly more important in clinical decision-making, as well as basic science. Diagnosing myocardial infarction (MI) is largely driven by detecting cardiac-specific proteins in patients' serum or plasma as an indicator of myocardial injury. Having recently shown that cardiac myosin binding protein-C (cMyBP-C) is detectable in the serum after MI, we have proposed it as a potential biomarker for MI. Biomarkers are typically detected by traditional sandwich enzyme-linked immunosorbent assays. However, this technique requires a large sample volume, has a small dynamic range, and can measure only one protein at a time. Here we show a multiplex immunoassay in which three cardiac proteins can be measured simultaneously with high sensitivity. Measuring cMyBP-C in uniplex or together with creatine kinase MB and cardiac troponin I showed comparable sensitivity. This technique uses the Meso Scale Discovery (MSD) method of multiplexing in a 96-well plate combined with electrochemiluminescence for detection. While only small sample volumes are required, high sensitivity and a large dynamic range are achieved. Using this technique, we measured cMyBP-C, creatine kinase MB, and cardiac troponin I levels in serum samples from 16 subjects with MI and compared the results with 16 control subjects. We were able to detect all three markers in these samples and found all three biomarkers to be increased after MI. This technique is, therefore, suitable for the sensitive detection of cardiac biomarkers in serum samples. PMID:23963065

  5. Serum leptin and C-reactive protein levels in the physiological spontaneous menstrual cycle in reproductive age women

    Microsoft Academic Search

    D M Wunder; M Yared; N A Bersinger; D Widmer; R Kretschmer; M H Birkhauser

    2006-01-01

    Objective: Only a few studies have investigated variations of different markers for inflammatory processes during the physiological menstrual cycle. The results are conflicting, particularly concerning the correlation between the marker leptin and steroid hormones. The aim of the study was to investigate the inflammatory markers C-reactive protein (CRP) and leptin in the serum of healthy, normally ovulating women and to

  6. Values of markers of early and advanced glycation and lipoxidation in serum proteins of children with diabetes mellitus

    Microsoft Academic Search

    Jakus V; Bauerova K; Michalkova D; Carsky J

    2000-01-01

    Jakus V, Bauerova K, Michalkova D, Carsky J: Values of markers of early and advanced glycation and lipo- xidation in serum proteins of children with diabetes mellitus Bratisl Lek Listy 2000; 101 (9): 484ñ489 Background: Advanced glycation endproducts (AGEs) have been established as one of the major factors responsible for the multi-organ damage seen in diabetes. AGEs and lipoxida- tion

  7. Comparison of serum phospholipase A 2 , polymorphonuclear granulocyte elastase, C-reactive protein and serum amyloid A with the APACHE II score in the prognosis of multiple injured patients

    Microsoft Academic Search

    R. Ensenauer; M. Püttmann; M. Quintel; R. Kattermann; J. Aufenanger

    1994-01-01

    This prospective study of 35 multitraumatized intensive care unit patients requiring mechanical ventilation examined the relative utility of four biochemical parameters with a physiological scoring system for predicting lethal outcome. Levels of serum phospholipase A2 (PLA2), serum amyloid A (SAA), polymorphonuclear granulocyte elastase (PMN elastase), and C-reactive protein (CRP) were determined at short intervals during the patient's hospitalization. The first

  8. Biophysical Insights into How Surfaces, Including Lipid Membranes, Modulate Protein Aggregation Related to Neurodegeneration

    PubMed Central

    Burke, Kathleen A.; Yates, Elizabeth A.; Legleiter, Justin

    2013-01-01

    There are a vast number of neurodegenerative diseases, including Alzheimer’s disease (AD), Parkinson’s disease (PD), and Huntington’s disease (HD), associated with the rearrangement of specific proteins to non-native conformations that promotes aggregation and deposition within tissues and/or cellular compartments. These diseases are commonly classified as protein-misfolding or amyloid diseases. The interaction of these proteins with liquid/surface interfaces is a fundamental phenomenon with potential implications for protein-misfolding diseases. Kinetic and thermodynamic studies indicate that significant conformational changes can be induced in proteins encountering surfaces, which can play a critical role in nucleating aggregate formation or stabilizing specific aggregation states. Surfaces of particular interest in neurodegenerative diseases are cellular and subcellular membranes that are predominately comprised of lipid components. The two-dimensional liquid environments provided by lipid bilayers can profoundly alter protein structure and dynamics by both specific and non-specific interactions. Importantly for misfolding diseases, these bilayer properties can not only modulate protein conformation, but also exert influence on aggregation state. A detailed understanding of the influence of (sub)cellular surfaces in driving protein aggregation and/or stabilizing specific aggregate forms could provide new insights into toxic mechanisms associated with these diseases. Here, we review the influence of surfaces in driving and stabilizing protein aggregation with a specific emphasis on lipid membranes. PMID:23459674

  9. [Quantitative study of serum protein loss into the alimentary tract in patients with gastric cancer].

    PubMed

    Nakatani, N

    1994-09-01

    Quantitative study of serum protein loss into the alimentary tract from the tumor of gastric cancer using 111Indium-transferrin (111In-Tf) was performed. Gamma counting of 111In-Tf excreted in feces and 111In-Tf scintigram were performed in 24 patients with gastric cancer and 10 controls. Transferrin was labelled by incubating 111MBq (3mCi) of 111In chloride with approximately 13ml of patient plasma in vitro. After intravenous injection of 111In-Tf, an aliquot was weighed and its radioactivity was measured in a gamma-counter. Feces were collected every 24hrs up to 72hrs. Then 111In in the feces was calculated as a percentage of the injection dose. 111In excreted in the feces within 72hrs in patients with gastric cancer was 3.71 +/- 3.87% (mean +/- SD), which was significantly higher than the 0.48 +/- 0.26% in 10 controls. 111In in feces correlated the area of the tumor in the stomach (p < 0.001). In 18 patients positive scan was recognized and was localized the protein loss in the cavity of the stomach within 10 minutes. Positive scan was found to move along the intestine successively. 111In-Tf can be useful in assessing of hypoproteinemia of gastric cancer. PMID:7933636

  10. Animal Model Evaluation of Dairy Goats for Milk, Fat, and Protein Yields with Crossbred Animals Included

    Microsoft Academic Search

    G. R. Wiggans

    1989-01-01

    Genetic evaluation of dairy goats was extended to include evaluation of protein yield and evaluation of Oberhasli and experimental breeds. Diverse genetic background of parents of crossbred ani- mals can be accounted for with an animal model that includes all relationships. The animal model system implemented for dairy goats differed from the one for dairy cattle in that all breeds

  11. Correlation between Serum Levels of Protein-Bound Uremic Toxins in Hemodialysis Patients Measured by LC/MS/MS.

    PubMed

    Itoh, Yoshiharu; Ezawa, Atsuko; Kikuchi, Kaori; Tsuruta, Yoshinari; Niwa, Toshimitsu

    2013-01-01

    Uremic toxins are involved in a variety of symptoms in advanced chronic kidney disease. Especially, the accumulation of protein-bound uremic toxins in the blood of dialysis patients might play an important role in the development of cardiovascular disease. Serum concentration of protein-bound uremic toxins such as indoxyl sulfate, indoxyl glucuronide, indoleacetic acid, p-cresyl sulfate, p-cresyl glucuronide, phenyl sulfate, phenyl glucuronide, phenylacetic acid, phenylacetylglutamine, hippuric acid, 4-ethylphenyl sulfate, and 3-carboxy-4-methyl-5-propyl-2-furanpropionic acid (CMPF) in hemodialysis patients were simultaneously measured by liquid chromatography/tandem mass spectrometry. Serum levels of these protein-bound uremic toxins were increased in hemodialysis patients. Indoxyl sulfate, p-cresyl sulfate, and CMPF could not be removed efficiently by hemodialysis due to their high protein-binding ratios. Serum level of total indoxyl sulfate did not show any significant correlation with total p-cresyl sulfate. However, free indoxyl sulfate correlated with free p-cresyl sulfate, and reduction rate by hemodialysis of indoxyl sulfate correlated with that of p-cresyl sulfate. Serum levels of total and free indoxyl sulfate showed significantly positive correlation with those of indoxyl glucuronide, phenyl sulfate, and phenyl glucuronide. Serum levels of total and free p-cresyl sulfate showed significantly positive correlation with those of p-cresyl glucuronide, phenylacetylglutamine, and phenylacetic acid. Indoxyl sulfate and indoxyl glucuronide are produced from indole which is produced in the intestine from tryptophan by intestinal bacteria. p-Cresyl sulfate and p-cresyl glucuronide are produced from p-cresol which is produced in the intestine from tyrosine by intestinal bacteria. Thus, intestinal bacteria play an important role in the metabolism of protein-bound uremic toxins. PMID:24349936

  12. Acute phase proteins in serum and milk from dairy cows with clinical mastitis

    Microsoft Academic Search

    P. D. Eckersall; F. J. Young; C. McComb; C. J. Hogarth; S. Safi; J. L. Fitzpatrick; A. M. Nolan; A Weber; T. McDonald

    2001-01-01

    The serum concentrations of haptoglobin, serum amyloid A and ? acid glycoprotein were determined in serum collected from healthy dairy cows and cows with clinical mastitis, graded as mild (clots in milk) or moderate (clots in milk and visible signs of inflammation in the mammary gland\\/s) to assess their relative diagnostic value in detecting the disease. The concentrations of haptoglobin

  13. Correlation of Bone Morphogenetic Protein-2 Levels in Serum and Synovial Fluid with Disease Severity of Knee Osteoarthritis

    PubMed Central

    Liu, Yan; Hou, Ruizhi; Yin, Ruofeng; Yin, Weitian

    2015-01-01

    Background This study aimed to investigate the bone morphogenetic protein-2 (BMP-2) levels in serum and synovial fluid (SF) of patients with primary knee osteoarthritis (OA) and to exam its correlation with radiographic and symptomatic severity of the disease. Material/Methods A total of 37 knee OA patients and 20 healthy controls were enrolled in this study. Knee OA radiographic grading was performed according to the Kellgren-Lawrence (KL) grading system by evaluating X-ray changes observed in anteroposterior knee radiography. Symptomatic severity of the disease was evaluated according to the Western Ontario McMaster University Osteoarthritis Index (WOMAC) scores. BMP-2 levels in serum and SF were determined using enzyme-linked immunosorbent assay. Results Serum BMP-2 level in patients with knee OA was higher than that in healthy controls. Knee OA patients with KL grade 4 showed significantly elevated BMP-2 levels in the serum and SF compared with those with KL grade 2 and 3. Knee OA patients with KL grade 3 had significant higher SF levels of BMP-2 than those with KL grade 2. BMP-2 levels in the serum and SF of knee OA patients were both positively correlated with KL grades and WOMAC scores. Conclusions BMP2 levels in serum and SF were closely related to the radiographic and symptomatic severity of knee OA and may serve as an alternative biochemical parameter to determine disease severity of primary knee OA. PMID:25644704

  14. Identification of serum sirtuins as novel noninvasive protein markers for frailty

    PubMed Central

    Kumar, Rahul; Mohan, Navinath; Upadhyay, Ashish Datt; Singh, Amrendra Pratap; Sahu, Vishal; Dwivedi, Sadanand; Dey, Aparajit B; Dey, Sharmistha

    2014-01-01

    Frailty has emerged as a major health issue among older patients. A consensus on definition and diagnosis is yet to be achieved. Various biochemical abnormalities have been reported in frailty. Activation of sirtuins, a conserved family of NAD-dependent proteins, is one of the many mimics of calorie restriction which improves lifespan and health in experimental animals. In this cross-sectional study, we assessed the circulating sirtuin levels in 119 (59.5%) nonfrail and 81 (40.5%) frail individuals, diagnosed by Fried's criteria. Serum SIRT1, SIRT2, and SIRT3 were estimated by surface plasmon resonance (SPR) and Western blot. Serum sirtuins level in mean+SD; SIRT1 (nonfrail –4.67 ± 0.48 ng/?L; frail – 3.72 ± 0.48 ng/?L; P < 0.0001), SIRT2 (nonfrail – 15.18 ± 2.94 ng/?L; frail – 14.19 ± 2.66 ng/?L; P = 0.016), and SIRT3 (nonfrail-7.72 ± 1.84 ng/?L; frail – 6.12 ± 0.97 ng/?L; P < 0.0001) levels were significantly lower among frail patients compared with the nonfrail. In multivariable regression analysis, lower sirtuins level were significantly associated with frailty after adjusting age, gender, diabetes mellitus, hypertension, cognitive status (Mini Mental State Examination scores) and number of comorbidities. For detecting the optimum diagnostic cutoff value a ROC analysis was carried out. The area under curve for SIRT1 was 0.9037 (cutoff – 4.29 ng/?L; sensitivity – 81.48%; specificity – 79.83%) and SIRT3 was 0.7988 (cutoff – 6.61 ng/?L; sensitivity – 70.37%; specificity – 70.59%). This study shows that lower circulating SIRT1 and SIRT3 levels can be distinctive marker of frailty. PMID:25100619

  15. Multifunctionality of Acidulated Serum Albumin on Inhibiting Zn(2+)-Mediated Amyloid ?-Protein Fibrillogenesis and Cytotoxicity.

    PubMed

    Xie, Baolong; Dong, Xiaoyan; Wang, Yongjian; Sun, Yan

    2015-07-01

    Fibrillogenesis of amyloid ?-proteins (A?) mediated by transition-metal ions such as Zn(2+) in neuronal cells plays a causative role in Alzheimer's disease. Hence, it is highly desired to design multifunctional agents capable of inhibiting A? aggregation and modulating metal-A? species. In this study, we fabricated acidulated human serum albumin (A-HSA) as a multifunctional agent for binding Zn(2+) and modulating Zn(2+)-mediated A? fibrillogenesis and cytotoxicity. On average, 19.5 diglycolic anhydrides were modified onto the surface of human serum albumin (HSA). It was confirmed that A-HSA kept the stability and biocompatibility of native HSA. Moreover, it could inhibit A?42 fibrillogenesis and change the pathway of Zn(2+)-mediated A?42 aggregation, as demonstrated by extensive biophysical assays. In addition, upon incubation with A-HSA, the cytotoxicity presented by Zn(2+)-A?42 aggregates was significantly mitigated in living cells. The results showed that A-HSA had much stronger inhibitory effect on Zn(2+)-mediated A?42 fibrillogenesis and cytotoxicity than equimolar HSA. Isothermal titration calorimetry and stopped-flow fluorescence measurements were then performed to investigate the working mechanism of A-HSA. The studies showed that the A-HSA surface, with more negative charges, not only had stronger affinity for Zn(2+) but also might decrease the binding affinity of A?42 for Zn(2+). Moreover, hydrophobic binding and electrostatic repulsion could work simultaneously on the bound A?42 on the A-HSA surface. As a result, A?42 conformations could be stretched, which avoided the formation of toxic Zn(2+)-A?42 aggregates. The research thus revealed that A-HSA is a multifunctional agent capable of altering the pathway of Zn(2+)-mediated A?42 aggregation and greatly mitigating the amyloid cytotoxicity. PMID:26070334

  16. Transcortin and vitamin D-binding protein levels in mouse serum.

    PubMed

    Faict, D; De Moor, P; Bouillon, R; Heyns, W; Heiniger, H J; Corrow, D; Lesaffre, E

    1986-05-01

    The influence of age, sex and strain on the serum concentration of transcortin (corticosteroid-binding globulin) and vitamin D-binding protein (DBP) in mice was investigated. The effect of age was studied in two strains, C57BL/6JPfd and BALB/cmHeAPfd. The concentration of transcortin and DBP increased with age. In young animals the concentration of each protein showed a significant strain difference, which disappeared in older mice for DBP, but not for transcortin. In 7-day-old animals, no sex difference was observed for either protein, but in older animals a clear sex difference was found for transcortin. Adult males tended to have somewhat higher levels of DBP than adult females, but this difference was significant only on day 70. The variation in transcortin and DBP levels was further investigated in a large number of mouse strains. The DBP concentration did not markedly vary among strains (5.98-9.65 mumol/l in males and 5.08-8.85 mumol/l in females). Transcortin, however, showed marked strain variations, ranging from 0.72 to 2.06 mumol/l in males and from 1.02 to 4.55 mumol/l in females and there was a significant correlation (r = 0.66, n = 26, P less than 0.001) between the mean transcortin levels in males and females of different strains. Interstrain variation was much higher than intrastrain variation or variation among related strains, suggesting that the transcortin concentration is largely controlled by genetically determined factors. There was a significant correlation (r = 0.82, n = 9, P less than 0.01) between the mean corticosterone and transcortin concentrations (measured at 21.00 h).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:3711757

  17. Insight into the Interaction of Graphene Oxide with Serum Proteins and the Impact of the Degree of Reduction and Concentration.

    PubMed

    Wei, Xue-Qin; Hao, Li-Ying; Shao, Xiao-Ru; Zhang, Quan; Jia, Xiao-Qin; Zhang, Zhi-Rong; Lin, Yun-Feng; Peng, Qiang

    2015-06-24

    As novel applied nanomaterials, both graphene oxide (GO) and its reduced form (rGO) have attracted global attention, because of their excellent properties. However, the lack of comprehensive understanding of their interactions with biomacromolecules highly limits their biomedical applications. This work aims to initiate a systematic study on the property changes of GO/rGO upon interaction with serum proteins and on how their degree of reduction and exposure concentration affect this interaction, as well as to analyze the possible biomedical impacts of the interaction. We found that the adsorption of proteins on GO/rGO occurred spontaneously and rapidly, leading to significant changes in size, zeta potential, and morphology. Compared to rGO, GO showed a higher ability in quenching intrinsic fluorescence of serum proteins in a concentration-dependent manner. The protein adsorption efficiency and the types of associated proteins varied, depending on the degree of reduction and concentration of graphene. Our findings indicate the importance of evaluating the potential protein adsorption before making use of GO/rGO in drug delivery, because the changed physicochemical properties after protein adsorption will have significant impacts on safety and effectiveness of these delivery systems. On the other hand, this interaction can also be used for the separation, purification, or delivery of certain proteins. PMID:26029973

  18. The Effect of Animal Protein and Vegetable Protein Diets Having the Same Fat Content on the Serum Lipid Levels of Young Women1

    Microsoft Academic Search

    GEORGIANNA R. WALKER; ELLEN H. MORSE; VIRGINIA A. OVERLEY

    In nutritional studies of vegetarians, Hardinge and Stare ('54) found lower serum cholesterol levels among those who were pure vegetarians than among lacto- ovovegetarians. Both groups had signifi cantly lower levels than nonvegetarians. Olson et al. ('58), by reducing the protein content of the diet from 100 gm (17% of total calories), mainly from animal sources, to 25 gm (4%

  19. Serum bone Gla protein and carboxyterminal cross-linked telopeptide of type I collagen in patients with Cushing's syndrome.

    PubMed Central

    Sartorio, A.; Conti, A.; Ferrario, S.; Passini, E.; Re, T.; Ambrosi, B.

    1996-01-01

    Serum bone Gla protein, a marker of bone formation, and carboxyterminal cross-linked telopeptide of type I collagen levels, an index of bone resorption, were evaluated in eight patients with active Cushing's syndrome and in four with 'preclinical' Cushing's syndrome, before and after surgery. In basal conditions, serum bone Gla protein levels were significantly lower (p < 0.0001) in patients with active Cushing's syndrome (1.0 +/- 0.35 ng/ml) than in controls (5.4 +/- 0.15 ng/ml); two out of four patients with the 'preclinical' form had reduced bone Gla protein levels, while in the other two cases levels were in the normal range. Serum levels of carboxyterminal cross-linked telopeptide of type I collagen (3.0 +/- 0.4 ng/ml), although slightly reduced, were similar to those recorded in controls (4.1 +/- 0.3 ng ml), both in patients with active and with preclinical Cushing's syndrome. After surgery serum levels of both marker proteins significantly increased in seven out of eight patients with active Cushing's syndrome; in one patient, who was not cured after surgery, bone Gla proteins levels remained lower than in normals, while levels of carboxyterminal cross-linked telopeptide of type I collagen had a transient increase after six months. In the two patients with a 'preclinical' Cushing's syndrome who underwent surgery, a significant rise of the levels of both marker proteins was observed, similar to that observed in patients with active Cushing's syndrome. It was concluded that serial determinations of these new markers of bone formation and resorption may be usefully employed to follow-up the clinical course of Cushing's syndrome and provide information on the rate of bone turnover in response to medical and/or surgical therapies. Moreover, the evaluation of these markers in preclinical states of Cushing's syndrome might suggest the need for surgery. PMID:8935602

  20. Flavorase, a novel non-haemorrhagic metalloproteinase in Protobothrops flavoviridis venom, is a target molecule of small serum protein-3.

    PubMed

    Shioi, Narumi; Nishijima, Ayumi; Terada, Shigeyuki

    2015-07-01

    Some venomous snakes possess anti-toxic proteins in their sera that may play a role in neutralizing the haemorrhagic factors or toxins in their own venom. Five small serum proteins (SSP-1-SSP-5) were isolated from the serum of Japanese viper (Protobothrops flavoviridis), and were found to act as self-defence proteins against the viper's own toxic components. However, the physiological function of SSP-3 has not been completely elucidated. Affinity chromatography of the venom on an SSP-3-immobilized column identified a novel 55-kDa protein as the target molecule of SSP-3. Sequences of internal fragments of this SSP-3-binding protein showed high homology to those of metalloproteinases from the P. flavoviridis venom. The cDNA sequence revealed that this protein, termed flavorase, is a P-III class metalloproteinase consisting of 423 amino acid residues. The purified protein did not show haemorrhagic and cytotoxic activity. Biacore measurements revealed that SSP-3 was bound to flavorase with a dissociation constant of 6.4 × 10(-9) M. SSP-3 non-competitively inhibited the peptidase activity of flavorase with an inhibition constant of 6.6 × 10(-9) M. PMID:25681613

  1. Microfiltration: Effect of channel diameter on limiting flux and serum protein removal.

    PubMed

    Hurt, E E; Adams, M C; Barbano, D M

    2015-06-01

    Our objective was to determine the limiting flux and serum protein (SP) removal at 8, 9 and 10% true protein (TP) in the retentate recirculation loop using 0.1-µm ceramic graded permeability (GP) microfiltration (MF) membranes with 3mm channel diameters (CD). An additional objective was to compare the limiting flux and SP removal between 0.1-µm ceramic GP membranes with 3mm CD and previous research using 4-mm CD membranes. The MF system was operated at 50°C, using a diluted milk protein concentrate with 85% protein on a total solids basis (MPC85) as the MF feed. The limiting flux for the MF of diluted MPC85 was determined at 8, 9, and 10% TP concentration in the recirculation loop. The experiment using the 3-mm CD membranes was replicated 3 times for a total of 9 runs. On the morning of each run MPC85 was diluted with reverse osmosis water to a MF feed TP concentration of 5.4%. In all runs the starting flux was 55kg/m(2) per hour, the flux was then increased in steps until the limiting flux was reached. For the 3-mm CD membranes, the limiting flux was 128±0.3, 109±4, and 97±0.5kg/m(2) per hour at recirculation loop TP concentrations of 8.1±0.07, 9.2±0.04, and 10.2±0.03%, respectively. For the 3-mm CD membranes, increasing the flux from the starting to the limiting flux decreased the SP removal factor from 0.72±0.02 to 0.67±0.01; however, no difference in SP removal factor among the target recirculation loop TP concentrations was detected. The limiting flux at each recirculation loop target TP concentration was lower for the 3- compared with the 4-mm CD membranes. The differences in limiting fluxes between the 3- and 4-mm CD membranes were explained in part by the difference in cross-flow velocity (5.5±0.03 and 7.0±0.03 m/s for the 3- and 4-mm CD membranes, respectively). The SP removal factor was also lower for the 3- compared with the 4-mm CD membranes, indicating that more membrane fouling may have occurred in the 3- versus 4-mm CD membranes. PMID:25892692

  2. Cyr61 Is Regulated by cAMP-Dependent Protein Kinase With Serum Levels Correlating With Prostate Cancer Aggressiveness

    PubMed Central

    Terada, Naoki; Shiraishi, Takumi; Zeng, Yu; Mooney, Steven M.; Yeater, David B.; Mangold, Leslie A.; Partin, Alan W.; Kulkarni, Prakash; Getzenberg, Robert H.

    2012-01-01

    BACKGROUND Cysteine-rich angiogenic inducer 61 (Cyr61) is an extracellular matrix protein involved in the transduction of growth factor and hormone signaling. Previously, we demonstrated that Cyr61 was highly expressed in prostate cancer (PCa) but that the expression levels were associated with a lower risk of PCa recurrence. In the present study, we demonstrate that serum Cyr61 is a potential biomarker that correlates with PCa aggressiveness. Furthermore, we also explore the potential mechanism underlying the changes in Cyr61 expression during PCa progression. METHODS Cyr61 concentrations in the medium from PCa cell lines and in serum samples obtained from PCa patients were measured by sandwich ELISA. Serum Cyr61 levels were correlated with disease characteristics and the association between Cyr61 expression changes by several types of stimulation or stress and cAMP/cAMP-dependent protein kinase (PKA) pathway were examined. RESULTS There was a positive correlation between Cyr61 levels in cell supernatants and mRNA expression in these cell lines. Serum Cyr61 levels were significantly higher in non-organ-confined PCa patients (116.3 ± 140.2 ng/ml) than in organ-confined PCa patients (79.7 ± 56.1 ng/ml) (P = 0.031). Cyr61 expression was up-regulated in response to both lysophosphatidic acid and androgen treatments which promoted PCa cell invasion. Serum starvation and phosphoinositide-3-kinase inhibition also resulted in Cyr61 up-regulation; however, they suppressed cell proliferation. Cyr61 up-regulation was correlated with an increase in cAMP and suppressed by PKA inhibition. CONCLUSIONS These findings suggest that Cyr61 expression in PCa is regulated by the cAMP/PKA pathway and that circulating Cyr61 levels are a potential serum-based biomarker for characterizing PCa. PMID:22025384

  3. Direct detection of C-reactive proteins in human serum using nanoparticle-enhanced surface plasmon resonance biosensing

    NASA Astrophysics Data System (ADS)

    Lin, H.-Y.; Tsang, K. Y.; Hu, W. P.; Hsu, H.-Y.; Chiou, A.; Chang, G.-L.; Chen, S.-J.

    2006-08-01

    C-reactive protein (CRP) produced by the liver is one of the most characteristic acute-phase proteins. It has been suggested that the level of CRP in human serum may be a significant tool of detecting risks of developing cardiovascular disease and atherosclerosis. Here we propose an advanced plasmonic surface plasmon resonance (SPR) bioassay with Au nanoparticles embedded in the dielectric film that demonstrates a 10X improvement in resolution compared to the conventional SPR biosensor. The co-sputtered film was modified with (3-Aminopropyl)triethoxysilane to sequentially immobilize protein G, monoclonal anti-CRP antibody (C8), and human serum albumins (HSA). After blocked by ethanolamine, the sensor was used to detect CRP. Using this extremely sensitive biochip, the lowest reliable concentration of CRP without any exterior labeling is simplified to human physiological level. The novel assay has the latent capability of not only eliminating the disturbances coming from serum proteins resulting in false signals, but is also able to be applied in rapid and label-free clinical detections of CRP with large improved sensitivity.

  4. Influence of transportation on serum concentrations of acute phase proteins in horse.

    PubMed

    Casella, S; Fazio, F; Giannetto, C; Giudice, E; Piccione, G

    2012-10-01

    The modifications of Haptoglobin (Hp), Serum Amyloid A (SAA), Fibrinogen (Fbg) and White Blood Cells (WBCs) were evaluated in 15 Saddle Italian horses. Ten horses were transported covering a distance of about 320 km within 4 h with an average speed of 80 km/h (experimental group) and five horses were not subject to transportation (control group). Blood was collected via jugular venipuncture before the transportation (T0), immediately after the transportation (T1), 12 (T12), 24 (T24) and 48 (T48)hours after the transportation in experimental group and at the same time point in control group. For each parameter statistical analysis of different groups and sampling time was performed using a two-way analysis of covariance, with the data before the transportation (T0) as the covariate, by the GLM procedure of SAS. For all parameters the interaction (Group × Time) was tested and it was resulted no significant. The application of statistical analysis showed significant differences between the control group and horses subjected to transportation (P<0.01), and the influence of sampling time (P<0.05) on Hp, SAA and WBCs. These modifications appeared to be innovative showing that equine Hp, generally considered as moderate acute phase protein, increases more rapidly than the SAA after transportation-induced stress. PMID:22296939

  5. Chemical structure of the arabinogalactan protein from gum ghatti and its interaction with bovine serum albumin.

    PubMed

    Ghosh, Kanika; Ray, Sayani; Ghosh, Debjani; Ray, Bimalendu

    2015-03-01

    Exudate gums, because of their beneficial properties, have been significant items of international trade in various industries for centuries. This manuscript sets out to gain insight into the fine structural details of an arabinogalactan protein (AGP) of gum ghatti (Anogeissus latifolia gum). The presence of a highly branched 554 kDa AGP having 1,6-linked Galp, 1,2-linked Manp, 1,3-linked Araf and 1,4-linked GlcpA main chain, substituted at O-4,6 of 1,2-linked Manp, and O-3/O-3,4 of 1,6-linked Galp residues by Araf, Arap and Galp units was revealed by chemical, chromatographic, ESMS, and NMR analyses. In particular, ESMS analysis of per acetylated oligomeric fragments derived from AGP by Smith degradation followed by acetylation was described as a commanding tool for providing critical structural information on a spectrum of glycerol tagged oligosaccharides. In addition, formation of an electrostatically driven complex between the isolated AGP and bovine serum albumin resulting in changes in the microenvironment around the tryptophan residues of BSA was established. A moderate radical scavenging activity comparable with those of standard antioxidants was observed from the AGP fraction (?94% at 1 mg/mL) that could be valuable in foods or pharmaceutical products as alternatives to synthetic antioxidants. PMID:25498648

  6. Recombinant Protein Production by the Baculovirus-Insect Cell System in Basal Media Without Serum Supplementation

    Microsoft Academic Search

    Norikatsu Nishikawa; Hideki Yamaji; Hideki Fukuda

    2003-01-01

    The production of ?-galactosidase by Sf9 cells infected with recombinant Autographa californica nucleopolyhedrovirus (AcNPV) was investigated in shake-flask culture using two serum-free basal media: Grace's medium and\\u000a TNM-FH (Grace's medium supplemented with lactalbumin hydrolysate and yeast extract). At the time of infection, cells grown\\u000a in serum-supplemented TNM-FH were transferred into fresh basal media without adaptation. The absence of serum depressed

  7. Correlation between Serum Levels of High Mobility Group Box-1 Protein and Pancreatitis: A Meta-Analysis

    PubMed Central

    Lin, Yan; Lin, Lian-Jie; Jin, Yu; Cao, Yong; Zhang, Ying; Zheng, Chang-Qing; Liu, Jia-Li; Yang, Sheng-Li

    2015-01-01

    Background. Aberrant expression of high mobility group box-1 protein (HMGB1) contributes to the progression of various inflammatory diseases. This meta-analysis focused on the clinical significance of serum HMGB1 levels in pancreatitis patients, with the goal of building a novel diagnostic score model. Method. We conducted a meta-analysis by searching in the PubMed, Embase, Web of Science, Cochrane Library, CISCOM, CINAHL, Google Scholar, China BioMedicine (CBM), and China National Knowledge Infrastructure (CNKI) databases without any language restrictions. Studies were pooled and standard mean difference (SMD) and its corresponding 95% confidence intervals (95% CIs) were calculated. Version 12.0 STATA software was used for statistical analysis. Results. We performed a final analysis of 841 subjects from 12 clinical case-control studies. The meta-analysis results showed a positive association between serum HMGB1 levels and the progression of pancreatitis. In the subgroup analysis by country, high serum level of HMGB1 may be related to pancreatitis progression in China, Korea, Hungary, and Japan populations (all P < 0.05). Conclusion. The present meta-analysis indicated that serum HMGB1 level was statistically elevated in patients with pancreatitis, and thus serum levels of HMGB1 could be determined to be a useful biomarker for pancreatitis patients. PMID:25695079

  8. Correlation of salivary and serum IgG, IgA levels with total protein in oral submucous fibrosis.

    PubMed

    Patidar, Kalpana A; Parwani, Rajkumar N; Wanjari, Sangeeta Panjab

    2011-03-01

    Oral submucous fibrosis (OSMF) is a disabling, potentially malignant condition of the oral cavity. The aetiology of OSMF is multifactorial but remains obscure. Although arecanut is considered to be the most important causative agent, responses observed in individuals using arecanut vary in relation to quantity and duration. It is considered that an immunological process is responsible for the pathogenesis of disease. We correlated salivary immunoglobulin A (IgA), salivary immunoglobulin G (IgG) and serum immunoglobulin A (IgA), serum immunoglobulin G (IgG), levels by turbidometric immunoassay. We estimated the levels of total serum protein (TSP) and haemoglobin (Hb) to determine the role of nutritional deficiency. The study population comprised 30 cases of OSMF and 10 controls. Five milliliters of blood and 2 ml of saliva were collected. Quantitative analysis of serum and salivary IgG, IgA was done by turbidometric immunoassay. TSP and Hb were estimated by Biuret and cyanmethaemoglobin methods, respectively. All patients showed significant (P < 0.01) increase in serum and salivary IgG, IgA levels as compared to controls. TSP patients showed significant (P < 0.01) decrease as compared to controls. Results of Hb in patients were not significant. The estimation of immunoglobulin levels is important to support the concept of autoimmune basis. Estimation of TSP and Hb suggests that nutrition has a definite role in OSMF. PMID:21467820

  9. Efficacy of wheat germ lectin-precipitated alkaline phosphatase in serum as an estimator of bone mineralization rate: Comparison to serum total alkaline phosphatase and serum bone Gla-protein

    Microsoft Academic Search

    Kim Brixen; Henning K. Nielsen; Erik F. Eriksen; Peder Charles; Leif Mosekilde

    1989-01-01

    Summary  Serum levels of total alkaline phosphatase activity (S-T-AP), wheat germ lectin-precipitated alkaline phosphatase activity\\u000a (S-L-AP), and bone Gla-protein immunoreactivity (S-BGP) were measured in 26 patients (23 females and 3 males) aged 35–73 years\\u000a (mean 59 years) with primary hyperparathyroidism (n=7), hyperthyroidism (n=9), and hypothyroidism (n=10) in whom the bone\\u000a mineralization rate (m) was determined by47Ca-kinetics (continuously expanding calcium pool model).

  10. MATERNAL SERUM INTERLEUKIN-6, C-REACTIVE PROTEIN, AND MATRIX METALLOPROTEINASE-9 CONCENTRATIONS AS RISK FACTORS FOR PRETERM BIRTH < 32 WEEKS AND ADVERSE NEONATAL OUTCOMES

    PubMed Central

    Sorokin, Yoram; Romero, Roberto; Mele, Lisa; Wapner, Ronald J.; Iams, Jay D.; Dudley, Donald J.; Spong, Catherine Y.; Peaceman, Alan M.; Leveno, Kenneth J.; Harper, Margaret; Caritis, Steve N.; Miodovnik, Menachem; Mercer, Brian M.; Thorp, John M.; O’Sullivan, Mary Jo; Ramin, Susan M.; Carpenter, Marshall W.; Rouse, Dwight J.; Sibai, Baha

    2010-01-01

    OBJECTIVE Elevated concentrations of Interleukin-6 (IL-6), C-reactive protein (CRP), and Matrix Metalloproteinase-9 (MMP-9) in fetal and neonatal compartments have been associated with an increased risk for preterm birth (PTB) and/or neonatal morbidity. The purpose of this study was to determine if the maternal serum concentration of IL-6, CRP, and MMP-9 in women at risk for preterm birth (PTB) who are not in labor, and have intact membranes, are associated with an increased risk for preterm birth < 32 weeks and/or neonatal morbidity. STUDY DESIGN Maternal serum samples collected from 475 patients enrolled in a multicenter randomized controlled trial of single versus weekly corticosteroids for women at increased risk for preterm delivery were assayed. Serum was collected at randomization (24-32 weeks gestation). Maternal serum concentrations of IL-6, CRP, and MMP-9 were subsequently determined using enzyme-linked immunoassays. Multivariate logistic regression analysis was performed to explore the relationship between maternal serum concentrations of IL-6, CRP and MMP-9, and preterm birth < 32 weeks, Respiratory Distress Syndrome (RDS), Chronic Lung Disease (CLD), Intraventricular Hemorrhage (IVH), Necrotizing Enterocolitis (NEC) and Any Sepsis (S). RESULTS Maternal serum concentrations of IL-6 and CRP, but not MMP-9, above the 90th percentile, at the time of randomization, were associated with preterm birth < 32 weeks. In contrast, there was no significant relationship between RDS and NEC and the maternal serum concentration of IL-6, CRP, or MMP-9 (univariate analysis). The development of CLD was associated with a high (above 90th percentile) IL-6 and CRP in maternal serum, even after adjustment for gestational age (GA) at randomization, and treatment group. However, when GA at delivery was added to the model, this finding was non-significant. Neonatal sepsis was more frequent in neonates born to mothers with a high maternal serum concentration of CRP (above >90th percentile). However, there was no significant association after adjustment for GA at randomization, and treatment group. Logistic regression analysis for each analyte indicated that high maternal serum concentrations of IL-6 and CRP, but not MMP-9, were associated with an increased risk of IVH (O.R. 4.60, 95% C.I. 1.86-10.68; O.R. 4.07, 95% C.I. 1.63-9.50), after adjusting for GA at randomization and treatment group. Most babies (25/30) had grade I IVH. When GA at delivery was included, elevated IL-6 remained significantly associated with IVH (O.R. 2.77, 95% C.I. 1.02-7.09). CONCLUSION An elevated maternal serum concentration of IL-6 and CRP are risk factors for preterm birth < 32 weeks and subsequent development of neonatal IVH. An elevated maternal serum IL-6 appears to confer additional risk for IVH even after adjusting for gestational age at delivery. PMID:20195952

  11. A fluorescence-based high throughput assay for the determination of small molecule–human serum albumin protein binding

    PubMed Central

    McCallum, Megan M.; Pawlak, Alan J.; Shadrick, William R.; Simeonov, Anton; Jadhav, Ajit; Yasgar, Adam; Maloney, David J.; Arnold, Leggy A.

    2014-01-01

    Herein, we describe the development of a fluorescence-based high throughput assay to determine the small molecule binding towards human serum albumin (HSA). This innovative competition assay is based on the use of a novel fluorescent small molecule Red Mega 500 with unique spectroscopic and binding properties. The commercially available probe displays a large fluorescence intensity difference between the protein-bound and protein-unbound state. The competition of small molecules for HSA binding in the presence of probe resulted in low fluorescence intensities. The assay was evaluated with the LOPAC small molecule library of 1280 compounds identifying known high protein binders. The small molecule competition of HSA–Red Mega 500 binding was saturable at higher compound concentrations and exhibited IC50 values between 3–24 ?M. The compound affinity towards HSA was confirmed by isothermal titration calorimetry indicating that the new protein binding assay is a valid high throughput assay to determine plasma protein binding. PMID:24390461

  12. Comparing serum responses to acute feedings of an extensively hydrolyzed whey protein concentrate versus a native whey protein concentrate in rats: a metabolomics approach.

    PubMed

    Roberts, Michael D; Cruthirds, Clayton L; Lockwood, Christopher M; Pappan, Kirk; Childs, Thomas E; Company, Joseph M; Brown, Jacob D; Toedebusch, Ryan G; Booth, Frank W

    2014-02-01

    We examined how gavage feeding extensively hydrolyzed whey protein (WPH) versus a native whey protein concentrate (WPC) transiently affected serum biochemical profiles in rodents. Male Wistar rats (250-300 g) were 8 h fasted and subsequently fed isonitrogenous amounts of WPH or WPC, or remained unfed (control). Animals were sacrificed 15 min, 30 min, and 60 min post-gavage for serum extraction, and serum was analyzed using untargeted global metabolic profiling via gas chromatography/mass spectrometry (MS) and liquid chromatography/MS/MS platforms. We detected 333 serum metabolites amongst the experimental and control groups. Both WPH and WPC generally increased amino acids (1.2-2.8-fold), branched-chain amino acids (1.2-1.7-fold), and serum di- and oligo-peptides (1.1-2.7-fold) over the 60 min time course compared with control (q < 0.05). However, WPH increased lysine (false discovery rate using a q-value <0.05) and tended to increase isoleucine and valine 15 min post-feeding (q < 0.10) as well as aspartylleucine 30 min post-feeding compared with WPC (q < 0.05). While both protein sources led to a dramatic increase in free fatty acids compared with control (up to 6-fold increases, q < 0.05), WPH also uniquely resulted in a 30 min post-feeding elevation in free fatty acids compared with WPC (q < 0.05), an effect which may be due to the robust 30 min postprandial increase in epinephrine in the WPH cohort. These data provide a unique postprandial time-course perspective on how WPH versus WPC feedings affect circulating biochemicals and will guide future research comparing these 2 protein sources. PMID:24476471

  13. A chloroplast transgenic approach to hyper-express and purify Human Serum Albumin, a protein highly susceptible to proteolytic degradation

    PubMed Central

    Millán, Alicia Fernández-San; Mingo-Castel, Angel; Miller, Michael; Daniell, Henry

    2012-01-01

    Summary Human Serum Albumin (HSA) accounts for 60% of the total protein in blood serum and it is the most widely used intravenous protein in a number of human therapies. HSA, however, is currently extracted only from blood because of a lack of commercially feasible recombinant expression systems. HSA is highly susceptible to proteolytic degradation in recombinant systems and is expensive to purify. Expression of HSA in transgenic chloroplasts using Shine-Dalgarno sequence (SD), which usually facilitates hyper-expression of transgenes, resulted only in 0.02% HSA in total protein (tp). Modification of HSA regulatory sequences using chloroplast untranslated regions (UTRs) resulted in hyper-expression of HSA (up to 11.1% tp), compensating for excessive proteolytic degradation. This is the highest expression of a pharmaceutical protein in transgenic plants and 500-fold greater than previous reports on HSA expression in transgenic leaves. Electron micrographs of immunogold labelled transgenic chloroplasts revealed HSA inclusion bodies, which provided a simple method for purification from other cellular proteins. HSA inclusion bodies could be readily solubilized to obtain a monomeric form using appropriate reagents. The regulatory elements used in this study should serve as a model system for enhancing expression of foreign proteins that are highly susceptible to proteolytic degradation and provide advantages in purification, when inclusion bodies are formed. PMID:17147744

  14. Fatty acid-binding site environments of serum vitamin D-binding protein and albumin are different

    PubMed Central

    Swamy, Narasimha; Ray, Rahul

    2008-01-01

    Vitamin D-binding protein (DBP) and albumin (ALB) are abundant serum proteins and both possess high-affinity binding for saturated and unsaturated fatty acids. However, certain differences exist. We surmised that in cases where serum albumin level is low, DBP presumably can act as a transporter of fatty acids. To explore this possibility we synthesized several alkylating derivatives of 14C-palmitic acid to probe the fatty acid binding pockets of DBP and ALB. We observed that N-ethyl-5-phenylisooxazolium-3?-sulfonate-ester (WRK ester) of 14C-palmitic acid specifically labeled DBP; but p-nitrophenyl- and N-hydroxysuccinimidyl-esters failed to do so. However, p-nitrophenyl ester of 14C-palmitic acid specifically labeled bovine ALB, indicating that the micro-environment of the fatty acid-binding domains of DBP and ALB may be different; and DBP may not replace ALB as a transporter of fatty acids. PMID:18374965

  15. Is there a relationship between serum S-100? protein and neuropsychologic dysfunction after cardiopulmonary bypass?

    Microsoft Academic Search

    Stephen Westaby; Kjell Saatvedt; Samantha White; Takahiro Katsumata; Willem van Oeveren; Narendra K. Bhatnagar; Stuart Brown; Peter W. Halligan

    2000-01-01

    Objectives: Over the past decade, the glial protein S-100? has been used to detect cerebral injury in a number of clinical settings including cardiac surgery. Previous investigations suggest that S-100? is capable of identifying patients with cerebral dysfunction after cardiopulmonary bypass. Whether detection of elevated levels S-100? reflects long-term cognitive impairment remains to be shown. The present study evaluated whether

  16. Serum concentration of cartilage oligomeric matrix protein (COMP) is sensitive to physiological cyclic loading in healthy adults1

    Microsoft Academic Search

    Chris O. Dyrby Eng; Karen B. King

    Summary Objective: To test the hypothesis that physiological cyclic loading during a 30-min walking exercise causes an increase in serum cartilage oligomeric matrix protein (COMP) concentration in a healthy population. Methods: Blood samples (5 ml) were drawn from 10 physically active adults immediately before and after, and 0.5 h, 1.5 h, 3.5 h and 5.5 h after a 30-min walking

  17. Serial Changes in Surfactant-associated Proteins in Lung and Serum before and after Onset of ARDS

    Microsoft Academic Search

    KELLY E. GREENE; JO RAE WRIGHT; KENNETH P. STEINBERG; JOHN T. RUZINSKI; ELLEN CALDWELL; WES B. WONG; WILLIAM HULL; JEFFREY A. WHITSETT; TOYOAKI AKINO; YOSHIO KUROKI; HISATO NAGAE; LEONARD D. HUDSON; THOMAS R. MARTIN

    1999-01-01

    The goal of this study was to determine the changes that occur in surfactant-associated proteins in bronchoalveolar lavage fluid (BAL) and serum of patients at risk for ARDS and during the course of ARDS. We found that the concentrations of SP-A and SP-B were low in the BAL of patients at risk for ARDS before the onset of clinically defined

  18. Effects of various dietary animal and vegetable proteins on serum and biliary lipids and on gallstone formation in the hamster

    Microsoft Academic Search

    Mary Anne Sullivan; Anne Duffy; Nancy Dimarco; George Liepa

    1985-01-01

    The objective of this study was to determine the effects of various dietary animal (casein, bovine albumin and egg albumin)\\u000a and vegetable (soy, cottonseed and peanut) proteins on serum and biliary constitutents and gallstone formation in the hamster.\\u000a Eighty-four hamsters (60±5g) were assigned to either a control group (Purina rat chow) or to one of the 6 experimental groups.\\u000a Experimental

  19. Preoperative serum retinol-binding protein 4 is associated with the prognosis of patients with hepatocellular carcinoma after curative resection

    Microsoft Academic Search

    Dan-Dan Wang; Yi-Ming Zhao; Lu Wang; Guang Ren; Fei Wang; Zu-Guang Xia; Xi-Long Wang; Tao Zhang; Qi Pan; Zhi Dai; Ju-Ping Chen; Hai-Yan Dai; Wei Zhang; Hong-Wei He; Jia-Min Zhou; Guang-Yu Tang; Jian Zhou; Jia Fan; Zhao-You Tang

    2011-01-01

    Purpose  Metabolic syndrome and insulin resistance have been linked to increased risk of occurrence and mortality of hepatocellular\\u000a carcinoma (HCC). Recently, retinol-binding protein 4 (RBP4) was clarified as a specific serological marker of insulin resistance.\\u000a The aim of this study was to determine whether serum RBP4 could be used as a potential marker for predicting prognosis in\\u000a patients with HCC after

  20. Femtomolar detection of a cancer biomarker protein in serum with ultralow background current by anodic stripping voltammetry.

    PubMed

    Shiddiky, Muhammad J A; Kithva, Prakash H; Rauf, Sakandar; Trau, Matt

    2012-05-22

    An electrochemical immunosensor for the detection of a cancer biomarker protein in serum at femtomolar concentrations with ultralow background response has been developed, which consists of (i) a hydrophilic polyacrylic acid brush-modified indium tin oxide substrate as an antifouling substrate and (ii) a graphene-quantum dots-antibody 'bionanoconjugate' as a signal amplification label in voltammetric detection of targets in a glassy carbon electrode. PMID:22618633

  1. Outer Membrane Protein A, Peptidoglycan-Associated Lipoprotein, and Murein Lipoprotein Are Released by Escherichia coli Bacteria into Serum

    Microsoft Academic Search

    JUDITH HELLMAN; PAUL M. LOISELLE; MEGAN M. TEHAN; JENNIFER E. ALLAIRE; LENORA A. BOYLE; JAMES T. KURNICK; DAVID M. ANDREWS; KWANG SIK KIM; H. SHAW WARREN

    2000-01-01

    Complexes containing lipopolysaccharide (LPS) and three outer membrane proteins (OMPs) are released by gram-negative bacteria incubated in human serum and into the circulation in an experimental model of sepsis. The same OMPs are bound by immunoglobulin G (IgG) in the cross-protective antiserum raised to Escherichia coli J5 (anti-J5 IgG). This study was performed to identify the three OMPs. The 35-kDa

  2. Clinical significances of preoperative serum interleukin-6 and C-reactive protein level in operable gastric cancer

    Microsoft Academic Search

    Do-Kyong Kim; Sung Yong Oh; Hyuk-Chan Kwon; Suee Lee; Kyung A Kwon; Byung Geun Kim; Seong-Geun Kim; Sung-Hyun Kim; Jin Seok Jang; Min Chan Kim; Kyeong Hee Kim; Jin-Yeong Han; Hyo-Jin Kim

    2009-01-01

    BACKGROUND: The interleukin-6 (IL-6) pathway is one of the mechanisms that link inflammation and angiogenesis to malignancy. Because the C-reactive protein (CRP) is a representative marker for inflammation, CRP has recently been associated with the progression of disease in many cancer types. The principal objective of this study was to determine the preoperative serum levels of IL-6 and CRP in

  3. Clara Cell Secretory Protein in Tracheobronchial Aspirates and Umbilical Cord Serum of Extremely Premature Infants with Systemic Inflammation

    Microsoft Academic Search

    Wolfgang Thomas; Silvia Seidenspinner; Maria Chmielnicka-Kopaczyk; Alexander Marx; Johannes Wirbelauer; Marta Szymankiewicz; Christian P. Speer

    2010-01-01

    Background: A systemic fetal inflammatory response, reflected by chorioamnionitis with funisitis, is a risk factor for bronchopulmonary dysplasia. Clara cell secretory protein (CC10), a product of pulmonary Clara cells, has anti-inflammatory properties. Local down-regulation of CC10 has been associated with inflammatory lung disease. Increased serum levels of CC10 can indicate injury to alveolar-capillary integrity. Objective: We hypothesized that extremely premature

  4. The Effect of Simulated Microgravity Environment of RWV Bioreactors on Surface Reactions and Adsorption of Serum Proteins on Bone-bioactive Microcarriers

    NASA Technical Reports Server (NTRS)

    Radin, Shula; Ducheyne, P.; Ayyaswamy, P. S.

    2003-01-01

    Biomimetically modified bioactive materials with bone-like surface properties are attractive candidates for use as microcarriers for 3-D bone-like tissue engineering under simulated microgravity conditions of NASA designed rotating wall vessel (RWV) bioreactors. The simulated microgravity environment is attainable under suitable parametric conditions of the RWV bioreactors. Ca-P containing bioactive glass (BG), whose stimulatory effect on bone cell function had been previously demonstrated, was used in the present study. BG surface modification via reactions in solution, resulting formation of bone-like minerals at the surface and adsorption of serum proteins is critical for obtaining the stimulatory effect. In this paper, we report on the major effects of simulated microgravity conditions of the RWV on the BG reactions surface reactions and protein adsorption in physiological solutions. Control tests at normal gravity were conducted at static and dynamic conditions. The study revealed that simulated microgravity remarkably enhanced reactions involved in the BG surface modification, including BG dissolution, formation of bone-like minerals at the surface and adsorption of serum proteins. Simultaneously, numerical models were developed to simulate the mass transport of chemical species to and from the BG surface under normal gravity and simulated microgravity conditions. The numerical results showed an excellent agreement with the experimental data at both testing conditions.

  5. Photoactivable analogs for labeling 25-hydroxyvitamin D3 serum binding protein and for 1,25-dihydroxyvitamin D3 intestinal receptor protein

    NASA Technical Reports Server (NTRS)

    Kutner, A.; Link, R. P.; Schnoes, H. K.; DeLuca, H. F.

    1986-01-01

    3-Azidobenzoates and 3-azidonitrobenzoates of 25-hydroxyvitamin D3 as well as 3-deoxy-3-azido-25-hydroxyvitamin D3 and 3-deoxy-3-azido-1,25-dihydroxyvitamin D3 were prepared as photoaffinity labels for vitamin D serum binding protein and 1,25-dihydroxyvitamin D3 intestinal receptor protein. The compounds prepared were easily activated by short- or long-wavelength uv light, as monitored by uv and ir spectrometry. The efficacy of the compounds to compete with 25-hydroxyvitamin D3 or 1,25-dihydroxyvitamin D3 for the binding site of serum binding protein and receptor, respectively, was studied to evaluate the vitamin D label with the highest affinity for the protein. The presence of an azidobenzoate or azidonitrobenzoate substituent at the C-3 position of 25-OH-D3 significantly decreased (10(4)- to 10(6)-fold) the binding activity. However, the labels containing the azido substituent attached directly to the vitamin D skeleton at the C-3 position showed a high affinity, only 20- to 150-fold lower than that of the parent compounds with their respective proteins. Therefore, 3-deoxy-3-azidovitamins present potential ligands for photolabeling of vitamin D proteins and for studying the structures of the protein active sites.

  6. Integrative Proteomics and Tissue Microarray Profiling Indicate the Association between Overexpressed Serum Proteins and Non-Small Cell Lung Cancer

    PubMed Central

    Hu, Haichuan; Wang, Rui; Sun, Yihua; Zeng, Rong; Chen, Haiquan

    2012-01-01

    Lung cancer is the leading cause of cancer deaths worldwide. Clinically, the treatment of non-small cell lung cancer (NSCLC) can be improved by the early detection and risk screening among population. To meet this need, here we describe the application of extensive peptide level fractionation coupled with label free quantitative proteomics for the discovery of potential serum biomarkers for lung cancer, and the usage of Tissue microarray analysis (TMA) and Multiple reaction monitoring (MRM) assays for the following up validations in the verification phase. Using these state-of-art, currently available clinical proteomic approaches, in the discovery phase we confidently identified 647 serum proteins, and 101 proteins showed a statistically significant association with NSCLC in our 18 discovery samples. This serum proteomic dataset allowed us to discern the differential patterns and abnormal biological processes in the lung cancer blood. Of these proteins, Alpha-1B-glycoprotein (A1BG) and Leucine-rich alpha-2-glycoprotein (LRG1), two plasma glycoproteins with previously unknown function were selected as examples for which TMA and MRM verification were performed in a large sample set consisting about 100 patients. We revealed that A1BG and LRG1 were overexpressed in both the blood level and tumor sections, which can be referred to separate lung cancer patients from healthy cases. PMID:23284758

  7. Jun is phosphorylated by several protein kinases at the same sites that are modified in serum-stimulated fibroblasts.

    PubMed Central

    Baker, S J; Kerppola, T K; Luk, D; Vandenberg, M T; Marshak, D R; Curran, T; Abate, C

    1992-01-01

    c-jun is a member of the family of immediate-early genes whose expression is induced by factors such as serum stimulation, phorbol ester, and differentiation signals. Here we show that increased Jun synthesis after serum stimulation is accompanied by a concomitant increase in phosphorylation. Several serine-threonine kinases were evaluated for their ability to phosphorylate Jun in vitro. p34cdc2, protein kinase C, casein kinase II, and pp44mapk phosphorylated Jun efficiently, whereas cyclic AMP-dependent protein kinase and glycogen synthase kinase III did not. The sites phosphorylated by p34cdc2 were similar to those phosphorylated in vivo after serum induction. The major sites of phosphorylation were mapped to serines 63, 73, and 246. Phosphorylation of full-length Jun with several kinases did not affect the DNA-binding activity of Jun homodimers or Fos-Jun heterodimers. Comparison of the DNA binding and in vitro transcription properties of wild-type and mutated proteins containing either alanine or aspartic acid residues in place of Ser-63, -73, and -246 revealed only minor differences among homodimeric complexes and no differences among Fos-Jun heterodimers. Thus, phosphorylation of Jun did not produce a significant change in dimerization, DNA-binding, or in vitro transcription activity. The regulatory role of phosphorylation in the modulation of Jun function is likely to be considerably more complex than previously suggested. Images PMID:1328860

  8. Implant debris particle size affects serum protein adsorption which may contribute to particle size-based bioreactivity differences

    PubMed Central

    Reddy, Anand; Caicedo, Marco; Samelko, Lauryn; Jacobs, Joshua J; Hallab, Nadim James

    2014-01-01

    Biologic reactivity to orthopedic implant debris mediates long-term clinical performance of total joint arthroplasty implants. However, why some facets of implant debris are more pro-inflammatory remains controversial such as particle size, shape, base material etc. This precludes accurate prediction and optimal design of modern total joint replacements. We hypothesized that debris particle size can influence adsorbed protein film composition and affect subsequent bioreactivity. We measured size-dependent protein film-adsorption, and adsorbed protein film-dependent cytokine release using equal surface areas of different sized cobalt-chromium-alloy (CoCr-alloy) particle and in vitro challenge of human macrophages (THP-1 and human primary). Smaller 5?m vs 70?m sized particles preferentially adsorbed more serum protein in general (p<0.03), where higher molecular weight serum proteins consistent with IgG were identified. Additionally, 5?m CoCr-alloy particles pre-coated with different protein biofilms (IgG vs albumin) resulted in differential cytokine expression where albumin-coated particles induced more TNF-? and IgG-coated particles induced more IL-1? release from human monocyte/macrophages. In these preliminary in vitro studies we demonstrated the capability of equal surface areas of different particle sizes to influence adsorbed protein composition and that adsorbed protein differences on identical particles can translate into complex differences in bioreactivity. Together this suggests adsorbed protein differences on different sized particles of the same material may be a contributing mechanism by which different sized particles induce differences in reactivity. PMID:24941408

  9. Quantitation of Serum Free Light Chains in Combination with Protein Electrophoresis and Clinical Information for Diagnosing Multiple Myeloma in a General Hospital Population

    Microsoft Academic Search

    Armin P. Piehler; Nina Gulbrandsen; Peter Kierulf; Petter Urdal

    BACKGROUND: Serum free light chain (SFLC) measure- ments have recently come into use as an aid for diag- nosing monoclonal gammopathy. We evaluated SFLC measurements in combination with serum protein electrophoresis (SPE) and clinical information for di- agnosing multiple myeloma (MM) in a hospital population.

  10. Cross flow diafiltration of serum with basal medium suitable for growth of hybridomas, sterilization and protein reduction.

    PubMed

    Jungbauer, A; Wenisch, E; Steindl, F; Himmler, G; Reiter, S; Rüker, F; Wagner, K; Katinger, H

    1987-01-01

    Fetal Calf Serum (FCS) was extensively extracted by cross flow diafiltration (Pellicon, Millipore) and sterilized using the basal growth medium (DMEM) for the extraction. Ultrafiltration membranes of 10(5) and 3 X 10(5) Dalton cut off were used respectively. The diafiltrates were used for hybridoma cultivation and the results of growth and mAb-production were compared with standard medium (DMEM + 5% FCS). Slightly reduced mAb-titers were achieved. These were, however, compensated by decreased concentration of contaminating protein and higher specific mAb/protein ratio as examined by SDS-PAGE and enzyme linked immuno electro transfer blot (EITB). PMID:3582774

  11. Isolation and characterization of pharmaceutical grade human pentraxins, serum amyloid P component and C?reactive protein, for clinical use

    PubMed Central

    Pepys, Mark B.; Gallimore, J. Ruth; Lloyd, Joanne; Li, Zhanhong; Graham, David; Taylor, Graham W.; Ellmerich, Stephan; Mangione, Palma P.; Tennent, Glenys A.; Hutchinson, Winston L.; Millar, David J.; Bennett, Gary; More, John; Evans, David; Mistry, Yogesh; Poole, Stephen; Hawkins, Philip N.

    2012-01-01

    The human pentraxin proteins, serum amyloid P component (SAP) and C?reactive protein (CRP) are important in routine clinical diagnosis, SAP for systemic amyloidosis and CRP for monitoring the non?specific acute phase response. They are also targets for novel therapies currently in development but their roles in health and disease are controversial. Thus, both for clinical use and to rigorously elucidate their functions, structurally and functionally intact, pharmaceutical grade preparations of the natural, authentic proteins are required. We report here the production from normal human donor plasma and the characterization of the first such preparations. Importantly, we demonstrate that, contrary to reports using recombinant proteins and less well characterized preparations, neither CRP nor SAP stimulate the release by human peripheral blood mononuclear cells in vitro of any TNF?, IL?6 or IL?8, nor does SAP cause release of IL?1? or IL?10. Furthermore neither of our preparations was pro?inflammatory in mice in vivo. PMID:22867744

  12. Isolation and characterization of pharmaceutical grade human pentraxins, serum amyloid P component and C-reactive protein, for clinical use.

    PubMed

    Pepys, Mark B; Gallimore, J Ruth; Lloyd, Joanne; Li, Zhanhong; Graham, David; Taylor, Graham W; Ellmerich, Stephan; Mangione, Palma P; Tennent, Glenys A; Hutchinson, Winston L; Millar, David J; Bennett, Gary; More, John; Evans, David; Mistry, Yogesh; Poole, Stephen; Hawkins, Philip N

    2012-10-31

    The human pentraxin proteins, serum amyloid P component (SAP) and C-reactive protein (CRP) are important in routine clinical diagnosis, SAP for systemic amyloidosis and CRP for monitoring the non-specific acute phase response. They are also targets for novel therapies currently in development but their roles in health and disease are controversial. Thus, both for clinical use and to rigorously elucidate their functions, structurally and functionally intact, pharmaceutical grade preparations of the natural, authentic proteins are required. We report here the production from normal human donor plasma and the characterization of the first such preparations. Importantly, we demonstrate that, contrary to reports using recombinant proteins and less well characterized preparations, neither CRP nor SAP stimulate the release by human peripheral blood mononuclear cells in vitro of any TNF?, IL-6 or IL-8, nor does SAP cause release of IL-1? or IL-10. Furthermore neither of our preparations was pro-inflammatory in mice in vivo. PMID:22867744

  13. Associations of leisure time physical activity, self-rated physical fitness, and estimated aerobic fitness with serum C-reactive protein among 3803 adults

    Microsoft Academic Search

    Borodulin Katja; Tiina Laatikainen; Veikko Salomaa; Pekka Jousilahti

    2006-01-01

    ObjectiveSerum C-reactive protein (CRP), a marker of systemic inflammation, is a risk factor for cardiovascular disease. Obesity and physical activity are associated with CRP, though population studies are sparse.

  14. Larval serum proteins of the gypsy moth, Lymantria dispar: Allometric changes during development suggest several functions for arylphorin and lipophorin

    SciTech Connect

    Karpells, S.T.

    1989-01-01

    Storage proteins are the major nutritive intermediates in insects and although the serum storage proteins are relatively well studied, definitive roles for many of them have yet to be established. To further characterize their roles in development and to establish quantitative baselines for future studies, two serum proteins, arylphorin (Ap) and lipophorin (Lp), of the gypsy moth, Lymantria dispar, were studied. Ap and Lp, isolated from larval hemolymph, were partially characterized biochemically and immunologically. Hemolymph concentrations throughout larval development were determined using quantitative immunoelectrophoresis and absolute hemolymph amounts of protein were determined by measuring hemolymph volume. Cyclic fluctuations in hemolymph concentrations of Ap in particular correlated with each molting cycle and an increase in Lp levels just prior to pupation suggest a metamorphic change in the role or demand for the protein. Sexual dimorphism in protein concentrations are explained in part by the sexual dimorphism in the number of larval instars. In fact, an additional instar of Ap accumulation in the female gypsy moth is suggested to compensate for the lack of a female-specific storage protein in this species. The last two days of each instar were found to be the optimum time to sample protein concentration with minimum variance. Allometric relationships among Ap accumulation, Lp accumulation and weight gain were uncovered. Ap labelled with ({sup 14}C)-N-ethylmaleimide was shown to be incorporated into newly synthesized cuticle and setae during a larval-larval molt. The antiserum developed against L. dispar Ap was used to identify the Ap of Trichoplusia in and study Ap titers in parasitized T. in larvae. The antiserum was also used to determine the immunological relatedness of 5 species of Lepidoptera.

  15. Multifactorial analysis of affinity-mass spectrometry data from serum protein samples: A strategy to distinguish patients with preeclampsia from matching control individuals

    Microsoft Academic Search

    Ulrich Pecks; Franka Seidenspinner; Claudia Röwer; Toralf Reimer; Werner Rath; Michael O. Glocker

    2010-01-01

    A multifactorial differential analysis of serum proteins using mass spectrometry distinguished samples from pregnant women\\u000a with severe early-onset preeclampsia (n = 11) from those of control individuals with uneventful pregnancies (n = 13). Serum proteins were fractionated by either their affinities to reversed-phase material coated magnetic beads or by\\u000a fractionated precipitation. The on-average most abundant ion signals were observed at

  16. Longevity-associated mitochondrial DNA 5178 A\\/C polymorphism influences effects of cigarette smoking on serum protein fraction levels in Japanese men

    Microsoft Academic Search

    Akatsuki Kokaze; Mamoru Ishikawa; Naomi Matsunaga; Masao Yoshida; Yasuko Sekine; Kanako Sekiguchi; Masao Satoh; Matsuko Harada; Koji Teruya; Nobuo Takeda; Yoshiko Uchida; Yutaka Takashima

    2003-01-01

    Mitochondrial DNA 5178 adenine\\/cytosine (mt5178 A\\/C) polymorphism is reportedly associated with longevity and susceptibility to age-related diseases in Japanese individuals. We previously reported an association between mt5178 A\\/C polymorphism and serum protein fraction levels in healthy Japanese women. An association between habitual smoking and serum protein fraction levels has also been reported previously. The aim of this study was to

  17. Comparative Study of Somatostatin-Human Serum Albumin Fusion Proteins and Natural Somatostatin on Receptor Binding, Internalization and Activation

    PubMed Central

    Peng, Ying; Deng, Lili; Ding, Yuedi; Chen, Quancheng; Wu, Yu; Yang, Meilin; Wang, Yaping; Fu, Qiang

    2014-01-01

    Albumin fusion technology, the combination of small molecular proteins or peptides with human serum albumin (HSA), is an effective method for improving the medicinal values of natural small molecular proteins or peptides. However, comparative studies between HSA-fusion proteins or peptides and the parent small molecules in biological and molecular mechanisms are less reported. In this study, we examined the binding property of two novel somatostatin-HSA fusion proteins, (SST14)2-HSA and (SST28)2-HSA, to human SSTRs in stably expressing SSTR1-5 HEK 293 cells; observed the regulation of receptor internalization and internalized receptor recycling; and detected the receptors activation of HSA fusion proteins in stably expressing SSTR2- and SSTR3-EGFP cells. We showed that both somatostatin-HSA fusion proteins had high affinity to all five SSTRs, stimulated the ERK1/2 phosphorylation and persistently inhibited the accumulation of forskolin-stimulated cAMP in SSTR2- and SSTR3-expressing cells; but were less potent than the synthetic somatostatin-14 (SST-14). Our experiments also showed that somatostatin-HSA fusion proteins did not induce the receptors internalization; rather, they accelerated the recycling of the internalized receptors induced by SST-14 to the plasma membrane. Our results indicated that somatostatin-HSA fusion proteins, different from SST-14, exhibit some particular properties in binding, regulating, and activating somatostatin receptors. PMID:24587133

  18. Embryo culture in teratological surveillance and serum proteins in development. Final technical report

    SciTech Connect

    Klein, N.W.

    1986-01-01

    An overview of the authors research into teratogenesis of blood serum of patients on medication or rats injected with drugs is presented. In addition studies concerning the role of methionine in the developing fetus is given. 68 refs.

  19. Multiplexing of miniaturized planar antibody arrays for serum protein profiling--a biomarker discovery in SLE nephritis.

    PubMed

    Petersson, Linn; Dexlin-Mellby, Linda; Bengtsson, Anders A; Sturfelt, Gunnar; Borrebaeck, Carl A K; Wingren, Christer

    2014-06-01

    In the quest to decipher disease-associated biomarkers, miniaturized and multiplexed antibody arrays may play a central role in generating protein expression profiles, or protein maps, of crude serum samples. In this conceptual study, we explored a novel, 4-times larger pen design, enabling us to, in a unique manner, simultaneously print 48 different reagents (antibodies) as individual 78.5 ?m(2) (10 ?m in diameter) sized spots at a density of 38,000 spots cm(-2) using dip-pen nanolithography technology. The antibody array set-up was interfaced with a high-resolution fluorescent-based scanner for sensitive sensing. The performance and applicability of this novel 48-plex recombinant antibody array platform design was demonstrated in a first clinical application targeting SLE nephritis, a severe chronic autoimmune connective tissue disorder, as the model disease. To this end, crude, directly biotinylated serum samples were targeted. The results showed that the miniaturized and multiplexed array platform displayed adequate performance, and that SLE-associated serum biomarker panels reflecting the disease process could be deciphered, outlining the use of miniaturized antibody arrays for disease proteomics and biomarker discovery. PMID:24763547

  20. Barrel structures in proteins: automatic identification and classification including a sequence analysis of TIM barrels.

    PubMed Central

    Nagano, N.; Hutchinson, E. G.; Thornton, J. M.

    1999-01-01

    Automated methods for identifying and characterizing regular beta-barrels from coordinate data have been developed to analyze and classify various kinds of barrel structures based on geometric parameters such as the barrel strand number (n) and shear number (S). In total, we find 1,316 barrels in the January 1998 release of Protein Data Bank. Of 1,316 barrels, 1,277 barrels had an even shear number, corresponding to 50 nonhomologous families. The (beta alpha)8 triose phosphate isomerase (TIM) barrel (n = 8, S = 8) fold has the largest number of apparently nonhomologous entries, 16, although the trypsin like antiparallel (n = 6, S = 8) barrels (representing only three families) are the most common with 527 barrels. Of all the protein families that exhibit barrel structures, 68% are found to be various kinds of enzymes, the remainder being binding proteins and transport membrane proteins. In addition, the layers of side chains, which form the cores of barrels with S = n and S = 2n, are also analyzed. More sophisticated methods were developed for detecting TIM barrels specifically, including consideration of the amino acid propensities for the side chains that form the layers. We found that the residues on the outside of the eight stranded parallel beta-barrel, buried by the alpha-helices, are much more hydrophobic than the residues inside the barrel. PMID:10548053

  1. A highly sensitive targeted mass spectrometric assay for quantification of AGR2 protein in human urine and serum

    PubMed Central

    Shi, Tujin; Gao, Yuqian; Quek, Sue Ing; Fillmore, Thomas L.; Nicora, Carrie D.; Su, Dian; Zhao, Rui; Kagan, Jacob; Srivastava, Sudhir; Rodland, Karin D.; Liu, Tao; Smith, Richard D.; Chan, Daniel W.; Camp, David G.; Liu, Alvin Y.; Qian, Wei-Jun

    2014-01-01

    Anterior gradient 2 (AGR2) is a secreted, cancer-associated protein in many types of epithelial cancer cells. We developed a highly sensitive targeted mass spectrometric assay for quantification of AGR2 in urine and serum. Digested peptides from clinical samples were processed by PRISM (high pressure and high resolution separations coupled with intelligent selection and multiplexing), which incorporates high pH reversed-phase LC separations to fractionate and select target fractions for follow-on LC-SRM analyses. The PRISM-SRM assay for AGR2 showed a reproducibility of <10% CV and LOQ values of ~130 pg/mL in serum and ~10 pg per 100 ?g total protein mass in urine, respectively. A good correlation (R2 = 0.91) was observed for the measurable AGR2 concentrations in urine between SRM and ELISA. Based on an initial cohort of 37 subjects, urinary AGR2/PSA concentration ratios showed a significant difference (P = 0.026) between non-cancer and cancer. Large clinical cohort studies are needed for the validation of AGR2 as a useful diagnostic biomarker for prostate cancer. Our work validated the approach of identifying candidate secreted protein biomarkers through genomics and measurement by targeted proteomics, especially for proteins where no immunoassays are available. PMID:24251762

  2. Phosphorylation of Heat Shock Protein 27 is Increased by Cast Immobilization and by Serum-free Starvation in Skeletal Muscles.

    PubMed

    Kim, Mee-Young; Lee, Jeong-Uk; Kim, Ju-Hyun; Lee, Lim-Kyu; Park, Byoung-Sun; Yang, Seung-Min; Jeon, Hye-Joo; Lee, Won-Deok; Noh, Ji-Woong; Kwak, Taek-Yong; Jang, Sung-Ho; Lee, Tae-Hyun; Kim, Ju-Young; Kim, Bokyung; Kim, Junghwan

    2014-12-01

    [Purpose] Cast immobilization- and cell starvation-induced loss of muscle mass are closely associated with a dramatic reduction in the structural muscle proteins. Heat shock proteins are molecular chaperones that are constitutively expressed in several eukaryotic cells and have been shown to protect against various stressors. However, the changes in the phosphorylation of atrophy-related heat shock protein 27 (HSP27) are still poorly understood in skeletal muscles. In this study, we examine whether or not phosphorylation of HSP27 is changed in the skeletal muscles after cast immobilization and serum-free starvation with low glucose in a time-dependent manner. [Methods] We undertook a HSP27 expression and high-resolution differential proteomic analysis in skeletal muscles. Furthermore, we used western blotting to examine protein expression and phosphorylation of HSP27 in atrophied gastrocnemius muscle strips and L6 myoblasts. [Results] Cast immobilization and starvation significantly upregulated the phosphorylation of HSP27 in a time-dependent manner, respectively. [Conclusion] Our results suggest that cast immobilization- and serum-free starvation-induced atrophy may be in part related to changes in the phosphorylation of HSP27 in rat skeletal muscles. PMID:25540511

  3. Phosphorylation of Heat Shock Protein 27 is Increased by Cast Immobilization and by Serum-free Starvation in Skeletal Muscles

    PubMed Central

    Kim, Mee-Young; Lee, Jeong-Uk; Kim, Ju-Hyun; Lee, Lim-Kyu; Park, Byoung-Sun; Yang, Seung-Min; Jeon, Hye-Joo; Lee, Won-Deok; Noh, Ji-Woong; Kwak, Taek-Yong; Jang, Sung-Ho; Lee, Tae-Hyun; Kim, Ju-Young; Kim, Bokyung; Kim, Junghwan

    2014-01-01

    [Purpose] Cast immobilization- and cell starvation-induced loss of muscle mass are closely associated with a dramatic reduction in the structural muscle proteins. Heat shock proteins are molecular chaperones that are constitutively expressed in several eukaryotic cells and have been shown to protect against various stressors. However, the changes in the phosphorylation of atrophy-related heat shock protein 27 (HSP27) are still poorly understood in skeletal muscles. In this study, we examine whether or not phosphorylation of HSP27 is changed in the skeletal muscles after cast immobilization and serum-free starvation with low glucose in a time-dependent manner. [Methods] We undertook a HSP27 expression and high-resolution differential proteomic analysis in skeletal muscles. Furthermore, we used western blotting to examine protein expression and phosphorylation of HSP27 in atrophied gastrocnemius muscle strips and L6 myoblasts. [Results] Cast immobilization and starvation significantly upregulated the phosphorylation of HSP27 in a time-dependent manner, respectively. [Conclusion] Our results suggest that cast immobilization- and serum-free starvation-induced atrophy may be in part related to changes in the phosphorylation of HSP27 in rat skeletal muscles. PMID:25540511

  4. Dietary total antioxidant capacity from different assays in relation to serum C-reactive protein among young Japanese women

    PubMed Central

    2012-01-01

    Background The association between dietary total antioxidant capacity (TAC) from different assays and serum C-reactive protein (CRP) has not been assessed in non-Western populations. We examined the association between dietary TAC and serum CRP concentration in young Japanese women using different four TAC assays. Methods The subjects were 443 young Japanese women aged 18–22?years. Dietary TAC was assessed with a self-administered diet history questionnaire and the TAC value of each food using the following four assays: ferric reducing ability of plasma (FRAP); oxygen radical absorbance capacity (ORAC); Trolox equivalent antioxidant capacity (TEAC); and total radical-trapping antioxidant parameter (TRAP). Serum CRP concentrations were measured by highly sensitive nephelometry. Results The major contributor to dietary TAC was green, barley, and oolong tea (FRAP: 53%, ORAC: 45%, TEAC: 36%, and TRAP: 44%). The prevalence of elevated CRP concentrations (? 1?mg/L) was 5.6%. TAC from FRAP was inversely associated with serum CRP concentrations (adjusted odds ratio [OR] for elevated CRP concentration in high [compared with low] dietary TAC group: 0.39 [95% confidence interval (CI): 0.16-0.98]; P?=?0.04). TAC from ORAC was inversely associated with CRP, although the association was not significant (OR: 0.48 [95% CI: 0.20-1.14]; P?=?0.10). TAC from TEAC was inversely associated with CRP (OR: 0.32 [95% CI: 0.12-0.82]; P?=?0.02), as was TAC from TRAP (OR: 0.31 [95% CI: 0.12-0.81]; P?=?0.02). Conclusions Dietary TAC was inversely associated with serum CRP concentration in young Japanese women regardless of assay. Further studies are needed in other populations to confirm these results. PMID:23110638

  5. Evidence That Kidney Function but Not Type 2 Diabetes Determines Retinol-Binding Protein 4 Serum Levels

    PubMed Central

    Henze, Andrea; Frey, Simone K.; Raila, Jens; Tepel, Martin; Scholze, Alexandra; Pfeiffer, Andreas F. H.; Weickert, Martin O.; Spranger, Joachim; Schweigert, Florian J.

    2008-01-01

    OBJECTIVE— It has been suggested that retinol-binding protein 4 (RBP4) links adiposity, insulin resistance, and type 2 diabetes. However, circulating RBP4 levels are also affected by kidney function. Therefore, the aim of this study was to test whether RBP4 serum levels are primarily associated with kidney function or type 2 diabetes. RESEARCH DESIGN AND METHODS— RBP4 serum concentration was determined by enzyme-linked immunosorbent assay in 126 nondiabetic and 104 type 2 diabetic subjects. The study population was divided according to estimated glomerular filtration rate (eGFR) into the following groups: eGFR >90 ml/min per 1.73 m2 (n = 53), 60–90 ml/min per 1.73 m2 (n = 90), 30–60 ml/min per 1.73 m2 (n = 38), and <30 ml/min per 1.73 m2 (n = 49). Each group was subdivided into nondiabetic and type 2 diabetic subjects. RESULTS— RBP4 serum concentration was elevated (2.65 vs. 2.01 ?mol/l; P < 0.001) and eGFR was reduced (56 vs. 74 ml/min per 1.73 m2; P < 0.001) in type 2 diabetic vs. nondiabetic subjects, respectively. By stratifying for eGFR, no more differences in RBP4 serum concentration were detectable between type 2 diabetic and nondiabetic subjects. A linear regression analysis revealed an influence of eGFR (r = ?0.477; P < 0.001) but not A1C (r = 0.093; P = 0.185) on RBP4 serum concentration. CONCLUSIONS— Existing human data showing elevated RBP4 levels in type 2 diabetic patients may be the result of moderate renal insufficiency rather than support for the suggestion that RBP4 links obesity to type 2 diabetes. PMID:18796616

  6. DECREASED HEART RATE IS ASSOCIATED WITH CARBAMATE-INDUCED ACTIVATION OF PRO-INFLAMMATORY SERUM PROTEINS.

    EPA Science Inventory

    Previously we reported that chlorpyrifos (CHP), an irreversible cholinesterase (ChE) inhibitor, induces hypertension in rats. Concomitant with hypertension, we found an increase in C-reactive protein, macrophage inflammatory protein-2 , monocyte chemotactic protein-5 and interfer...

  7. The possible link between the elevated serum levels of neurokinin A and anti-ribosomal P protein antibodies in children with autism

    PubMed Central

    2011-01-01

    Background Neurogenic inflammation is orchestrated by a large number of neuropeptides. Tachykinins (substance P, neurokinin A and neurokinin B) are pro-inflammatory neuropeptides that may play an important role in some autoimmune neuroinflammatory diseases. Autoimmunity may have a role in the pathogenesis of autism in some patients. We are the first to measure serum neurokinin A levels in autistic children. The relationship between serum levels of neurokinin A and anti-ribosomal P protein antibodies was also studied. Methods Serum neurokinin A and anti-ribosomal P protein antibodies were measured in 70 autistic children in comparison to 48 healthy-matched children. Results Autistic children had significantly higher serum neurokinin A levels than healthy controls (P < 0.001). Children with severe autism had significantly higher serum neurokinin A levels than patients with mild to moderate autism (P < 0.001). Increased serum levels of neurokinin A and anti-ribosomal P protein antibodies were found in 57.1% and 44.3%, respectively of autistic children. There was significant positive correlations between serum levels of neurokinin A and anti-ribosomal P protein antibodies (P = 0.004). Conclusions Serum neurokinin A levels were elevated in some autistic children and they were significantly correlated to the severity of autism and to serum levels of anti-ribosomal P protein antibodies. However, this is an initial report that warrants further research to determine the pathogenic role of neurokinin A and its possible link to autoimmunity in autism. The therapeutic role of tachykinin receptor antagonists, a potential new class of anti-inflammatory medications, should also be studied in autism. PMID:22189180

  8. Surface diffusion of interacting proteins. Effect of concentration on the lateral mobility of adsorbed bovine serum albumin.

    PubMed Central

    Tilton, R D; Gast, A P; Robertson, C R

    1990-01-01

    Surface diffusion of bovine serum albumin absorbed from aqueous solution to poly(methylmethacrylate) surfaces is significantly hindered by protein-protein lateral interactions. The long-time self diffusion coefficient measured by fluorescence recovery after pattern photobleaching decreases by approximately one order of magnitude as the surface area fraction occupied by protein increases from 0.10 to 0.69. Qualitative features of the surface concentration dependence of the self diffusion coefficient can be described by several recent models for lateral diffusion of interacting species. The mobile fraction is independent of the surface concentration, and both the self diffusion coefficient and the mobile fraction are constant between 15 min and 7 h of adsorption. PMID:2291948

  9. Prolonged prophylactic protection from botulism with a single adenovirus treatment promoting serum expression of a VHH-based antitoxin protein.

    PubMed

    Mukherjee, Jean; Dmitriev, Igor; Debatis, Michelle; Tremblay, Jacqueline M; Beamer, Gillian; Kashentseva, Elena A; Curiel, David T; Shoemaker, Charles B

    2014-01-01

    Current therapies for most acute toxin exposures are limited to administration of polyclonal antitoxin serum. We have shown that VHH-based neutralizing agents (VNAs) consisting of two or more linked, toxin-neutralizing heavy-chain-only VH domains (VHHs), each binding distinct epitopes, can potently protect animals from lethality in several intoxication models including Botulinum neurotoxin serotype A1 (BoNT/A1). Appending a 14 amino acid albumin binding peptide (ABP) to an anti-BoNT/A1 heterodimeric VNA (H7/B5) substantially improved serum stability and resulted in an effective VNA serum half-life of 1 to 2 days. A recombinant, replication-incompetent, adenoviral vector (Ad/VNA-BoNTA) was engineered that induces secretion of biologically active VNA, H7/B5/ABP (VNA-BoNTA), from transduced cells. Mice administered a single dose of Ad/VNA-BoNTA, or a different Ad/VNA, via different administration routes led to a wide range of VNA serum levels measured four days later; generally intravenous > intraperitoneal > intramuscular > subcutaneous. Ad/VNA-BoNTA treated mice were 100% protected from 10 LD50 of BoNT/A1 for more than six weeks and protection positively correlated with serum levels of VNA-BoNTA exceeding about 5 ng/ml. Some mice developed antibodies that inhibited VNA binding to target but these mice displayed no evidence of kidney damage due to deposition of immune complexes. Mice were also successfully protected from 10 LD50 BoNT/A1 when Ad/VNA-BoNTA was administered up to 1.5 hours post-intoxication, demonstrating rapid appearance of the protective VNA in serum following treatment. Genetic delivery of VNAs promises to be an effective method of providing prophylactic protection and/or acute treatments for many toxin-mediated diseases. PMID:25170904

  10. Association of serum Clara cell protein CC16 with respiratory infections and immune response to respiratory pathogens in elite athletes

    PubMed Central

    2014-01-01

    Background Respiratory epithelium integrity impairment caused by intensive exercise may lead to exercise-induced bronchoconstriction. Clara cell protein (CC16) has anti-inflammatory properties and its serum level reflects changes in epithelium integrity and airway inflammation. This study aimed to investigate serum CC16 in elite athletes and to seek associations of CC16 with asthma or allergy, respiratory tract infections (RTIs) and immune response to respiratory pathogens. Methods The study was performed in 203 Olympic athletes. Control groups comprised 53 healthy subjects and 49 mild allergic asthmatics. Serum levels of CC16 and IgG against respiratory viruses and Mycoplasma pneumoniae were assessed. Allergy questionnaire for athletes was used to determine symptoms and exercise pattern. Current versions of ARIA and GINA guidelines were used when diagnosing allergic rhinitis and asthma, respectively. Results Asthma was diagnosed in 13.3% athletes, of whom 55.6% had concomitant allergic rhinitis. Allergic rhinitis without asthma was diagnosed in 14.8% of athletes. Mean CC16 concentration was significantly lower in athletes versus healthy controls and mild asthmatics. Athletes reporting frequent RTIs had significantly lower serum CC16 and the risk of frequent RTIs was more than 2-fold higher in athletes with low serum CC16 (defined as equal to or less than 4.99 ng/ml). Athletes had significantly higher anti-adenovirus IgG than healthy controls while only non-atopic athletes had anti-parainfluenza virus IgG significantly lower than controls. In all athletes weak correlation of serum CC16 and anti-parainfluenza virus IgG was present (R?=?0.20, p?serum CC16 levels. Athletes with decreased CC16 are more susceptible to respiratory infections. Atopy may be an additional factor modifying susceptibility to infections in subjects performing regular high-load exercise. PMID:24735334

  11. Chemistry-dependent adsorption of serum proteins onto polyanhydride microparticles differentially influences dendritic cell uptake and activation.

    PubMed

    Carrillo-Conde, Brenda R; Ramer-Tait, Amanda E; Wannemuehler, Michael J; Narasimhan, Balaji

    2012-10-01

    The delivery of antigen-loaded microparticles to dendritic cells (DCs) may benefit from surface optimization of the microparticles themselves, thereby exploiting the material properties and introducing signals that mimic pathogens. Following in vivo administration microparticle surface characteristics are likely to be significantly modified as proteins are quickly adsorbed onto their surface. In this work we describe the chemistry-dependent serum protein adsorption patterns on polyanhydride particles and the implications for their molecular interactions with DCs. The enhanced expression of MHC II and CD40 on DCs after incubation with amphiphilic polyanhydride particles, and the increased secretion of IL-6, TNF-?, and IL-12p40 by hydrophobic polyanhydride particles exemplified the chemistry-dependent activation of DCs by sham-coated particles. The presence of proteins such as complement component 3 and IgG further enhanced the adjuvant properties of these vaccine carriers by inducing DC maturation (i.e. increased cell surface molecule expression and cytokine secretion) in a chemistry-dependent manner. Utilizing DCs derived from complement receptor 3-deficient mice (CR3(-/-) mice) identified a requirement for CR3 in the internalization of both sham- and serum-coated particles. These studies provide valuable insights into the rational design of targeted vaccine platforms aimed at inducing robust immune responses and improving vaccine efficacy. PMID:22684115

  12. Effect of storage time and temperature on the total protein concentration and electrophoretic fractions in equine serum

    PubMed Central

    Alberghina, Daniela; Casella, Stefania; Giannetto, Claudia; Marafioti, Simona; Piccione, Giuseppe

    2013-01-01

    Serum protein electrophoresis (SPE) is a technique that could be considered one of the most useful diagnostic aids available to the clinician. The effect of storage time and temperature on the total proteins and electrophoretic fractions (albumin, ?1-, ?2-, ?1-, ?2-, and ?-globulins) was assessed in 24 healthy horses. All samples, collected by jugular vein puncture, were centrifuged and divided into 4 aliquots. The 1st aliquot was analyzed within 3 h from collection (time 0), the 2nd was refrigerated at +4°C for 24 h, the 3rd was refrigerated at +4°C for 48 h, and the last was frozen at ?20°C for 48 h. One-way repeated-measures analysis of variance (ANOVA) showed a significant effect (P < 0.05) of the different storage conditions on the concentrations of all the parameters studied and significant variations in the percentages of albumin, ?1-globulins, ?2-globulins, and ?-globulins. Compared with time 0 the total protein concentration increased significantly after 48 h at ?20°C, the albumin percentage decreased after 48 h at ?20°C, the ?1-globulin percentage increased after 24 h at +4°C, the ?2-globulin percentage increased after 48 h at +4°C and at ?20°C, and the ?-globulin percentage increased after 48 h at ?20°C. The results should help veterinary practitioners handle and store equine serum samples appropriately. Further investigations at different storage times and temperatures could be useful. PMID:24124272

  13. Cryopreservation for bovine embryos in serum-free freezing medium containing silk protein sericin.

    PubMed

    Isobe, Tomohiro; Ikebata, Yoshihisa; Onitsuka, Takeshi; Do, Lanh Thi Kim; Sato, Yoko; Taniguchi, Masayasu; Otoi, Takeshige

    2013-10-01

    Because the use of serum in the embryo cryopreservation increases the probability of animal health problems such as bovine spongiform encephalopathy (BSE) and viral infections, this study was conducted to examine the effects of sericin supplementation for serum-free freezing medium on the survival and development of bovine embryos after freezing-thawing and direct transfer to recipients. When in vitro-produced bovine embryos were frozen conventionally in the freezing medium supplemented with various concentrations (0.1%, 0.5%, and 1.0%) of sericin, the percentages of damaged zona pellucida, survival, and development of frozen-thawed embryos were similar to those of embryos frozen in freezing medium supplemented with 0.4% bovine serum albumin (BSA) and 20% fetal bovine serum (FBS) (0.4BSA/20F; control). When in vivo-derived embryos were frozen with 0.4BSA/20F (control), 0.5% sericin +20% FBS (0.5S/20F) or 0.5% sericin (0.5S) and were subsequently transferred directly to recipients, the percentages of recipients with pregnancy and normal calving in the 0.5S/20F group were higher than those in the control group (47.3% vs. 40.1% and 94.6% vs. 87.3%, respectively). Moreover, the percentages of recipients with pregnancy and normal calving (42.2% and 92.4%, respectively) in the 0.5S group were similar with those of other groups. In conclusion, these results indicate that serum-free freezing medium supplemented with sericin is available for the cryopreservation of bovine embryos and that it is beneficial for the elimination of a potential source of biological contamination by serum or BSA. PMID:23850826

  14. Computational modeling of serum-binding proteins and clearance in extrapolations across life stages and species for endocrine active compounds.

    PubMed

    Teeguarden, Justin G; Barton, Hugh A

    2004-06-01

    One measure of the potency of compounds that lead to the effects through ligand-dependent gene transcription is the relative affinity for the critical receptor. Endocrine active compounds that are presumed to act principally through binding to the estrogen receptor (e.g., estradiol, genistein, bisphenol A, and octylphenol) comprise one class of such compounds. For making simple comparisons, receptor-binding affinity has been equated to in vivo potency, which consequently defines the dose-response characteristics for the compound. Direct extrapolation of in vitro estimated affinities to the corresponding in vivo system and to specific species or life stages (e.g., neonatal, pregnancy) can be misleading. Accurate comparison of the potency of endocrine active compounds requires characterization of biochemical and pharmacokinetic factors that affect their free concentration. Quantitative in vitro and in vivo models were developed for integrating pharmacokinetics factors (e.g., serum protein and receptor-binding affinities, clearance) that affect potency. Data for parameterizing these models for several estrogenic compounds were evaluated and the models exercised. While simulations of adult human or rat sera were generally successful, difficulties in describing early life stages were identified. Exogenous compounds were predicted to be largely ineffective at competing estradiol off serum-binding proteins, suggesting this was unlikely to be physiologically significant. Discrepancies were identified between relative potencies based upon modeling in vitro receptor-binding activity versus in vivo activity in the presence of clearance and serum-binding proteins. The examples illustrate the utility of this approach for integrating available experimental data from in vitro and in vivo studies to estimate the relative potency of these compounds. PMID:15209943

  15. Electrolyte effect on the phase behavior of silica nanoparticles with lysozyme and bovine-serum-albumin proteins

    NASA Astrophysics Data System (ADS)

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

    2015-05-01

    Small-angle neutron scattering (SANS) and dynamic light scattering (DLS) studies have been carried out to investigate the effect of an electrolyte on the phase behavior of anionic silica nanoparticles with two globular proteins—cationic lysozyme [molecular weight (MW) 14.7 kDa] and anionic bovine serum albumin (MW 66.4 kDa). The results are compared with our earlier published work on similar systems without any electrolyte [I. Yadav, S. Kumar, V. K. Aswal, and J. Kohlbrecher, Phys. Rev. E 89, 032304 (2014), 10.1103/PhysRevE.89.032304]. Both the nanoparticle-protein systems transform to two phase at lower concentration of protein in the presence of an electrolyte. The autocorrelation function in DLS suggests that the diffusion coefficient (D) of a nanoparticle-protein system decreases in approaching two phase with the increase in protein concentration. This variation in D can be attributed to increase in attractive interaction and/or overall increase in the size. Further, these two contributions (interaction and structure) are determined from the SANS data. The changes in the phase behavior of nanoparticle-protein systems in the presence of an electrolyte are explained in terms of modifications in both the repulsive and attractive components of interaction between nanoparticles. In a two-phase system individual silica nanoparticles coexist along with their fractal aggregates.

  16. Serum hormone and myocellular protein recovery after intermittent runs at the velocity associated with V? O 2max

    Microsoft Academic Search

    Timo Vuorimaa; Tommi Vasankari; Kari Mattila; Olli Heinonen; Keijo Häkkinen; Heikki Rusko

    1999-01-01

    The responses of serum myocellular proteins and hormones to exercise were studied in ten well-trained middle-distance runners\\u000a [maximal oxygen consumption (V?O2max)?=?69.4?(5.1)?ml?·?kg?1?·?min?1] during 3 recovery days and compared to various measures of physical performance. The purpose was to establish the duration\\u000a of recovery from typical intermittent middle-distance running exercises. The subjects performed, in random, order two 28-min\\u000a treadmill running exercises at

  17. Regulation of heat shock protein message in Jurkat cells cultured under serum-starved and gravity-altered conditions

    NASA Technical Reports Server (NTRS)

    Lewis, M. L.; Hughes-Fulford, M.

    2000-01-01

    Although our understanding of effects of space flight on human physiology has advanced significantly over the past four decades, the potential contribution of stress at the cellular and gene regulation level is not characterized. The objective of this ground-based study was to evaluate stress gene regulation in cells exposed to altered gravity and environmentally suboptimal conditions. We designed primers to detect message for both the constitutive and inducible forms of the heat shock protein, HSP-70. Applying the reverse transcriptase-polymerase chain reaction (RT-PCR), we probed for HSP-70 message in human acute T-cell leukemia cells, Jurkat, subjected to three types of environmental stressors: (1) altered gravity achieved by centrifugation (hypergravity) and randomization of the gravity vector in rotating bioreactors, (2) serum starvation by culture in medium containing 0.05% serum, and (3) temperature elevation (42 degrees C). Temperature elevation, as the positive control, significantly increased HSP-70 message, while centrifugation and culture in rotating bioreactors did not upregulate heat shock gene expression. We found a fourfold increase in heat shock message in serum-starved cells. Message for the housekeeping genes, actin and cyclophilin, were constant and comparable to unstressed controls for all treatments. We conclude that gravitational perturbations incurred by centrifugal forces, exceeding those characteristic of a Space Shuttle launch (3g), and culture in rotating bioreactors do not upregulate HSP-70 gene expression. In addition, we found RT-PCR useful for evaluating stress in cultured cells. Copyright 2000 Wiley-Liss, Inc.

  18. A Rapid Lateral Flow Immunoassay for the Detection of Tyrosine Phosphatase-Like Protein IA-2 Autoantibodies in Human Serum

    PubMed Central

    Kikkas, Ingrid; Mallone, Roberto; Larger, Etienne; Volland, Hervé; Morel, Nathalie

    2014-01-01

    Type 1 diabetes (T1D) results from the destruction of pancreatic insulin-producing beta cells and is strongly associated with the presence of islet autoantibodies. Autoantibodies to tyrosine phosphatase-like protein IA-2 (IA-2As) are considered to be highly predictive markers of T1D. We developed a novel lateral flow immunoassay (LFIA) based on a bridging format for the rapid detection of IA-2As in human serum samples. In this assay, one site of the IA-2As is bound to HA-tagged-IA-2, which is subsequently captured on the anti-HA-Tag antibody-coated test line on the strip. The other site of the IA-2As is bound to biotinylated IA-2, allowing the complex to be visualized using colloidal gold nanoparticle-conjugated streptavidin. For this study, 35 serum samples from T1D patients and 44 control sera from non-diabetic individuals were analyzed with our novel assay and the results were correlated with two IA-2A ELISAs. Among the 35 serum samples from T1D patients, the IA-2A LFIA, the in-house IA-2A ELISA and the commercial IA-2A ELISA identified as positive 21, 29 and 30 IA-2A-positive sera, respectively. The major advantages of the IA-2A LFIA are its rapidity and simplicity. PMID:25047039

  19. Determination of the Interday and Intraday Reliability of Serum Cartilage Oligomeric Matrix Protein in a Physically Active Population

    PubMed Central

    Hoch, Johanna M.; Mattacola, Carl G.; Medina-McKeon, Jennifer M.; Shah, Jay N.; Lattermann, Christian

    2011-01-01

    Objective: To determine the intraday and interday reliability of serum cartilage oligomeric matrix protein (sCOMP) in a physically active population with no history of lower extremity surgery. Design: A repeated-measures reliability study was employed to determine the intraday and interday reliability of sCOMP in a physically active cohort. A total of 23 subjects were recruited to the laboratory on 3 separate occasions for nonfasting serum collection. Subjects had no history of lower extremity surgery and were free from acute injury within the last 3 months. Results: Our results indicate strong reliability for both intraday intraclass correlation coefficient (ICC) (0.76) and interday ICC (0.74) sCOMP values. Conclusion: Our results demonstrate that following 30 minutes of inactivity, nonfasting serum samples remain stable over the course of 1 day and between 2 consecutive days in a healthy population with no history of lower extremity surgery. Future research studies are needed to further investigate the magnitude of change in this biomarker for patients with acute articular cartilage damage to determine its appropriateness for use in this population and for varying degrees of articular cartilage severity.

  20. The effects of temperature on thyroid hormone binding to serum proteins in sea turtles

    E-print Network

    Haynes, Shane Patrick

    1990-01-01

    ) Chelonia ~m das 1 4. 89 ~te ideche1 ~ki 1 6, 29 Caret ta ~carett 1 7. 81 4. 73 2. 48 0. 604 ~trace ~acri ta 1 Homo ~sa iens 83. 9 255. 7 8. 43 0. 188 0. 0484 0. 665 56 Table 6, Scatchard analysis of 3, 3', 5-triiodothyronine binding to serum...

  1. EFFECTS OF DIETARY CRUDE PROTEIN ON SERUM AND URINE UREA NITROGEN IN FEEDLOT STEERS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We evaluated the effects of dietary CP concentration and source on serum urea N (SUN) and urine urea N (UUN). A metabolism trial with three collection periods (approximately d 35, 95, and 155 on feed) was conducted using twenty seven crossbred steers (average BW = 353.2 ± 8.4 kg). Treatments were ...

  2. Serum Proteins and Alkaline Phosphatase Levels in Patients with Tuberous Sclerosis

    ERIC Educational Resources Information Center

    Fischer, M. H.; And Others

    1974-01-01

    Six 4- to 37-year-old patients with tuberosis sclerosis (a chronic condition characterized by siezures, intercranial calcification, a reddish-yellow sebaceous glandular mass on the face, and frequent crises in early years), did not exhibit an elevation of the (alpha + beta) globulin fraction in their serum. (Author/MC)

  3. Prognostic Significance of Initial Serum Albumin and 24 Hour Daily Protein Excretion before Treatment in Multiple Myeloma

    PubMed Central

    Chen, Jia-Hong; Hsu, Shun-Neng; Huang, Tzu-Chuan; Wu, Yi-Ying; Lin, Chin; Chang, Ping-Ying; Chen, Yeu-Chin; Ho, Ching-Liang

    2015-01-01

    Renal failure is a common morbidity in multiple myeloma (MM). Although proteinuria has been increasingly reported in malignancies, it is not routinely used to refine risk estimates of survival outcomes in patients with MM. Here we aimed to investigate initial serum albumin and 24-hour daily protein excretion (24-h DPE) before treatment as prognostic factors in patients with MM. We conducted a retrospective analysis of 102 patients with myeloma who were ineligible for haematopoietic stem cell transplantation between October 2000 and December 2012. Initial proteinuria was assessed before treatment by quantitative analysis of 24-hour urine samples. The demographic and laboratory characteristics, survival outcome, and significance of pre-treatment 24-h DPE and albumin in the new staging system of MM were analyzed. Pre-treatment proteinuria (>300 mg/day) was present in 66 patients (64.7%). The optimal cut-off value of 24-h DPE before treatment was 500 mg/day. Analysis of the time-dependent area under the curve showed that the serum albumin and 24-h DPE before treatment were better than 24-h creatinine clearance rate and ?2-microglobulin. A subgroup analysis showed that an initial excess proteinuria (24-h DPE ? 500 mg) was associated with poor survival status (17.51 vs. 34.24 months, p = 0.002). Furthermore, initial serum albumin was an independent risk factor on multivariate analysis (<2.8 vs. ?2.8, hazard ratio = 0.486, p = 0.029). Using the A-DPE staging system, there was a significant survival difference among patients with stage I, II, and III MM (p < 0.001). Initial serum albumin and 24-h DPE before treatment showed significant prognostic factors in patients with MM, and the new A-DPE staging system may be utilized instead of the International Staging System. Its efficacy should be evaluated by further large prospective studies. PMID:26053837

  4. Effect of serum growth factors on intracellular protein degradation in cell culture 

    E-print Network

    Bodner, John Bernard

    1980-01-01

    of the degradative process to EGF was observed in a human fibroblast line. The inhibirory action of these factors on protein degradation is additive, not synergistic, at low concentrations Studies of protein turnover in a selective normal and trans- formed cell... synthesis 18 21 RESULTS AND DISCUSSION 23 Inhibition of intracellular protein degradation Comparative studies of protein synthesis and degradation 23 40 Receptor mediation of response CONCLUSION REFERENCES APPENDIX I APPENDIX II VITA 53 68...

  5. N-Glycomic Changes in Serum Proteins in Type 2 Diabetes Mellitus Correlate with Complications and with Metabolic Syndrome Parameters

    PubMed Central

    Bonfigli, Anna Rita; Boemi, Massimo; Olivieri, Fabiola; Ceriello, Antonio; Genovese, Stefano; Spazzafumo, Liana; Borelli, Vincenzo; Bacalini, Maria Giulia; Salvioli, Stefano; Garagnani, Paolo; Dewaele, Sylviane; Libert, Claude; Franceschi, Claudio

    2015-01-01

    Background Glycosylation, i.e the enzymatic addition of oligosaccharides (or glycans) to proteins and lipids, known as glycosylation, is one of the most common co-/posttranslational modifications of proteins. Many important biological roles of glycoproteins are modulated by N-linked oligosaccharides. As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. Methods We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). N-glycome was evaluated in serum glycoproteins. Results We found significant changes in N-glycan composition in the sera of T2DM patients. In particular, ?(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) were significantly reduced in T2DM compared with CTR subjects. Importantly, they were equally reduced in diabetic patients with and without complications (P<0.001) compared with CTRs. Macro vascular-complications were found to be related with decreased levels of NG1(6)A2F. In addition, NG1(6)A2F and NG1(3)A2F, identifying, respectively, monogalactosylated N-glycans with ?(1,6)- and ?(1,3)-antennary galactosylation, resulted strongly correlated with most MS parameters. The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme “unhealthy” phenotype (T2DM+ with MS). Conclusions Imbalance of glycosyltransferases, glycosidases and sugar nucleotide donor levels is able to cause the structural changes evidenced by our findings. Serum N-glycan profiles are thus sensitive to the presence of diabetes and MS. Serum N-glycan levels could therefore provide a non-invasive alternative marker for T2DM and MS. PMID:25793407

  6. Selective binding of naphthoquinone derivatives to serum albumin proteins and their effects on cytotoxicity.

    PubMed

    Jali, Bigyan R; Kuang, Yuting; Neamati, Nouri; Baruah, Jubaraj B

    2014-05-01

    Naphthoquinone derivatives such as lapachol, plumbagin, dichloroallyl lawsone show anticancer activity and generally cytotoxicity measurements are carried out in presence of bovine serum albumin; so understanding on the ability of serum albumin binding with such derivatives are essential. We have investigated cytotoxicity and serum albumin binding of a series of structurally related naphthoquinone derivatives. Substrate dependency and high selectivity in binding of naphthoquinone tethered carboxylic acids or pyridines with bovine serum albumin (BSA) and human serum albumin (HSA) are observed. For example, the binding constant of BSA with 3-(1,4-dihydro-2-methyl-1,4-dioxonaphthalen-3yl-thio)propanoic acid is ?594 times higher than 3-(1,4-dioxo-1,4-dihydronaphthalen-2-yl-amino)benzoic acid; whereas 4-(1,4-dioxo-1,4-dihydronaphthalen-2-yl-amino)benzoic acid shows ?367 times higher binding constant than the latter compound. The BSA weakly bind to pyridine tethered naphthoquinones, whereas HSA does not binds with them. The binding constant of HSA with 2-(1,4-dihydro-2-methyl-1,4-dioxonaphthalene-3-ylthio)benzoic acid is 134 times higher than the HSA binding constant with 2,2'-(1,4-dihydro-1,4-dioxo-naphthalen-2,3-diylthio)dipropanoic acid. Among the naphthoquinone carboxylic acids, the 3-(1,4-dioxo-1,4-dihydronaphthalen-2-yl-amino)benzoic acid binds selectively to BSA, but it does not bind to HSA. The 2-hydroxybenzoic acid or 4-mercaptobenzoic acid strongly binds to BSA. The binding of BSA with 4-hydroxybenzoic acid or 2-mercaptobenzoic acid are insignificant. We have not observed clear relationships of structure of naphthoquinone derivatives versus serum albumin binding, but could identify the compound having the best IC50 values of cytotoxicity among the twelve naphthoquinone compounds. The compound 3-(1,2-dihydro-1,2-dioxonaphthalen-4-yl-thio)propanoic acid in four cancer cell lines has IC50 values in the range 2.7-7.6?M. This compound also has optimum binding constant with BSA (35.042×10(3)Lmol(-1)) or HSA (21.427×10(3)Lmol(-1)). The cytotoxicity values of the compounds were influenced by concentration of BSA. PMID:24560625

  7. A novel protein from the serum of Python sebae, structurally homologous with type-? phospholipase A(2) inhibitor, displays antitumour activity.

    PubMed

    Donnini, Sandra; Finetti, Federica; Francese, Simona; Boscaro, Francesca; Dani, Francesca R; Maset, Fabio; Frasson, Roberta; Palmieri, Michele; Pazzagli, Mario; De Filippis, Vincenzo; Garaci, Enrico; Ziche, Marina

    2011-12-01

    Cytotoxic and antitumour factors have been documented in the venom of snakes, although little information is available on the identification of cytotoxic products in snake serum. In the present study, we purified and characterized a new cytotoxic factor from serum of the non-venomous African rock python (Python sebae), endowed with antitumour activity. PSS (P. sebae serum) exerted a cytotoxic activity and reduced dose-dependently the viability of several different tumour cell lines. In a model of human squamous cell carcinoma xenograft (A431), subcutaneous injection of PSS in proximity of the tumour mass reduced the tumour volume by 20%. Fractionation of PSS by ion-exchange chromatography yielded an active protein fraction, F5, which significantly reduced tumour cell viability in vitro and, strikingly, tumour growth in vivo. F5 is composed of P1 (peak 1) and P2 subunits interacting in a 1:1 stoichiometric ratio to form a heterotetramer in equilibrium with a hexameric form, which retained biological activity only when assembled. The two peptides share sequence similarity with PIP {PLI-? [type-? PLA(2) (phospholipase A(2)) inhibitor] from Python reticulatus}, existing as a homohexamer. More importantly, although PIP inhibits the hydrolytic activity of PLA(2), the anti-PLA(2) function of F5 is negligible. Using high-resolution MS, we covered 87 and 97% of the sequences of P1 and P2 respectively. In conclusion, in the present study we have identified and thoroughly characterized a novel protein displaying high sequence similarity to PLI-? and possessing remarkable cytotoxic and antitumour effects that can be exploited for potential pharmacological applications. PMID:21834793

  8. Centromere protein F includes two sites that couple efficiently to depolymerizing microtubules.

    PubMed

    Volkov, Vladimir A; Grissom, Paula M; Arzhanik, Vladimir K; Zaytsev, Anatoly V; Renganathan, Kutralanathan; McClure-Begley, Tristan; Old, William M; Ahn, Natalie; McIntosh, J Richard

    2015-06-22

    Firm attachments between kinetochores and dynamic spindle microtubules (MTs) are important for accurate chromosome segregation. Centromere protein F (CENP-F) has been shown to include two MT-binding domains, so it may participate in this key mitotic process. Here, we show that the N-terminal MT-binding domain of CENP-F prefers curled oligomers of tubulin relative to MT walls by approximately fivefold, suggesting that it may contribute to the firm bonds between kinetochores and the flared plus ends of dynamic MTs. A polypeptide from CENP-F's C terminus also bound MTs, and either protein fragment diffused on a stable MT wall. They also followed the ends of dynamic MTs as they shortened. When either fragment was coupled to a microbead, the force it could transduce from a shortening MT averaged 3-5 pN but could exceed 10 pN, identifying CENP-F as a highly effective coupler to shortening MTs. PMID:26101217

  9. Protein signature-based estimation of metagenomic abundances including all domains of life and viruses

    PubMed Central

    Klingenberg, Heiner; Aßhauer, Kathrin Petra; Lingner, Thomas; Meinicke, Peter

    2013-01-01

    Motivation: Metagenome analysis requires tools that can estimate the taxonomic abundances in anonymous sequence data over the whole range of biological entities. Because there is usually no prior knowledge about the data composition, not only all domains of life but also viruses have to be included in taxonomic profiling. Such a full-range approach, however, is difficult to realize owing to the limited coverage of available reference data. In particular, archaea and viruses are generally not well represented by current genome databases. Results: We introduce a novel approach to taxonomic profiling of metagenomes that is based on mixture model analysis of protein signatures. Our results on simulated and real data reveal the difficulties of the existing methods when measuring achaeal or viral abundances and show the overall good profiling performance of the protein-based mixture model. As an application example, we provide a large-scale analysis of data from the Human Microbiome Project. This demonstrates the utility of our method as a first instance profiling tool for a fast estimate of the community structure. Availability: http://gobics.de/TaxyPro. Contact: pmeinic@gwdg.de Supplementary information: Supplementary Material is available at Bioinformatics online. PMID:23418187

  10. The relationship between serum levels of lipoprotein(a) and proteins associated with the acute phase response.

    PubMed

    Ledue, T B; Neveux, L M; Palomaki, G E; Ritchie, R F; Craig, W Y

    1993-12-31

    The association of serum lipoprotein(a) (Lp(a)) with inflammation was investigated in a primarily rheumatologic study group (n = 570; 202 males and 368 females) by studying the relationship between serum levels of Lp(a) and a panel of acute phase proteins (C-reactive protein (CRP), alpha 1-antitrypsin (AAT), alpha 1-acid glycoprotein (AGP), haptoglobin (HPT), complement components 3 and 4 (C3, C4), prealbumin (PAL), albumin (ALB) and transferrin (TRF)). Lp(a) data were adjusted for age and sex, but not clinical condition as no significant differences in Lp(a) levels were observed, using analysis of variance, among the 15 diagnostic categories in the study group. Univariate analyses revealed significant positive associations between Lp(a) levels and levels of C4, AGP, C3 and HPT. Multivariate analysis revealed that C4 and AGP (in descending order of significance) were significant independent predictors of Lp(a) concentration, together accounting for 2.9% of the variability in Lp(a) concentration in the present study group. The data indicate that confounding effects of an acute phase response should be considered in epidemiologic studies, if a high prevalence of inflammation is suspected. PMID:7511489

  11. Short communication: serum immunoglobulin G and total protein concentrations in dairy calves fed a colostrum-replacement product.

    PubMed

    Fidler, A P; Alley, M L; Smith, G W

    2011-07-01

    Neonatal calf health is largely dependent on the ingestion and absorption of maternally derived antibodies via colostrum administration. The objective of this study was to evaluate the efficacy of a commercially available plasma-derived colostrum-replacement (CR) product as compared with bovine colostrum. Holstein calves were removed from the dam immediately after birth and randomly allocated to 1 of 3 groups. Group 1 calves (n=22) were fed 1 package of the CR product; group 2 calves (n=22) were fed 2 packages of the CR product; and group 3 calves (n=22) were fed 3 L of bovine colostrum. Blood samples were collected from all calves 24h after colostrum or CR feeding and analyzed for serum IgG and total protein concentrations. Calves fed bovine colostrum had significantly higher serum IgG and total protein concentration than calves in either group fed the CR product. Group 1 calves (1 package of CR product) had a significantly higher incidence of failure of transfer of passive immunity than calves in groups 2 or 3. The results of this study indicated that 2 packages of this CR product achieved adequate IgG concentrations in calves. However, calves fed 1 package of this CR product consistently had failure of transfer of passive immunity. PMID:21700049

  12. Quantification of two basic proteins related to a thymic hormone (LSH) and their occurrence in serum of normal and cancerous adults.

    PubMed

    Luckey, T D; Venugopal, B; LeGrand, R; Militzer, E; Say, C

    1976-08-01

    A quantitative assay for a thymic hormone (LSHr) was developed and utilized for biologic materials. The protein was isolated from tissue extracts or biologic fluids by precipitation with 20% (NH4)2SO4 and reprecipitation with 75% methanol with subsequent disc-electrophoresis on polyacrylamide gel. The proteins on the gel were stained with amido black. One of the two protein bands was readily identified with LSHr by its electrophoretic mobility and agar immunodiffusion tests. Comparison of the intensity of the protein bands obtained from biologic tissues with those of graded amounts of the pure LSHr allowed quantitative estimation. Another basic protein of the serum is found along with LSHr by the above fractionation procedure; it was tentatively named GEM 126 on the basis of its electrophoretic mobility. The total quantity of each of these serum basic proteins and their ratios to each other showed no correlation to any of 20 clinical and laboratory parameters in either healthy or cancerous adults. The lack of correlation between the quantity of serum protein presumed to be LSHr and cancer susceptibility suggests this thymic hormone is not deficient in patients with cancer. Treatment of female cancer patients with radiation and chemotherapy caused a decrease in serum GEM 126 when compared to healthy adults, and untreated or surgically treated cancer patients. PMID:975024

  13. The fat and protein fractions of freshwater clam ( Corbicula fluminea) extract reduce serum cholesterol and enhance bile acid biosynthesis and sterol excretion in hypercholesterolaemic rats fed a high-cholesterol diet.

    PubMed

    Chijimatsu, Takeshi; Umeki, Miki; Okuda, Yuji; Yamada, Koji; Oda, Hiroaki; Mochizuki, Satoshi

    2011-02-01

    We investigated whether the fat and protein fractions of freshwater clam (Corbicula fluminea) extract (FCE) could ameliorate hypercholesterolaemia in rats fed a high-cholesterol diet. We also explored the mechanism and the components that exert the hypocholesterolaemic effect of FCE. The doses of the fat and protein fractions were equivalent to those in 30 % FCE. The fat and protein fractions of FCE, two major components of FCE, significantly reduced the serum and hepatic cholesterol levels. The fat fraction more strongly reduced serum cholesterol levels than the same level of total FCE. The excretion of faecal neutral sterols increased in rats fed the total the FCE and the fat fraction of FCE. On the other hand, faecal bile acid levels were greater in rats fed the total FCE and the fat and protein fractions of FCE than in control animals. The hepatic gene expression of ATP-binding cassette transporter G5 and cholesterol 7?-hydroxylase was up-regulated by the administration of the total FCE and both the fat and protein fractions of FCE. These results showed that the fat and protein fractions of FCE had hypocholesterolaemic properties, and that these effects were greater with the fat fraction than with the protein fraction. The present study indicates that FCE exerts its hypocholesterolaemic effects through at least two different mechanisms, including enhanced excretion of neutral sterols and up-regulated biosynthesis of bile acids. PMID:20932354

  14. Serum measurements of testosterone, insulin-like growth factor 1, and insulin-like growth factor binding protein-3 in the diagnosis of prostate cancer among Korean men

    Microsoft Academic Search

    Sung Kyu Hong; Byung Kyu Han; Jae Seung Jeong; Seong Jin Jeong; Ki Hyuk Moon; Seok Soo Byun; Sang Eun Lee

    2008-01-01

    Aim:To investigate the relationships of serum testosterone, insulin-like growth factor (IGF)-1 and IGF-binding protein (IGFBP)-3 levels with prostate cancer risk and also with known prognostic parameters of prostate cancer in Korean men who received radical retropubic prostatectomy (RRP) for clinically-localized prostate cancer.Methods:Serum levels of total testosterone, free testosterone, IGF-1 and IGFBP-3 were determined in 592 patients who subsequently received prostate

  15. Longitudinal analysis of serum insulin-like growth factor-I and insulin-like growth factor binding protein-1 in antiphospholipid syndrome and in healthy pregnancy

    Microsoft Academic Search

    Sophia Stone; Kate Langford; Paul T. Seed; Munther A. Khamashta; Beverley J. Hunt; Lucilla Poston

    2003-01-01

    Objective: The purpose of this study was to determine the concentrations of serum insulin-like growth factor-I and insulin-like growth factor binding protein-1 in pregnancies that are complicated with primary antiphospholipid syndrome. Study Design: Longitudinal blood samples were collected from 8 weeks of gestation in 28 women with treated primary antiphospholipid syndrome and 19 control women. Serum insulin-like growth factor-I and

  16. Viscomagnetic Study of Bovine Serum Albumin and Egg Albumin Proteins in Aqueous Solutions

    Microsoft Academic Search

    Man Singh

    2007-01-01

    Intrinsic viscosities [?] for 0.2, 0.4, 0.6, and 0.8 mg% aqueous of each Bovine serum albumin (BSA) and egg albumin (EA) solutions under zero, 0.5 and 0.7 magnetic flux at 298.15 K and 1 atmosphere are reported. The [?] data for BSA are higher than EA and values of both increase with magnetic flux strengthening molecular interactions.

  17. CD4 T Cell Tolerance to Human C-reactive Protein, an Inducible Serum Protein, Is Mediated by Medullary Thymic Epithelium

    PubMed Central

    Klein, Ludger; Klein, Thomas; Rüther, Ulrich; Kyewski, Bruno

    1998-01-01

    Inducible serum proteins whose concentrations oscillate between nontolerogenic and tolerogenic levels pose a particular challenge to the maintenance of self-tolerance. Temporal restrictions of intrathymic antigen supply should prevent continuous central tolerization of T cells, in analogy to the spatial limitation imposed by tissue-restricted antigen expression. Major acute-phase proteins such as human C-reactive protein (hCRP) are typical examples for such inducible self-antigens. The circulating concentration of hCRP, which is secreted by hepatocytes, is induced up to 1,000-fold during an acute-phase reaction. We have analyzed tolerance to hCRP expressed in transgenic mice under its autologous regulatory regions. Physiological regulation of basal levels (<10?9 M) and inducibility (>500-fold) are preserved in female transgenics, whereas male transgenics constitutively display induced levels. Surprisingly, crossing of hCRP transgenic mice to two lines of T cell receptor transgenic mice (specific for either a dominant or a subdominant epitope) showed that tolerance is mediated by intrathymic deletion of immature thymocytes, irrespective of widely differing serum levels. In the absence of induction, hCRP expressed by thymic medullary epithelial cells rather than liver-derived hCRP is necessary and sufficient to induce tolerance. Importantly, medullary epithelial cells also express two homologous mouse acute-phase proteins. These results support a physiological role of “ectopic” thymic expression in tolerance induction to acute-phase proteins and possibly other inducible self-antigens and have implications for delineating the relative contributions of central versus peripheral tolerance. PMID:9653079

  18. Association between serum adipocyte fatty-acid binding protein concentrations, left ventricular function and myocardial perfusion abnormalities in patients with coronary artery disease

    PubMed Central

    2013-01-01

    Background Adipokines, including adipocyte fatty acid-binding protein (A-FABP), have been demonstrated to be involved in the pathogenesis of atherosclerosis. In the present study, we investigated the association of circulating A-FABP level with severity of myocardial perfusion abnormalities analyzed by Tl-201 dipyridamole single-photon emission computed tomography. Methods A total of 170 patients with coronary artery disease (CAD) from cardiovascular clinics were enrolled in the study. Serum A-FABP levels, echocardiography, and stress myocardial perfusion imaging results were analyzed. Results Compared with the patients with mild CAD (summed stress score [SSS] ? 8), those with moderate to severe CAD (SSS > 8) had significantly higher A-FABP concentrations. However, the difference was attenuated in the subgroup of patients with heart failure. In the correlation analyses, A-FABP level was correlated with age, body mass index, waist circumference, levels of creatinine, fasting glucose, high-sensitivity C-reactive protein, N-terminal pro-brain natriuretic peptide, adiponectin, and several echocardiographic parameters, including left ventricular ejection fraction. Multivariate logistic regression analysis demonstrated that the A-FABP level was not only associated with higher SSS (odds ratio, 1.30; 95% confidence interval [CI], 1.01–1.69; P = 0.048), but also an independent risk factor for heart failure (odds ratio 2.71, 95% CI, 1.23–5.94; P = 0.013). Conclusions Serum A-FABP levels not only were associated with myocardial perfusion abnormalities and left ventricular function, but also predicted the presence of heart failure in our patients with CAD. PMID:23866022

  19. High-Level Expression of Single-Chain Fv-Fc Fusion Protein in Serum and Egg White of Genetically Manipulated Chickens by Using a Retroviral Vector

    PubMed Central

    Kamihira, Masamichi; Ono, Ken-ichiro; Esaka, Kazuhisa; Nishijima, Ken-ichi; Kigaku, Ryoko; Komatsu, Hiroyuki; Yamashita, Takashi; Kyogoku, Kenji; Iijima, Shinji

    2005-01-01

    We report here the generation of transgenic chickens using a retroviral vector for the production of recombinant proteins. It was found that the transgene expression was suppressed when a Moloney murine leukemia virus-based retroviral vector was injected into chicken embryos at the blastodermal stage. When a concentrated viral solution was injected into the heart of developing embryos after 50 to 60 h of incubation, transgene expression was observed throughout the embryo, including the gonads. For practical production, a retroviral vector encoding an expression cassette of antiprion single-chain Fv fused with the Fc region of human immunoglobulin G1 (scFv-Fc) was injected into chicken embryos. The birds that hatched stably produced scFv-Fc in their serum and eggs at high levels (?5.6 mg/ml). We obtained transgenic progeny from a transgenic chicken generated with this procedure. The transgene was stably integrated into the chromosomes of transgenic progeny. The transgenic progeny also expressed scFv-Fc in the serum and eggs. PMID:16103139

  20. Spectroscopic probe analysis for exploring probe-protein interaction: a mapping of native, unfolding and refolding of protein bovine serum albumin by extrinsic fluorescence probe.

    PubMed

    Samanta, Anuva; Paul, Bijan Kumar; Guchhait, Nikhil

    2011-07-01

    Steady state and dynamic fluorescence measurements have been used to investigate interaction between Bovine Serum Albumin (BSA) and fluorescence probe para-N,N-dimethylamino orthohydroxy benzaldehyde (PDOHBA), a structurally important molecule exhibiting excited state coupled proton transfer (PT) and charge transfer (CT) reaction. Fluorescence anisotropy, acrylamide quenching, and time resolved fluorescence measurements corroborate the binding nature of the probe with protein. The binding constant between BSA and PDOHBA has been determined by using Benesi-Hildebrand and Stern-Volmer equations. The negative value of ?G indicates the spontaneity of this probe-protein complexation process. Observations from synchronous, three dimensional fluorescence spectra and circular dichroism spectra point toward the fact that the hydrophobicity as well as ?-helix content of BSA are altered in presence of probe PDOHBA. The PT band of PDOHBA is found to be an excellent reporter for the mapping of destructive and protective behavior of SDS with variation of chaotrope concentration. PMID:21514035

  1. [Protein losing enteropathy (PLE) detected by Tc99m-labelled human serum albumin abdominal scintigraphy--case report].

    PubMed

    Hubalewska-Ho?a, Alicja; Sowa-Staszczak, Anna; Szczerbi?ski, Tomasz; Lis, Grzegorz; Huszno, Bohdan; Szybi?ski, Zbigniew

    2003-01-01

    Protein losing enteropathy (PLE) is a gastrointestinal disorder that is associated with excessive loss of plasma protein into the gut resulting from abnormal mucosal permeability. The disease is usually caused by inflammation. The loss of protein in PLE is a nonselective process affecting albumin, globulin and transferrin. Abdominal scintigraphy with human serum albumin marked by Tc99m seems to be an easy and sensitive method for diagnosing PLE. An 4-year-old girl was presented to an outside Pediatric Department due to hypoproteinemia and recurrent pneumonia which had caused several prior hospitalizations. The laboratory tests revealed hypoproteinemia, hypoalbuminemia, low level of IgG, sideropenia, and a decreased level of T lymphocytes. The loss of protein into the gut was confirmed by fecal clearance of alfa-1 antitrypsin. Only nonspecific inflammation was detected by biopsy of the small intestine. These clinical and laboratory findings, quickly decreasing IgG and albumin levels in spite of i.v. supplementation and the lack of proteinuria permitted PLE diagnosis. The abdominal scintigraphy was planned to assess and localise protein losing through GIT and for strategy of possible surgical treatment. Abdominal dynamic scintigraphy was performed immediately after the injection of 300 MBq Tc99m human albumin. 90 images were taken within 180 minutes. Delayed abdominal images were obtained 6 and 24 hours after the tracer injection. Anterior abdominal scintigraphy showed pathological activity of Tc99m-albumin in small bowel in the upper left segment of the abdomen in the 40th minute after injection. Extensive accumulation of albumin was seen in the 160th minute. Delayed images, after 3 and 6 hours, revealed translocation of the tracer into the lower right abdominal segment. The further passage and tracer concentration was detected in ascendant and transverse colon. Based on the laboratory tests and scintigraphic images the girl was suspected to have segmental lymphangiectasia of small intestine and was qualified for laparatomy and, possibly, for surgical resection of the pathologically changed bowel. However, diffuse pathological changes revealed in the guts during the operation, rendered the operation impossible. Tc99m-labelled human serum albumin scintigraphy may be considered the method-of-choice in the diagnosis of protein-losing enteropathy. However, in this test the assessment of the enteropathy dimension is difficult and surgical treatment should be planned with caution. PMID:15065343

  2. IgG antibody against formaldehyde human serum proteins: A comparison with other IgG antibodies against inhalant proteins and reactive chemicals

    SciTech Connect

    Patterson, R.; Dykewicz, M.S.; Evans, R. 3d.; Grammer, L.C.; Greenberger, P.A.; Harris, K.E.; Lawrence, I.D.; Pruzansky, J.J.; Roberts, M.; Shaughnessy, M.A. (Northwestern Univ. Medical School, Chicago, IL (USA))

    1989-09-01

    Immune responses to formaldehyde (F) have been recorded for seven decades. More recently, sensitive assays for antibody against F-human serum albumin (HSA) have been reported. IgG antibody against F-HSA has been said to correlate with symptoms against F-HSA. We report on 61 serum samples analyzed for IgG antibodies against F-HSA. IgG antibodies against F-HSA were most prevalent in subjects who had received intravenous F. In no case (either presumed symptomatic to F or with IgG antibody against F-HSA) was there a correlation of serologic results with symptoms. We also reviewed inhalation disease caused by chemicals and proteins acting as immunogens and report that at this time there is no evidence that gaseous F meets the criteria for causation of inhalational IgG-mediated lung disease by clinical or serologic studies. Very high IgG antibody levels occur in respiratory immunologic inhalational disease, and the absence of these high IgG levels against F is strong evidence against F or F proteins being an inhalational antigen of significance.

  3. C-reactive protein, haptoglobin, serum amyloid A and pig major acute phase protein response in pigs simultaneously infected with H1N1 swine influenza virus and Pasteurella multocida

    PubMed Central

    2013-01-01

    Background Swine influenza (SI) is an acute respiratory disease caused by swine influenza virus (SIV). Swine influenza is generally characterized by acute onset of fever and respiratory symptoms. The most frequent complications of influenza are secondary bacterial pneumonia. The objective of this work was to study the acute phase proteins (APP) responses after coinfection of piglets with H1N1 swine influenza virus (SwH1N1) and Pasteurella multocida (Pm) in order to identify whether the individual APP response correlate with disease severity and whether APP could be used as markers of the health status of coinfected pigs. Results In all coinfected pigs clinical sings, including fever, coughing and dyspnea, were seen. Viral shedding was observed from 2 to 7 dpi. The mean level of antibodies against Pm dermonecrotoxin in infected piglets increase significantly from 7 dpi. Anti-SwH1N1 antibodies in the serum were detected from 7 dpi. The concentration of C-reactive protein (CRP) increased significantly at 1 dpi as compared to control pigs, and remained significantly higher to 3 dpi. Level of serum amyloid A (SAA) was significantly higher from 2 to 3 dpi. Haptoglobin (Hp) was significantly elevated from 3 dpi to the end of study, while pig major acute phase protein (Pig-MAP) from 3 to 7 dpi. The concentrations of CRP, Hp and SAA significantly increased before specific antibodies were detected. Positive correlations were found between serum concentration of Hp and SAA and lung scores, and between clinical score and concentrations of Pig-MAP and SAA. Conclusions The results of current study confirmed that monitoring of APP may revealed ongoing infection, and in this way may be useful in selecting clinically healthy pigs (i.e. before integration into an uninfected herd). Present results corroborated our previous findings that SAA could be a potentially useful indicator in experimental infection studies (e.g. vaccine efficiency investigations) or as a marker for disease severity, because of correlation observed between its concentration in serum and disease severity (lung scores, clinical scores). PMID:23332090

  4. Soybean protein lowers serum cholesterol levels in hamsters: Effect of debittered undigested fraction

    Microsoft Academic Search

    Marites Gatchalian-Yee; Yoko Arimura; Eiko Ochiai; Koji Yamada; Michihiro Sugano

    1997-01-01

    the undigested fraction (UDF) of soybean protein exerts a marked hypocholesterolemic effect in relation to soybean protein (SOY) in rats. The present study was conducted to confirm whether UDF was effective in hamsters as in rats in combination with different fat sources, either perilla oil (PER) or safflower oil (SAF). Because the hamster, unlike the rat, disliked the bitter taste

  5. Separation of ?-Lactoglobulin from Other Milk Serum Proteins by Trichloroacetic Acid

    Microsoft Academic Search

    K. K. Fox; V. H. Holsinger; L. P. Posati; M. J. Pallansch

    1967-01-01

    fl-Lactoglobulin can be easily separated from milk because it is the whey protein most resistant to precipitation by tri- chloroacetie acid. After removing casein from milk by acid precipitation, the re- sidual whey is made up to contain 3% triehloroacetie acid. All proteins other than fl-lactoglobulin precipitate and can be fil- tered. The filtrate is concentrated by nega- tive pressure

  6. Elevated serum insulin-like growth factor (IGF)-II and IGF binding protein-2 in patients with colorectal cancer

    PubMed Central

    Renehan, A G; Jones, J; Potten, C S; Shalet, S M; O'Dwyer, S T

    2000-01-01

    This study explored the relationships of serum insulin-like growth factors, IGF-I and IGF-II, and their binding proteins (IGFBP)-2 and IGFBP-3, with key clinicopathological parameters in 92 patients with colorectal cancer (cases). Comparisons were made with 57 individuals who had a normal colonoscopy (controls). Serial changes were examined in 27 cases. As IGF-related peptides are age- and sex-dependent, absolute concentrations were converted to standard deviation scores (SDS). Mean IGF-II SDS were elevated in Dukes A (n= 12 P< 0.001) and Dukes B (n= 25 P< 0.001) cases compared with controls, but not in advanced disease. Compared with controls, mean IGFBP-2 SDS were significantly elevated in patients with Dukes B (P< 0.001), Dukes C (n= 13 P< 0.001) and advanced disease (n= 42 P< 0.0001), with a significant trend from early to advanced disease (one-way ANOVA P< 0.001). Furthermore, IGFBP-2 SDS were positively related to tumour size (P= 0.01) and fell significantly in patients following curative resection (P= 0.04), suggesting that circulating levels reflect tumour load. We tested the potential tumour marker characteristics of IGFBP-2 SDS against three endpoints: metastasis alone; local pelvic recurrence alone; and metastasis and recurrence combined. The sensitivities for IGFBP-2 alone (? + 2SD) were modest at 55%, 46%, and 52%, but in combination with CEA, increased substantially to 90%, 77% and 86%, respectively. We conclude that the serum IGF-II and IGFBP-2 profiles may provide insights into underlying biological mechanisms, and that serum IGFBP-2 may have an adjunct role in cancer surveillance in patients with colorectal cancer. © 2000 Cancer Research Campaign PMID:11044360

  7. Serum-free hybridoma culture: ethical, scientific and safety considerations

    Microsoft Academic Search

    Megha S. Even; Chad B. Sandusky; Neal D. Barnard

    2006-01-01

    Despite considerable progress in the development of cell culture techniques, including the development of the serum- and protein-free media that now routinely support hybridoma and mammalian cell growth, fetal bovine serum (FBS) supplemented media are still commonly used: a practice that raises ethical, scientific and safety concerns. The use of FBS in hybridoma culture media is examined here, with regards

  8. Serum levels of acute phase proteins: SAA, Hp and progesterone (P4) in mares with early embryonic death.

    PubMed

    Krakowski, L; Krawczyk, C H; Kostro, K; Stefaniak, T; Novotny, F; Obara, J

    2011-08-01

    The study involved 46 healthy purebred Arabian mares exhibiting regular oestrous cycles that underwent artificial insemination (AI). Pregnancy was detected ultrasonographically (US) in 40 mares. In 15 mares in foal, early embryonic death (EED) was observed during the pregnancy days 14-21. Blood for determinations of serum acute phase proteins (SAA and Hp) and progesterone (P4) was sampled 12-24 h before ovulation and the first insemination, at 12, 24, 72, 96 h and on day 7, 10, 14, 21, 35 and 55 after ovulation. The results revealed that in 25 mares without EED, the serum levels of P4, SAA and Hp were within physiological limits; in 15 mares with EED, the levels of SAA and Hp were significantly increased. In seven mares with EED, high levels of SAA and Hp were already found before ovulation and at 12, 24, 72, 96 h as well as on day 7 and 10 post-ovulation, whereas the level of P4 was normal for early pregnancy. In the remaining eight mares with EED, increased levels of SAA and Hp were found at 72 h after ovulation and maintained until day 55. In this group, the level of P4 decreased since 96 h after ovulation. Determinations of SAA, Hp and P4 in mares in early pregnancy (EP) are useful for monitoring normal development of pregnancy and for diagnosis of subclinical genital inflammations, which may lead to EED. PMID:21241377

  9. Interaction of the anticancer gallium(III) complexes of 8-hydroxyquinoline and maltol with human serum proteins.

    PubMed

    Enyedy, Éva A; Dömötör, Orsolya; Bali, Krisztina; Hetényi, Anasztázia; Tuccinardi, Tiziano; Keppler, Bernhard K

    2015-01-01

    Tris(8-quinolinolato)gallium(III) (KP46) and tris(maltolato)gallium(III) (GaM) are promising orally active antitumor metallodrugs currently undergoing clinical trials. Their interaction with human serum albumin (HSA) and transferrin (Tf) was studied in detail in aqueous solution by the combination of various methods such as spectrofluorometry, UV-vis spectrophotometry, (1)H and saturation transfer difference NMR spectroscopy, and ultrafiltration-UV-vis spectrophotometry. Binding data were evaluated quantitatively. Tf was found to replace the original ligand much less efficiently in KP46 than in GaM, whereas a significant noncovalent binding of KP46 with HSA (log K' = 4.04) retaining the coordination environment around gallium(III) was found. The interaction between HSA and KP46 was also confirmed by protein-complex modeling calculations. On the basis of the conditional stability constants, the distribution of gallium(III) in serum was computed and compared for these metallodrugs under physiological conditions, and revealed the prominent role of HSA in the case of KP46 and that of Tf for GaM. PMID:25398250

  10. Detection of hepatitis C virus core protein in serum by atomic force microscopy combined with mass spectrometry

    PubMed Central

    Ivanov, Yuri D; Kaysheva, Anna L; Frantsuzov, Pavel A; Pleshakova, Tatyana O; Krohin, Nikolay V; Izotov, Alexander A; Shumov, Ivan D; Uchaikin, Vasiliy F; Konev, Vladimir A; Ziborov, Vadim S; Archakov, Alexander I

    2015-01-01

    A method for detection and identification of core antigen of hepatitis C virus (HCVcoreAg)-containing particles in the serum was proposed, with due account taken of the interactions of proteotypic peptides with Na+, K+, and Cl? ions. The method is based on a combination of reversible biospecific atomic force microscopy (AFM)-fishing and mass spectrometry (MS). AFM-fishing enables concentration, detection, and counting of protein complexes captured on the AFM chip surface, with their subsequent MS identification. Biospecific AFM-fishing of HCVcoreAg-containing particles from serum samples was carried out using AFM chips with immobilized antibodies against HCVcoreAg (HCVcoreAgim). Formation of complexes between anti-HCVcoreAgim and HCVcoreAg-containing particles on the AFM chip surface during the fishing process was demonstrated. These complexes were registered and counted by AFM. Further MS analysis allowed reliable identification of HCVcoreAg within the complexes formed on the AFM chip surface. It was shown that MS data processing, with account taken of the interactions between HCVcoreAg peptides and Na+, K+ cations, and Cl? anions, allows an increase in the number of peptides identified. PMID:25759582

  11. Recombinant nucleocapsid protein based single serum dilution ELISA for the detection of antibodies to infectious bronchitis virus in poultry.

    PubMed

    Pradhan, Sunil K; Kamble, Nitin M; Pillai, Aravind S; Gaikwad, Satish S; Khulape, Sagar A; Reddy, M R; Mohan, C Madhan; Kataria, Jag Mohan; Dey, Sohini

    2014-12-01

    Avian infectious bronchitis is ubiquitous and highly contagious disease of poultry, with profound effect on commercial poultry production. For effective control of infectious bronchitis virus (IBV), quick and specific diagnosis is of utmost importance. In this study, the virus was isolated from clinical samples from India and the full length nucleocapsid (N) gene was amplified, cloned and expressed in a prokaryotic system. The purified recombinant N protein based single serum dilution enzyme linked immunosorbent assay (ELISA) was developed for IBV to measure specific antibody in the sera of chickens. A total of 310 chicken sera samples were tested using the commercial IDEXX kit along with the assay developed. A linear correlation was obtained between predicted antibody titres at a single working dilution of 1:100 and the corresponding serum titres observed as determined by the standard serial dilution method. Regression analysis was used to construct a standard curve from which an equation was derived which confirmed their correlation. The developed equation was then used to extrapolate predicated ELISA antibody titer from corrected absorbance readings of the single working dilution. The assay proved to be specific (95.8%) and sensitive (96.8%) when compared to the commercial IDEXX ELISA test. PMID:25173423

  12. Detection of the inflammation biomarker C-reactive protein in serum samples: towards an optimal biosensor formula.

    PubMed

    Fakanya, Wellington M; Tothill, Ibtisam E

    2014-12-01

    The development of an electrochemical immunosensor for the biomarker, C-reactive protein (CRP), is reported in this work. CRP has been used to assess inflammation and is also used in a multi-biomarker system as a predictive biomarker for cardiovascular disease risk. A gold-based working electrode sensor was developed, and the types of electrode printing inks and ink curing techniques were then optimized. The electrodes with the best performance parameters were then employed for the construction of an immunosensor for CRP by immobilizing anti-human CRP antibody on the working electrode surface. A sandwich enzyme-linked immunosorbent assay (ELISA) was then constructed after sample addition by using anti-human CRP antibody labelled with horseradish peroxidase (HRP). The signal was generated by the addition of a mediator/substrate system comprised of 3,3,5',5'-Tetramethylbenzidine dihydrochloride (TMB) and hydrogen peroxide (H2O2). Measurements were conducted using chronoamperometry at -200 mV against an integrated Ag/AgCl reference electrode. A CRP limit of detection (LOD) of 2.2 ng·mL(-1) was achieved in spiked serum samples, and performance agreement was obtained with reference to a commercial ELISA kit. The developed CRP immunosensor was able to detect a diagnostically relevant range of the biomarker in serum without the need for signal amplification using nanoparticles, paving the way for future development on a cardiac panel electrochemical point-of-care diagnostic device. PMID:25587427

  13. Detection of follicular fluid and serum antibodies by protein microarrays in women undergoing in vitro fertilization treatment.

    PubMed

    Ardizzoni, Andrea; Manca, Lidia; Capodanno, Francesco; Baschieri, Maria Cristina; Rondini, Ilaria; Peppoloni, Samuele; Righi, Elena; La Sala, Giovanni Battista; Blasi, Elisabetta

    2011-04-01

    A protein microarray serological assay was used to assess the antibody profile of 102 women subjected to in vitro fertilization treatment. The studies were conducted on pairs of serum and follicular fluid samples, collected from each woman on the same day at the time of oocyte recovery. The samples, stored as frozen aliquotes, were assessed by both microarray and ELISA. Follicular fluids and sera were screened to detect the presence of specific IgG and IgM antibodies against seven vertically transmitted pathogens. The IgG reactivity of follicular fluids closely mirrored that of serum in all the patients and for all the antigens, with an agreement of more than 85%. IgM antibodies were undetectable in follicular fluids. The antibody patterns were subsequently related to the biological and clinical outcomes of in vitro fertilization cycles. The results showed that varicella zoster virus (VZV) IgG positive women and cytomegalovirus (CMV) IgG negative women had on average a higher number of inseminated, good quality oocytes compared to VZV IgG negative and CMV IgG positive women. In addition, the rate of successful embryo transfers was significantly higher in Toxoplasma gondii IgG negative women than in their positive counterparts. Overall, the microarray was proven to be a suitable tool for detecting analytes in follicular fluids, therefore supporting its application in a wide spectrum of investigations. PMID:21477867

  14. Diagnostic usefulness of serum interleukin 6 (IL-6) and C-reactive protein (CRP) in the differentiation between pancreatic cancer and chronic pancreatitis.

    PubMed

    Mroczko, Barbara; Groblewska, Magdalena; Gryko, Mariusz; Kedra, Bogus?aw; Szmitkowski, Maciej

    2010-01-01

    Previous studies have shown elevated serum levels of interleukin 6 (IL-6) and C-reactive protein (CRP) in patients with pancreatic cancer (PC). The aim of this study was to assess the diagnostic usefulness of pretreatment serum levels of IL-6 and CRP to differentiate between PC and chronic pancreatitis (CP) patients. Serum levels of CRP, IL-6, carcinoembryonic antigen (CEA), and carbohydrate antigen 19-9 (CA 19-9) were determined in 78 patients with PC before surgery, in 45 patients with CP, and in 70 healthy controls. Serum levels of all the proteins tested were significantly higher in cancer patients when compared with CP and healthy subjects, and increased in more advanced tumor stages. Concentrations of IL-6 were significantly higher in nonresectable tumors and in patients who died during the 2-year observation period. Area under receiver operating characteristic curve for IL-6 was higher than for other substances tested in the differentiation between PC and CP. Cox's univariate analysis revealed serum IL-6 as a significant prognostic factor of patients' survival. Our findings suggest higher diagnostic usefulness of serum IL-6 than CRP, CEA, and CA 19-9 in the diagnosis and prognosis of patients with PC and in the differentiation with CP. PMID:20626020

  15. Nonstructural Protein 2 (NS2) of Respiratory Syncytial Virus (RSV) Detected by an Antipeptide Serum

    Microsoft Academic Search

    Elke Weber; Barbara Humbert; Hans-Jürgen Streckert; Hermann Werchau

    1995-01-01

    The human respiratory syncytial virus (RSV) is often associated with airway obstruction and is suspected to induce bronchial hyperreactivity. Interactions of viral proteins with cellular components may be responsible for epithelial damage leading to bronchial hyperreactivity. In this study, we describe the localization of the 14.7-kD nonstructural protein 2 (NS2) in RSV-infected cells. The detection of NS2 was performed using

  16. Serum Proteomic Signature of Human Chagasic Patients for the Identification of Novel Potential Protein Biomarkers of Disease*

    PubMed Central

    Wen, Jian-Jun; Zago, M. Paola; Nuñez, Sonia; Gupta, Shivali; Burgos, Federico Nuñez; Garg, Nisha Jain

    2012-01-01

    Chagas disease is initiated upon infection by Trypanosoma cruzi. Among the health consequences is a decline in heart function, and the pathophysiological mechanisms underlying this manifestation are not well understood. To explore the possible mechanisms, we employed IgY LC10 affinity chromatography in conjunction with ProteomeLab PF2D and two-dimensional gel electrophoresis to resolve the proteome signature of high and low abundance serum proteins in chagasic patients. MALDI-TOF MS/MS analysis yielded 80 and 14 differentially expressed proteins associated with cardiomyopathy of chagasic and other etiologies, respectively. The extent of oxidative stress-induced carbonyl modifications of the differentially expressed proteins (n = 26) was increased and coupled with a depression of antioxidant proteins. Functional annotation of the top networks developed by ingenuity pathway analysis of proteome database identified dysregulation of inflammation/acute phase response signaling and lipid metabolism relevant to production of prostaglandins and arachidonic acid in chagasic patients. Overlay of the major networks identified prothrombin and plasminogen at a nodal position with connectivity to proteome signature indicative of heart disease (i.e., thrombosis, angiogenesis, vasodilatation of blood vessels or the aorta, and increased permeability of blood vessel and endothelial tubes), and inflammatory responses (e.g., platelet aggregation, complement activation, and phagocyte activation and migration). The detection of cardiac proteins (myosin light chain 2 and myosin heavy chain 11) and increased levels of vinculin and plasminogen provided a comprehensive set of biomarkers of cardiac muscle injury and development of clinical Chagas disease in human patients. These results provide an impetus for biomarker validation in large cohorts of clinically characterized chagasic patients. PMID:22543060

  17. Serum secreted frizzled-related protein 5 levels differentially decrease in patients with hepatitis B virus-associated chronic infection and hepatocellular carcinoma

    PubMed Central

    PENG, CHUAN; XIAO, XIAOQIU; KANG, BING; HE, SONG; LI, JIBIN

    2014-01-01

    The aim of this study was to investigate the characteristics of serum secreted frizzled-related protein 5 (SFRP5), an inhibitor of Wnt signaling, in hepatitis B virus (HBV)-associated infections and hepatocellular carcinoma (HCC) patients. Serum SFRP5 levels were detected in 147 patients with HBV-associated chronic infection or HCC. Compared with the non-HBV-infected and non-HCC group, the HBV-associated chronic infection and HCC groups exhibited decreased serum SFRP5 levels. A significant inverse correlation between serum SFRP5 levels and HBV DNA levels was identified in the HBV-associated chronic infection and HCC groups. Furthermore, SFRP5 levels differentially decreased in patients with HBV-associated diseases, in a manner which was dependent on liver disease status. Compared with patients exhibiting HBV-associated chronic infection, patients with HCC were found to exhibit lower serum SFRP5 levels. The results of the present study indicated that patients with HBV-associated liver infection and HCC exhibited significantly deceased serum SFRP5 levels, which were found to negatively correlate with HBV DNA levels. Serum SFRP5 levels may present a biomarker for the severity of HBV-associated liver infection, and the risk of HCC initiation and progression. PMID:25120720

  18. A 28,000-dalton protein of normal mouse serum binds specifically to the inner core region of bacterial lipopolysaccharide.

    PubMed Central

    Brade, L; Brade, H

    1985-01-01

    Normal mouse serum was found to contain a protein, referred to here as factor, which binds to the inner core region of lipopolysaccharides (LPSs) of various bacterial families. Since factor-LPS interactions resulted in activation of guinea pig complement, factor activity could be assayed by a passive hemolysis test with sheep erythrocytes coated with LPS or lipid A from Acinetobacter calcoaceticus (which was found earlier to bind particularly well to factor). Factor was purified by G-50 and hydroxyapatite chromatography whereby the specific hemolytic activity was enriched 1,675-fold. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions revealed the presence of a 28,000-dalton protein as the main band. The identity of this band was determined by absorption experiments with LPS-coated sheep erythrocytes or latex beads, whereby the 28,000-dalton band disappeared after specific absorption and could be recovered from the absorbent. The binding specificity of factor was determined in a passive hemolysis inhibition assay with defined oligosaccharides representative for the inner core region of LPS. Thus, the di- and trisaccharides alpha-D-mannoheptopyranosyl-(1----5)-2-keto-3-deoxy-D-mannoocto nic acid and alpha-D-mannoheptopyranosyl-(1----3)-alpha-D-mannoheptopyranosy l-(1----5)-2- keto-3-deoxy-D-mannooctonic acid, respectively, were able to inhibit binding of factor to LPS. The results are in accordance with our earlier observation that the heptose-2-keto-3-deoxy-D-mannooctonic acid region represents a common antigen of bacterial LPS. Rabbit hyperimmune serum directed against this common antigen and purified factor was found to exhibit the same specificity for LPS. Factor activity was followed in mice in vivo after injection of LPS; it disappeared completely 15 min after the injection of LPS and reappeared within 1 h. Images PMID:4066028

  19. Comparative examination of adsorption of serum proteins on HSA- and PLGA-based nanoparticles using SDS-PAGE and LC-MS.

    PubMed

    Gossmann, R; Fahrländer, E; Hummel, M; Mulac, D; Brockmeyer, J; Langer, K

    2015-06-01

    The behavior of nanosized drug carrier systems under cell culture conditions and therefore also the destiny in the body are highly influenced by the protein corona, which is formed upon entering a biological environment. Some of the adsorbed proteins, named opsonins, lead to a shortened plasma circulation half-life of the nanoparticles. Others are attributed to promote the transport of nanoparticles into other compartments of the body, just to mention two examples. Hence, detailed knowledge concerning the composition of the protein corona is of great importance. The aim of this work was to investigate the influence of the nanoparticle starting material and the surface modification on the composition of the adsorbed serum proteins in a cell culture environment. Therefore, positively charged nanoparticles based on the biodegradable polymer poly(dl-lactide-co-glycolide) (PLGA) stabilized with didodecyldimethylammonium bromide (DMAB) and negatively charged nanoparticles based on human serum albumin (HSA) were prepared and modified with hydrophilic polymers. By incubating the nanoparticles with fetal bovine serum (FBS) the adsorption of serum proteins on the colloidal system was investigated. Using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) a semi-quantitative analysis of the protein corona was performed and after enzymatic in-solution-digestion the adsorbed proteins were identified using high resolution LC-MS. Our study accentuates the influence of the core material, surface charge, and surface modification on the amount and nature of the adsorbed proteins. The combination of SDS-PAGE and LC-MS turns out to be a simple and reliable method to investigate the protein corona of nanoparticles. PMID:25813886

  20. Concentrations of serum constituents in cold-stressed calves from heifers fed inadequate protein and(or) energy.

    PubMed

    Bull, R C; Everson, D O; Olson, D P; Kelley, K W; Curtis, S; Tzou, G

    1991-02-01

    A study with neonatal calves was conducted to determine the effects of maternal crude protein (CP) and(or) metabolizable energy (ME) malnutrition, cold stress (0 or 21 degrees C), and age on concentrations of selected serum constituents. For each of 2 yr, 60 artificially bred Angus heifers were assigned randomly to a 2 x 2 factorial nutritional plan 150 d before predicted parturition. The diets provided each heifer with either .32 or .96 kg/d of CP and 8.7 or 12.6 Mcal/d of ME. Blood samples were obtained from heifers at parturition and from their calves at birth and at 12, 24, 36, 48, and 72 h of age. Sera were analyzed for concentrations of blood urea nitrogen (BUN), creatinine (Creat), iron, total protein (TProt), alkaline phosphatase (AlkPhos), total bilirubin (TBil), and cholesterol (Chol). Mean correlations of these constituents in calf sera between 12-h adjacency intervals were high, but those between longer times (48 or 60 h) were low. Simple correlations of serum constituents between cows and calves at birth were low except for BUN (r = .578 and .295 for yr 1 and 2, respectively). There were significant main treatment effects for maternal CP consumption on BUN levels, for environmental temperature on BUN, Creat, and TBil levels, and for years on BUN, Creat, iron, and AlkPhos levels in calves. Significant polynomial relationships were found over hours of age for all variables. Blood urea N decreased in normal calves but remained relatively constant at a low level in deficient calves. Year x hour of age interactions occurred for iron, TProt, AlkPhos, TBil, and Chol. Protein x year x hour of age interactions were found for iron and Chol. These results suggest that random sampling times are not useful for decision making during the first 72 h after birth. Consideration must be given to multiple samples taken at specific calf ages, to environmental temperatures, and to maternal protein nutritional levels when interpreting calf blood sera data. PMID:1901848

  1. Characterization of cDNA clones encoding rabbit and human serum paraoxonase: the mature protein retains its signal sequence.

    PubMed

    Hassett, C; Richter, R J; Humbert, R; Chapline, C; Crabb, J W; Omiecinski, C J; Furlong, C E

    1991-10-22

    Serum paraoxonase hydrolyzes the toxic metabolites of a variety of organophosphorus insecticides. High serum paraoxonase levels appear to protect against the neurotoxic effects of organophosphorus substrates of this enzyme [Costa et al. (1990) Toxicol. Appl. Pharmacol. 103, 66-76]. The amino acid sequence accounting for 42% of rabbit paraoxonase was determined by (1) gas-phase sequencing of the intact protein and (2) peptide fragments from lysine and arginine digests. From these data, two oligonucleotide probes were synthesized and used to screen a rabbit liver cDNA library. A clone was isolated and sequenced, and contained a 1294-bp insert encoding an open reading frame of 359 amino acids. Northern blot hybridization with RNA isolated from various rabbit tissues indicated that paraoxonase mRNA is synthesized predominately, if not exclusively, in the liver. Southern blot experiments suggested that rabbit paraoxonase is coded by a single gene and is not a family member of closely related genes. Human paraoxonase clones were isolated from a liver cDNA library by using the rabbit cDNA as a hybridization probe. Inserts from three of the longest clones were sequenced, and one full-length clone contained an open reading frame encoding 355 amino acids, four less than the rabbit paraoxonase protein. Each of the human clones appeared to be polyadenylated at a different site, consistent with the absence of the canonical polyadenylation signal sequence. Of potential significance with respect to the paraoxonase polymorphism, the derived amino acid sequence from one of the partial human cDNA clones differed at two positions from the full-length clone. Amino-terminal sequences derived from purified rabbit and human paraoxonase proteins suggested that the signal sequence is retained, with the exception of the initiator methionine residue [Furlong et al. (1991) Biochemistry (preceding paper in this issue)]. Characterization of the rabbit and human paraoxonase cDNA clones confirms that the signal sequences are not processed, except for the N-terminal methionine residue. The rabbit and human cDNA clones demonstrate striking nucleotide and deduced amino acid similarities (greater than 85%), suggesting an important metabolic role and constraints on the evolution of this protein. PMID:1657140

  2. Soybean protein lowers serum cholesterol levels in hamsters: effect of debittered undigested fraction.

    PubMed

    Gatchalian-Yee, M; Arimura, Y; Ochiai, E; Yamada, K; Sugano, M

    1997-01-01

    The undigested fraction (UDF) of soybean protein exerts a marked hypocholesterolemic effect in relation to soybean protein (SOY) in rats. The present study was conducted to confirm whether UDF was effective in hamsters as in rats in combination with different fat sources, either perilla oil (PER) or safflower oil (SAF). Because the hamster, unlike the rat, disliked the bitter taste of UDF, the effect of debittering UDF also was studied. Cholesterol-enriched (0.2%) diets containing 20% protein and 10% fat were fed to hamsters for 4 wk. UDF was more hypocholesterolemic than soybean protein in hamsters regardless of the dietary fat source. The ratio of high-density lipoprotein to total cholesterol essentially remained unchanged. The debittered UDF, without influencing food intake and thereby weight gain, exerted a significant hypocholesterolemic effect comparable with UDF in relation to SOY accompanying stimulation of fecal neutral and acidic steroid excretion. The fatty acid composition of liver phospholipids was influenced by the type of the dietary protein, and both UDF seemed to interfere characteristically more than SOY with the desaturation systems between linoleic acid and arachidonic acid. Thus, the debittered UDF, similar to UDF, exerted a distinct influence on the various parameters of lipid metabolism in relation to SOY. PMID:9263255

  3. Multiplexed Activity-based Protein Profiling of the Human Pathogen Aspergillus fumigatus Reveals Large Functional Changes upon Exposure to Human Serum

    SciTech Connect

    Wiedner, Susan D.; Burnum, Kristin E.; Pederson, Leeanna M.; Anderson, Lindsey N.; Fortuin, Suereta; Chauvigne-Hines, Lacie M.; Shukla, Anil K.; Ansong, Charles; Panisko, Ellen A.; Smith, Richard D.; Wright, Aaron T.

    2012-08-03

    Environmental and metabolic adaptability is critical for survival of the fungal human pathogen Aspergillus fumigatus in the immunocompromised lung. We employed an activity-based protein profiling (ABPP) approach utilizing a new aryl vinyl sulfonate probe and a serine hydrolase probe combined with quantitative LC-MS based accurate mass and time (AMT) tag proteomics for the identification of functional pathway adaptation of A. fumigatus to environmental variability relevant to pulmonary Invasive Aspergillosis. When the fungal pathogen was grown with human serum, metabolism and energy processes were markedly decreased compared to no serum culture. Additionally, functional pathways associated with amino acid and protein biosynthesis were limited as the fungus scavenged from the serum to obtain essential nutrients. Our approach revealed significant metabolic adaptation by A. fumigatus, and provides direct insight into this pathogen’s ability to survive and proliferate.

  4. Animal protein intake, serum insulin-like growth factor I, and growth in healthy 2.5-y-old Danish children1-3

    Microsoft Academic Search

    Camilla Hoppe; Tina Rovenna Udam; Lotte Lauritzen; Christian Mølgaard; Anders Juul; Kim Fleischer Michaelsen

    Background: Studies from developing countries indicate that in- take of animal protein, especially of milk, is associated with greater velocity of linear growth in childhood. Whether the same association exists in industrialized countries, where protein intake is high, is not clear. Objective: Our objective was to examine associations between pro- tein intake, serum insulin-like growth factor I (sIGF-I) concentra- tions,

  5. Demonstration of membrane estrogen binding proteins in rat brain by ligand blotting using a 17?-estradiol-[ 125I]bovine serum albumin conjugate

    Microsoft Academic Search

    Jianbiao Zheng; Victor D. Ramirez

    1997-01-01

    This paper describes a ligand blotting procedure to visualize membrane estrogen receptors\\/binding proteins immobilized on nitrocellulose membranes. Using 17?-estradiol covalently linked with [125I]bovine serum albumin (BSA) at the C-6 position (17?-E-6-[125I]BSA) as a ligand, three major binding proteins with molecular masses of approximately 23, 28, and 32 kDa were identified from crude synaptosomal fractions (P2) of female rat brains. The

  6. Matrix-Assisted Laser Desorption\\/Ionization Time-of-Flight Mass Spectrometry Combined with Magnetic Beads for Detecting Serum Protein Biomarkers in Parkinson’s Disease

    Microsoft Academic Search

    Yao-Hua Li; Jian Wang; Xiao-Li Zheng; Yan-Li Zhang; Xin Li; Shun Yu; Xin He; Piu Chan

    2011-01-01

    Background: Biomarkers for neurodegenerative diseases are essential to facilitate disease diagnosis. Application of proteomics has greatly hastened the search for novel biomarkers. In this study, new potential biomarkers were discovered, and a diagnostic pattern was established for idiopathic Parkinson’s disease (PD) by using proteomic technology. Methods: Serum proteins from PD patients and controls were captured by magnetic bead-based weak cation

  7. Low serum levels of ApoE associate with hippocampal atrophy in the ADNI cohort We analyzed the imaging and plasma protein biomarker data from

    E-print Network

    Thompson, Paul

    Low serum levels of ApoE associate with hippocampal atrophy in the ADNI cohort Methods: We analyzed 244 were apolipoprotein E4 gene (APOE4) carriers and 231 APOE4 non-carriers. Baseline EDTA plasma (ApoE), apolipoprotein J (ApoJ), brain derived neurotrophic factor (BDNF), heat shock protein 40 (HSP40

  8. Antibody-free, targeted mass-spectrometric approach for quantification of proteins at low picogram per milliliter levels in human plasma/serum

    SciTech Connect

    Shi, Tujin; Fillmore, Thomas L.; Sun, Xuefei; Zhao, Rui; Schepmoes, Athena A.; Hossain, Mahmud; Xie, Fang; Wu, Si; Kim, Jong Seo; Jones, Nathaniel J.; Moore, Ronald J.; Pasa-Tolic, Ljiljana; Kagan, Jacob; Rodland, Karin D.; Liu, Tao; Tang, Keqi; Camp, David G.; Smith, Richard D.; Qian, Weijun

    2012-09-18

    The detection and quantification of proteins at sub-ng/mL concentrations in plasma/serum is of critical importance for candidate biomarker verification. Sensitive detection of serum proteins has typically been achieved by immunoassays; however, de novo development of antibodies is associated with high cost and long lead time. To address this challenge, we developed an antibody-free strategy, termed high-pressure high-resolution separations with intelligent selection and multiplexing (PRISM), for achieving accurate detection of proteins at ~50 pg/mL level in plasma/serum using selected reaction monitoring (SRM). This strategy is based on high resolution reversed phase liquid chromatographic separations for analyte enrichment along with intelligent selection of target fractions via on-line SRM monitoring of internal standards and fraction multiplexing prior to SRM quantification. Our work represents a major technological advance for achieving pg/mL level of serum protein quantification without specific affinity reagents and holds great promise for broad applications in biomarker verification and systems biology studies.

  9. Multiple phosphorylated forms of the Saccharomyces cerevisiae Mcm1 protein include an isoform induced in response to high salt concentrations.

    PubMed Central

    Kuo, M H; Nadeau, E T; Grayhack, E J

    1997-01-01

    The Saccharomyces cerevisiae Mcm1 protein is an essential multifunctional transcription factor which is highly homologous to human serum response factor. Mcm1 protein acts on a large number of distinctly regulated genes: haploid cell-type-specific genes, G2-cell-cycle-regulated genes, pheromone-induced genes, arginine metabolic genes, and genes important for cell wall and cell membrane function. We show here that Mcm1 protein is phosphorylated in vivo. Several (more than eight) isoforms of Mcm1 protein, resolved by isoelectric focusing, are present in vivo; two major phosphorylation sites lie in the N-terminal 17 amino acids immediately adjacent to the conserved MADS box DNA-binding domain. The implications of multiple species of Mcm1, particularly the notion that a unique Mcm1 isoform could be required for regulation of a specific set of Mcm1's target genes, are discussed. We also show here that Mcm1 plays an important role in the response to stress caused by NaCl. G. Yu, R. J. Deschenes, and J. S. Fassler (J. Biol. Chem. 270:8739-8743, 1995) showed that Mcm1 function is affected by mutations in the SLN1 gene, a signal transduction component implicated in the response to osmotic stress. We find that mcm1 mutations can confer either reduced or enhanced survival on high-salt medium; deletion of the N terminus or mutation in the primary phosphorylation site results in impaired growth on high-salt medium. Furthermore, Mcm1 protein is a target of a signal transduction system responsive to osmotic stress: a new isoform of Mcm1 is induced by NaCl or KCl; this result establishes that Mcm1 itself is regulated. PMID:9001236

  10. Introducing Proteomics in the Undergraduate Curriculum: A Simple 2D Gel Electrophoresis Exercise with Serum Proteins

    ERIC Educational Resources Information Center

    Kim, Thomas D.; Craig, Paul A.

    2010-01-01

    Two-dimensional gel electrophoresis (2DGE) remains an important tool in the study of biological systems by proteomics. While the use of 2DGE is commonplace in research publications, there are few instructional laboratories that address the use of 2DGE for analyzing complex protein samples. One reason for this lack is the fact that the preparation…

  11. Head-to-Head Comparison of Serum Fractionation Techniques

    SciTech Connect

    Whiteaker, Jeffrey R.; Zhang, Heidi; Eng, Jimmy K.; Fang, Ruihua; Piening, Brian D.; Feng, Li-Chia; lorentzen, Travis D.; Schoenherr, Regine M.; Keane, John F.; Holzman, Ted; Fitzgibbon, Matthew; Lin, Chen Wei; Zhang, Hui; Cooke, Kelly; Liu, Tao; Camp, David G.; Anderson, Leigh N.; Watts, Julian; Smith, Richard D.; McIntosh, Martin; Paulovich, Amanda G.

    2007-02-01

    Multiple approaches for simplifying the serum proteome have been described. These techniques are generally developed across different laboratories, samples, mass spectrometry platforms, and analysis tools. Hence, comparing the available schemes is impossible from the existing literature because of confounding variables. We describe a head-to-head comparison of several serum fractionation schemes, including N-linked glycopeptide enrichment, cysteinyl-peptide enrichment, magnetic bead separation (C3, C8, and WCX), size fractionation, Protein A/G depletion, and immunoaffinity column depletion of abundant serum proteins. Each technique was compared to results obtained from unfractionated human serum. The results show immunoaffinity subtraction is the most effective means for simplifying the serum proteome and maintaining reasonable sample throughput. The reported dataset is publicly available and provides a standard against which emergent technologies can be compared and evaluated for their contribution to serum-based biomarker discovery.

  12. Effect of Crude Protein on Reproductive Efficiency, Serum Total Protein, and Albumin in the High-Producing Dairy Cow1

    Microsoft Academic Search

    E. R. Jordan; L. V. Swanson

    1979-01-01

    Forty-five high-producing dairy cows were assigned randomly in equal numbers and fed one of three isocaloric rations of 12.7, 16.3, or 19.3% crude protein starting at 4 days postpartum and con- tinuing for 91 days. The 19.3% group had fewer days to first observed estrus (27 days) than the 16.3 and 12.7% groups (41 days). The 12.7 and 16.3% groups

  13. Maturation and fertilisation of sheep oocytes cultured in serum-free medium containing silk protein sericin.

    PubMed

    Yasmin, Cut; Otoi, Takeshige; Setiadi, Mohamad Agus; Karja, Ni Wayan Kurniani

    2015-03-01

    Sericin is a water-soluble component of silk and has been used as a biomaterial due to its antibacterial and ultraviolet radiation-resistant properties. This study was designed to evaluate the effect of sericin supplementation in a maturation medium on the meiotic competence and fertilisability of sheep oocytes. Cumulus-oocyte complexes (COCs) were cultured in TCM199 supplemented with sericin at various concentrations of 0 (control), 0.1, 0.25 and 0.5%, either with or without bovine serum albumin (BSA). When the COCs were matured without BSA, the supplementation of 0.1% sericin significantly increased the rates of maturation to metaphase II and the total fertilisation of oocytes compared with the other concentrations of sericin. When the COCs were matured with BSA, the beneficial effects of 0.1% sericin supplementation on the maturation and fertilisation of oocytes were not observed. Our findings indicate that supplementation with 0.1% sericin during maturation culture may improve the nuclear maturation and fertilisability of sheep oocytes. Moreover, it may be possible to replace BSA with sericin in chemically defined media without the risk of disease transmission. PMID:25655418

  14. The glass transition behavior of the globular protein bovine serum albumin.

    PubMed

    Brownsey, Geoffrey J; Noel, Timothy R; Parker, Roger; Ring, Stephen G

    2003-12-01

    The glass-like transition behavior of concentrated aqueous solutions of bovine serum albumin was examined using rheological techniques. At mass fractions >0.4, there was a marked concentration dependence of viscosity with a glass-like kinetic arrest observed at mass fractions in the region of 0.55. At mass fractions >0.6 the material behaved as a solid with a Young's modulus rising from approximately 20 MPa at a mass fraction of 0.62-1.1 GPa at 0.86. The solid was viscoelastic and exhibited stress relaxation with relaxation times increasing from 33 to 610 s over the same concentration range. The concentration dependence of the osmotic pressure was measured, at intermediate concentrations, using an osmotic stress technique and could be described using a hard sphere model, indicating that the intermolecular interactions were predominantly repulsive. In summary, a major structural relaxation results from the collective motion of the globules at the supra-globule length scale and, at 20 degrees C, this is arrested at water contents of 40% w/w. This appears to be analogous to the glass transition in colloidal hard spheres. PMID:14645083

  15. Label-free and homogeneous aptamer proximity binding assay for fluorescent detection of protein biomarkers in human serum.

    PubMed

    Wei, Yulian; Zhou, Wenjiao; Liu, Jun; Chai, Yaqin; Xiang, Yun; Yuan, Ruo

    2015-08-15

    By using the aptamer proximity binding assay strategy, the development of a label-free and homogeneous approach for fluorescent detection of human platelet-derived growth factor BB (PDGF-BB) is described. Two G-quadruplex forming sequence-linked aptamers bind to the PDGF-BB proteins, which leads to the increase in local concentration of the aptamers and promotes the formation of the G-quadruplex structures. Subsequently, the fluorescent dye, N-methylmesoporphyrin IX, binds to these G-quadruplex structures and generates enhanced fluorescence emission signal for sensitive detection of PDGF-BB. The association of the aptamers to the PDGF-BB proteins is characterized by using native polyacrylamide gel electrophoresis. The experimental conditions are optimized to reach an estimated detection limit of 3.2nM for PDGF-BB. The developed method is also selective and can be applied for monitoring PDGF-BB in human serum samples. With the advantages of label-free and homogeneous detection, the demonstrated approach can be potentially employed to detect other biomarkers in a relatively simple way. PMID:25966407

  16. Competitive adsorption of human serum albumin and gamma-globulin from a binary protein mixture onto hexadecyltrichlorosilane coated glass.

    PubMed

    Tremsina, Y S; Sevastianov, V I; Petrash, S; Dando, W; Foster, M D

    1998-01-01

    The kinetics of competitive adsorption of proteins onto hexadecyltrichlorosilane coated glass (HTS-glass) from model solutions containing fluorescein isothiocyanate (FITC)-labeled human serum albumin (HSA-FITC) and gamma-globulin (HGG-FITC) were studied by total internal reflection fluorescence (TIRF) spectroscopy. The processes of displacement of HSA-FITC by HGG are independent of the conformational state of HSA adsorbed onto glass. On HTS-glass, displacement of protein is hindered by the presence of large numbers of CH3-terminated alkyl tails which induce conformational (reorientational) changes in HSA-FITC and HGG-FITC adsorbed from simple solutions. In contrast to HSA, adsorption of HGG onto HTS-glass from a simple solution is characterized by the absence of irreversible adsorption in the initial portion of the kinetic curve. Competition between HSA and HGG-FITC induces replacement of end-on adsorbed HGG-FITC on HTS-glass surface with subsequent desorption of the HGG-FITC into solution. Upon further increase in the HSA concentration in solution the competition of HSA for adsorption sites prevails, which leads to a decrease in the amount of adsorbed HGG-FITC and, consequently, to a decrease in the rate of its displacement. PMID:9493842

  17. Expression and purification of recombinant human serum albumin fusion protein with VEGF165b in Pichia pastoris.

    PubMed

    Zhu, Rui-Yu; Xin, Xin; Dai, Hui-yun; Li, Qi; Lei, Jian-Yong; Chen, Yun; Jin, Jian

    2012-09-01

    VEGF165b is an endogenous transcriptional splice variant of VEGF and has been shown to have a therapeutic potency as an anti-cancer agent. In this report, a fusion gene consisting of a human VEGF165b and a human albumin (HSA) gene was constructed and then inserted into a pPIC9k vector. The recombinant fusion protein, rhHSA-VEGF165b, was over expressed in the methylotrophic yeast Pichia pastoris under the control of AOX1 promoter. After induction with methanol, the expression level of rhHSA-VEGF165b was 275 mg/L in broth. The fusion protein rhHSA-VEGF165b was purified to more than 95% purity by using Blue Sepharose Fast Flow and SP Sepharose Fast Flow. Biological activity of the prepared rhHSA-VEGF165b was characterized by transwell migration assay, retaining about 9% of that of unmodified rhVEGF165b on a molar basis. Data from mice show that the serum half-life time of rhHSA-VEGF165b was nearly 20 times longer than that of rhVEGF165b. PMID:22750397

  18. Human Serum Amyloid A3 (SAA3) Protein, Expressed as a Fusion Protein with SAA2, Binds the Oxidized Low Density Lipoprotein Receptor

    PubMed Central

    Tomita, Takeshi; Ieguchi, Katsuaki; Sawamura, Tatsuya; Maru, Yoshiro

    2015-01-01

    Serum amyloid A3 (SAA3) possesses characteristics distinct from the other serum amyloid A isoforms, SAA1, SAA2, and SAA4. High density lipoprotein contains the latter three isoforms, but not SAA3. The expression of mouse SAA3 (mSAA3) is known to be up-regulated extrahepatically in inflammatory responses, and acts as an endogenous ligand for the toll-like receptor 4/MD-2 complex. We previously reported that mSAA3 plays an important role in facilitating tumor metastasis by attracting circulating tumor cells and enhancing hyperpermeability in the lungs. On the other hand, human SAA3 (hSAA3) has long been regarded as a pseudogene, which is in contrast to the abundant expression levels of the other isoforms. Although the nucleotide sequence of hSAA3 is very similar to that of the other SAAs, a single oligonucleotide insertion in exon 2 causes a frame-shift to generate a unique amino acid sequence. In the present study, we identified that hSAA3 was transcribed in the hSAA2-SAA3 fusion transcripts of several human cell lines. In the fusion transcript, hSAA2 exon 3 was connected to hSAA3 exon 1 or hSAA3 exon 2, located approximately 130kb downstream from hSAA2 exon 3 in the genome, which suggested that it is produced by alternative splicing. Furthermore, we succeeded in detecting and isolating hSAA3 protein for the first time by an immunoprecipitation-enzyme linked immune assay system using monoclonal and polyclonal antibodies that recognize the hSAA3 unique amino acid sequence. We also demonstrated that hSAA3 bound oxidized low density lipoprotein receptor (oxLDL receptor, LOX-1) and elevated the phosphorylation of ERK, the intracellular MAP-kinase signaling protein. PMID:25738827

  19. Increased pentosidine, an advanced glycation end product, in serum and synovial fluid from patients with knee osteoarthritis and its relation with cartilage oligomeric matrix protein

    PubMed Central

    Senolt, L; Braun, M; Olejarova, M; Forejtova, S; Gatterova, J; Pavelka, K

    2005-01-01

    Background: Pentosidine, an advanced glycation end product, increasingly accumulates in articular cartilage with age, and contributes to the pathogenesis of osteoarthritis (OA). Increased pentosidine concentrations are associated with inflammatory disorders—for example, rheumatoid arthritis. Objective: To compare pentosidine serum concentrations in patients with knee OA and in healthy volunteers and to determine a relationship between pentosidine and cartilage oligomeric matrix protein (COMP)—a marker of articular cartilage destruction. Methods: Paired serum and synovial fluid samples were obtained by arthrocentesis from 38 patients with knee OA and from 38 healthy volunteers. Pentosidine concentration was measured by reverse phase high performance liquid chromatography with fluorescent detection and COMP was determined by sandwich ELISA. Results: Significantly increased serum pentosidine (p<0.01) and COMP (p<0.05) levels were detected in the patients with OA compared with the control group. Serum pentosidine correlated significantly with synovial fluid pentosidine (p<0.001). Pentosidine in synovial fluid (p<0.05) and in serum (p<0.05) correlated significantly with synovial fluid COMP. Pentosidine and COMP concentrations did not correlate significantly with the radiological stage of the disease. Conclusion: Increased pentosidine serum concentration in patients with OA and its correlation with the cartilage destruction marker COMP in synovial fluid suggests that pentosidine may be important in OA pathology and is a new potential OA marker. PMID:15897309

  20. Serum insulin-like growth factor binding protein 6 (IGFBP6) is increased in patients with type 1 diabetes and its complications

    PubMed Central

    Lu, Shangsu; Purohit, Sharad; Sharma, Ashok; Zhi, Wenbo; He, Mingfang; Wang, Yiqian; Li, Chao-Jun; She, Jin-Xiong

    2012-01-01

    Insulin-like growth factor binding proteins (IGFBPs) are associated with insulin resistance and accelerated micro- and macro-vascular complications of diabetes. We investigated the relationship between serum levels of IGFBP6 in type-1 diabetes (T1D) patients and diabetic complications. In this study, IGFBP6 was measured in the sera from 697 T1D patients and 681 healthy controls using a Luminex assay. Mean serum levels of IGFBP6 were higher in T1D patients than controls matched for sex and age (119.7 vs 130.6 ng/ml, p < 10-4). Subject age, sex and duration of disease have a significant impact on serum IGFBP6 levels in both T1D patients and healthy controls. Patients with complications have significantly higher mean serum IGFBP6 than patients without any complication (p = 3.5x10-6). More importantly, conditional logistic regression analysis suggested that T1D patients are more likely to have very high levels of serum IGFBP6 (in the 4th quartile) (OR = 1.7) than healthy controls. Furthermore, T1D patients with various complications are more likely to have very high levels of serum IGFBP6 (in the 4th quartile) than patients without any complication (OR = 1.7 - 22.9). These results indicate the clinical importance of measuring IGFBP6 to the better management of T1D patients. PMID:22837797

  1. Simultaneous separation and quantitation of the major bovine whey proteins including proteose peptone and caseinomacropeptide by reversed-phase high-performance liquid chromatography on polystyrene–divinylbenzene

    Microsoft Academic Search

    David F. Elgar; Carmen S. Norris; John S. Ayers; Mark Pritchard; Don E. Otter; Kate P. Palmano

    2000-01-01

    A precise, sensitive and reliable RP-HPLC method was developed to enable not only unequivocal determination of ?-lactalbumin and ?-lactoglobulin in bovine whey samples, but also simultaneous measurement of proteose peptone, caseinomacropeptide, bovine serum albumin and immunoglobulin G. The optimised method on the Resource RPC column allowed separation of the proteins in 30 min and could be applied to the analysis

  2. Elevated Serum Triglyceride and Retinol-Binding Protein 4 Levels Associated with Fructose-Sweetened Beverages in Adolescents

    PubMed Central

    Chan, Te-Fu; Lin, Wei-Ting; Chen, Yi-Ling; Huang, Hsiao-Ling; Yang, Wei-Zeng; Lee, Chun-Ying; Chen, Meng-Hsueh; Wang, Tsu-Nai; Huang, Meng-Chuan; Chiu, Yu-Wen; Huang, Chun-Chi; Tsai, Sharon; Lin, Chih-Lung; Lee, Chien-Hung

    2014-01-01

    Background The metabolic effect of fructose in sugar-sweetened beverages (SSB) has been linked to de novo lipogenesis and uric acid (UA) production. Objectives This study investigated the biological effects of SSB consumption on serum lipid profiles and retinol-binding protein 4 (RBP4) among Taiwanese adolescents. Methods We evaluated the anthropometric parameters and biochemical outcomes of 200 representative adolescents (98 boys and 102 girls) who were randomly selected from a large-scale cross-sectional study. Data were analyzed using multiple regression models adjusted for covariates. Results Increased SSB consumption was associated with increased waist and hip circumferences, body mass index (BMI) values and serum UA, triglyceride (TG) and RBP4 levels. Adolescents who consumed >500 ml/day of beverages half-to-heavily sweetened with high-fructose corn syrup (HFCS) exhibited TG and RBP4 levels 22.7 mg/dl and 13.92 ng/ml higher than non-drinkers, respectively. HFCS drinkers with hyperuricemia had higher TG levels than HFCS drinkers with normal UA levels (98.6 vs. 81.6 mg/dl). The intake of HFCS-rich SSBs and high value of BMI (?24) interactively reinforced RBP4 levels among overweight/obese adolescents. Circulating RBP4 levels were significantly correlated with weight-related outcomes and TG and UA concentration among HFCS drinkers (r?=?0.253 to 0.404), but not among non-drinkers. Conclusions High-quantity HFCS-rich beverage consumption is associated with higher TG and RBP4 levels. Hyperuricemia is likely to intensify the influence of HFCS-rich SSB intake on elevated TG levels, and in overweight and obese adolescents, high BMI may modify the action of fructose on higher circulating levels of RBP4. PMID:24475021

  3. Acute-phase protein serum amyloid A3 is a novel paracrine coupling factor that controls bone homeostasis.

    PubMed

    Thaler, Roman; Sturmlechner, Ines; Spitzer, Silvia; Riester, Scott M; Rumpler, Monika; Zwerina, Jochen; Klaushofer, Klaus; van Wijnen, Andre J; Varga, Franz

    2015-04-01

    Serum amyloid A (A-SAA/Saa3) was shown before to affect osteoblastic metabolism. Here, using RT-quantitative PCR and/or immunoblotting, we show that expression of mouse Saa3 and human SAA1 and SAA2 positively correlates with increased cellular maturation toward the osteocyte phenotype. Expression is not detected in C3H10T1/2 embryonic fibroblasts but is successively higher in preosteoblastic MC3T3-E1 cells, late osteoblastic MLO-A5 cells, and MLO-Y4 osteocytes, consistent with findings using primary bone cells from newborn mouse calvaria. Recombinant Saa3 protein functionally inhibits osteoblast differentiation as reflected by reductions in the expression of osteoblast markers and decreased mineralization in newborn mouse calvaria. Yet, Saa3 protein enhances osteoclastogenesis in mouse macrophages/monocytes based on the number of multinucleated and tartrate-resistant alkaline phosphatase-positive cells and Calcr mRNA expression. Depletion of Saa3 in MLO osteocytes results in the loss of the mature osteocyte phenotype. Recombinant osteocalcin, which is reciprocally regulated with Saa3 at the osteoblast/osteocyte transition, attenuates Saa3 expression in MLO-Y4 osteocytes. Mechanistically, Saa3 produced by MLO-Y4 osteocytes is integrated into the extracellular matrix of MC3T3-E1 osteoblasts, where it associates with the P2 purinergic receptor P2rx7 to stimulate Mmp13 expression via the P2rx7/MAPK/ERK/activator protein 1 axis. Our data suggest that Saa3 may function as an important coupling factor in bone development and homeostasis. PMID:25491310

  4. [Immunogenicity of recombinant proteins including ectodomain of M2 influenza virus A].

    PubMed

    Stepanova, L A; Kovaleva, A A; Potapchuk, M V; Korotkov, A V; Kupriianov, V V; Blokhina, E A; Kotliarov, R Iu; Tsybalova, L M

    2013-01-01

    Two recombinant proteins with three copies of the ectodomain of the conserved influenza protein M2 (M2e) of influenza viruses were developed: A (H1N1)pdm09, A/Kurgan/05/05 (H5N1), and M2e consensus sequence of the human influenza A virus (H1N1, H2N2, H3N2) based on flagellin and core antigen of hepatitis B (HBc). The first recombinant protein comprised flagellin fused to three tandem copies of M2e, the second preparation was based on non-covalent interaction between M2e peptides and HBc. The immunogenicity of two preparations was comparatively tested. A covalent linkage of flagellin with M2e significant increased the immunogenicity of the target antigen compared with non-covalent interaction M2e and HBc. Flagellin as a protein carrier of M2e induced mainly IgG1 subclass, whereas HBc stimulated more balanced Th1/Th2 response. Our study showed a decrease in the viral titers in lung tissues of immunized mice after lethal challenge of A/PR/8/34 (H1N1). The study revealed a possibility to obtain a vaccine preparation with equal immunogenicity both against human influenza viruses and highly pathogenic avian influenza viruses. PMID:24006628

  5. Microfiltration: Effect of retentate protein concentration on limiting flux and serum protein removal with 4-mm-channel ceramic microfiltration membranes.

    PubMed

    Hurt, E E; Adams, M C; Barbano, D M

    2015-04-01

    The objective of our study was to determine if the limiting flux and serum protein (SP) removal were different at 8, 9, or 10% true protein (TP) in the microfiltration (MF) retentate recirculation loop using 0.1-µm ceramic graded permeability membranes with 4-mm-channel diameters operated at 50°C using a diluted milk protein concentrate with 85% protein on a total solids basis (MPC85) as the MF feed. The limiting flux for the MF of diluted MPC85 was determined at 3 TP concentrations in the recirculation loop (8, 9, and 10%). The experiment was replicated 3 times for a total of 9 runs. On the morning of each run, MPC85 was diluted with reverse osmosis water to an MF feed TP concentration of 5.4%. In all runs, the starting flux was 55kg/m(2) per hour, the flux was increased in steps until the limiting flux was reached. The minimum flux increase was 10kg/m(2) per hour. The limiting flux decreased as TP concentration in the recirculation loop increased. The limiting flux was 154±0.3, 133±0.7, and 117±3.3kg/m(2) per hour at recirculation loop TP concentrations of 8.2±0.07, 9.2±0.04, and 10.2±0.09%, respectively. No effect of recirculation loop TP concentration on the SP removal factor was detected. However, the SP removal factor decreased from 0.80±0.02 to 0.75±0.02 as flux was increased from the starting flux of 55kg/m(2) per hour to the limiting flux, with a similar decrease seen at all recirculation loop TP concentrations. PMID:25682139

  6. Structural analogs of human insulin-like growth factor I with reduced affinity for serum binding proteins and the type 2 insulin-like growth factor receptor

    SciTech Connect

    Bayne, M.L.; Applebaum, J.; Chicchi, G.G.; Hayes, N.S.; Green, B.G.; Cascieri, M.A.

    1988-05-05

    Four structural analogs of human insulin-like growth factor I (hIGF-I) have been prepared by site-directed mutagenesis of a synthetic IGF-I gene and subsequent expression and purification of the mutant protein from the conditioned media of transformed yeast. (Phe/sup -1/, Val/sup 1/, Asn/sup 2/, Gln/sup 3/, His/sup 4/, Ser/sup 8/, His/sup 9/, Glu/sup 12/, Tyr/sup 15/, Leu/sup 16/)IGF-I (B-chain mutant), in which the first 16 amino acids of hIGF-I were replaced with the first 17 amino acids of the B-chain of insulin, has >1000-, 100-, and 2-fold reduced potency for human serum binding proteins, the rat liver type 2 IGF receptor, and the human placental type 1 IGF receptor, respectively. The B-chain mutant also has 4-fold increased affinity for the human placental insulin receptor. (Gln/sup 3/, Ala/sup 4/) IGF-I has 4-fold reduced affinity for human serum binding proteins, but is equipotent to hIGF-I at the types 1 and 2 IGF and insulin receptors. (Tyr/sup 15/, Leu/sup 16/) IGH-I has 4-fold reduced affinity for human serum binding proteins and 10-fold increased affinity for the insulin receptor. The peptide in which these four-point mutations are combined, (Gln/sup 3/, Ala/sup 4/, Tyr/sup 15/,Leu/sup 16/)IGF-I, has 600-fold reduced affinity for the serum binding proteins. All four of these mutants stimulate DNA synthesis in the rat vascular smooth muscle cell line A10 with potencies reflecting their potency at the type 1 IGF receptor. These studies identify some of the domains of hIGF-I which are responsible for maintaining high affinity binding with the serum binding protein and the type 2 IGF receptor. In addition, These peptides will be useful in defining the role of the type 2 IGF receptor and serum binding proteins in the physiological actions of hIGF-I.

  7. Defense proteins from seed of Cassia fistula include a lipid transfer protein homologue and a protease inhibitory plant defensin.

    PubMed

    Wijaya; Neumann; Condron; Hughes; Polya

    2000-11-01

    A novel trypsin inhibitor was extracted from the seeds of Cassia fistula by a process successively involving soaking seeds in water, extraction of the seeds in methanol, and extraction of the cell wall material at high ionic strength. The protease inhibitor (PI) was subsequently purified by chromatography on carboxymethylcellulose, gel filtration and reversed phase HPLC (RP-HPLC). Electrospray ionization mass spectrometry (ESMS) of the oxidized from of the PI yielded an average molecular mass of 5458.6+/-0.8 Da. Edman sequencing of the PI yielded a full-length 50 amino acid sequence inferred to contain eight cysteines and with a calculated average molecular mass (fully oxidized form) of 5459.3 Da, in agreement with the observed mass. The C. fistula seed PI is homologous to the family of plant defensins (gamma-thionins), which have four disulfide linkages at highly conserved locations. The C. fistula PI inhibits trypsin (IC(50) 2 µM), and is the first known example of a plant defensin with protease inhibitory activity, suggesting a possible additional function for some members of this class of plant defensive proteins. C. fistula seeds also contain a 9378 Da lipid transfer protein (LTP) homologue, other LTPs, a 7117 Da protein copurifying with PI activity and a 5144 Da defensin which does not inhibit trypsin. The complete sequence of the 5144 Da defensin was determined by Edman sequencing, yielding a calculated average molecular mass (oxidized form) of 5144.1 Da, in agreement with the mass observed by ESMS. The likely trypsin inhibitory residue on the 5459 Da defensin is Lysine-25, the corresponding amino acid being Tyrosine-25 in the homologous 5144 Da defensin that is not a trypsin inhibitor. PMID:11074277

  8. The Intrinsic Antiviral Defense to Incoming HSV1 Genomes Includes Specific DNA Repair Proteins and Is Counteracted by the Viral Protein ICP0

    Microsoft Academic Search

    Caroline E. Lilley; Mira S. Chaurushiya; Chris Boutell; Roger D. Everett; Matthew D. Weitzman

    2011-01-01

    Cellular restriction factors responding to herpesvirus infection include the ND10 components PML, Sp100 and hDaxx. During the initial stages of HSV-1 infection, novel sub-nuclear structures containing these ND10 proteins form in association with incoming viral genomes. We report that several cellular DNA damage response proteins also relocate to sites associated with incoming viral genomes where they contribute to the cellular

  9. Radiation-induced changes in levels of selected proteins in peripheral blood serum of breast cancer patients as a potential triage biodosimeter for large-scale radiological emergencies.

    PubMed

    Deperas-Kaminska, Marta; Bajinskis, Ainars; Marczyk, Michal; Polanska, Joanna; Wersäll, Peter; Lidbrink, Elisabet; Ainsbury, Elizabeth A; Guipaud, Oliver; Benderitter, Marc; Haghdoost, Siamak; Wojcik, Andrzej

    2014-12-01

    The threat of a large scale radiological emergency, where thousands of people may require fast biological dosimetry for the purpose of triage, makes it necessary to search for new, high throughput biological dosimeters. The authors tested an assay based on the quantitative analysis of selected proteins in peripheral blood serum. They were particularly interested in testing proteins that are specific to irradiation of skin, as these can be used in cases of partial body exposure. Candidate proteins were identified in an earlier study with mice, where skin of the animals was exposed to different doses of radiation and global expression of serum proteins was analyzed. Eight proteins were found, the expression of which showed a consistent dose-response relationship. Human analogues of these proteins were identified, and their expression was measured in peripheral blood serum of 16 breast cancer patients undergoing external beam radiotherapy. The proteins were Apolipoprotein E; Apolipoprotein H; Complement protein 7; Prothrombinase; Pantothenate Kinase 4; Alpha-2-macroglobulin; Fetuin B and Alpha-1-Anti-Chymotrypsin. Measurements were carried out in blood samples collected prior to exposure (control), on the day after one fraction (2 Gy), on the day after five fractions (10 Gy), on the day after 10 fractions (20 Gy), and 1 mo after 23-25 fractions (total dose of 46-50 Gy). Multivariate analysis was carried out, and a multinomial logistic regression model was built. The results indicate that the combined analysis of Apolipoprotein E, Factor X, and Pantothenate Kinase 4 allows discriminating between exposure to 2 Gy and lower and between 10 Gy and higher. The discrimination is possible up to 1 mo after exposure. PMID:25353241

  10. A method for including protein flexibility in protein-ligand docking: improving tools for database mining and virtual screening.

    PubMed

    Broughton, H B

    2000-06-01

    Second-generation methods for docking ligands into their biological receptors, such as FLOG, provide for flexibility of the ligand but not of the receptor. Molecular dynamics based methods, such as free energy perturbation, account for flexibility, solvent effects, etc., but are very time consuming. We combined the use of statistical analysis of conformational samples from short-run protein molecular dynamics with grid-based docking protocols and demonstrated improved performance in two test cases. Our statistical analysis explores the importance of the average strength of a potential interaction with the biological target and optionally applies a weighting depending on the variability in the strength of the interaction seen during dynamics simulation. Using these methods, we improved the num-top-ranked 10% of a database of drug-like molecules, in searches based on the three-dimensional structure of the protein. These methods are able to match the ability of manual docking to assess likely inactivity on steric grounds and indeed to rank order ligands from a homologous series of cyclooxygenase-2 inhibitors with good correlation to their true activity. Furthermore, these methods reduce the need for human intervention in setting up molecular docking experiments. PMID:11021541

  11. Proteins excreted by primary human colon carcinoma cells (SW 480) promote spreading and growth of metastatic human colon carcinoma cells (SW 620) in serum-free medium.

    PubMed

    Grimmond, H E; Hill, G J; Zirvi, K A

    1989-10-01

    The growth behavior of the two human colon tumor cell lines (SW 480, primary and SW 620, metastatic), originating from the same patient, was studied in six different serum-free media (SFM) [GF3, Chee's essential medium plus insulin, transferrin and selenium; GF3F, GF3 plus fetuin; GF4, GF3 plus linoleic acid-BSA; GF5, GF4 plus fetuin; GF5E, GF5 plus EGF; GF5T, GF5 plus triiodothyronine]. SW 480 grew in all of the SFM. In contrast, SW 620 grew only in four SFM. The cells did not grow in GF3 and GF4. When grown in SFM, SW 480 attached much more firmly to the dishes than SW 620 as determined by the time required to detach the cells with trypsin-EDTA (SW 480, greater than 20 min and SW 620, less than 5 min). It was speculated that SW 480 cells excrete proteins in SFM which influence attachment and growth of the cells. Growth behavior of SW 480 cells which did not grow in GF3, was studied using GF3 medium and SW 480 substratum dishes. SW 620 cells readily attached to the SW 480 substratum dishes and grew. Furthermore, when SW 620 cells were grown on substratum prepared from serum-supplemented medium incubated in the absence of cells (serum substratum), the cell growth was comparable to the cell growth on SW 480 substratum in GF3. Substratum from SW 480 cells and the serum substratum were compared for their components using SDS-PAGE system. The SW 480 substratum contains many more components than serum substratum. A protein band at 60 kD appears to be common in both SW 480 and serum substrata. PMID:2620293

  12. Evaluation of intrathecal serum amyloid P (SAP) and C-reactive protein (CRP) synthesis in Alzheimer's disease with the use of index values.

    PubMed

    Mulder, Sandra D; Hack, C Erik; van der Flier, Wiesje M; Scheltens, Philip; Blankenstein, Marinus A; Veerhuis, Robert

    2010-01-01

    Serum amyloid P (SAP) and C-reactive protein (CRP) are proteins involved in innate immunity. The expression of SAP and CRP is increased in Alzheimer's disease (AD) brain tissue, compared to healthy controls. Although both proteins are found in cerebrospinal fluid (CSF), their origin is unclear. We investigated if increased local production of SAP and CRP in AD brain results in higher levels in CSF with the use of index values. To study this, SAP, CRP, and albumin levels were determined in CSF and serum samples of 30 control (65 ± 11 years; 57% female) and 140 AD subjects (65 ± 9 years; 53% female). To correct for inter-individual differences in protein diffusion from blood to CSF, quotients (Q =CSF/serum) of SAP, CRP, and albumin and index values (Qprotein/Qalb) were calculated. The results showed no significant differences in SAP and CRP index values between control and AD subjects, although eight percent of individual AD patients showed evidence of intrathecal SAP or CRP production using the Reiber hyperbolic model. Interestingly, the SAP index value was much lower than expected, based on its molecular size. In conclusion, these data suggest that local production of SAP and CRP in the AD brain does not substantially contribute to the CSF levels. PMID:20930309

  13. Comparison of Serum Concentrations of ?-Trace Protein, ?2-Microglobulin, Cystatin C, and Creatinine in the US Population

    PubMed Central

    Juraschek, Stephen P.; Coresh, Josef; Inker, Lesley A.; Levey, Andrew S.; Köttgen, Anna; Foster, Meredith C.; Astor, Brad C.; Eckfeldt, John H.

    2013-01-01

    Summary Background and objectives ?-trace protein (?TP), ?2-microglobulin (?2M), and cystatin C (CysC) have advantages over creatinine for estimating GFR and prognosis. This study compares the distribution of all four markers in the general population and their associations with possible determinants of GFR. Design, setting, participants, & measurements ?TP and ?2M were measured in 7596 participants (aged ?12 years) of the Third National Health and Nutrition Examination Survey (1988–1994). ?TP and ?2M concentrations and the proportion of persons with elevated (?99th percentile for young healthy participants) ?TP (?0.81 mg/L), ?2M (?2.80 mg/L), standardized CysC (?1.03 mg/L), and creatinine (?1.2 mg/dl for men and ?1.0 mg/dl for women) were compared across demographic and clinical factors. Results Elevated ?TP, ?2M, and CysC showed stronger associations with age than elevated serum creatinine, the prevalence of elevated levels reaching 47%, 44%, 58%, and 26%, respectively, by age 80 years. ?TP, CysC, and creatinine were higher in men but ?2M was not associated with sex. Mexican Americans had lower ?TP, ?2M, CysC, and creatinine compared with non-Hispanic whites. Hypertension and higher C-reactive protein were associated with elevations in all markers, whereas non-Hispanic black race, body mass index, diabetes, smoking status, triglycerides, HDL cholesterol, and education were not associated in a consistent manner across the different markers. Conclusions ?TP, ?2M, CysC, and creatinine differ in their associations with demographic and clinical factors, suggesting variation in their non-GFR determinants. Future studies should examine these markers with measured GFR to determine their diagnostic and prognostic utility. PMID:23335043

  14. Up-Regulation of MUC2 Mucin Expression by Serum Amyloid A3 Protein in Mouse Colonic Epithelial Cells

    PubMed Central

    SHIGEMURA, Hiroaki; ISHIGURO, Naotaka; INOSHIMA, Yasuo

    2014-01-01

    ABSTRACT Serum amyloid A (SAA) proteins are acute-phase proteins and are classified into multiple isoforms; however, the biological functions of each SAA isoform are not fully understood. In this study, to clarify the roles of SAA3 in the intestine, we characterized mRNA expression in mouse colonic epithelial CMT-93 cells treated with rotavirus, Toxoplasma, Staphylococcus aureus, and Escherichia coli, as well as lipopolysaccharide (LPS) and recombinant murine SAAs (rSAAs). E. coli together with LPS, but not the other pathogens, enhanced SAA3 mRNA expression. The mRNA expression of SAA3 by dead E. coli was higher than that by living E. coli, and the mRNA expression by E. coli and LPS increased in a dose-dependent manner. In contrast, mRNA expressions of SAA1 and/or SAA2 were not stimulated by any of the treatments. In comparisons of cell treatments with rSAA1 or rSAA3, rSAA3 significantly up-regulated the mRNA expression of mucin 2 (MUC2), a major component of the mucus layer of the intestines that acts as an epithelial cell barrier against pathogens, while MUC2 mRNA expression was not significantly increased by E. coli and LPS. Furthermore, treatment with rSAAs intensively induced tumor necrosis factor-? mRNA expression. These results suggest that SAA3 plays a role in host innate immunity in the colon by up-regulating MUC2 mucin production, which builds a physiological barrier of colonic epithelia against bacterial invasion. PMID:24694941

  15. Human Serum Albumin and p53-Activating Peptide Fusion Protein Is Able to Promote Apoptosis and Deliver Fatty Acid-Modified Molecules

    PubMed Central

    Joshi, Michelle R.; Yao, Nianhuan; Myers, Kenneth A.; Li, Zhiyu

    2013-01-01

    Therapeutic peptides offer a high degree of specificity, potency, and low toxicity; making them promising candidates for cancer therapy. Despite these advantages, a number of hurdles, such as poor serum stability and inefficient cellular penetration, must be overcome. Fusing a therapeutic peptide to human serum albumin (HSA) is a common approach to extend the serum stability of a peptide that binds to extracellular receptors. However, no study has shown that this approach can be applied to target intracellular proteins. Here we demonstrate the feasibility of using a recombinant human serum albumin (rHSA) fusion protein to simultaneously deliver two types of molecules: a peptide capable of binding an intracellular target, as well as fatty acid (FA)-modified FITC (FA-FITC). Two peptides reported to disrupt the intracellular p53 and MDM2/MDMX interaction were fused to the C-terminal of HSA. Cellular and biochemical studies indicate that rHSA fusion proteins were efficiently taken up by SJSA-1 cells and retained MDM2- and MDMX-binding activity. By inducing the accumulation of p53, both fusion proteins promoted efficient cytotoxicity in SJSA-1 cells via caspase activation. Long chain fatty acid (LCFA) transportation is an essential endogenous function of HSA. This study also demonstrates that rHSA fusion proteins formed highly stable complexes with FA-FITC via non-covalent interactions. FA-FITC complexed with HSA could be internalized efficiently and rHSA-P53i and rHSA-PMI retained apoptotic activity as complex components. It is expected that such an approach can ultimately be used to facilitate intracellular delivery of two anticancer therapeutics, each with distinct but complimentary mechanisms, to achieve synergistic efficacy. PMID:24278348

  16. Preparation and characterization of bovine serum albumin surface-imprinted thermosensitive magnetic polymer microsphere and its application for protein recognition.

    PubMed

    Li, Xiangjie; Zhang, Baoliang; Li, Wei; Lei, Xingfeng; Fan, Xinlong; Tian, Lei; Zhang, Hepeng; Zhang, Qiuyu

    2014-01-15

    A novel bovine serum albumin surface-imprinted thermosensitive magnetic composite microsphere was successfully prepared by the surface grafting copolymerization method in the presence of temperature-sensitive monomer N-isopropylacrylamide (NIPAM), functional monomer methacrylic acid (MAA) and cross-linking agent N,N'-methylenebisacrylamide (MBA). The structure and component of the thermosensitive magnetic molecularly imprinted microsphere were investigated by transmission electron microscopy (TEM), Fourier transform infrared (FT-IR), vibrating sample magnetometer (VSM) and thermogravimetric analysis (TGA). The results of thermosensitivity, adsorption capacity, selectivity and reusability showed the formation of a thermosensitivity grafting polymer layer P(NIPAM-MAA-MBA) on the surface of Fe3O4@SiO2 and the good adsorption capacity and specific recognition for template protein. When the adsorption temperature was higher than the lower critical solution temperature (LCST) of poly(N-isopropylacrylamide) (PNIPAM), shape memory effect of imprinted cavities would be more effective. In other words, it was more conducive to capture template molecules under this condition and the imprinting factor would be higher. On the other hand, when the desorption temperature was lower than LCST of PNIPAM, the decrease of shape memory effect between imprinted cavities and template molecules would facilitate the release of template molecules from the imprinted cavities. Based on this property, the adsorption and desorption of template molecules could be regulated by system temperature indirectly which benefited from the existence of thermosensitivity imprinting layer. PMID:23973936

  17. Elevated Serum C-Reactive Protein Relates to Increased Cerebral Myoinositol Levels in Middle-Aged Adults

    PubMed Central

    Eagan, Danielle E.; Gonzales, Mitzi M.; Tarumi, Takashi; Tanaka, Hirofumi; Stautberg, Sandra; Haley, Andreana P.

    2012-01-01

    C-reactive protein (CRP), a systemic marker of inflammation, is a risk factor for late life cognitive impairment and dementia, yet the mechanisms that link elevated CRP to cognitive decline are not fully understood. In this study we examined the relationship between CRP and markers of neuronal integrity and cerebral metabolism in middle-aged adults with intact cognitive function, using proton magnetic resonance spectrocospy. We hypothesized that increased levels of circulating CRP would correlate with changes in brain metabolites indicative of early brain vulnerability. Thirty-six individuals, aged 40 to 60, underwent neuropsychological assessment, a blood draw for CRP quantification, and 1H MRS examining N-acetyl-aspartate, myo-inositol, creatine, choline, and glutamate concentrations in occipito-parietal grey matter. Independent of age, sex and education, serum CRP was significantly related to higher cerebral myo-inositol/creatine ratio (F(4,31) = 4.74, P = 0.004), a relationship which remained unchanged after adjustment for cardiovascular risk (F(5,30) = 4.356, CRP ? = 0.322, P = 0.045). Because these biomarkers are detectable in midlife they may serve as useful indicators of brain vulnerability during the preclinical period when mitigating intervention is still possible. PMID:22461977

  18. Ultrasensitive electrochemical immunosensor based on orderly oriented conductive wires for the detection of human monocyte chemotactic protein-1 in serum.

    PubMed

    Li, Yuliang; He, Junlin; Xia, Chunyong; Gao, Liuliu; Yu, Chao

    2015-08-15

    For the first time, a simple, ultrasensitive and label-free electrochemical monocyte chemotactic protein-1 (MCP-1) immunosensor based on orderly oriented conductive wires has been developed. A conductive wire, which is similar to an electron-conducting tunnel, was designed with Au nanoparticles (AuNPs) joined to Au@Pt core-shell microspheres via a cysteamine (CA) crosslinker. To enhance the sensitivity of the immunosensor, Au nanoparticles were electrodeposited onto the gold electrode, and CA was self-assembled via strong Au-S covalent bonds, providing an appropriate surface and promoting electron transfer. Next, Au@Pt core-shell microspheres with large surface area were grafted onto the modified electrode to immobilize more MCP-1 antibodies. MCP-1 is an initiating factor and biomarker of atherosclerotic diseases. Under optimal experimental conditions, differential pulse voltammetry (DPV) current changes were used to detect MCP-1 with a broad linear range of 0.09-360pgmL(-1) and a low detection limit of 0.03pgmL(-1) (S/N=3). The proposed immunosensor exhibited good selectivity, reproducibility and reusability. When applied to spiked serum samples, the data for the developed immunosensor were in agreement with an enzyme linked immunosorbent assay, suggesting that the electrochemical immunosensor would be suitable for practical detection. PMID:25845330

  19. Next-generation sequencing and protein mass spectrometry for the comprehensive analysis of human cellular and serum antibody repertoires.

    PubMed

    Lavinder, Jason J; Horton, Andrew P; Georgiou, George; Ippolito, Gregory C

    2015-02-01

    Recent developments of high-throughput technologies are enabling the molecular-level analysis and bioinformatic mining of antibody-mediated (humoral) immunity in humans at an unprecedented level. These approaches explore either the sequence space of B-cell receptor repertoires using next-generation deep sequencing (BCR-seq), or the amino acid identities of antibody in blood using protein mass spectrometry (Ig-seq), or both. Generalizable principles about the molecular composition of the protective humoral immune response are being defined, and as such, the field could supersede traditional methods for the development of diagnostics, vaccines, and antibody therapeutics. Three key challenges remain and have driven recent advances: (1) incorporation of innovative techniques for paired BCR-seq to ascertain the complete antibody variable-domain VH:VL clonotype, (2) integration of proteomic Ig-seq with BCR-seq to reveal how the serum antibody repertoire compares with the antibody repertoire encoded by circulating B cells, and (3) a demand to link antibody sequence data to functional meaning (binding and protection). PMID:25461729

  20. Serum Amyloid A, C-Reactive Protein, and Retinal Microvascular Changes in Hypertensive Diabetic and Nondiabetic Individuals

    PubMed Central

    Stettler, Christoph; Witt, Nicholas; Tapp, Robyn J.; Thom, Simon; Allemann, Sabin; Tillin, Therese; Stanton, Alice; O'Brien, Eoin; Poulter, Neil; Gallimore, J. Ruth; Hughes, Alun D.; Chaturvedi, Nish

    2009-01-01

    OBJECTIVE To study the association of the inflammatory markers serum amyloid A (SAA) and C-reactive protein (CRP) with retinal microvascular parameters in hypertensive individuals with and without type 2 diabetes. RESEARCH DESIGN AND METHODS This cross-sectional analysis was a substudy in 711 patients (159 with and 552 without diabetes) of the Anglo-Scandinavian Cardiac Outcomes Trial (ASCOT) based on digital 30-degree images of superior and inferior temporal retinal fields. RESULTS SAA was associated with arteriolar length-to-diameter ratio positively in nondiabetic patients (Ptrend= 0.028) but negatively in diabetic patients (Ptrend= 0.005). The difference was unlikely to be a chance finding (P = 0.007 for interaction). Similar results were found for the association of SAA with arteriolar tortuosity (P = 0.05 for interaction). Associations were less pronounced for CRP and retinal parameters. CONCLUSIONS Inflammatory processes are differentially involved in retinal microvascular disease in diabetic compared with nondiabetic hypertensive individuals. PMID:19244088

  1. Relationship Between Spontaneous Passage Rates of Ureteral Stones Less Than 8 mm and Serum C-Reactive Protein Levels and Neutrophil Percentages

    PubMed Central

    Park, Chang Hyun; Ha, Ji Yong; Park, Choal Hee; Kim, Chun Il; Kim, Kwang Se

    2013-01-01

    Purpose A ureter obstruction caused by a ureteral stone results in inflammatory changes in the proximal submucosal layer and prevents the spontaneous passage of the ureteral stone. Accordingly, we analyzed the relationship between the spontaneous passage rates of ureteral stones less than 8 mm in size and serum C-reactive protein (CRP) levels and neutrophil percentages. Materials and Methods A total of 187 patients who were diagnosed with ureteral stones less than 8 mm in size and were managed consecutively at Keimyung University Dongsan Medical Center from January 2001 to January 2011 were retrospectively analyzed. Ureteral stone removal was defined as no ureteral stone shown in an imaging test without any treatment for 8 weeks after diagnosis. The patients were divided into three groups according to the levels of serum CRP and into two groups according to neutrophil percentage. The associations between these factors and ureteral stone passage rates were then examined. Results The ureteral stone passage rates of the low serum CRP level group, the medium serum CRP level group, and the high serum CRP level group were 94.1% (159/169), 70% (7/10), and 50.0% (4/8), respectively. The passage rates of ureteral stones in the group with a normal neutrophil percentage and in the group with a higher neutrophil percentage were 94.5% (121/128) and 83.1% (49/59), respectively (p=0.011). Conclusions Measuring serum CRP levels and neutrophil percentages in patients with small ureteral stones of less than 8 mm is useful in predicting whether the stone will be spontaneously passed. When the serum CRP level and neutrophil percentage of a patient are high, aggressive treatment such as extracorporeal shock wave lithotripsy should be considered. PMID:24044096

  2. Spectrum of activity, mutation rates, synergistic interactions, and the effects of pH and serum proteins for fusidic acid (CEM-102).

    PubMed

    Biedenbach, Douglas J; Rhomberg, Paul R; Mendes, Rodrigo E; Jones, Ronald N

    2010-03-01

    Fusidic acid (CEM-102) is a steroidal antimicrobial agent with focused Gram-positive activity that acts by preventing bacterial protein synthesis via interacting with elongation factor G. A collection of 114 wild-type isolates (> 80 species) was used to define the contemporary limits of fusidic acid spectrum against Gram-positive and Gram-negative species. Reference broth microdilution and anaerobic agar dilution methods were performed. Modifications of standardized test methods included adding 10% human serum and adjusting the medium pH to 5, 6, and 8. Synergy was assessed by the checkerboard method and time-kill studies. Mutational rates to resistance were determined at 4 x, 8 x, and 16 x MIC. Against Gram-positive pathogens, fusidic acid MIC values ranged from 0.06 to 32 microg/mL with the greatest potency against Staphylococcus aureus, Corynebacterium spp., and Micrococcus luteus (MIC results, 0.25, < or = 0.12, and < or = 0.5 microg/mL, respectively). Enterococci and streptococci were less susceptible (MIC ranges, 2-8 and 16-32 microg/mL, respectively). Fusidic acid activity against Gram-negative species was more limited (all MIC values, > or = 2 microg/mL) except for Empedobacter brevis, Moraxella catarrhalis and Neisseria meningitidis. A 4-fold increase in fusidic acid MIC results was observed when 10% serum was added to the broth. Decreasing medium pH to 5.0 to 6.0 negated the protein binding effects. Among the 8 antimicrobial combinations tested, gentamicin and rifampin enhanced the activity when combined with fusidic acid (no antagonism). Fusidic acid in vitro activity was most improved when combined with rifampin. Single-step mutational rates ranged from 1.2 x 10(-6) for 4x MIC to 9.8 x 10(-8) for 16 x MIC. In conclusion, these in vitro results for fusidic acid tested against contemporary strains confirm a persisting antimicrobial spectrum, especially against staphylococci and some other Gram-positive species. PMID:20159376

  3. Interaction between casein micelles and whey protein/?-casein complexes during renneting of heat-treated reconstituted skim milk powder and casein micelle/serum mixtures.

    PubMed

    Kethireddipalli, Prashanti; Hill, Arthur R; Dalgleish, Douglas G

    2011-02-23

    Casein micelles were separated from unheated reconstituted skim milk powder (RSMP) and were resuspended in the serum of RSMP that had been heated, with and without dialysis of this serum against unheated RSMP. Using size-exclusion chromatography, it was found that the soluble complexes of whey protein (WP) with ?-casein in the serum of the heated milk bind progressively to unheated casein micelles during renneting, even prior to the onset of clotting. Similar trends were noted when casein micelles from RSMP heated at pH values of 6.7, 7.1, or 6.3, each with different amounts of WP coating the micelles, were renneted in the presence of soluble WP/?-casein complexes. No matter what was the initial load of micelle-bound WP complexes, all micelle types were capable of binding additional serum protein complexes during renneting. However, it is not clear that this binding of WP/?-casein complexes to the micellar surface is a direct cause of the impaired rennet clotting of the RSMP. PMID:21287987

  4. Comparison of serum concentrations of symmetric dimethylarginine and creatinine as kidney function biomarkers in healthy geriatric cats fed reduced protein foods enriched with fish oil, L-carnitine, and medium-chain triglycerides.

    PubMed

    Hall, J A; Yerramilli, M; Obare, E; Yerramilli, M; Yu, S; Jewell, D E

    2014-12-01

    The purpose of this study was to determine whether feeding cats reduced protein and phosphorus foods with added fish oil, L-carnitine, and medium-chain triglycerides (MCT) altered serum biomarkers of renal function. Thirty-two healthy cats, mean age 14.0 (8.3-19.6) years, were fed control food or one of two experimental foods for 6 months. All foods had similar concentrations of moisture, protein, and fat (approximately 8.0%, 26.5%, and 20.0%, respectively). Both experimental foods contained added fish oil (1.5%) and L-carnitine (500?mg/kg). Experimental-food 2 also contained increased MCT (10.5% from coconut oil), 1.5% added corn oil, and reduced animal fat. Glomerular filtration rate (GFR), serum biochemistries, renal function biomarkers including serum creatinine (sCr) and symmetrical dimethylarginine (SDMA), and plasma metabolomic profiles were measured at baseline, and at 1.5, 3, and 6 months. Body composition was determined by dual-energy X-ray absorptiometry. Although both experimental foods altered plasma fatty acids, carnitine and related metabolites, and lysophospholipid concentrations, there were no changes in renal function biomarkers. There was, however, a benefit in using SDMA versus sCr to assess renal function in older cats with less total lean mass. Compared with cats <12 years, those >15 years had lower total lean mass (P?serum SDMA concentration is more highly correlated with GFR than sCr concentration, and, unlike sCr, which declines with age because of muscle wasting, SDMA increases as GFR declines with age. PMID:25458884

  5. Reduction of blood serum cholesterol

    NASA Technical Reports Server (NTRS)

    Winitz, M. (inventor)

    1974-01-01

    By feeding a human subject as the sole source of sustenance a defined diet wherein the carbohydrate consists substantially entirely of glucose, maltose or a polysaccharide of glucose, the blood serum cholesterol level of the human subject is substantially reduced. If 25 percent of the carbohydrate is subsequently supplied in the form of sucrose, an immediate increase from the reduced level is observed. The remainder of the defined diet normally includes a source of amino acids, such as protein or a protein hydrolysate, vitamins, minerals and a source of essential fatty acid.

  6. The CALMODULIN-BINDING PROTEIN60 family includes both negative and positive regulators of plant immunity.

    PubMed

    Truman, William; Sreekanta, Suma; Lu, You; Bethke, Gerit; Tsuda, Kenichi; Katagiri, Fumiaki; Glazebrook, Jane

    2013-12-01

    Two members of the eight-member CALMODULIN-BINDING PROTEIN60 (CBP60) gene family, CBP60g and SYSTEMIC ACQUIRED RESISTANCE DEFICIENT1 (SARD1), encode positive regulators of plant immunity that promote the production of salicylic acid (SA) and affect the expression of SA-dependent and SA-independent defense genes. Here, we investigated the other six family members in Arabidopsis (Arabidopsis thaliana). Only cbp60a mutations affected growth of the bacterial pathogen Pseudomonas syringae pv maculicola ES4326. In contrast to cbp60g and sard1 mutations, cbp60a mutations reduced pathogen growth, indicating that CBP60a is a negative regulator of immunity. Bacterial growth was increased by cbp60g only in the presence of CBP60a, while the increase in growth due to sard1 was independent of CBP60a, suggesting that the primary function of CBP60g may be to counter the repressive effect of CBP60a. In the absence of pathogen, levels of SA as well as of several SA-dependent and SA-independent pathogen-inducible genes were higher in cbp60a plants than in the wild type, suggesting that the enhanced resistance of cbp60a plants may result from the activation of immune responses prior to pathogen attack. CBP60a bound calmodulin, and the calmodulin-binding domain was defined at the C-terminal end of the protein. Transgenes encoding mutant versions of CBP60a lacking the ability to bind calmodulin failed to complement null cbp60a mutations, indicating that calmodulin-binding ability is required for the immunity-repressing function of CBP60a. Regulation at the CBP60 node involves negative regulation by CBP60a as well as positive regulation by CBP60g and SARD1, providing multiple levels of control over the activation of immune responses. PMID:24134885

  7. Glycomic Alterations in the Highly-abundant and Lesser-abundant Blood Serum Protein Fractions for Patients Diagnosed with Hepatocellular Carcinoma.

    PubMed

    Kang, Pilsoo; Madera, Milan; Alley, William R; Goldman, Radoslav; Mechref, Yehia; Novotny, Milos V

    2011-08-15

    Hepatocellular cancer is a serious human disease with an unfortunately low survival rate. It further poses a significant epidemic threat to our society through its viral vectors associated with cirrhosis conditions preceding the cancer. A search for biomarkers of these diseases enlists analytical glycobiology, in general, and quantitative biomolecular mass spectrometry (MS), in particular, as valuable approaches to cancer research. The recent advances in quantitative glycan permethylation prior to MALDI-MS oligosaccharide profiling has enabled us to compare the glycan quantitative proportions in the small serum samples of cancer and cirrhotic patients against control individuals. We were further able to fractionate the major serum proteins from the minor components and compare statistically their differential glycosylation, elucidating some causes of quantitatively unusual glycosylation events. Numerous glycan structures were tentatively identified and connected with the origin proteins, with a particular emphasis on sialylated and fucosylated glycans. PMID:23788846

  8. Intake of n-3 fatty acids from fish does not lower serum concentrations of C-reactive protein in healthy subjects

    Microsoft Academic Search

    A. Geelen; I. A. Brouwer; E. G. Schouten; C Kluft; M B Katan; P L Zock

    2004-01-01

    Objective: High-sensitivity C-reactive protein (CRP), a marker of systemic inflammation, is a powerful predictor of cardiovascular risk. We hypothesised that n-3 fatty acids reduce underlying inflammatory processes and consequently CRP concentrations in healthy middle-aged subjects.Design: Placebo-controlled, double-blind study.Subjects: A total of 43 men and 41 postmenopausal women aged 50–70 y. Before and after intervention, we measured serum CRP concentrations with

  9. Reduced serum insulin-like growth factor-1 (IGF-1) and IGF-binding protein-3 levels in adults with inflammatory bowel disease

    Microsoft Academic Search

    Konstantinos H Katsanos; Agathocles Tsatsoulis; Dimitrios Christodoulou; Anna Challa; Afroditi Katsaraki; Epameinondas V Tsianos

    2001-01-01

    In the present study, the changes in circulating IGF-1 and its binding protein IGFBP-3 were determined in adult patients with active inflammatory bowel disease (IBD) in order to assess the effect of this inflammatory condition on the IGF system. IGF-1 and IGFBP-3, as well as interleukin-6 (IL-6) were measured in serum obtained from 22 consecutive newly diagnosed patients (mean age

  10. The relation between the psychological functioning of children with Down syndrome and their urine peptide levels and levels of serum antibodies to food proteins

    Microsoft Academic Search

    Egil Nygaard; Karl Ludvig Reichelt; Joseph F. Fagan

    2001-01-01

    Aims: To investigate the relation between psychological functioning of subjects with Down syndrome, and their levels of urine peptide and serum antibodies to food proteins. Meth- ods: 55 children with Down syndrome in a cross-sectional study. Psychological functioning was measured by the Stanford-Binet Intelligence Scale: Fourth Edition, McCarthy Scales of Children's Abilities and Fagan's computer based test of novelty preference.

  11. Study on acute phase proteins (haptoglobin, serum amyloid A, fibrinogen, and ceruloplasmin) changes and their diagnostic values in bovine tropical theileriosis

    Microsoft Academic Search

    S. Nazifi; S. M. Razavi; Z. Esmailnejad; H. Gheisari

    2009-01-01

    This study was conducted to assess the pattern of changes and the relative value of acute phase proteins (haptoglobin (Hp),\\u000a serum amyloid A (SAA), ceruloplasmin and fibrinogen) in bovine tropical theileriosis. The diseased group comprised 50 dairy\\u000a Holstein cattle, 2?3 years old, naturally infected with Theileria annulata. Infected animals were divided into four subgroups with different parasitemia rates (5%). As a

  12. Comparisons of Serum Somatotropin, 3,5,3?Triiodothyronine, Thyroxine, Total Protein and Free Fatty Acid Levels in Newborn Sakiz Lambs Separated from or Suckling Their Dams

    Microsoft Academic Search

    Behiç Serpek

    2005-01-01

    Objective: To determine the effects on serum somatotropin, 3,5,3?-triiodothyronine (T3), thyroxine (T4), free fatty acids (FFAs) and total protein levels of different feedings and age in Sakiz sheep that have a high twin-bearing rate supported by estrous synchronization. Methods: 20 newborn lambs were used in the study. Lambs were divided into 2 equal groups. The lambs in 1 group were

  13. Serum levels of autoantibodies against C-reactive protein correlate with renal disease activity and response to therapy in lupus nephritis

    Microsoft Academic Search

    Christopher Sjöwall; Agneta Zickert; Thomas Skogh; Jonas Wetterö; Iva Gunnarsson

    2009-01-01

    INTRODUCTION: Serum levels of C-reactive protein (CRP) seldom reflect disease activity in systemic lupus erythematosus (SLE). We have previously shown that autoantibodies against neo-epitopes of CRP often occur in SLE, but that this does not explain the modest CRP response seen in flares. However, we have repeatedly found that anti-CRP levels parallel lupus disease activity, with highest levels in patients

  14. A single intravenous dose of endotoxin rapidly alters serum lipoproteins and lipid transfer proteins in normal volunteers

    Microsoft Academic Search

    Lisa C. Hudgins; Thomas S. Parker; Daniel M. Levine; Bruce R. Gordon; Stuart S. Saal; Xian-cheng Jiang; Cindy E. Seidman; Jolanta D. Tremaroli; Julie Lai; Albert L. Rubin

    2003-01-01

    Endotoxemia is associated with rapid and marked declines in serum levels of LDL and HDL by unknown mechanisms. Six normal volunteers received a single, small intravenous (iv) dose of endotoxin ( Escherichia coli 0113, 2 ng\\/ kg) or saline in a random order, cross-over design. After en- dotoxin treatment, volunteers had mild, transient flu-like symptoms and markedly increased serum levels

  15. Evaluation of high-performance liquid chromatography laser-induced fluorescence for serum protein profiling for early diagnosis of oral cancer

    NASA Astrophysics Data System (ADS)

    Patil, Ajeetkumar; Prabhu, Vijendra; Choudhari, K. S.; Unnikrishnan, V. K.; George, Sajan D.; Ongole, Ravikiran; Pai, Keerthilatha M.; Shetty, Jayarama K.; Bhat, Sujatha; Kartha, Vasudevan Bhaskaran; Chidangil, Santhosh

    2010-11-01

    The present work deals with the evaluation of a high-performance liquid chromatography laser-induced fluorescence (HPLC-LIF) technique developed in our laboratory for early detection of oral cancer from protein profiles of body fluids. The results show that protein profiles of serum samples from a given class of samples, say, normal, premalignant, or malignant, are statistically very close to each other, while profiles of members of any class are significantly different from other classes. The performance of the technique is evaluated by the use of sensitivity and specificity pairs, receiver operating characteristic (ROC) analysis, and Youden's Index. The technique uses protein profile differences in serum samples, registered by the HPLC-LIF technique. The study is carried out using serum samples from volunteers diagnosed as normal or premalignant clinically, and as malignant by histopathology. The specificities and sensitivities of the HPLC-LIF method at an ideal threshold (M-distance = 2) for normal, malignant, and premalignant classes are 100, 69.5, and 61.5%, and 86.5, 87.5, and 87.5% respectively.

  16. Soy protein supplementation increases serum insulin-like growth factor-I in young and old men but does not affect markers of bone metabolism.

    PubMed

    Khalil, Dania A; Lucas, Edralin A; Juma, Shanil; Smith, Brenda J; Payton, Mark E; Arjmandi, Bahram H

    2002-09-01

    Recent studies suggest that soy protein (SP) protects bone in women; however, its effects on bone metabolism in men have not been investigated. Healthy men (59.2 +/- 17.6 y) were assigned to consume 40 g of either SP or milk-based protein (MP) daily for 3 mo in a double-blind, randomized, controlled, parallel design. Serum insulin-like growth factor-I (IGF-I), which is associated with higher rates of bone formation, was greater (P < 0.01) in men supplemented with SP than in those consuming MP. Serum alkaline phosphatase and bone-specific alkaline phosphatase activities, markers of bone formation, and urinary deoxypyridinoline excretion, a specific marker of bone resorption, were not different between the SP and MP groups. Furthermore, because substantial reductions in bone density occur in men at approximately 65 y of age, data were analyzed separately for men >/=65 y and those <65 y of age. The response to protein supplementation was consistent in the two age groups. The effects of SP on serum IGF-I levels suggest that SP may positively influence bone in men. Longer-duration studies examining the effects of SP or its isoflavones on bone turnover and bone mineral density and content in men are warranted. PMID:12221217

  17. Association between serum insulin-like growth factor I or IGF-binding protein 3 and estimated glomerular filtration rate: results of a population-based sample

    PubMed Central

    2012-01-01

    Background Insulin-like growth factor I (IGF-I), which is mostly carried in blood by IGF-binding protein 3 (IGFBP-3), was associated to the glomerular filtration rate and chronic kidney disease in a multiethnic study among US adults. The aim of the present study was to investigate whether serum IGF-I or IGFBP-3 are associated with estimated glomerular filtration rate (eGFR) in a population-based study of Caucasian adults. Methods Data from 4028 subjects (2048 women) aged 20 to 81 years from the Study of Health in Pomerania (SHIP) were analyzed. Total serum IGF-I and IGFBP-3 concentrations were determined by chemiluminescence immunoassays and categorized into sex- and age-specific quartiles. Results After adjusting for age, waist circumference and type 2 diabetes mellitus, analysis of variance (ANOVA) revealed inverse associations between serum IGF-I concentrations and eGFR in men as well as between serum IGFBP-3 concentrations and eGFR in men and women. Logistic regression analyses confirmed these findings and showed that high IGF-I or IGFBP-3 concentrations were associated with an increased risk of decreased eGFR (<60 mL/min/1.73 m2) in men or women. These relations became stronger when lower eGFR cut-offs were used for the analyses. Conclusion Our data revealed associations of increased serum IGF-I concentrations and decreased eGFR in men but not in women and an association of increased serum IGFBP-3 concentrations and decreased eGFR in both sexes. PMID:23237568

  18. Serum Alkaline Phosphatase and Risk of Incident Cardiovascular Disease: Interrelationship with High Sensitivity C-Reactive Protein

    PubMed Central

    Kunutsor, Setor K.; Bakker, Stephan J. L.; Kootstra-Ros, Jenny E.; Gansevoort, Ronald T.; Gregson, John; Dullaart, Robin P. F.

    2015-01-01

    Background Alkaline phosphatase (ALP) has been suggested to be associated with cardiovascular disease (CVD) risk, however, important aspects of the association, such as shape and independence from established risk factors, have yet to be characterized in detail. We assessed the association of ALP with CVD risk and determined its utility for CVD risk prediction. Methods Alkaline phosphatase activity was measured at baseline in the PREVEND prospective cohort involving 6,974 participants aged 28-75 years without pre-existing CVD. Hazard ratios (95% confidence intervals [CI]) and measures of risk discrimination and reclassification were assessed. Results During a median follow-up of 10.5 years, 737 participants developed CVD. Serum ALP was correlated with several risk markers for CVD, with strongest correlations for age (r = 0.30; P < 0.001), gamma-glutamyltransferase (r = 0.26; P < 0.001), and C-reactive protein (CRP) (r = 0.25; P < 0.001). There was a non-linear “J-shaped” relationship between ALP and CVD risk. In analyses adjusted for conventional risk factors, the hazard ratio (95% CI) for CVD in a comparison of the top quintile versus bottom quintiles 1-4 of ALP values was 1.34 (1.14 to 1.56; P<0.001), which persisted after additional adjustment for potential confounders 1.33 (1.13 to 1.55; P<0.001). However, the association was somewhat attenuated after adjustment for CRP 1.24 (1.05 to 1.45; P=0.009). Addition of information on ALP to a CVD risk prediction model containing established risk factors did not improve the C-index or net reclassification. Conclusions Available evidence suggests a non-linear association between ALP activity and CVD risk, which is partly dependent on CRP. Taking account of conventional risk factors, additional information on ALP does not improve CVD risk assessment. PMID:26167920

  19. Deglycosylation of serum vitamin D3-binding protein by alpha-N-acetylgalactosaminidase detected in the plasma of patients with systemic lupus erythematosus.

    PubMed

    Yamamoto, N; Naraparaju, V R; Moore, M; Brent, L H

    1997-03-01

    A serum glycoprotein, Gc protein (vitamin D3-binding protein), can be converted by beta-galactosidase of B cells and sialidase of T cells to a potent macrophage-activating factor (MAF), a protein with N-acetylgalactosamine as the remaining sugar moiety. Thus, Gc protein is the precursor for MAF. Treatment of Gc protein with immobilized beta-galactosidase and sialidase generates a remarkably high titered macrophage-activating factor (GcMAF). When peripheral blood monocytes/ macrophages (designated macrophages) of 33 systemic lupus erythematosus patients were incubated with GcMAF (100 pg/ml), the macrophages of all patients were activated as determined by superoxide generation. However, the precursor activity of patient plasma Gc protein was lost or reduced in these patients. Loss of the precursor activity was the result of deglycosylation of plasma Gc protein by alpha-N-acetylgalactosaminidase activity found in the patient plasma. Levels of plasma alpha-N-acetylgalactosaminidase activity in individual patients had an inverse correlation with the MAF precursor activity of their plasma Gc protein. Deglycosylated Gc protein cannot be converted to macro-phage-activating factor. The resulting defect in macro-phage activation may lead to an inability to clear pathogenic immune complexes. Thus, elevated plasma alpha-N-acetylgalactosaminidase activity resulting in the loss of MAF precursor activity and reduced macro-phage activity may play a role in the pathogenesis of systemic lupus erythematosus. PMID:9073553

  20. Associations of serum insulin-like growth factor-I and insulin-like growth factor-binding protein 3 levels with biomarker-calibrated protein, dairy product and milk intake in the Women's Health Initiative.

    PubMed

    Beasley, Jeannette M; Gunter, Marc J; LaCroix, Andrea Z; Prentice, Ross L; Neuhouser, Marian L; Tinker, Lesley F; Vitolins, Mara Z; Strickler, Howard D

    2014-03-14

    It is well established that protein-energy malnutrition decreases serum insulin-like growth factor (IGF)-I levels, and supplementation of 30 g of whey protein daily has been shown to increase serum IGF-I levels by 8 % after 2 years in a clinical trial. Cohort studies provide the opportunity to assess associations between dietary protein intake and IGF axis protein levels under more typical eating conditions. In the present study, we assessed the associations of circulating IGF axis protein levels (ELISA, Diagnostic Systems Laboratories) with total biomarker-calibrated protein intake, as well as with dairy product and milk intake, among postmenopausal women enrolled in the Women's Health Initiative (n 747). Analyses were carried out using multivariate linear regression models that adjusted for age, BMI, race/ethnicity, education, biomarker-calibrated energy intake, alcohol intake, smoking, physical activity and hormone therapy use. There was a positive association between milk intake and free IGF-I levels. A three-serving increase in milk intake per d (approximately 30 g of protein) was associated with an estimated average 18·6 % higher increase in free IGF-I levels (95 % CI 0·9, 39·3 %). However, total IGF-I and insulin-like growth factor-binding protein 3 (IGFBP-3) levels were not associated with milk consumption and nor were there associations between biomarker-calibrated protein intake, biomarker-calibrated energy intake, and free IGF-I, total IGF-I or IGFBP-3 levels. The findings of the present study carried out in postmenopausal women are consistent with clinical trial data suggesting a specific relationship between milk consumption and serum IGF-I levels, although in the present study this association was only statistically significant for free, but not total, IGF-I or IGFBP-3 levels. PMID:24094144

  1. Serum protein removal from skim milk with a 3-stage, 3× ceramic Isoflux membrane process at 50°C.

    PubMed

    Adams, Michael C; Barbano, David M

    2013-04-01

    Small pore microfiltration (MF) can be used to remove serum proteins (SP) from skim milk. The process's SP removal efficiency directly influences the technology's economic feasibility. Our objective was to quantify the capacity of 0.14?m ceramic Isoflux MF membranes (TAMI, Nyons, France) to remove SP from skim milk. A 3-stage, 3×, feed-and-bleed MF study with diafiltration in the latter 2 stages was conducted at 50°C using Isoflux membranes to determine cumulative SP removal percentages and SP removal rates at each processing stage. The experiment was replicated 3 times starting with 3 separate lots of raw milk. In contrast to 3× MF theoretical cumulative SP removal percentages of 68, 90, and 97% after 1, 2, and 3 stages, respectively, the 3× Isoflux MF process removed only 39.5, 58.4, and 70.2% of SP after 1, 2, and 3 stages, respectively. Previous research has been published that provides the skim milk SP removal capacities of 3-stage, 3× 0.1?m ceramic Membralox (Pall Corp., Cortland, NY) uniform transmembrane pressure (UTP), 0.1?m ceramic Membralox graded permeability (GP), and 0.3?m polymeric polyvinylidene fluoride spiral-wound (PVDF-SW) MF systems (Parker-Hannifin, Process Advanced Filtration Division, Tell City, IN) at 50°C. No difference in cumulative SP removal percentage after 3 stages was detected between the Isoflux and previously published PVDF-SW values (70.3%), but SP removal was lower than published GP (96.5%) and UTP (98.3%) values. To remove 95% of SP from 1,000kg of skim milk in 12h it would take 7, 3, 3, and 7 stages with 6.86, 1.91, 2.82, and 17.98m(2) of membrane surface area for the Isoflux, GP, UTP, and PVDF-SW systems, respectively. The MF systems requiring more stages would produce additional permeate at lower protein concentrations. The ceramic MF systems requiring more surface area would incur higher capital costs. The authors hypothesize that SP removal with the Isoflux membranes was lower than theoretical for the following reasons: a range of membrane pore sizes existed (i.e., some pores were too small to pass SP), the selective layer modification and reverse flow conditions at the membrane outlet combined to reduce the effective membrane surface area, and the geometric shape of the Isoflux flow channels promoted early fouling of the membrane and rejection of SP by the foulant. PMID:23415524

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

    NASA Astrophysics Data System (ADS)

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

    2014-03-01

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

  3. Loss of Ypk1, the yeast homolog to the human serum- and glucocorticoid-induced protein kinase, accelerates phospholipase B1-mediated phosphatidylcholine deacylation.

    PubMed

    Surlow, Beth A; Cooley, Benjamin M; Needham, Patrick G; Brodsky, Jeffrey L; Patton-Vogt, Jana

    2014-11-01

    Ypk1, the yeast homolog of the human serum- and glucocorticoid-induced kinase (Sgk1), affects diverse cellular activities, including sphingolipid homeostasis. We now report that Ypk1 also impacts the turnover of the major phospholipid, phosphatidylcholine (PC). Pulse-chase radiolabeling reveals that a ypk1? mutant exhibits increased PC deacylation and glycerophosphocholine production compared with wild type yeast. Deletion of PLB1, a gene encoding a B-type phospholipase that hydrolyzes PC, in a ypk1? mutant curtails the increased PC deacylation. In contrast to previous data, we find that Plb1 resides in the ER and in the medium. Consistent with a link between Ypk1 and Plb1, the levels of both Plb1 protein and PLB1 message are elevated in a ypk1? strain compared with wild type yeast. Furthermore, deletion of PLB1 in a ypk1? mutant exacerbates phenotypes associated with loss of YPK1, including slowed growth and sensitivity to cell wall perturbation, suggesting that increased Plb1 activity buffers against the loss of Ypk1. Because Plb1 lacks a consensus phosphorylation site for Ypk1, we probed other processes under the control of Ypk1 that might be linked to PC turnover. Inhibition of sphingolipid biosynthesis by the drug myriocin or through utilization of a lcb1-100 mutant results in increased PLB1 expression. Furthermore, we discovered that the increase in PLB1 expression observed upon inhibition of sphingolipid synthesis or loss of Ypk1 is under the control of the Crz1 transcription factor. Taken together, these results suggest a functional interaction between Ypk1 and Plb1 in which altered sphingolipid metabolism up-regulates PLB1 expression via Crz1. PMID:25258318

  4. Application of a Serum\\/Protein-Free Medium for the Growth of Mammalian Cell Lines and High Level Production of Classical, Variant and Very Virulent Strain of Infectious Bursal Disease Virus (IBDV)

    Microsoft Academic Search

    Ane Franciele Tapparo; Paloma Oliveira Tonietti; Maria Cecília; R. Luvizotto

    2008-01-01

    The aim of this study was to verify the capacity of IBDV strains, Lukert, F52\\/70 and very virulent Brazilian genotype (G11) to infect and replicate on cell culture maintained without serum or protein supplementation. BHK21 and CER cells were directed adapted to growth under absence of protein and\\/or fetal bovine serum with addition of 5 ?g mL-1 of insulin-like growth

  5. An albumin-like protein in the serum of non-parasitic brook lamprey ( Lampetra appendix) is restricted to preadult phases of the life cycle in contrast to the parasitic species Petromyzon marinus

    Microsoft Academic Search

    Mehmet H. Danis; Michael F. Filosa; John H. Youson

    2000-01-01

    Previous work showed that the parasitic sea lamprey,Petromyzonmarinus, has two different albumin-like serum proteins during the course of its life cycle. One of these, AS, is the predominant protein in the serum of larval and metamorphosing intervals but is absent in the upstream migration phase of the life cycle; the other, SDS-1, is found at its highest level in the

  6. Serum Levels of Adipocyte Fatty Acid-Binding Protein Are Associated with the Severity of Coronary Artery Disease in Chinese Women

    PubMed Central

    Zhou, Mi; Li, Huating; Wang, Ye; Gao, Meifang; Wei, Meng; Jia, Weiping

    2011-01-01

    Background Adipocyte fatty acid-binding protein (A-FABP) has been described as a novel adipokine, playing an important role in the development of metabolic syndrome, type 2 diabetes and atherosclerosis. In this study, we investigated the relationship between serum levels of A-FABP and the presence and severity of coronary artery disease (CAD) in Chinese subjects. Methodology/Principal Findings Circulating A-FABP level was determined by ELISA in 341 Chinese subjects (221 men, 120 women) who underwent coronary angiography. A-FABP levels in patients with CAD were significantly higher compared with non-CAD subjects (P?=?0.029 in men; P?=?0.031 in women). Serum A-FABP increased significantly in multi-vessel diseased patients than in non-CAD subjects (P?=?0.011 in men, P?=?0.004 in women), and showed an independent correlation with coronary atherosclerosis index (standardized ??=?0.173, P?=?0.025). In multiple logistic regression analysis, serum A-FABP was an independent risk factor for CAD in women (OR?=?5.637, 95%CI: 1.299-24.457, P?=?0.021). In addition, amino terminal pro-brain natriuretic peptide (NT-proBNP) was demonstrated to be positively and independently correlated with A-FABP (standardized ??=?0.135, P?=?0.027). Conclusions/Significance Serum A-FABP is closely associated with the presence and severity of CAD in Chinese women. PMID:21552513

  7. FOAMING PROPERTIES OF PORCINE SERUM AND PORCINE SERUM ALBUMIN

    Microsoft Academic Search

    M. G. Ramos-Clamont; L. Vázquez-Moreno

    2006-01-01

    The purpose of this study was to compare the foam capacity (FC) and foam stability (FS) of freeze-dried porcine serum (PS) and porcine serum albumin (PSA) with egg white protein (EW). PSA was obtained by hydrophobic interaction chromatography in a single chromatographic step. Foaming properties were determined at 2, 4, 6, 8, and 10 % of protein solutions in buffer

  8. UMBILICAL CORD SERUM INTERLEUKIN-6, C-REACTIVE PROTEIN, AND MYELOPEROXIDASE CONCENTRATIONS AT BIRTH AND ASSOCIATION WITH NEONATAL MORBIDITIES AND LONG TERM NEURODEVELOPMENTAL OUTCOMES

    PubMed Central

    Sorokin, Yoram; Romero, Roberto; Mele, Lisa; Iams, Jay D.; Peaceman, Alan M.; Leveno, Kenneth J.; Harper, Margaret; Caritis, Steve N.; Mercer, Brian M.; Thorp, John M.; O’Sullivan, Mary Jo; Ramin, Susan M.; Carpenter, Marshall W.; Rouse, Dwight J.; Sibai, Baha

    2015-01-01

    OBJECTIVE To determine if umbilical cord serum concentrations of interleukin-6 (IL-6), C-reactive protein (CRP), and myeloperoxidase (MPO), in pregnancies at risk for preterm birth (PTB), are associated with neonatal morbidities and/or altered neurodevelopmental outcomes in the children. STUDY DESIGN Umbilical cord serum samples were collected at birth from 400 newborns delivered within a multicenter randomized controlled trial of repeated versus single course of antenatal corticosteroids (ACs), in women at increased risk for PTB. Newborns were followed through discharge and were evaluated between 36 and 42 months corrected age with neurological examination and Bayley Scales of Infant Development. Umbilical cord serum concentrations of IL-6, CRP, and MPO were determined using enzyme-linked immunoassays. Multivariate logistic regression analyses explored the relationship between umbilical cord serum IL-6, CRP and MPO levels, adverse newborn outcomes and PTB < 32 weeks of gestational age (GA). RESULTS Univariate analysis revealed that umbilical cord IL-6 above the 75th percentile was associated with increased respiratory distress syndrome (RDS) and chronic lung disease (CLD), but not with necrotizing enterocolitis (NEC), intraventricular hemorrhage (IVH), or neonatal sepsis; however, this association was not significant after adjusting for gestational age at delivery and treatment group. No significant associations between CRP or MPO, and RDS, CLD, NEC, sepsis or IVH were evident. Regression analysis revealed that CRP above the 75th percentile was associated with a decreased risk of CLD (O.R. 0.10, 95% C.I. 0.02–0.41). No associations between umbilical cord IL-6, CRP or MPO, and MDI < 70 or PDI < 70 were evident. Umbilical cord serum IL-6, CRP, and MPO, above the 75th percentile, were associated with more frequent PTB < 32 weeks GA. CONCLUSION Elevated umbilical cord serum concentration of CRP is associated with reduced risk for CLD even after adjusting for gestational age at delivery. Occurrence of levels > 75th percentile of IL-6, CRP, and MPO in umbilical cord serum were associated with PTB < 32 weeks GA. Elevated umbilical cord serum concentrations of IL-6, CRP, and MPO at birth were not associated with poor neurodevelopmental outcomes. PMID:24338120

  9. An extended gene protein/products boolean network model including post-transcriptional regulation

    PubMed Central

    2014-01-01

    Background Networks Biology allows the study of complex interactions between biological systems using formal, well structured, and computationally friendly models. Several different network models can be created, depending on the type of interactions that need to be investigated. Gene Regulatory Networks (GRN) are an effective model commonly used to study the complex regulatory mechanisms of a cell. Unfortunately, given their intrinsic complexity and non discrete nature, the computational study of realistic-sized complex GRNs requires some abstractions. Boolean Networks (BNs), for example, are a reliable model that can be used to represent networks where the possible state of a node is a boolean value (0 or 1). Despite this strong simplification, BNs have been used to study both structural and dynamic properties of real as well as randomly generated GRNs. Results In this paper we show how it is possible to include the post-transcriptional regulation mechanism (a key process mediated by small non-coding RNA molecules like the miRNAs) into the BN model of a GRN. The enhanced BN model is implemented in a software toolkit (EBNT) that allows to analyze boolean GRNs from both a structural and a dynamic point of view. The open-source toolkit is compatible with available visualization tools like Cytoscape and allows to run detailed analysis of the network topology as well as of its attractors, trajectories, and state-space. In the paper, a small GRN built around the mTOR gene is used to demonstrate the main capabilities of the toolkit. Conclusions The extended model proposed in this paper opens new opportunities in the study of gene regulation. Several of the successful researches done with the support of BN to understand high-level characteristics of regulatory networks, can now be improved to better understand the role of post-transcriptional regulation for example as a network-wide noise-reduction or stabilization mechanisms. PMID:25080304

  10. Once Phosphorylated, Tyrosines in Carboxyl Terminus of Protein-tyrosine Kinase Syk Interact with Signaling Proteins, Including TULA-2, a Negative Regulator of Mast Cell Degranulation*

    PubMed Central

    de Castro, Rodrigo Orlandini; Zhang, Juan; Groves, Jacqueline R.; Barbu, Emilia Alina; Siraganian, Reuben P.

    2012-01-01

    Activation of the high affinity IgE-binding receptor (Fc?RI) results in the tyrosine phosphorylation of two conserved tyrosines located close to the COOH terminus of the protein-tyrosine kinase Syk. Synthetic peptides representing the last 10 amino acids of the tail of Syk with these two tyrosines either nonphosphorylated or phosphorylated were used to precipitate proteins from mast cell lysates. Proteins specifically precipitated by the phosphorylated peptide were identified by mass spectrometry. These included the adaptor proteins SLP-76, Nck-1, Grb2, and Grb2-related adaptor downstream of Shc (GADS) and the protein phosphatases SHIP-1 and TULA-2 (also known as UBASH3B or STS-1). The presence of these in the precipitates was further confirmed by immunoblotting. Using the peptides as probes in far Western blots showed direct binding of the phosphorylated peptide to Nck-1 and SHIP-1. Immunoprecipitations suggested that there were complexes of these proteins associated with Syk especially after receptor activation; in these complexes are Nck, SHIP-1, SLP-76, Grb2, and TULA-2 (UBASH3B or STS-1). The decreased expression of TULA-2 by treatment of mast cells with siRNA increased the Fc?RI-induced tyrosine phosphorylation of the activation loop tyrosines of Syk and the phosphorylation of phospholipase C-?2. There was parallel enhancement of the receptor-induced degranulation and activation of nuclear factor for T cells or nuclear factor ?B, indicating that TULA-2, like SHIP-1, functions as a negative regulator of Fc?RI signaling in mast cells. Therefore, once phosphorylated, the terminal tyrosines of Syk bind complexes of proteins that are positive and negative regulators of signaling in mast cells. PMID:22267732

  11. Once phosphorylated, tyrosines in carboxyl terminus of protein-tyrosine kinase Syk interact with signaling proteins, including TULA-2, a negative regulator of mast cell degranulation.

    PubMed

    de Castro, Rodrigo Orlandini; Zhang, Juan; Groves, Jacqueline R; Barbu, Emilia Alina; Siraganian, Reuben P

    2012-03-01

    Activation of the high affinity IgE-binding receptor (Fc?RI) results in the tyrosine phosphorylation of two conserved tyrosines located close to the COOH terminus of the protein-tyrosine kinase Syk. Synthetic peptides representing the last 10 amino acids of the tail of Syk with these two tyrosines either nonphosphorylated or phosphorylated were used to precipitate proteins from mast cell lysates. Proteins specifically precipitated by the phosphorylated peptide were identified by mass spectrometry. These included the adaptor proteins SLP-76, Nck-1, Grb2, and Grb2-related adaptor downstream of Shc (GADS) and the protein phosphatases SHIP-1 and TULA-2 (also known as UBASH3B or STS-1). The presence of these in the precipitates was further confirmed by immunoblotting. Using the peptides as probes in far Western blots showed direct binding of the phosphorylated peptide to Nck-1 and SHIP-1. Immunoprecipitations suggested that there were complexes of these proteins associated with Syk especially after receptor activation; in these complexes are Nck, SHIP-1, SLP-76, Grb2, and TULA-2 (UBASH3B or STS-1). The decreased expression of TULA-2 by treatment of mast cells with siRNA increased the Fc?RI-induced tyrosine phosphorylation of the activation loop tyrosines of Syk and the phosphorylation of phospholipase C-?2. There was parallel enhancement of the receptor-induced degranulation and activation of nuclear factor for T cells or nuclear factor ?B, indicating that TULA-2, like SHIP-1, functions as a negative regulator of Fc?RI signaling in mast cells. Therefore, once phosphorylated, the terminal tyrosines of Syk bind complexes of proteins that are positive and negative regulators of signaling in mast cells. PMID:22267732

  12. Effects of Cyclooxygenase2 Inhibition on Serum and Tumor Gastrins and Expression of Apoptosis-Related Proteins in Colorectal Cancer

    Microsoft Academic Search

    Peter C. Konturek; Kazimierz Rembiasz; Grzegorz Burnat; Stanis?aw J. Konturek; Marcin Tusinela; W?adys?aw Bielanski; Jens Rehfeld; Danuta Karcz; Eckhart Hahn

    2006-01-01

    The objective of the present study was to determine the influence of cyclooxygenase-2 (COX-2) inhibition by Celecoxib (CLX)\\u000a in humans with distal colorectal adenocarcinoma (CRC) on serum and tumor levels of progastrin and gastrin and serum levels\\u000a of proinflammatory cytokines (IL-8, TNF-?). In addition, the effects of this CLX treatment on tumor and adjacent mucosa expression\\u000a of gastrin, its receptors

  13. The mechanisms of uremic serum-induced expression of bone matrix proteins in bovine vascular smooth muscle cells

    Microsoft Academic Search

    N X Chen; D Duan; K D O'Neill; G O Wolisi; J J Koczman; R LaClair; S M Moe

    2006-01-01

    We have previously found that uremic human serum upregulates RUNX2 in vascular smooth muscle cells (VSMCs), and that RUNX2 is upregulated in areas of vascular calcification in vivo. To confirm the role of RUNX2, we transiently transfected a dominant-negative RUNX2 (?RUNX2) construct in bovine vascular smooth muscle cells (BVSMCs). Blocking RUNX2 transcriptional activity significantly decreased uremic serum induced alkaline phosphatase

  14. The associations of diet with serum insulin-like growth factor I and its main binding proteins in 292 women meat-eaters, vegetarians, and vegans.

    PubMed

    Allen, Naomi E; Appleby, Paul N; Davey, Gwyneth K; Kaaks, Rudolf; Rinaldi, Sabina; Key, Timothy J

    2002-11-01

    The lower rates of some cancers in Asian countries than in Western countries may be partly because of diet, although the mechanisms are unknown. The aim of this cross-sectional study was to determine whether a plant-based (vegan) diet is associated with a lower circulating level of insulin-like growth factor I (IGF-I) compared with a meat-eating or lacto-ovo-vegetarian diet among 292 British women, ages 20-70 years. The mean serum IGF-I concentration was 13% lower in 92 vegan women compared with 99 meat-eaters and 101 vegetarians (P = 0.0006). The mean concentrations of both serum IGF-binding protein (IGFBP)-1 and IGFBP-2 were 20-40% higher in vegan women compared with meat-eaters and vegetarians (P = 0.005 and P = 0.0008 for IGFBP-1 and IGFBP-2, respectively). There were no significant differences in IGFBP-3, C-peptide, or sex hormone-binding globulin concentrations between the diet groups. Intake of protein rich in essential amino acids was positively associated with serum IGF-I (Pearson partial correlation coefficient; r = 0.27; P < 0.0001) and explained most of the differences in IGF-I concentration between the diet groups. These data suggest that a plant-based diet is associated with lower circulating levels of total IGF-I and higher levels of IGFBP-1 and IGFBP-2. PMID:12433724

  15. Characterization of serum protein components from Donryu rats which were immune to hepatoma AH-66 tumor cells.

    PubMed

    Wepsic, H T; Tsukada, Y; Kobayashi, K; Hara, A; Hirai, H

    1982-08-01

    Immune serum from Donryu rats which resisted rechallenge with 5 X 10(6) AH-66 tumor cells (1000 X Threshold) was examined for reactivity with these cells using three assays: 1) tumor cell agglutination 2) immunofluorescence and 3) complement mediated cytotoxicity. Whole immune serum and its components had different levels of activities in the various assays and differences in their antigenic specificities were also observed. Whole immune serum showed some activity in all assays to AH-66 tumor cells. Its specificity and that of the IgG "enriched" fraction (insoluble in 40% sat. AmSO4) was studied using the agglutination assay. A general cross-reactivity was seen with all tumor cell lines. These reactions could be eliminated by absorption of the serum or serum fraction with tumor lines other than AH-66 or with embryonic tissue. An IgG "free" derived from the 40% sat. AmSO4 precipitate showed only agglutinin activity but this reactivity, even to AH-66 tumor cells, was abrogated by absorption with embryonic tissue. Incubation of the IgG "free" fraction with tumor target cells blocked the complement mediated cytotoxicity and the immunofluorescence reactivities of the whole immune serum. The alpha 2 macroglobulin component of the IgG "free" fraction appeared to be responsible for its agglutinin activity. PMID:6184506

  16. Characterization of a monoclonal antibody to human serum vitamin D binding protein (Gc globulin): recognition of an epitope hidden in membranes of circulating monocytes.

    PubMed

    McLeod, J F; Kowalski, M A; Haddad, J G

    1986-07-01

    We have developed a murine hybridoma cell line that secretes a monoclonal antibody directed to the serum human vitamin D binding protein (hDBP), a 58,000-dalton alpha-globulin with a high avidity for 25-hydroxycholecalciferol and globular actin. This immunoglobulin G1 kappa-light chain antibody was produced by the fusion of the spleen cells from BALB/c mice, immunized with purified hDBP, with SP2/0-AG4 myeloma cells. The antibody was easily removed from the supernatant of hybridoma cultures or mouse ascites fluid by Protein A affinity chromatography. Apparent serum monospecificity was demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and isoelectric focusing gels transblotted to nylon membranes and overlayed with purified MAK 89 antibody and radioiodinated Protein A. The affinity of the antibody is high [dissociation constant (Kd) = 2.6 X 10(-11) M]. Parallel displacement of tracer by hDBP and human serum was observed. The sera from various species displaced the hDBP tracer in the following potency: monkey more than cat more than dog more than guinea pig. RIAs for DBP from several species are feasible with this antibody. This antibody does not, in contrast to polyclonal anti-hDBP antiserum, bind to viable monocytes. However, the MAK 89 antibody does bind to the membranes of well washed, fixed, and permeant circulating monocytes. Surface membrane radioiodination of monocytes and immunoprecipitation of the detergent lysates with the antibody demonstrates a protein with molecular weight equivalent to hDBP. The epitope recognized, therefore, appears to be hidden in the viable cells, suggesting an intimate and intricate association of the hDBP and monocyte plasma membrane. PMID:2424747

  17. Canine distemper virus neutralization activity is low in human serum and it is sensitive to an amino acid substitution in the hemagglutinin protein.

    PubMed

    Zhang, Xinsheng; Wallace, Olivia L; Domi, Arban; Wright, Kevin J; Driscoll, Jonathan; Anzala, Omu; Sanders, Eduard J; Kamali, Anatoli; Karita, Etienne; Allen, Susan; Fast, Pat; Gilmour, Jill; Price, Matt A; Parks, Christopher L

    2015-08-01

    Serum was analyzed from 146 healthy adult volunteers in eastern Africa to evaluate measles virus (MV) and canine distemper virus (CDV) neutralizing antibody (nAb) prevalence and potency. MV plaque reduction neutralization test (PRNT) results indicated that all sera were positive for MV nAbs. Furthermore, the 50% neutralizing dose (ND50) for the majority of sera corresponded to antibody titers induced by MV vaccination. CDV nAbs titers were low and generally were detected in sera with high MV nAb titers. A mutant CDV was generated that was less sensitive to neutralization by human serum. The mutant virus genome had 10 nucleotide substitutions, which coded for single amino acid substitutions in the fusion (F) and hemagglutinin (H) glycoproteins and two substitutions in the large polymerase (L) protein. The H substitution occurred in a conserved region involved in receptor interactions among morbilliviruses, implying that this region is a target for cross-reactive neutralizing antibodies. PMID:25880113

  18. Serum factors involved in human microvascular endothelial cell morphogenesis.

    PubMed

    Harvey, Kevin; Siddiqui, Rafat A; Sliva, Daniel; Garcia, Joe G N; English, Denis

    2002-09-01

    Our previous studies have demonstrated that lipid and protein angiogenic factors operate in tandem to induce optimal angiogenic responses in vivo. This study was undertaken to clarify the nature of the substances in human serum that are responsible for its remarkable ability to promote capillary morphogenesis in vitro. The ability of dilute (2%) human serum to promote the morphogenic differentiation of human dermal microvascular endothelial cells on Matrigel supports was depleted by more than 50% by treatment of the serum with activated charcoal, a procedure that effectively removes biologically active lipid growth factors. The remainder of the activity within serum was lost on heating to 60 degrees C for 60 minutes, indicating the involvement of a protein in the response. The ability of charcoal-treated serum to promote capillary morphogenesis was completely restored by the addition of sphingosine 1-phosphate (SPP, 500 nmol/L), but other lipids thought to be released into serum during clotting were ineffective. In addition, basic fibroblast growth factor (bFGF) effectively restored the ability of heat-treated serum to promote endothelial cell morphogenesis, but other protein growth factors, including vascular endothelial growth factor and platelet-derived growth factor, were ineffective. Together, SPP and bFGF were as effective as whole serum in promoting capillary morphogenesis. Responses to purified SPP were entirely sensitive to the effects of preexposure of the cells to pertussis toxin, whereas responses to bFGF were entirely pertussis toxin-resistant. Consistent with our hypothesis that two distinct factors in serum play a role in promoting capillary morphogenesis, responses induced by serum were inhibited approximately 50% by preexposure of endothelial cells to pertussis toxin. We conclude that platelet-released SPP acts in conjunction with circulating bFGF to promote capillary formation by microvascular endothelial cells. Lipid and protein growth factors apparently exert complementary roles in the angiogenic response, as demonstrated by their ability to promote chemotaxis, angiogenic differentiation, and angiogenesis in vivo. PMID:12271276

  19. Effect of annatto addition and bleaching treatments on ultrafiltration flux during production of 80% whey protein concentrate and 80% serum protein concentrate.

    PubMed

    Adams, Michael C; Zulewska, Justyna; Barbano, David M

    2013-04-01

    The goals of this study were to determine if adding annatto color to milk or applying a bleaching process to whey or microfiltration (MF) permeate influenced ultrafiltration (UF) flux, diafiltration (DF) flux, or membrane fouling during production of 80% whey protein concentrate (WPC80) or 80% serum protein concentrate (SPC80). Separated Cheddar cheese whey (18 vats using 900 kg of whole milk each) and MF permeate of skim milk (18 processing runs using 800 kg of skim milk each) were produced to make WPC80 and SPC80, respectively. The 6 treatments, replicated 3 times each, that constituted the 18 processing runs within either whey or MF permeate UF were as follows: (1) no annatto; (2) no annatto+benzoyl peroxide (BPO); (3) no annatto+hydrogen peroxide (H2O2); (4) annatto; (5) annatto+BPO; and (6) annatto+H2O2. Approximately 700 kg of whey or 530 kg of MF permeate from each treatment were heated to 50°C and processed in 2 stages (UF and DF) with the UF system in batch recirculation mode using a polyethersulfone spiral-wound UF membrane with a molecular weight cutoff of 10,000 Da. Addition of annatto color had no effect on UF or DF flux. The processes of bleaching whey or MF permeate with or without added color improved flux during processing. Bleaching with H2O2 usually produced higher flux than bleaching with BPO. Bleaching with BPO increased WPC80 flux to a greater extent than it did SPC80 flux. Though no differences in mean flux were observed for a common bleaching treatment between the WPC80 and SPC80 production processes during the UF stage, mean flux during WPC80 DF was higher than mean flux during SPC80 DF for each bleaching treatment. Water flux values before and after processing were used to calculate a fouling coefficient that demonstrated differences in fouling which were consistent with flux differences among treatments. In both processes, bleaching with H2O2 led to the largest reduction in fouling. No effect of annatto on fouling was observed. The reasons for flux enhancement associated with bleaching treatments are unclear. PMID:23415538

  20. a p p l i c a t i o n n o t e Identification of human serum proteins detectable after Albumin removal with

    E-print Network

    Lebendiker, Mario

    in the analysis of blood proteins is the high dynamic range of the protein species, which can show differences effects, including masking many proteins located in the area of albumin itself and rendering the detection through. The concentrator was refilled with ultrapure water for desalting and centrifuged to a sample

  1. Effect of microfiltration concentration factor on serum protein removal from skim milk using spiral-wound polymeric membranes.

    PubMed

    Beckman, S L; Barbano, D M

    2013-10-01

    Our objective was to determine the effect of concentration factor (CF) on the removal of serum protein (SP) from skim milk during microfiltration (MF) at 50 °C using a 0.3-?m-pore-size spiral-wound (SW) polymeric polyvinylidene fluoride (PVDF) membrane. Pasteurized (72°C for 16 s) skim milk was MF (50 °C) at 3 CF (1.50, 2.25, and 3.00×), each on a separate day of processing starting with skim milk. Two phases of MF were used at each CF, with an initial startup-stabilization phase (40 min in full recycle mode) to achieve the desired CF, followed by a steady-state phase (90-min feed-and-bleed with recycle) where data was collected. The experiment was replicated 3 times, and SP removal from skim milk was quantified at each CF. System pressures, flow rates, CF, and fluxes were monitored during the 90-min run. Permeate flux increased (12.8, 15.3, and 19.0 kg/m(2) per hour) with decreasing CF from 3.00 to 1.50×, whereas fouled water flux did not differ among CF, indicating that the effect of membrane fouling on hydraulic resistance of the membrane was similar at all CF. However, the CF used when microfiltering skim milk (50°C) with a 0.3-?m polymeric SW PVDF membrane did affect the percentage of SP removed. As CF increased from 1.50 to 3.00×, the percentage of SP removed from skim milk increased from 10.56 to 35.57%, in a single stage bleed-and-feed MF system. Percentage SP removal from skim milk was lower than the theoretical value. Rejection of SP during MF of skim milk with SW PVDF membranes was caused by fouling of the membrane, not by the membrane itself and differences in the foulant characteristic among CF influenced SP rejection more than it influenced hydraulic resistance. We hypothesize that differences in the conditions near the surface of the membrane and within the pores during the first few minutes of processing, when casein micelles pass through the membrane, influenced the rejection of SP because more pore size narrowing and plugging occurred at low CF than at high CF due to a slower rate of gel layer formation at low CF. It is possible that percentage removal of SP from skim milk at 50 °C could be improved by optimization of the membrane pore size, feed solution composition and concentration, and controlling the rate of formation of the concentration polarization-derived gel layer at the surface of the membrane during the first few minutes of processing. PMID:23891300

  2. Lack of Relationship between Cord Serum Angiopoietin-Like Protein 4 (ANGPTL4) and Lipolytic Activity in Human Neonates Born by Spontaneous Delivery

    PubMed Central

    Ortega-Senovilla, Henar; Schaefer-Graf, Ute; Meitzner, Katrin; Graf, Kristof; Abou-Dakn, Michael; Herrera, Emilio

    2013-01-01

    Background Ligands of peroxisome-proliferator activated receptors (PPARs), such as non-esterified fatty acids (NEFAs), induce expression of angiopoietin-like protein 4 (ANGPTL4). Recently ANGPTL4 has been reported to be a mediator of intracellular adipose lipolysis induced by glucocorticoids. Objective To determine the concentrations of ANGPTL4 in cord serum of neonates born by spontaneous vaginal delivery (SVD) and by pre-labor cesarean section (CS) from healthy women, and to relate them to parameters of neonatal lipolytic activity at birth. Measurements In 54 neonates born by SVD and in 56 neonates born by CS, arterial cord blood was drawn to determine insulin, cortisol, triacylglycerols (TAGs), glycerol, non-esterified fatty acids (NEFAs), individual fatty acids, ANGPTL4, adiponectin, retinol binding protein 4 (RBP4) and leptin. Results Birth weight and neonatal fat mass in SVD and CS showed no difference, but the concentrations of glycerol, adiponectin, RBP4, NEFAs and most individual fatty acids were higher in cord serum of neonates born by SVD compared to CS, indicating a higher adipose tissue breakdown in the SVD group. The concentrations of TAG and cortisol were also higher and that of insulin was lower in cord serum of SVD compared to the CS group. However, the concentration in cord serum of ANGPTL4 did not differ between the two groups and no positive correlation with either NEFA or glycerol concentrations were detected. Conclusion ANGPTL4 is known to stimulate lipolysis in adults, but does not appear to mediate the increased activity in SVD, indicating the presence of different regulatory inputs. PMID:24324678

  3. A prediction method for the isoelectric point of binary protein mixtures of bovine serum albumin and lysozyme adsorbed on colloidal titania and alumina particles.

    PubMed

    Rezwan, Kurosch; Meier, Lorenz P; Gauckler, Ludwig J

    2005-04-12

    Bovine serum albumin and lysozyme mixtures of different mole fractions were adsorbed to colloidal alumina (116 nm) and titania particle (271 nm) suspensions of 2 vol % solid content for 16 h at pH 7.5. The total protein amount normalized to the powder surface area was 1000 ng/cm2. The zeta potential of the protein-treated suspensions was measured as a function of pH and the isoelectric point (IEP) obtained. A simple prediction model in two refinement steps was derived and evaluated for the obtained IEPs. The best model fit which takes into account moles of protein and surface fractions yielded an average prediction error of 7.5% and a maximum error of 16.7%. PMID:15807593

  4. Serum hyaluronic acid levels do not explain morning stiffness in patients with fibromyalgia

    Microsoft Academic Search

    Jong-Sun Kim; Shin-Seok Lee; Tae-Jong Kim; Yong-Wook Park

    2010-01-01

    To investigate the serum levels of hyaluronic acid (HA) in Korean female patients with fibromyalgia (FM) and correlate these\\u000a levels with variables of disease severity including morning stiffness, we measured HA serum levels in 69 FM patients, 72 rheumatoid\\u000a arthritis (RA) patients, and 71 healthy controls by enzyme-linked binding protein assay. The serum levels of HA in FM patients\\u000a did

  5. Elevated serum levels of Wisteria floribunda agglutinin-positive human Mac-2 binding protein predict the development of hepatocellular carcinoma in hepatitis C patients

    PubMed Central

    Yamasaki, Kazumi; Tateyama, Masakuni; Abiru, Seigo; Komori, Atsumasa; Nagaoka, Shinya; Saeki, Akira; Hashimoto, Satoru; Sasaki, Ryu; Bekki, Shigemune; Kugiyama, Yuki; Miyazoe, Yuri; Kuno, Atsushi; Korenaga, Masaaki; Togayachi, Akira; Ocho, Makoto; Mizokami, Masashi; Narimatsu, Hisashi; Yatsuhashi, Hiroshi

    2014-01-01

    The Wisteria floribunda agglutinin-positive human Mac-2-binding protein (WFA+-M2BP) was recently shown to be a liver fibrosis glycobiomarker with a unique fibrosis-related glycoalteration. We evaluated the ability of WFA+-M2BP to predict the development of hepatocellular carcinoma (HCC) in patients who were infected with the hepatitis C virus (HCV). A total of 707 patients who had been admitted to our hospital with chronic HCV infection without other potential risk factors were evaluated to determine the ability of WFA+-M2BP to predict the development of HCC; factors evaluated included age, sex, viral load, genotypes, fibrosis stage, aspartate and alanine aminotransferase levels, bilirubin, albumin, platelet count, alpha-fetoprotein (AFP), WFA+-M2BP, and the response to interferon (IFN) therapy. Serum WFA+-M2BP levels were significantly increased according to the progression of liver fibrosis stage (P?57 years, F4, AFP >20 ng/mL, WFA+-M2BP ?4, and WFA+-M2BP 1-4 as well as the response to IFN (no therapy vs. sustained virological response) as independent risk factors for the development of HCC. The time-dependent areas under the receiver operating characteristic curve demonstrated that the WFA+-M2BP assay predicted the development of HCC with higher diagnostic accuracy than AFP. Conclusion: WFA+-M2BP can be applied as a useful surrogate marker for the risk of HCC development, in addition to liver biopsy. (Hepatology 2014;60:1563–1570) PMID:25042054

  6. Serum antibody response to respiratory syncytial virus F and N proteins in two populations at high risk of infection: children and elderly.

    PubMed

    Sastre, P; Cusi, M G; Manoha, C; Schildgen, O; Ruiz, T; Vela, C; Rueda, P

    2010-09-01

    Human respiratory syncytial virus (hRSV) is the main viral cause of severe respiratory infections in children and a common cause of morbidity in the elderly. The nucleocapsid (N) and fusion (F) proteins of hRSV were expressed in insect cells and used as antigens in two independent enzyme-linked immunosorbent assays (ELISAs) to measure the serum antibody response in two populations at high risk of hRSV infection, children and the elderly. Fifty-seven serum specimens from children aged from 1 to 10 years old and 91 sera from adults over 60 years old were tested. The ELISA results were compared with those obtained by an immunofluorescence assay (IFA) based on hRSV-infected cells, which was considered as the reference technique. Sensitivity and specificity were 94% and 85% for the N-ELISA and 86% and 81% for the F-ELISA, respectively. When the immune responses of the two groups of individuals were compared, it appeared that almost 100% of the elderly had antibodies against the N or F protein whereas only 50% of the sera from children had antibodies against either of the two viral proteins. In conclusion, the F and N ELISAs can be used successfully for detecting a specific antibody response to hRSV. PMID:20488207

  7. Serum ceruloplasmin protein expression and activity increases in iron-deficient rats and is further enhanced by higher dietary copper intake

    PubMed Central

    Ranganathan, Perungavur N.; Lu, Yan; Jiang, Lingli; Kim, Changae

    2011-01-01

    Increases in serum and liver copper content are noted during iron deficiency in mammals, suggesting that copper-dependent processes participate during iron deprivation. One point of intersection between the 2 metals is the liver-derived, multicopper ferroxidase ceruloplasmin (Cp) that is important for iron release from certain tissues. The current study sought to explore Cp expression and activity during physiologic states in which hepatic copper loading occurs (eg, iron deficiency). Weanling rats were fed control or low iron diets containing low, normal, or high copper for ? 5 weeks, and parameters of iron homeostasis were measured. Liver copper increased in control and iron-deficient rats fed extra copper. Hepatic Cp mRNA levels did not change; however, serum Cp protein was higher during iron deprivation and with higher copper consumption. In-gel and spectrophotometric ferroxidase and amine oxidase assays demonstrated that Cp activity was enhanced when hepatic copper loading occurred. Interestingly, liver copper levels strongly correlated with Cp protein expression and activity. These observations support the possibility that liver copper loading increases metallation of the Cp protein, leading to increased production of the holo enzyme. Moreover, this phenomenon may play an important role in the compensatory response to maintain iron homeostasis during iron deficiency. PMID:21768302

  8. A targeted multiplexed proteomic investigation identifies ketamine-induced changes in immune markers in rat serum and expression changes in protein kinases/phosphatases in rat brain.

    PubMed

    Wesseling, Hendrik; Rahmoune, Hassan; Tricklebank, Mark; Guest, Paul C; Bahn, Sabine

    2015-01-01

    There is substantial interest in the N-methyl-d-aspartate (NMDA) receptor antagonist ketamine in psychiatric research because it exerts acute psychotomimetic and rapid antidepressant effects in rodents and humans. Here, we investigated proteomic changes in brain and serum after acute treatment of rats with ketamine using two targeted proteomic profiling methods. Multiplex immunoassay profiling of serum identified altered levels of interleukin 4, tumor necrosis factor alpha, and fibroblast growth factor 9, suggesting a link between ketamine exposure and peripheral inflammation and growth factor dysregulation. Selected reaction monitoring mass spectrometry profiling of rat brain tissue found that proteomic changes occurred in the frontal cortex and to a greater extent in the hippocampus. This involved changes in signaling kinases and proteases such as protein kinase C beta, neurochondrin (NCDN), calcineurin, extracellular signal-regulated kinsase 1 (ERK1), and mammalian target of rapamycin (MTOR). Furthermore, altered levels were found for proteins associated with neurotransmitter metabolism (mitochondrial aspartate aminotransferase, catechol O-methyl transferase, synaptic vesicle endo-/exocytosis (vesicle fusing ATPase (NSF), synapsin 1 (SYN1), syndapin-1 (PACN1)). Consistent with previous global proteomic studies, we confirmed known changes in mitochondrial complex I, prohibitin (PHB) and neurofilament proteins (neurofilament light chain and ?-internexin (AINX)). Taken together, the proteomic changes parallel those described in human psychiatric pathology. The results will help to elucidate ketamine's mechanism of action, which will facilitate development of novel drugs for the treatment of schizophrenia and major depressive disorder. PMID:25363195

  9. Exploring the affinity binding of alkylmaltoside surfactants to bovine serum albumin and their effect on the protein stability: A spectroscopic approach.

    PubMed

    Hierrezuelo, J M; Carnero Ruiz, C

    2015-08-01

    Steady-state and time-resolved fluorescence together with circular dichroism (CD) spectroscopic studies was performed to examine the interactions between bovine serum albumin (BSA) and two alkylmaltoside surfactants, i.e. n-decyl-?-d-maltoside (?-C10G2) and n-dodecyl-?-d-maltoside (?-C12G2), having identical structures but different tail lengths. Changes in the intrinsic fluorescence of BSA from static as well as dynamic measurements revealed a weak protein-surfactant interaction and gave the corresponding binding curves, suggesting that the binding mechanism of surfactants to protein is essentially cooperative in nature. The behavior of both surfactants is similar, so that the differences detected were attributed to the more hydrophobic nature of ?-C12G2, which favors the adsorption of micelle-like aggregates onto the protein surface. These observations were substantially demonstrated by data derived from synchronous, three-dimensional and anisotropy fluorescence experiments. Changes in the secondary structure of the protein induced by the interaction with surfactants were analyzed by CD to determine the contents of ?-helix and ?-strand. It was noted that whereas the addition of ?-C10G2 appears to stabilize the secondary structure of the protein, ?-C12G2 causes a marginal denaturation of BSA for a protein:surfactant molar ratio as high as 1 to 100. PMID:26042703

  10. Effects of dietary protein and amino acid levels on the expression of selected cationic amino acid transporters and serum amino acid concentration in growing pigs.

    PubMed

    García-Villalobos, Héctor; Morales-Trejo, Adriana; Araiza-Piña, Benedicto A; Htoo, John K; Cervantes-Ramírez, Miguel

    2012-08-01

    The absorption of lysine is facilitated by leucine, but there is no information regarding the effect of crude protein, lysine and leucine levels on the expression of cationic amino acid transporters in pigs. Therefore, an experiment was conducted with 20 pigs (14.9 +/- 0.62 kg initial body weight) to evaluate the effect of two protein levels, and the content of lysine, threonine, methionine and leucine in low crude protein diets on the expression of b(0,+) and CAT-1 mRNA in jejunum, Longissimus dorsi and Semitendinosus muscles and serum concentration of amino acids. Treatments were as follows: (i) wheat-soybean meal diet, 20% crude protein (Control); (ii) wheat diet deficient in lysine, threonine and methionine (Basal diet); (iii) Basal diet plus 0.70% L-lysine, 0.27% L-threonine, 0.10% DL-methionine (Diet LTM); (iv) Diet LTM plus 0.80% L-leucine (Diet LTM + Leu). Despite the Basal diet, all diets were formulated to meet the requirements of lysine, threonine and methionine; Diet LTM + Leu supplied 60% excess of leucine. The addition of lysine, threonine and methionine in Diet LTM increased the expression of b(0,+) in jejunum and CAT-1 in the Semitendinosus and Longissiums muscles and decreased CAT-1 in jejunum; the serum concentration of lysine was also increased (p < 0.01). Further addition of L-leucine (Diet LTM + Leu) decreased the b(0,+) expression in jejunum and CAT-1 in the Longissimus dorsi muscle (p < 0.05), increased the serum concentration ofleucine and arginine and decreased the concentration of isoleucine (p < 0.05). Pigs fed the Control diet expressed less b(0,+) in jejunum, and CAT-1 in the Semitendinosus and Longissiums muscles expressed more CAT-1 in jejunum (p < 0.05) and had lower serum concentration ofisoleucine, leucine and valine (p < 0.05), but higher lysine concentrations (p < 0.01) than those fed Diet LTM. These results indicated that both, the level and the source of dietary amino acids, affect the expression of cationic amino acid transporters in pigs fed wheat-based diets. PMID:22924173

  11. Effects of Leucine Supplementation and Serum Withdrawal on Branched-Chain Amino Acid Pathway Gene and Protein Expression in Mouse Adipocytes

    PubMed Central

    Vivar, Juan C.; Knight, Megan S.; Pointer, Mildred A.; Gwathmey, Judith K.; Ghosh, Sujoy

    2014-01-01

    The essential branched-chain amino acids (BCAA), leucine, valine and isoleucine, are traditionally associated with skeletal muscle growth and maintenance, energy production, and generation of neurotransmitter and gluconeogenic precursors. Recent evidence from human and animal model studies has established an additional link between BCAA levels and obesity. However, details of the mechanism of regulation of BCAA metabolism during adipogenesis are largely unknown. We interrogated whether the expression of genes and proteins involved in BCAA metabolism are sensitive to the adipocyte differentiation process, and responsive to nutrient stress from starvation or BCAA excess. Murine 3T3-L1 preadipocytes were differentiated to adipocytes under control conditions and under conditions of L-leucine supplementation or serum withdrawal. RNA and proteins were isolated at days 0, 4 and 10 of differentiation to represent pre-differentiation, early differentiation and late differentiation stages. Expression of 16 BCAA metabolism genes was quantified by quantitative real-time PCR. Expression of the protein levels of branched-chain amino acid transaminase 2 (Bcat2) and branched-chain alpha keto acid dehydrogenase (Bckdha) was quantified by immunoblotting. Under control conditions, all genes displayed induction of gene expression during early adipogenesis (Day 4) compared to Day 0. Leucine supplementation resulted in an induction of Bcat2 and Bckdha genes during early and late differentiation. Western blot analysis demonstrated condition-specific concordance between gene and protein expression. Serum withdrawal resulted in undetectable Bcat2 and Bckdha protein levels at all timepoints. These results demonstrate that the expression of genes related to BCAA metabolism are regulated during adipocyte differentiation and influenced by nutrient levels. These results provide additional insights on how BCAA metabolism is associated with adipose tissue function and extends our understanding of the transcriptomic response of this pathway to variations in nutrient availability. PMID:25050624

  12. C-reactive protein gene variants associated with recurrent pregnancy loss independent of CRP serum levels: A case-control study.

    PubMed

    Ahmed, Salma K; Mahmood, Naeema; Malalla, Zainab H; Alsobyani, Fanar M; Al-Kiyumi, Israa S; Almawi, Wassim Y

    2015-09-10

    The aim of this study is to investigate the association of recurrent pregnancy loss (RPL) with altered C-reactive protein (CRP) serum levels, and genetic variation in CRP gene. This was a retrospective case-control study, involving 275 women with three or more consecutive pregnancy losses, and 290 age-matched control women, who were recruited from outpatient obstetrics/gynecology clinics. CRP serum levels (hs-CRP) were determined by latex-enhanced nephelometry, and CRP genotyping was done by allelic discrimination. Mean serum CRP levels were higher in RPL cases than in control women, and carriage of the (minor) T allele of rs2794520 was associated with significant increase in CRP levels (P=0.017). Minor allele frequency (MAF) of rs7553007 was significantly different between RPL cases and control women, and was associated with reduced risk of RPL after adjusting for BMI and menarche. There was a significant enrichment of minor allele-carrying genotypes of rs1130864 and rs1417938 SNPs, and reduced frequency of minor allele-carrying genotypes of rs876537, rs2794520, and rs7553007 in RPL cases, thus assigning RPL-susceptible and -protective nature to these genotypes, respectively. Carriage of (minor) T allele of only rs2794520 was associated with significant increase in CRP levels. CRP variants that influenced circulating CRP levels in chronic inflammatory conditions are also associated with RPL, pointing to CRP as RPL candidate gene. PMID:26013044

  13. Neurokinin B induces c-fos transcription via protein kinase C and activation of serum response factor and Elk-1 in immortalized GnRH neurons.

    PubMed

    Glidewell-Kenney, Christine A; Trang, Crystal; Shao, Paul P; Gutierrez-Reed, Navarre; Uzo-Okereke, Adaku M; Coss, Djurdjica; Mellon, Pamela L

    2014-10-01

    Mutations in neurokinin B (NKB) and its receptor, NK3R, were identified in human patients with hypogonadotropic hypogonadism, a disorder characterized by lack of puberty and infertility. Further studies have suggested that NKB acts at the level of the hypothalamus to control GnRH neuron activity, either directly or indirectly. We recently reported that treatment with senktide, a NK3R agonist, induced GnRH secretion and expression of c-fos mRNA in GT1-7 cells. Here, we map the responsive region in the murine c-fos promoter to between -400 and -200 bp, identify the signal transducer and activator of transcription (STAT) (-345) and serum response element (-310) sites as required for induction, a modulatory role for the Ets site (-318), and show that induction is protein kinase C dependent. Using gel shift and Gal4 assays, we further show that phosphorylation of Elk-1 leads to binding to DNA in complex with serum response factor at serum response element and Ets sites within the c-fos promoter. Thus, we determine molecular mechanisms involved in NKB regulation of c-fos induction, which may play a role in modulation of GnRH neuron activation. PMID:25057795

  14. A novel multidimensional protein identification technology approach combining protein size exclusion prefractionation, peptide zwitterion-ion hydrophilic interaction chromatography, and nano-ultraperformance RP chromatography/nESI-MS2 for the in-depth analysis of the serum proteome and phosphoproteome: application to clinical sera derived from humans with benign prostate hyperplasia.

    PubMed

    Garbis, Spiros D; Roumeliotis, Theodoros I; Tyritzis, Stavros I; Zorpas, Kostas M; Pavlakis, Kitty; Constantinides, Constantinos A

    2011-02-01

    The current proof-of-principle study was aimed toward development of a novel multidimensional protein identification technology (MudPIT) approach for the in-depth proteome analysis of human serum derived from patients with benign prostate hyperplasia (BPH) using rational chromatographic design principles. This study constituted an extension of our published work relating to the identification and relative quantification of potential clinical biomarkers in BPH and prostate cancer (PCa) tissue specimens. The proposed MudPIT approach encompassed the use of three distinct yet complementary liquid chromatographic chemistries. High-pressure size-exclusion chromatography (SEC) was used for the prefractionation of serum proteins followed by their dialysis exchange and solution phase trypsin proteolysis. The tryptic peptides were then subjected to offline zwitterion-ion hydrophilic interaction chromatography (ZIC-HILIC) fractionation followed by their online analysis with reversed-phase nano-ultraperformance chromatography (RP-nUPLC) hyphenated to nanoelectrospray ionization-tandem mass spectrometry using an ion trap mass analyzer. For the spectral processing, the sequential use of the SpectrumMill, Scaffold, and InsPecT software tools was applied for the tryptic peptide product ion MS(2) spectral processing, false discovery rate (FDR) assessment, validation, and protein identification. This milestone serum analysis study allowed the confident identification of over 1955 proteins (p ? 0.05; FDR ? 5%) with a broad spectrum of biological and physicochemical properties including secreted, tissue-specific proteins spanning approximately 12 orders of magnitude as they occur in their native abundance levels in the serum matrix. Also encompassed in this proteome was the confident identification of 375 phosphoproteins (p ? 0.05; FDR ? 5%) with potential importance to cancer biology. To demonstrate the performance characteristics of this novel MudPIT approach, a comparison was made with the proteomes resulting from the immunodepletion of the high abundant albumin and IgG proteins with offline first dimensional tryptic peptide separation with both ZIC-HILIC and strong cation exchange (SCX) chromatography and their subsequent online RP-nUPLC-nESI-MS(2) analysis. PMID:21174401

  15. Molecular genetic analysis of a group A Streptococcus operon encoding serum opacity factor and a novel fibronectin-binding protein, SfbX.

    PubMed

    Jeng, Arthur; Sakota, Varja; Li, Zhongya; Datta, Vivekananda; Beall, Bernard; Nizet, Victor

    2003-02-01

    The group A Streptococcus (GAS) sof gene encodes the serum opacity factor protein, which is capable of opacifying mammalian sera and binding at least two host proteins, fibronectin and fibrinogen. The sof gene exists in approximately 50% of clinical isolates, and there is a classical association of so-called nephritogenic strains with the opacity factor-positive phenotype. In both a type emm49 strain and a type emm12 strain, the sequences upstream of the 5' end of sof and downstream of the putative terminator were determined to be nearly identical to a region in the M type 1 genome approximately 10 kb upstream of the emm1 gene. This close genetic linkage is likely reflected in the strict correlation of opacity factor phenotype with specific emm genotypes. A new fibronectin-binding protein gene, sfbX, was discovered immediately downstream of sof in emm12 and emm49 strains and in several other sof-positive strains. The sof and sfbX genes were found to be expressed on the same transcription unit, which was correlated with the putative promoter and rho-independant terminator sequences that flank these two genes. The sfbX genes from different emm types are predicted to encode approximately 650-residue surface-bound proteins sharing 89 to 92% sequence identity. SfbX residues approximately 1 to 480 are not highly similar to those of other known proteins, with the closest match being the Staphylococcus aureus coagulase protein. The remaining portions of these proteins (residues 481 to 650) contain four putative fibronectin-binding repeats highly similar to those of other streptococcal fibronectin-binding proteins and a potential LP(X)SG cell wall anchor motif. Targeted in-frame allelic-exchange mutagenesis, complementation, and heterologous-expression studies found that serum opacification is encoded by sof alone and that sfbX encodes a fibronectin-binding function. A recombinant SfbX protein was found to bind immobilized fibronectin and to partially inhibit GAS adherence to fibronectin. The sfbX gene was found to be present only in sof-positive strains, and together these genes could influence the spectrum of tissues colonized by sof-positive GAS. PMID:12562790

  16. Relation of Milk Serum Proteins and Milk Salts to the Effects of Heat Treatment on Rennet Clotting1

    Microsoft Academic Search

    A. Kannan; Robert Jenness

    1961-01-01

    SUMMARY A study was made of the effects of heat at 85 or 90°C. for 30 rain. on the rennet coagu)ability of skimmilk and of artificial systems composed of caseinate centrifuged from skimmilk and dispersed in milk dialysate with and without added milk serum pro- teins. Such heat treatments of skimmilk cause an immediate prolongation of the rennet clotting time

  17. Clinical significance of preoperative serum interleukin-6 and C-reactive protein level in breast cancer patients

    Microsoft Academic Search

    Praveen Ravishankaran; R Karunanithi

    2011-01-01

    BACKGROUND: Breast cancer is a disease that continues to plague females during their entire lifetime. IL-6 and CRP are found to be elevated in various inflammatory and malignant diseases and their levels are found to correlate with the extent of the disease. The primary objective of this study was to determine the preoperative serum levels of IL-6 and CRP in

  18. Value of detection of serum human epididymis secretory protein 4 and carbohydrate antigen 125 in diagnosis of early endometrial cancer of different pathological subtypes

    PubMed Central

    Liu, Xingui; Zhao, Fengting; Hu, Linli; Sun, Yingpu

    2015-01-01

    This study explored the value of detection of human epididymis secretory protein 4 (HE4) and carbohydrate antigen 125 (CA125) from serum in diagnosis of early endometrial cancer of different pathological subtypes and discussed the mechanism of HE4 and CA125 in diagnosis. In this study, enzyme-linked immunosorbent assay and chemiluminescent immunoassay were used to detect HE4 and CA125 from serum in endometrial cancer and control groups. Besides, the concentration of HE4 and CA125 was compared in these two groups, and the expression of CA125 and HE4 and clinicopathological characteristics in patients with endometrial cancer were also analyzed. Compared with the control group, the expression of HE4 was much higher in serum of patients with endometrial cancer, while there was no obvious change in the expression of CA125. The threshold detection value was acquired by receiver operating characteristic analysis method, that is, 141.5 pmol/L and 54.5 U/L, respectively. When comparing the concentration of HE4 in patients with endometrial cancer at the early stage (stage I) with healthy people, the difference therein had statistical significance, but there was no obvious difference in CA125. HE4 and CA125 in diagnosis of endometrial cancer in the stages I and II were found with no statistically significant difference. The difference of HE4 in the stages II and III had statistical significance while the difference of CA125 had no statistical significance. The specificity of both HE4 and CA125 was 95%, and the sensitivity of HE4 to uterine papillary serous carcinomas was higher than that to endometrioid adenocarcinoma. Thus, the serum HE4 is much better than CA125 in detecting the endometrial cancer at an early stage.

  19. Serum cleaved Tau protein and neurobehavioral battery of tests as markers of brain injury in experimental bacterial meningitis

    Microsoft Academic Search

    Jose E Irazuzta; Gabrielle de Courten-Myers; Frank P Zemlan; Marni Y. V Bekkedal; John Rossi

    2001-01-01

    Brain injury due to bacterial meningitis affects multiple areas of the brain with a heterogeneous distribution generating a challenge to assess severity. Tau proteins are microtubular binding proteins localized in the axonal compartment of neurons. Brain injury releases cleaved Tau proteins (C-tau) into the extracellular space where they are transported to the cerebral spinal fluid. We hypothesized that C-tau crosses

  20. Detection of serum antibodies to hepatitis E virus in domestic pigs in Italy using a recombinant swine HEV capsid protein

    PubMed Central

    2014-01-01

    Background The hepatitis E virus (HEV) has been detected in both humans and animals, particularly pigs, worldwide. Several evidences, including human infection following consumption of raw contaminated meat, suggest a zoonotic transmission of HEV. In Italy, large circulation of genotype 3 HEV has been reported in swine, and recent studies have confirmed the involvement of this genotype in autochthonous human cases. Result In this study 111 sera collected from healthy pigs in two Italian regions were tested for anti-HEV IgG antibodies. For specific HEV antibody detection in swine, we developed ELISA and Western blotting methods, using a truncated capsid (ORF2) protein lacking the first 111 amino acids of a swine HEV genotype 3 strain. The ORF2-based ELISA revealed anti-HEV antibodies in 104 out of 111 pigs compared with 102 detected with a commercial ELISA kit. A lower number of sera reacted with the recombinant ORF2 protein in a Western blotting format (81/111). Using a Latent class analysis (LCA), the estimated sensitivities for ELISA-ORF2 and ELISA-kit tests were 0.961 and 0.936, respectively, whereas specificities were 0.599 and 0.475. The estimated sensitivity of Western blotting was 0.775, and the specificity was 0.944. Conclusions The overall results confirm the high prevalence of HEV seropositive healthy pigs in Italy. Through comparisons with a commercial ELISA test, the swine genotype 3 HEV antigen produced in this study was proven suitable to detect anti-HEV antibodies in pig sera by both ELISA and Western Blotting. PMID:24934984

  1. Development of a highly sensitive immuno-PCR assay for the measurement of ?-galactosidase A protein levels in serum and plasma.

    PubMed

    Nakano, Sachie; Morizane, Yoshihito; Makisaka, Noriko; Suzuki, Toshihiro; Togawa, Tadayasu; Tsukimura, Takahiro; Kawashima, Ikuo; Sakuraba, Hitoshi; Shibasaki, Futoshi

    2013-01-01

    Fabry disease is an X-linked genetic disorder caused by defects in the ?-galactosidase A (GLA) gene, and heterogeneous mutations lead to quantitative and/or qualitative defects in GLA protein in male patients with Fabry disease. Random X-chromosomal inactivation modifies the clinical and biochemical features of female patients with Fabry disease. Functional polymorphisms have been frequently reported in recent times, and these increase the difficulty of understanding the pathogenetic basis of the disease. To date, GLA protein level has been measured using an enzyme-linked immunosorbent assay (ELISA). However, ELISA is not highly sensitive due to the high background noise. In this paper, we introduce a novel application of the immuno-polymerase chain reaction (PCR) method (termed Multiple Simultaneous Tag [MUSTag]) for measurement of the GLA protein level in blood samples. We compared the sensitivities of the MUSTag method with plates or magnetic beads with those of ELISA for recombinant human GLA and found that the apparent maximal sensitivity was higher for the former than for the latter. We then measured the GLA concentrations in serum and plasma from male patients with classic Fabry disease (Male Fabry), females with Fabry disease (Female Fabry), male subjects harboring the functional polymorphism p.E66Q (E66Q), and control (Control) subjects. Our results revealed that compared to the MUSTag plate and ELISA, the MUSTag beads assay afforded a clearer estimation of the GLA protein levels in the serum and plasma with minimal or no background noise, although all the methods could differentiate between the Male Fabry, E66Q, and Control groups. The Female Fabry group showed characteristic heterogeneity, which was consistent with the X-linked inheritance. This novel method is expected to be useful for the sensitive determination of GLA level in blood and elucidation of the pathogenetic basis of Fabry disease. PMID:24236025

  2. Yolk sac endoderm is the major source of serum proteins and lipids and is involved in the regulation of vascular integrity in early chick development.

    PubMed

    Nakazawa, Fumie; Alev, Cantas; Jakt, Lars M; Sheng, Guojun

    2011-08-01

    An important function of the vascular system is nutrient delivery. In adult animals, this is mediated through a close contact of the mesoderm-derived vasculature with the endoderm-derived enterocytes and hepatocytes. During embryonic development, the yolk sac (YS) endoderm has been suggested to play a similar role. Physiological and molecular nature of the contact between the YS endoderm and the vasculature is not well-understood. To understand roles of the YS endoderm in early development, we used the avian model and carried out a gene expression profiling analysis of isolated area vasculosa YS endoderm tissues from embryonic day 2-4 chick embryos, covering the first 48 hr of postcirculation development. Genes involved in lipid metabolism are highly enriched, indicating an active modification of lipid components during their transfer from the yolk to the circulatory system. We also uncovered genes encoding major serum proteins and key regulators of vascular integrity. In particular, PTGDS, an enzyme controlling the last step of prostaglandin D2 production, shows high expression in the YS endoderm. Experimental introduction of prostaglandin D2 into embryonic circulation led to intraembryonic vessel rupture. These data suggest that the YS endoderm is the major, if not exclusive, source of lipid and protein constituents of the early embryonic serum and plays an important role in the regulation of vascular integrity in developing embryo. PMID:21761483

  3. Comparative evaluation of serum C-reactive protein (CRP) levels in the different histological subtypes of esophageal cancer (squamous cell carcinoma and adenocarcinoma of esophagus).

    PubMed

    Lukaszewicz-Zaj?c, Marta; Mroczko, Barbara; Koz?owski, Miros?aw; Nikli?ski, Jacek; Lauda?ski, Jerzy; Siewko, Maria; Szmitkowski, Maciej

    2012-02-01

    Elevated C-reactive protein (CRP) levels have been found in patients with several malignancies. The aim of the present study was to analyze the diagnostic and prognostic values of CRP levels measurement in esophageal cancer (EC) patients in relation to its different histological subtypes (squamous cell carcinoma-ESCC and adenocarcinoma-AC of esophagus) and compared them with classic tumor markers-carcinoembryonic antigen (CEA) and squamous cell cancer antigen (SCC-Ag). The diagnostic sensitivity, specificity, and the areas under receiver operating characteristic curves (AUC) for all the proteins tested were defined. Serum CRP levels were statistically higher in EC, ESCC, and AC patients compared to healthy subjects and significantly increased in EC and ESCC patients with the presence of lymph node and distant metastases. The percentage of elevated CRP results in all the analyzed subgroups (EC, ESCC, and AC) was higher than CEA and SCC-Ag, similarly as AUC for CRP in comparison to SCC-Ag. Serum CRP level was a significant predictor of EC and ESCC patients' survival in univariate analysis. In conclusion, these results indicate that CRP can be used as an adjunct in evaluating the tumor markers-CEA and SCC-Ag and may improve the clinical diagnosis and follow-up of EC patients, especially for ESCC subgroup. PMID:22467322

  4. Enhancer 1 Binding Factor (E_1BF), a Ku-Related Protein, is a Growth-Regulated RNA Polymerase I Transcription Factor: Association of a Repressor Activity with Purified E_1BF from Serum-Deprived Cells

    Microsoft Academic Search

    Huifeng Niu; Samson T. Jacob

    1994-01-01

    Previous studies from this laboratory have demonstrated that the enhancer 1 binding factor (E_1BF), a Ku-related protein, purified from the serum-enriched cells functions as a positive factor in an RNA polymerase (pol I) transcription system. We have now shown that E_1BF purified from the serum-deprived cells (E_1BF_s) can inhibit rDNA transcription completely in a fractionated extract from the cells grown

  5. A screening program for trisomy 21 at 10-14 weeks using fetal nuchal translucency, maternal serum free ss-human chorionic gonadotropin and pregnancy-associated plasma protein-A

    Microsoft Academic Search

    K. Spencer; V. Souter; N. Tul; R. Snijders; K. H. Nicolaides

    1999-01-01

    Objective To examine the potential impact of combining maternal age with fetal nuchal translucency thickness and maternal serum free ?-human chorionic gonadotropin (?-hCG) and pregnancy-associated plasma protein-A (PAPP-A) in screening for trisomy 21 at 10-14 weeks of gestation. Methods Maternal serum free ?-hCG and PAPP-A were measured by Kryptor, a random access immunoassay ana- lyzer using time-resolved amplified cryptate emission,

  6. Monolith immuno-affinity enrichment liquid chromatography tandem mass spectrometry for quantitative protein analysis of recombinant bovine somatotropin in serum.

    PubMed

    Smits, Nathalie G E; Blokland, Marco H; Wubs, Klaas L; Nessen, Merel A; van Ginkel, Leen A; Nielen, Michel W F

    2015-08-01

    The use of recombinant bovine somatotropin (rbST) to enhance milk production is approved in several countries, but it is prohibited in the European Union. According to EU legislation, it is necessary to confirm positive screening results prior to enforcement. Although adequate screening assays are available nowadays, development of liquid chromatography tandem mass spectrometry (LC-MS/MS) confirmatory methods to detect low levels of rbST is still a challenge. Here, we present a novel approach using immuno-affinity enrichment on monolithic micro-columns in combination with state-of-the-art ultra-high pressure LC-MS/MS (UHPLC-MS/MS) detection. The developed approach enables detection and confirmation of rbST in serum at a decision limit (CC?) concentration of 0.8 ng mL(-1). Furthermore, the method is easy to handle, robust and reproducible. We successfully applied the confirmatory method to serum samples from rbST treated cows that were found suspect after immunoassay-based screening. The use of rbST could be confirmed over 1 week after treatment, and the developed method demonstrated the sensitivity needed for effective control. Graphical Abstract Graphical summary of the workflow, for serum preparation, enrichment with monolith microcolumns and LC-MS/MS measurement of rbST. PMID:26077745

  7. Alternatives to the Use of Fetal Bovine Serum: Serum-free Cell Culture

    Microsoft Academic Search

    Gerhard Gstraunthaler

    Summary Serum is commonly used as a supplement to cell culture media. It provides a broad spectrum of macromolecules, carrier proteins for lipoid substances and trace elements, attachment and spreading factors, low molecular weight nutrients, and hormones and growth factors. The most widely used animal serum supplement is fetal bovine serum, FBS. Since serum in general is an ill-defined component

  8. Association of Sarcopenic Obesity with Higher Serum High-Sensitivity C-Reactive Protein Levels in Chinese Older Males - A Community-Based Study (Taichung Community Health Study-Elderly, TCHS-E)

    PubMed Central

    Yang, Chuan-Wei; Li, Chia-Ing; Li, Tsai-Chung; Liu, Chiu-Shong; Lin, Chih-Hsueh; Lin, Wen-Yuan; Lin, Cheng-Chieh

    2015-01-01

    The prevalence of obesity and sarcopenia is high among the elderly. The simultaneous occurrence of these two disorders results in sarcopenic obesity. Research suggests that inflammation has an important role in the pathogenesis of obesity and sarcopenia. This study explores the impact of sarcopenic obesity on inflammatory markers, including interleukin-6 (IL-6), high-sensitivity C-reactive protein (hs-CRP), and tumor necrosis factor-alpha (TNF-?). This study is a community-based cross-sectional study. The study sample consisted of 844 community-dwelling people aged 65 years and older (448 men and 396 women). Sarcopenia was characterized by low muscle mass (skeletal muscle index < 6.87 and 5.46 kg/m2 for men and women, respectively), and obesity was characterized by excess body fat (body fat percentage greater than the 60th percentile of the study sample by sex [27.82% in men and 37.61% in women]). Older individuals identified with sarcopenic obesity were those who had both sarcopenia and obesity. Inflammatory markers such as IL-6, hs-CRP, and TNF-? were measured. The prevalence rates of obesity only, sarcopenia only, and sarcopenic obesity were 32.94%, 11.85%, and 7.23%, respectively. No difference was observed in the serum levels of IL-6 and TNF-? among the four groups of combined sarcopenia and obesity status. After multivariate adjustment, the serum hs-CRP levels in the obesity only and in the sarcopenic obesity groups were 0.14 and 0.16 mg/dL among males, respectively, which were significantly higher than that in the normal group (P=0.012 and 0.036). Our results provide evidence that obesity and sarcopenic obesity are associated with increased levels of serum hs-CRP among males. PMID:26177029

  9. Recent advances in the study of protein translocation across the membrane of the endoplasmic reticulum include insights

    E-print Network

    Nicchitta, Chris

    and compartmentalized protein synthesis has been suggested. Addresses Department of Cell Biology, Duke University of protein translocation have greatly benefited from recent structural biology studies. With the advent412 Recent advances in the study of protein translocation across the membrane of the endoplasmic

  10. Development and Characterization of a Novel Fusion Protein of a Mutated Granulocyte Colony-Stimulating Factor and Human Serum Albumin in Pichia pastoris

    PubMed Central

    Huang, Yan-Shan; Wen, Xiao-Fang; Yang, Zhi-Yu; Wu, Yi-Liang; Lu, You; Zhou, Lin-Fu

    2014-01-01

    The purpose of the present work was to develop a novel, long-acting and potent human serum albumin/granulocyte colony stimulating factor (HSA/G-CSF) therapeutic fusion protein. The novel fusion protein, called HMG, was constructed by genetically fusing mutated human derived G-CSF (mG-CSF) to the C-terminal of HSA and then prepared in Pichia pastoris. The molecular mass of HMG was about 85 kDa and the isoelectric point was 5.3. Circular dichroism spectroscopy suggested that mG-CSF retained nearly all of its native secondary structure, regardless of fusion. The binding capabilities of mG-CSF moiety to G-CSF receptor and HSA moiety to warfarin showed very little change after fusing. The bioactivity of HMG (11.0×106 IU/mg) was more than twice that of rHSA/G-CSF (4.6×106 IU/mg). A mutation was made at the 718th amino acid of HMG, substituting Ala for Thr, to investigate the glycosylation of HMG expressed in P. pastoris. Data indicated that HMG was modified at Thr718, speculatively with the addition of a mannose chain. In conclusion, a novel HSA/G-CSF fusion protein was successfully constructed based on a mutated G-CSF. This protein showed more potent bioactivity than rHSA/G-CSF and thus may be a suitable long-acting G-CSF. PMID:25535738

  11. Effects of heat stress on serum insulin, adipokines, AMP-activated protein kinase, and heat shock signal molecules in dairy cows.

    PubMed

    Min, Li; Cheng, Jian-Bo; Shi, Bao-Lu; Yang, Hong-Jian; Zheng, Nan; Wang, Jia-Qi

    2015-06-01

    Heat stress affects feed intake, milk production, and endocrine status in dairy cows. The temperature-humidity index (THI) is employed as an index to evaluate the degree of heat stress in dairy cows. However, it is difficult to ascertain whether THI is the most appropriate measurement of heat stress in dairy cows. This experiment was conducted to investigate the effects of heat stress on serum insulin, adipokines (leptin and adiponectin), AMP-activated protein kinase (AMPK), and heat shock signal molecules (heat shock transcription factor (HSF) and heat shock proteins (HSP)) in dairy cows and to research biomarkers to be used for better understanding the meaning of THI as a bioclimatic index. To achieve these objectives, two experiments were performed. The first experiment: eighteen lactating Holstein dairy cows were used. The treatments were: heat stress (HS, THI average=81.7, n=9) and cooling (CL, THI average=53.4, n=9). Samples of HS were obtained on August 16, 2013, and samples of CL were collected on April 7, 2014 in natural conditions. The second experiment: HS treatment cows (n=9) from the first experiment were fed for 8 weeks from August 16, 2013 to October 12, 2013. Samples for moderate heat stress, mild heat stress, and no heat stress were obtained, respectively, according to the physical alterations of the THI. Results showed that heat stress significantly increased the serum adiponectin, AMPK, HSF, HSP27, HSP70, and HSP90 (P<0.05). Adiponectin is strongly associated with AMPK. The increases of adiponectin and AMPK may be one of the mechanisms to maintain homeostasis in heat-stressed dairy cows. When heat stress treatment lasted 8 weeks, a higher expression of HSF and HSP70 was observed under moderate heat stress. Serum HSF and HSP70 are sensitive and accurate in heat stress and they could be potential indicators of animal response to heat stress. We recommend serum HSF and HSP70 as meaningful biomarkers to supplement the THI and evaluate moderate heat stress in dairy cows in the future. PMID:26055916

  12. Effects of heat stress on serum insulin, adipokines, AMP-activated protein kinase, and heat shock signal molecules in dairy cows*

    PubMed Central

    Min, Li; Cheng, Jian-bo; Shi, Bao-lu; Yang, Hong-jian; Zheng, Nan; Wang, Jia-qi

    2015-01-01

    Heat stress affects feed intake, milk production, and endocrine status in dairy cows. The temperature-humidity index (THI) is employed as an index to evaluate the degree of heat stress in dairy cows. However, it is difficult to ascertain whether THI is the most appropriate measurement of heat stress in dairy cows. This experiment was conducted to investigate the effects of heat stress on serum insulin, adipokines (leptin and adiponectin), AMP-activated protein kinase (AMPK), and heat shock signal molecules (heat shock transcription factor (HSF) and heat shock proteins (HSP)) in dairy cows and to research biomarkers to be used for better understanding the meaning of THI as a bioclimatic index. To achieve these objectives, two experiments were performed. The first experiment: eighteen lactating Holstein dairy cows were used. The treatments were: heat stress (HS, THI average=81.7, n=9) and cooling (CL, THI average=53.4, n=9). Samples of HS were obtained on August 16, 2013, and samples of CL were collected on April 7, 2014 in natural conditions. The second experiment: HS treatment cows (n=9) from the first experiment were fed for 8 weeks from August 16, 2013 to October 12, 2013. Samples for moderate heat stress, mild heat stress, and no heat stress were obtained, respectively, according to the physical alterations of the THI. Results showed that heat stress significantly increased the serum adiponectin, AMPK, HSF, HSP27, HSP70, and HSP90 (P<0.05). Adiponectin is strongly associated with AMPK. The increases of adiponectin and AMPK may be one of the mechanisms to maintain homeostasis in heat-stressed dairy cows. When heat stress treatment lasted 8 weeks, a higher expression of HSF and HSP70 was observed under moderate heat stress. Serum HSF and HSP70 are sensitive and accurate in heat stress and they could be potential indicators of animal response to heat stress. We recommend serum HSF and HSP70 as meaningful biomarkers to supplement the THI and evaluate moderate heat stress in dairy cows in the future. PMID:26055916

  13. May the evaluation of nitrosative stress through selective increase of 3-nitrotyrosine proteins other than nitroalbumin and dominant tyrosine-125/136 nitrosylation of serum ?-synuclein serve for diagnosis of sporadic Parkinson's disease?

    PubMed

    Fernández, Emilio; García-Moreno, José-Manuel; Martín de Pablos, Angel; Chacón, José

    2013-09-20

    Nitrosative stress, where nitrosylation of tyrosine (Tyr) leading to 3-nitrotyrosine proteins or free 3-nitrotyrosine is the most prominent change, has been proposed as a pathogenic mechanism in Parkinson's disease (PD). Levels of 3-nitrotyrosine proteins in serum and cerebrospinal fluid (CSF) of patients with PD have not been studied. Nitrosative stress-induced protein changes in serum and CSF were analyzed in patients with PD (n=54) and controls (n=40). Herein, we demonstrate the presence of nitrosative stress in serum and CSF of patients with early PD leading to selective increase of 3-nitrotyrosine proteins other than nitroalbumin, without free 3-nitrotyrosine (Hoehn-Yahr stage 1, p<0.05; stage 2, p<0.01). Among 3-nitrotyrosine proteins, nitro-?-synuclein (N-?Syn) was detected in serum, not CSF, and the sites of Tyr nitrosylation were observed to be modified in patients with early PD. Thus, the intensity of nitrosylation of Tyr125/136 residues is enhanced (stage 1, p<0.05; stage 2, p<0.01), and that of the Tyr39 site is reduced (stage 1, p<0.05), and the ratio between both parameters (?-synuclein with nitrosylated tyrosines 125 and 136 [N-?Syn-Tyr125/136]:?-synuclein with nitrosylated tyrosine 39 [N-?Syn-Tyr39] ratio) is significantly higher in patients with early PD (p<0.01). These observations lead to the hypothesis that evaluating nitrosative stress through enhanced levels of 3-nitrotyrosine proteins in serum and CSF without changes in nitroalbumin, together with the profile of tyrosine nitrosylation of serum ?Syn characterized by dominant nitrosylation of Tyr125/136, could serve for the diagnosis of sporadic PD. PMID:23418747

  14. Effect of fibroblastic growth factor (FGF) or serum on protein degradation, the migration of non-histone proteins (NHP) to the nucleus and DNA in synthesis WI-38 cells

    SciTech Connect

    Polet, H.

    1986-03-01

    WI-38 cells were induced into G/sub 0/-G/sub 1/ by limiting serum in the medium to 1% for 2 days, while proteins were labeled with /sup 3/H-leucine. Subsequent stimulation of cell growth by FGF or fetal calf serum (FCS) caused increased migration of /sup 3/H-NHP to the nucleus and DNA synthesis and a parallel decrease of proteolysis. /sup 3/H-HNP migration and DNA synthesis were not affected by blocking protein synthesis by 91% with cycloheximid. Fractionation of the nuclear /sup 3/H-NHP by IEF pH 2.5-6.5 in gels, showed that protein fractions with the highest rate of degradation in unstimulated cells correspond to the fractions with the highest rate of migration in stimulated cells, suggesting that degradation and migration of NHP are linked. FGF inhibited the cellular accumulation of /sup 3/H-chloroquine, suggesting that FGF inhibits NHP degradation via lysosomes. The lysosomotropic amine eserine had similar effects as FGF. These effects are similar to the ones reported for lectin stimulated lymphocytes. These data suggest that lysosomes play a role in cellular growth control; NHP play an important role in growth control. It is proposed that FGF induces migration of NHP to the nucleus by inhibiting their degradation in lysosomes. FGF also induced migration of /sup 3/H-histones, however its mechanism is unknown.

  15. Comparative spectroscopic studies on drug binding characteristics and protein surface hydrophobicity of native and modified forms of bovine serum albumin: Possible relevance to change in protein structure/function upon non-enzymatic glycation

    NASA Astrophysics Data System (ADS)

    Khodarahmi, Reza; Karimi, Seyyed Arash; Ashrafi Kooshk, Mohammad Reza; Ghadami, Seyyed Abolghasem; Ghobadi, Sirous; Amani, Mojtaba

    2012-04-01

    The interaction between serum albumin (SA) and drugs has provided an interesting ground for understanding of drug effects, especially in drug distribution and drug-drug interaction on SA, in the case of multi-drug therapy. Determination of the impact of various factors on drug-protein interaction is especially important upon significant binding of drug to albumin. In the present study, the interaction of two drugs (furosemide and indomethacin) with native and modified albumins were investigated by using various spectroscopic methods. Fluorescence data indicated that 1:1 binding of drugs to bovine serum albumin (BSA) is associated with quenching of albumin intrinsic fluorescence. The Job's plot also confirmed that drug binds to BSA via mentioned stoichiometry. Analysis of the quenching and thermodynamic parameters indicated that intermolecular interactions between drug and albumin may change upon protein modification. The theoretical analyses also suggested some conformational changes of interacting side chains in subdomain IIA binding site (at the vicinity of W237), which were in good agreement with experimental data. Decrease of protein surface hydrophobicity (PSH) was also observed upon both albumin modification and drug binding.

  16. Serum C-reactive Protein Level Is Associated with Renal Function and It Affects Echocardiographic Cardiovascular Disease in Pre-Dialysis Patients

    Microsoft Academic Search

    Özgür Yilmaz; Sim Kutlay

    2005-01-01

    Background: Relevance of serum CRP for morbidity and mortality in pre-dialysis patients has not been assessed extensively. The aim of this study was to determine the incidence of elevated serum CRP in pre-dialysis patients and to identify the factors that associate with serum CRP. We were also evaluated the effects of serum CRP on cardiovascular disease, anemia, and nutritional markers.

  17. Factor H-binding protein is important for meningococcal survival in human whole blood and serum and in the presence of the antimicrobial peptide LL-37.

    PubMed

    Seib, K L; Serruto, D; Oriente, F; Delany, I; Adu-Bobie, J; Veggi, D; Aricò, B; Rappuoli, R; Pizza, M

    2009-01-01

    Factor H-binding protein (fHBP; GNA1870) is one of the antigens of the recombinant vaccine against serogroup B Neisseria meningitidis, which has been developed using reverse vaccinology and is the basis of a meningococcal B vaccine entering phase III clinical trials. Binding of factor H (fH), an inhibitor of the complement alternative pathway, to fHBP enables N. meningitidis to evade killing by the innate immune system. All fHBP null mutant strains analyzed were sensitive to killing in ex vivo human whole blood and serum models of meningococcal bacteremia with respect to the isogenic wild-type strains. The fHBP mutant strains of MC58 and BZ83 (high fHBP expressors) survived in human blood and serum for less than 60 min (decrease of >2 log(10) CFU), while NZ98/254 (intermediate fHBP expressor) and 67/00 (low fHBP expressor) showed decreases of >1 log(10) CFU after 60 to 120 min of incubation. In addition, fHBP is important for survival in the presence of the antimicrobial peptide LL-37 (decrease of >3 log(10) CFU after 2 h of incubation), most likely due to electrostatic interactions between fHBP and the cationic LL-37 molecule. Hence, the expression of fHBP by N. meningitidis strains is important for survival in human blood and human serum and in the presence of LL-37, even at low levels. The functional significance of fHBP in mediating resistance to the human immune response, in addition to its widespread distribution and its ability to induce bactericidal antibodies, indicates that it is an important component of the serogroup B meningococcal vaccine. PMID:18852235

  18. Effects of Different Anesthetic Techniques on Serum Leptin, C-reactive Protein, and Cortisol Concentrations in Anorectal Surgery

    PubMed Central

    Buyukkocak, Unase; Daphan, Cagatay; Caglayan, Osman; Aydinuraz, Kuzey; Kaya, Tahsin; Saygun, Oral; Agalar, Fatih

    2006-01-01

    Aim To compare the effects of intratracheal general anesthesia (ITGA) and regional (saddle block) anesthesia on leptin, C-reactive protein (CRP), and cortisol blood concentrations during anorectal surgery. Methods Fifty-eight patients suffering from hemorrhoidal disease, pilonidal sinus, anal fissure, or anal fistula were included the study. Patients were randomly assigned into one of the two groups (n?=?29). Patients in one group received ITGA. After thiopental and fentanyl induction, vecuronium was used as a muscle relaxant. Anesthesia was maintained with sevoflurane. In the other group we applied saddle block, injecting hyperbaric bupivacaine into the subarachnoid space, through the L3-L4 intervertebral space, in the sitting position. Blood samples were collected for leptin, CRP, and cortisol analysis before the induction of anesthesia at 3 and 24 hours postoperatively. Results Preoperative leptin, CRP, and cortisol concentrations were comparable between the groups. There was no significant difference in postoperative levels of leptin and CRP in both groups. Although not significant, leptin and CRP concentrations were lower in the saddle block group at three hours postoperatively (mean?±?SD, 6.95?±?8.59 and 6.02?±?12.25, respectively) than in the ITGA group (mean?±?SD, 9.04?±?9.89 and 8.40?±?15.75, respectively). During early postoperative period, cortisol increased slightly in the ITGA group and remained at similar level in the saddle block group, but later decreased in both groups. Cortisol levels in the saddle block group were significantly lower than in the ITGA group at 3 hours postoperatively (343.7?±?329.6 vs 611.4?±?569.8; P?=?0.034). Conclusion Saddle block, a regional anesthetic technique, may attenuate stress response in patients undergoing anorectal surgery, by blocking afferent neural input during early postoperative period. PMID:17167859

  19. Prevalence and Isotypic Complexity of the Anti-Chinese Hamster Ovary Host Cell Protein Antibodies in Normal Human Serum

    Microsoft Academic Search

    Li Xue; Rasheeda Johnson; Boris Gorovits

    2010-01-01

    Host cell-derived protein impurities may be present at low levels in biopharmaceutical products. Antibodies to host cell proteins\\u000a are present in individuals with no known exposure to these products. In this study, antibodies to drug process-specific Chinese\\u000a hamster ovary host cell-derived proteins (CHO-HCP) were measured in unexposed individuals using a validated enzyme-linked\\u000a immunosorbent assay. Samples that tested positive for anti-CHO-HCP

  20. The role of fibronectin in the development of experimental amyloidosis. Evidence of immunohistochemical codistribution and binding property with serum amyloid protein A.

    PubMed Central

    Kawahara, E.; Shiroo, M.; Nakanishi, I.; Migita, S.

    1989-01-01

    Azocasein-induced amyloid A (AA) amyloidosis in CBA/K1Jms mice was investigated to elucidate a preference of serum amyloid A (SAA) deposition in the spleen. By indirect immunofluorescence using anti-SAA/AA antibodies the initial deposition of SAA/AA was recognized in the marginal zone of spleen at 20 days after azocasein injection. Indirect immunofluorescence using anti-fibronectin antibodies also showed meshwork positivity in the corresponding area more intensely than that in controls. Immunoelectron microscopy using anti-SAA/AA revealed the presence of positively stained flocculent materials on cell surfaces of macrophages in the marginal area in addition to amyloid fibril. The tissue fibronectin rapidly increased in the spleen and maintained 10 times more than that of controls until the 20th day. Binding assay of SAA on frozen sections revealed the presence of SAA-binding substances in the perifollicular area. Affinity chromatographic assay showed fibronectin have a binding capacity to SAA1 and SAA2. By binding assay on the microtiter plate, SAA had more affinity to fibronectin than those of heparan sulfate, collagen type I, or serum amyloid P component. These results indicate that fibronectin plays an important role in the development of amyloidosis by working as a linking protein between SAA and the cell surface of macrophages. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 7 Figure 8 PMID:2757120

  1. Serum proteins, indirect fluorescent antibody titers and complement fixing antibody titers in dexamethasone exacerbated anaplasmosis carrier calves

    E-print Network

    Johnson, Lloran Wesley

    1979-01-01

    proteins were compared with other serological and parasitological parameters of infection. LITERATURE REVIEW Anaplasmosis is a highly infectious disease of cattle recognized principally by clinical signs of fever, anemia, and icterus. Other...

  2. A filtered database search algorithm for endogenous serum protein carbonyl modifications in a mouse model of inflammation

    E-print Network

    Slade, Peter G.

    During inflammation, the resulting oxidative stress can damage surrounding host tissue, forming protein-carbonyls. The SJL mouse is an experimental animal model used to assess in vivo toxicological responses to reactive ...

  3. Folate/homocysteine phenotypes and MTHFR 677C>T genotypes are associated with serum levels of monocyte chemoattractant protein-1

    PubMed Central

    Hammons, Andrea L.; Summers, Carolyn M.; Woodside, Jayne V.; McNulty, Helene; Strain, J.J.; Young, Ian S.; Murray, Liam; Boreham, Colin A.; Scott, John M.; Mitchell, Laura E.; Whitehead, Alexander S.

    2014-01-01

    Monocyte chemoattractant protein-1 (MCP-1) is a chemokine that recruits monocytes into the subendothelial cell layer in atherosclerotic lesions. Elevated homocysteine (hyper-homocysteinemia), which is usually associated with low-folate status, is a known risk factor for many pathologies with inflammatory etiologies. The present study was undertaken to examine whether there are associations between MCP-1 concentrations and folate/Hcy phenotype or methylenetetrahydrofolate reductase (MTHFR) 677C>T genotype in healthy young adults. In females, MCP-1 concentrations were positively correlated with Hcy and negatively correlated with both serum and red blood cell folate; female smokers and MTHFR 677T carriers had particularly elevated MCP-1 concentrations. Similar relationships were not seen in males. These findings may have implications for understanding the female predominance observed for a range of autoimmune diseases, such as systemic lupus erythematosus and rheumatoid arthritis. PMID:19625220

  4. Folate/homocysteine phenotypes and MTHFR 677C>T genotypes are associated with serum levels of monocyte chemoattractant protein-1.

    PubMed

    Hammons, Andrea L; Summers, Carolyn M; Woodside, Jayne V; McNulty, Helene; Strain, J J; Young, Ian S; Murray, Liam; Boreham, Colin A; Scott, John M; Mitchell, Laura E; Whitehead, Alexander S

    2009-10-01

    Monocyte chemoattractant protein-1 (MCP-1) is a chemokine that recruits monocytes into the subendothelial cell layer in atherosclerotic lesions. Elevated homocysteine (hyperhomocysteinemia), which is usually associated with low-folate status, is a known risk factor for many pathologies with inflammatory etiologies. The present study was undertaken to examine whether there are associations between MCP-1 concentrations and folate/Hcy phenotype or methylenetetrahydrofolate reductase (MTHFR) 677C>T genotype in healthy young adults. In females, MCP-1 concentrations were positively correlated with Hcy and negatively correlated with both serum and red blood cell folate; female smokers and MTHFR 677T carriers had particularly elevated MCP-1 concentrations. Similar relationships were not seen in males. These findings may have implications for understanding the female predominance observed for a range of autoimmune diseases, such as systemic lupus erythematosus and rheumatoid arthritis. PMID:19625220

  5. Goodpasture antigen-binding protein/ceramide transporter binds to human serum amyloid P-component and is present in brain amyloid plaques.

    PubMed

    Mencarelli, Chiara; Bode, Gerard H; Losen, Mario; Kulharia, Mahesh; Molenaar, Peter C; Veerhuis, Robert; Steinbusch, Harry W M; De Baets, Marc H; Nicolaes, Gerry A F; Martinez-Martinez, Pilar

    2012-04-27

    Serum amyloid P component (SAP) is a non-fibrillar glycoprotein belonging to the pentraxin family of the innate immune system. SAP is present in plasma, basement membranes, and amyloid deposits. This study demonstrates, for the first time, that the Goodpasture antigen-binding protein (GPBP) binds to human SAP. GPBP is a nonconventional Ser/Thr kinase for basement membrane type IV collagen. Also GPBP is found in plasma and in the extracellular matrix. In the present study, we demonstrate that GPBP specifically binds SAP in its physiological conformations, pentamers and decamers. The START domain in GPBP is important for this interaction. SAP and GPBP form complexes in blood and partly colocalize in amyloid plaques from Alzheimer disease patients. These data suggest the existence of complexes of SAP and GPBP under physiological and pathological conditions. These complexes are important for understanding basement membrane, blood physiology, and plaque formation in Alzheimer disease. PMID:22396542

  6. Brain protein kinase C phosphorylating poly(arginine,serine) or lamin B is stimulated by anions and by an activator purified from bovine serum albumin preparations.

    PubMed Central

    Abdel-Ghany, M; el-Gendy, K; Zhang, S; Raden, D; Racker, E

    1989-01-01

    The phosphorylation of histone by purified protein kinase C (PK-C) from rat brain is dependent on the presence of Ca2+ and lipids. Phosphorylation of a synthetic random polymer of arginine and serine (3:1) is only moderately enhanced by Ca2+ and lipids, but it is greatly enhanced in the absence of Ca2+ and lipids by a contaminant in crystalline bovine serum albumin or by heated cellular fractions. The phosphorylation ratio of histone to poly(arginine,serine) varies between different PK-C fractions from brains of rat, pig, or lamb. These variations are partly caused by a PK-C isozyme that prefers poly(arginine,serine) over histone as substrate. The kinase activator (KA) was partly purified from bovine serum albumin and from extracts of plasma membranes of human placenta. KA is also present in mitochondria, nuclei, and the cytosol. Sulfates and phosphates at 10 mM substitute for KA with poly(arginine,serine) as substrate. The phosphorylation of histone III in the presence of Ca2+ and lipids is moderately stimulated by KA, but the phosphorylation of lamin B and some other endogenous proteins is greatly enhanced by KA. With histones as substrates, inorganic anions do not stimulate phosphorylation. The phosphorylation of poly-(arginine,serine) is very sensitive to low concentrations of staurosporin and is inhibited by PK-C antibody, but, in contrast to histone phosphorylation, it is resistant to sphingosine and polymyxin B. The poly(arginine,serine) phosphorylating activity is more stable at 4 degrees C than the histone phosphorylating activity, but the latter is stabilized by 0.05% Triton X-100. Images PMID:2928301

  7. Vitamin D Binding Protein Genotype Is Associated with Serum 25-Hydroxyvitamin D and PTH Concentrations, as Well as Bone Health in Children and Adolescents in Finland

    PubMed Central

    Viljakainen, Heli T.; Kokkonen, Elina; Jakobsen, Jette; Cashman, Kevin; Mäkitie, Outi; Lamberg-Allardt, Christel

    2014-01-01

    Vitamin D binding protein (DBP)/group-specific component (Gc), correlates positively with serum vitamin D metabolites, and phenotype influences serum 25-hydroxyvitamin D (S-25(OH)D) concentration. The protein isoform has been associated with decreased bone mineral density (BMD) and increased fracture risk. We examined the role of GC genotypes in S-25(OH)D status and BMD in 231 Finnish children and adolescents aged 7?19 yr. BMD was measured with DXA from lumbar spine (LS), total hip, and whole body, and for 175 subjects, radial volumetric BMD was measured with pQCT. Background characteristic and total dietary intakes of vitamin D and calcium were collected. The concentrations of 25(OH)D, parathyroid hormone (PTH), calcium and other markers of calcium homeostasis were determined from blood and urine. Genotyping was based on single-nucleotide polymorphism (rs4588) in the GC gene. The genotype distribution was: GC 1/1 68%, GC 1/2 26% and GC 2/2 6%. A significant difference emerged in 25(OH)D and PTH concentrations between the genotypes, (p?=?0.001 and 0.028 respectively, ANCOVA). There was also a linear trend in: Gc 2/2 had the lowest 25(OH)D and PTH concentrations (p?=?0.025 and 0.012, respectively). Total hip bone mineral content was associated with GC genotype (BMC) (p?=?0.05, ANCOVA) in boys. In regression analysis, after adjusting for relevant covariates, GC genotype was associated with LS BMC and strength and strain index (SSI) Z-score in both genders, and LS BMD in boys. In conclusion, the present study demonstrates the association between GC genotypes and S-25(OH)D and PTH concentrations. The results show the influence of DBP genetic variation on bone mass accrual in adolescence. PMID:24498064

  8. Sericin, a protein derived from silkworms, accelerates the proliferation of several mammalian cell lines including a hybridoma

    Microsoft Academic Search

    Satoshi Terada; Taeko Nishimura; Masahiro Sasaki; Hideyuki Yamada; Masao Miki

    2002-01-01

    Sericin, a constituent of the silkworm cocoon, was added to the culture of four mammalian cell lines: murine hybridoma 2E3-O,human\\u000a hepatoblastoma HepG2, human epithelial HeLa and human embryonal kidney 293 cells. The proliferation of all cell lineswas accelerated\\u000a in the presence of sericin. The hybridoma cellline was further studied. The 2E3-O cell line was so well adapted to serum-free\\u000a medium

  9. Cephalosporin binding sites to human serum albumin and the relation with the N-B transition of this protein.

    PubMed

    Nerli, B; García, F; Ballán, C; Picó, G

    1995-05-01

    The binding of some cephalosporins to human serum albumin was studied using probes for the so-called I, II, bilirubin and fatty acids binding sites. The results showed that cephradine and cefsulodin bind to site II, cefaclor, cefamandole, cefsulodin, cephaloglycin and cefadroxil bind to the bilirubin binding site, while cefaclor does it to the fatty acid binding site. No binding of these cephalosporins to site I of albumin was found. The binding produced a perturbation on the N-B equilibrium of albumin, stabilizing the N conformational form, which suggests that the N form of albumin has more affinity with the cephalosporins than the B form. This finding gives support to the assumption that the binding of cephalosporins to site II, bilirubin and fatty acids binding sites affects the N-B transition of albumin. PMID:7663413

  10. Development of a rapid, single-step procedure using protein G affinity chromatography to deplete fetal calf serum of its IgG and to isolate murine IgG1 monoclonal antibodies from supernatants of hybridoma cells

    Microsoft Academic Search

    Cemalettin Aybay; Turgut Imir

    2000-01-01

    Fetal calf serum (FCS) was depleted of its immunoglobulin G (IgG) in a rapid procedure using protein G affinity chromatography. 20 ml of FCS was depleted of its IgG in less than 80 min by applying 5 ml of FCS to a 1 ml HiTrap protein G Sepharose column followed by appropriate elution. Various concentrations of IgG-depleted FCS (G-FCS) were

  11. The silk protein, sericin, protects against cell death caused by acute serum deprivation in insect cell culture

    Microsoft Academic Search

    Masakazu Takahashi; Kazuhisa Tsujimoto; Hideyuki Yamada; Hiroshi Takagi; Shigeru Nakamori

    2003-01-01

    Sericin is the silk protein that covers fibroin fibers and functions as a `glue' in the cocoons of silkworms, and its most abundant component, Ser1, contains repeats of Ser- and Thr-rich 38 amino acid residues. The viability of Sf9 insect cells was 20, 57 and 49% on the fifth day and 41, 91 and 70% on the ninth day after

  12. Divergent antibody isotype responses induced in mice by systemic exposure to proteins: a comparison of ovalbumin with bovine serum albumin

    Microsoft Academic Search

    R. J Dearman; H Caddick; D. A Basketter; I Kimber

    2000-01-01

    Whereas many foreign proteins are immunogenic, only a proportion is associated commonly with allergy, having the potential to induce the quality of immune response necessary for IgE antibody production and the development of immediate type hypersensitivity reactions in the gastrointestinal and\\/or respiratory tracts. In the context of toxicological evaluations there is a need to identify those properties that confer on

  13. Evaluation of Multiplexed Foot-and-Mouth Disease Nonstructural Protein Antibody Assay Against Standardized Bovine Serum Panel

    SciTech Connect

    Perkins, J; Parida, S; Clavijo, A

    2007-05-14

    Liquid array technology has previously been used to show proof-of-principle of a multiplexed non structural protein serological assay to differentiate foot-and-mouth infected and vaccinated animals. The current multiplexed assay consists of synthetically produced peptide signatures 3A, 3B and 3D and recombinant protein signature 3ABC in combination with four controls. To determine diagnostic specificity of each signature in the multiplex, the assay was evaluated against a naive population (n = 104) and a vaccinated population (n = 94). Subsequently, the multiplexed assay was assessed using a panel of bovine sera generated by the World Reference Laboratory for foot-and-mouth disease in Pirbright, UK. This sera panel has been used to assess the performance of other singleplex ELISA-based non-structural protein antibody assays. The 3ABC signature in the multiplexed assay showed comparative performance to a commercially available non-structural protein 3ABC ELISA (Cedi test{reg_sign}) and additional information pertaining to the relative diagnostic sensitivity of each signature in the multiplex is acquired in one experiment. The encouraging results of the evaluation of the multiplexed assay against a panel of diagnostically relevant samples promotes further assay development and optimization to generate an assay for routine use in foot-and-mouth disease surveillance.

  14. The Use of Bovine Serum Protein as an Oral Support Therapy Following Coronavirus Challenge in Calves1

    Microsoft Academic Search

    J. D. Arthington; C. A. Jaynes; H. D. Tyler; S. Kapil; J. D. Quigley III

    2002-01-01

    The objective of this experiment was to investigate thetherapeuticefficacyofasupplementalbovineserum protein blend fed to calves challenged with virulent coronavirus. Twelve Holstein bull calves (approxi- mately3wkofage)wereallocatedbyinitialbodyweight to Control (n = 5) and treated (n = 7) groups. On d 0, all calves were orally challenged with 1 × 107 plaque- forming units of virulent coronavirus isolate. Infection was allowed to progress for

  15. Clinical significance of preoperative serum vascular endothelial growth factor, interleukin-6, and C-reactive protein level in colorectal cancer

    Microsoft Academic Search

    Kyung A Kwon; Sung Hyun Kim; Sung Yong Oh; Suee Lee; Jin-Yeong Han; Kyeong Hee Kim; Ri Young Goh; Hong Jo Choi; Ki Jae Park; Mee Sook Roh; Hyo-Jin Kim; Hyuk-Chan Kwon; Jong Hoon Lee

    2010-01-01

    BACKGROUND: Angiogenesis is a multistep process in which many growth factors and cytokines have an essential role. Vascular endothelial growth factor (VEGF) is a potent angiogenic agent that acts as a specific mitogen for vascular endothelial cells through specific cell surface receptors. The interleukin-6 (IL-6) pathway is another mechanism linking angiogenesis to malignancy. C-reactive protein (CRP), a representative marker for

  16. Obesity Is an Important Determinant of Baseline Serum C-Reactive Protein Concentration in Monozygotic Twins, Independent of Genetic Influences

    Microsoft Academic Search

    Jerry R. Greenfield; Katherine Samaras; Arthur B. Jenkins; Paul J. Kelly; Tim D. Spector; J. Ruth Gallimore; Mark B. Pepys; Lesley V. Campbell

    Background—C-reactive protein (CRP) values predict atherothrombotic cardiovascular disease and type 2 diabetes mellitus. Associations between CRP and obesity, predominantly assessed anthropometrically, may partly explain these observations. Previous studies have been unable to control for genetic influences on CRP and obesity. The aim of this study was to examine the relationship between CRP and accurately measured body fat, lipids, apolipoproteins, blood

  17. Effects of Protein-Free Diet and Food Deprivation on Hepatic Rhodanese Activity, Serum Proteins and Acute Cyanide Lethality in Mice12

    Microsoft Academic Search

    JOSEPH V. RUTKOWSKI; B. D. ROEBUCK; ROGER P. SMITH

    This study was undertaken to evaluate changes in hepatic and extra- hepatic cyanide detoxication resulting from nutritional deficits and any parallel changes in cyanide sensitivity. Male CD-I mice were fed control diet, control diet in reduced quantities or protein-free diet. After 14 d body weight and relative liver size were lower in the protein-free diet and the food-deprived groups than

  18. The Cytoskeleton-associated PDZ-LIM Protein, ALP, Acts on Serum Response Factor Activity to Regulate Muscle Differentiation

    PubMed Central

    Pashmforoush, Mohammad; Vegezzi, Cristina; Chien, Kenneth R.; Auffray, Charles; Beckerle, Mary C.

    2007-01-01

    In this report, an antisense RNA strategy has allowed us to show that disruption of ALP expression affects the expression of the muscle transcription factors myogenin and MyoD, resulting in the inhibition of muscle differentiation. Introduction of a MyoD expression construct into ALP-antisense cells is sufficient to restore the capacity of the cells to differentiate, illustrating that ALP function occurs upstream of MyoD. It is known that MyoD is under the control of serum response factor (SRF), a transcriptional regulator whose activity is modulated by actin dynamics. A dramatic reduction of actin filament bundles is observed in ALP-antisense cells and treatment of these cells with the actin-stabilizing drug jasplakinolide stimulates SRF activity and restores the capacity of the cells to differentiate. Furthermore, we show that modulation of ALP expression influences SRF activity, the level of its coactivator, MAL, and muscle differentiation. Collectively, these results suggest a critical role of ALP on muscle differentiation, likely via cytoskeletal regulation of SRF. PMID:17332502

  19. Studies of the Interaction between Isoimperatorin and Human Serum Albumin by Multispectroscopic Method: Identification of Possible Binding Site of the Compound Using Esterase Activity of the Protein

    PubMed Central

    Ranjbar, Samira; Shokoohinia, Yalda; Ghobadi, Sirous; Gholamzadeh, Saeed; Moradi, Nastaran; Ashrafi-Kooshk, Mohammad Reza; Aghaei, Abbas

    2013-01-01

    Isoimperatorin is one of the main components of Prangos ferulacea as a linear furanocoumarin and used as anti-inflammatory, analgesic, antispasmodic, and anticancer drug. Human serum albumin (HSA) is a principal extracellular protein with a high concentration in blood plasma and carrier for many drugs to different molecular targets. Since the carrying of drug by HSA may affect on its structure and action, we decided to investigate the interaction between HSA and isoimperatorin using fluorescence and UV spectroscopy. Fluorescence data indicated that isoimperatorin quenches the intrinsic fluorescence of the HSA via a static mechanism and hydrophobic interaction play the major role in the drug binding. The binding average distance between isoimperatorin and Trp 214 of HSA was estimated on the basis of the theory of Förster energy transfer. Decrease of protein surface hydrophobicity (PSH) was also documented upon isoimperatorin binding. Furthermore, the synchronous fluorescence spectra show that the microenvironment of the tryptophan residues does not have obvious changes. Site marker compettive and fluorescence experiments revealed that the binding of isoimperatorin to HSA occurred at or near site I. Finally, the binding details between isoimperatorin and HSA were further confirmed by molecular docking and esterase activity inhibition studies which revealed that drug was bound at subdomain IIA. PMID:24319355