Sample records for based serum protein

  1. Screening serum hepatocellular carcinoma-associated proteins by SELDI-based protein spectrum analysis

    Institute of Scientific and Technical Information of China (English)


    AIM: To find out potential serum hepatocellular carcinoma (HCC)-associated proteins with low molecular weight and low abundance by SELDI-based serum protein spectra analysis, that will have much application in the diagnosis or differentiated diagnosis of HCC, as well as giving a better undersyanding of the mechanism of hepato-carcinogenesis. METHODS: Total serum samples were collected with informed consent from 81 HCC patients with HBV (+)/cirrhosis(+),36 cirrhosis patients and 43 chronic hepatitis B patients. Serum protein fingerprint profiles were first generated by selected WCX2 protein chip capture integrating with SELDI-TOF-MS, then normalized and aligned by Ciphergen SELDI Software 3.1.1 with Biomarker Wizard.. Comparative analysis of the intensity of corresponding protein fingerprint peaks in normalized protein spectra, some protein peaks with significant difference between H.CC and cirrhosis or chronic hepatitis B were found. RESULTS: One hundred and twenty-eight serum protein peaks between.2000 and 30000Da were identified under the condition of signal-to-noise>5 and minimum threshold for cluster>20%.Eight-seven of these proteins were showed significant differences in intensity between HCC and cirrhosis (P<0.05=.Of the above differential proteins, 45 proteins had changes greater than two-fold, including 15 upregulated proteins and 30downregulated proteins in HCC serum. Between HCC and chronic hepatitis B,9 of 52 differential proteins (P<0.05= had intensities of more than two-fold, including 2 upregulated proteins and 7 downregulated proteins in HCC serum. Between cirrhosis and chronic hepatitis B,28 of 79 significant differential proteins (P<0.05= changes greater than two-fold in intensity, including 17 upregulated proteins and 11 downregulated proteins in cirrhosis serum. For the analysis of these leading differential proteins in subtraction difference mode among three diseases, the five common downregulated proteins in HCC serum (M/Z 2870

  2. Identification of serum protein biomarkers for utrophin based DMD therapy (United States)

    Guiraud, Simon; Edwards, Benjamin; Squire, Sarah E.; Babbs, Arran; Shah, Nandini; Berg, Adam; Chen, Huijia; Davies, Kay E.


    Despite promising therapeutic avenues, there is currently no effective treatment for Duchenne muscular dystrophy (DMD), a lethal monogenic disorder caused by the loss of the large cytoskeletal protein, dystrophin. A highly promising approach to therapy, applicable to all DMD patients irrespective to their genetic defect, is to modulate utrophin, a functional paralogue of dystrophin, able to compensate for the primary defects of DMD restoring sarcolemmal stability. One of the major difficulties in assessing the effectiveness of therapeutic strategies is to define appropriate outcome measures. In the present study, we utilised an aptamer based proteomics approach to profile 1,310 proteins in plasma of wild-type, mdx and Fiona (mdx overexpressing utrophin) mice. Comparison of the C57 and mdx sera revealed 83 proteins with statistically significant >2 fold changes in dystrophic serum abundance. A large majority of previously described biomarkers (ANP32B, THBS4, CAMK2A/B/D, CYCS, CAPNI) were normalised towards wild-type levels in Fiona animals. This work also identified potential mdx markers specific to increased utrophin (DUS3, TPI1) and highlights novel mdx biomarkers (GITR, MYBPC1, HSP60, SIRT2, SMAD3, CNTN1). We define a panel of putative protein mdx biomarkers to evaluate utrophin based strategies which may help to accelerate their translation to the clinic. PMID:28252048

  3. Protein electrophoresis - serum (United States)

    ... this page: // Protein electrophoresis - serum To use the sharing features on ... JavaScript. This lab test measures the types of protein in the fluid (serum) part of a blood ...

  4. Combined experimental and statistical strategy for mass spectrometry based serum protein profiling for diagnosis of breast cancer

    DEFF Research Database (Denmark)

    Callesen, Anne Kjærgaard; Vach, Werner; Jørgensen, Per E


    Serum protein profiling by mass spectrometry is a promising method for early detection of cancer. We have implemented a combined strategy based on matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) and statistical data analysis for serum protein profiling and applied...... of nine mass spectrometric protein profiles were obtained for each serum sample. A total of 533 common peaks were defined and represented a 'reference protein profile'. Among these 533 common peaks, we identified 72 peaks exhibiting statistically significant intensity differences ( p ... and specificity. We conclude that optimized serum sample handling and mass spectrometry data acquisition strategies in combination with statistical analysis provide a viable platform for serum protein profiling in cancer diagnosis....

  5. Intact-protein analysis system for discovery of serum-based disease biomarkers. (United States)

    Wang, Hong; Hanash, Samir


    Profiling of serum and plasma proteins has substantial relevance to the discovery of circulating disease biomarkers. However, the extreme complexity and vast dynamic range of protein abundance in serum and plasma present a formidable challenge for protein analysis. Thus, integration of multiple technologies is required to achieve high-resolution and high-sensitivity proteomic analysis of serum or plasma. In this chapter, we describe an orthogonal multidimensional intact-protein analysis system (IPAS) (Wang et al., Mol Cell Proteomics 4:618-625, 2005) coupled with protein tagging (Faca et al., J Proteome Res 5:2009-2018, 2006) to profile the serum and plasma proteomes quantitatively, which we have applied in our biomarker discovery studies (Katayama et al., Genome Med 1:47, 2009; Faca et al., PLoS Med 5:e123, 2008; Zhang et al. Genome Biol 9:R93, 2008).

  6. Monte carlo method-based QSAR modeling of penicillins binding to human serum proteins. (United States)

    Veselinović, Jovana B; Toropov, Andrey A; Toropova, Alla P; Nikolić, Goran M; Veselinović, Aleksandar M


    The binding of penicillins to human serum proteins was modeled with optimal descriptors based on the Simplified Molecular Input-Line Entry System (SMILES). The concentrations of protein-bound drug for 87 penicillins expressed as percentage of the total plasma concentration were used as experimental data. The Monte Carlo method was used as a computational tool to build up the quantitative structure-activity relationship (QSAR) model for penicillins binding to plasma proteins. One random data split into training, test and validation set was examined. The calculated QSAR model had the following statistical parameters: r(2)  = 0.8760, q(2)  = 0.8665, s = 8.94 for the training set and r(2)  = 0.9812, q(2)  = 0.9753, s = 7.31 for the test set. For the validation set, the statistical parameters were r(2)  = 0.727 and s = 12.52, but after removing the three worst outliers, the statistical parameters improved to r(2)  = 0.921 and s = 7.18. SMILES-based molecular fragments (structural indicators) responsible for the increase and decrease of penicillins binding to plasma proteins were identified. The possibility of using these results for the computer-aided design of new penicillins with desired binding properties is presented.

  7. Mink serum amyloid A protein. Expression and primary structure based on cDNA sequences. (United States)

    Marhaug, G; Husby, G; Dowton, S B


    The nucleotide sequences of two mink serum amyloid A (SAA) cDNA clones have been analyzed, one (SAA1) 776 base pairs long and the other (SAA2) 552 base pairs long. Significant differences were discovered when derived amino acid sequences were compared with data for apoSAA isolated from high density lipoprotein. Previous studies of mink protein SAA and amyloid protein A (AA) suggest that only one SAA isotype is amyloidogenic. The cDNA clone for SAA2 defines the "amyloid prone" isotype while SAA1 is found only in serum. Mink SAA1 has alanine in position 10, isoleucine in positions 24, 67, and 71, lysine in position 27, and proline in position 105. Residue 10 in mink SAA2 is valine while arginine and asparagine are at positions 24 and 27, respectively, all characteristics of protein AA isolated from mink amyloid fibrils. Mink SAA2 also has valine in position 67, phenylalanine in position 71, and amino acid 105 is serine. It remains unknown why these six amino acid substitutions render SAA2 more amyloidogenic than SAA1. Eighteen hours after lipopolysaccharide stimulation, mink SAA mRNA is abundant in liver with relatively minor accumulations in brain and lung. Genes encoding both SAA isotypes are expressed in all three organs while no SAA mRNA was detectable in amyloid prone organs, including spleen and intestine, indicating that deposition of AA from locally synthesized SAA is unlikely. A third mRNA species (2.2 kilobases) was identified and hybridizes with cDNA probes for mink SAA1 and SAA2. In addition to a major primary translation product (molecular mass 14,400 Da) an additional product with molecular mass 28,000 Da was immunoprecipitable.

  8. Protections of bovine serum albumin protein from damage on functionalized graphene-based electrodes by flavonoids

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Bolu [School of Pharmacy, Lanzhou University, Lanzhou 730000 (China); Gou, Yuqiang [Lanzhou Military Command Center for Disease Prevention and Control, Lanzhou 730000 (China); Xue, Zhiyuan; Zheng, Xiaoping; Ma, Yuling [School of Pharmacy, Lanzhou University, Lanzhou 730000 (China); Hu, Fangdi, E-mail: [School of Pharmacy, Lanzhou University, Lanzhou 730000 (China); Zhao, Wanghong, E-mail: [Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou 51515 (China)


    A sensitive electrochemical sensor based on bovine serum albumin (BSA)/poly (diallyldimethylammonium chloride) (PDDA) functionalized graphene nanosheets (PDDA-G) composite film modified glassy carbon electrode (BSA/PDDA-G/GCE) had been developed to investigate the oxidative protein damage and protections of protein from damage by flavonoids. The performance of this sensor was remarkably improved due to excellent electrical conductivity, strong adsorptive ability, and large effective surface area of PDDA-G. The BSA/PDDA-G/GCE displayed the greatest degree of BSA oxidation damage at 40 min incubation time and in the pH 5.0 Fenton reagent system (12.5 mM FeSO{sub 4}, 50 mM H{sub 2}O{sub 2}). The antioxidant activities of four flavonoids had been compared by fabricated sensor based on the relative peak current ratio of SWV, because flavonoids prevented BSA damage caused by Fenton reagent and affected the BSA signal in a solution containing Co(bpy){sub 3}{sup 3+}. The sensor was characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and scanning electron microscopy (SEM). UV–vis spectrophotometry and FTIR were also used to investigate the generation of hydroxyl radical and BSA damage, respectively. On the basis of results from electrochemical methods, the order of the antioxidant activities of flavonoids is as follows: (+)-catechin > kaempferol > apigenin > naringenin. A novel, direct SWV analytical method for detection of BSA damage and assessment of the antioxidant activities of four flavonoids was developed and this electrochemical method provided a simple, inexpensive and rapid detection of BSA damage and evaluation of the antioxidant activities of samples. - Highlights: • Hydroxyl radicals were produced by Fenton reagents. • An electrochemical bovine serum albumin (BSA) damage sensor was successfully fabricated. • The proposed biosensor can assess the antioxidant capacity of four flavonoids. • The order of antioxidant

  9. Characterization of protein degradation in serum-based lubricants during simulation wear testing of metal-on-metal hip prostheses. (United States)

    Maskiewicz, Victoria K; Williams, Paul A; Prates, Sarah J; Bowsher, John G; Clarke, Ian C


    A size exclusion high performance liquid chromatography (SEC-HPLC) method has been developed which is capable of separation and quantitation of bovine serum albumin (BSA) and bovine serum globulin (BSG) components of serum-based lubricant (SBL) solutions. This allowed characterization of the stability profiles of these proteins when acting as lubricants during hip wear simulation, and identification of wear-specific mechanisms of degradation. Using cobalt-chromium metal-on-metal (MOM) hip joints, it was observed that BSA remained stable for up to 3 days (215K cycles) of wear testing after which the protein degraded in a fairly linear fashion. BSG on the other hand, began to degrade immediately and in a linear fashion with a rate constant of 5% per day. Loss of both proteins occurred via the formation of high molecular weight aggregates which precipitated out of solution. No fragmentation of the polypeptide backbone of either protein was observed. Data obtained suggest that protein degradation was not due to microbial contamination, denaturation at the air-water interface, or frictional heating of articulating joint surfaces in these studies. We conclude that the primary source of protein degradation during MOM simulation testing occurs via high shear rates experienced by SBL solutions at articulating surfaces, possibly coupled with metal-protein interactions occurring as new and reactive metal surfaces are generated during wear testing. The development of this analytical methodology will allow new studies to clarify the role of SBL solutions in wear simulation studies and the interactions and lubricating properties of serum proteins with prosthetic surfaces other than MOM.

  10. Protein Crystal Serum Albumin (United States)


    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

  11. Identification of novel biomarkers in chronic immune thrombocytopenia (ITP) by microarray-based serum protein profiling. (United States)

    Bal, Gürkan; Futschik, Matthias E; Hartl, Daniela; Ringel, Frauke; Kamhieh-Milz, Julian; Sterzer, Viktor; Hoheisel, Jörg D; Alhamdani, Mohamed S S; Salama, Abdulgabar


    The pathological mechanisms underlying the development of immune thrombocytopenia (ITP) are unclear and its diagnosis remains a process of exclusion. Currently, there are no known specific biomarkers for ITP to support differential diagnosis and treatment decisions. Profiling of serum proteins may be valuable for identifying such biomarkers. Sera from 46 patients with primary chronic ITP and 34 healthy blood donors were analysed using a microarray of 755 antibodies. We identified 161 differentially expressed proteins. In addition to oncoproteins and tumour-suppressor proteins, including apoptosis regulator BCL2, breast cancer type 1 susceptibility protein (BRCA1), Fanconi anaemia complementation group C (FANCC) and vascular endothelial growth factor A (VEGFA), we detected six anti-nuclear autoantibodies in a subset of ITP patients: anti-PCNA, anti-SmD, anti-Ro/SSA60, anti-Ro/SSA52, anti-La/SSB and anti-RNPC antibodies. This finding may provide a rational explanation for the association of ITP with malignancies and other autoimmune diseases. While RUNX1mRNA expression in the peripheral blood mononuclear cells (PBMC) of patients was significantly downregulated, an accumulation of RUNX1 protein was observed in the platelets of ITP patients. This may indicate dysregulation of RUNX1 expression in PBMC and megakaryocytes and may lead to an imbalanced immune response and impaired thrombopoiesis. In conclusion, we provide novel insights into the pathogenic mechanisms of ITP that warrant further exploration.

  12. Off surface matrix based on-chip electrochemical biosensor platform for protein biomarker detection in undiluted serum. (United States)

    Arya, Sunil K; Kongsuphol, Patthara; Park, Mi Kyoung


    The manuscript describes a concept of using off surface matrix modified with capturing biomolecule for on-chip electrochemical biosensing. 3D matrix made by laser engraving of polymethyl methacrylate (PMMA) sheet as off surface matrix was integrated in very close vicinity of the electrode surface. Laser engraving and holes in PMMA along with spacing from surface provide fluidic channel and incubation chamber. Covalent binding of capturing biomolecule (anti-TNF-α antibody) on off-surface matrix was achieved via azide group activity of 4-fluoro-3-nitro-azidobenzene (FNAB), which act as cross-linker and further covalently binds to anti-TNF-α antibody via thermal reaction. Anti-TNF-α/FNAB/PMMA matrix was then integrated over comb structured gold electrode array based sensor chip. Separate surface modification followed by integration of sensor helped to prevent the sensor chip surface from fouling during functionalization. Nonspecific binding was prevented using starting block T20 (PBS). Results for estimating protein biomarker (TNF-α) in undiluted serum using Anti-TNF-α/FNAB/PMMA/Au reveal that system can detect TNF-α in 100pg/ml to 100ng/ml range with high sensitivity of 119nA/(ng/ml), with negligible interference from serum proteins and other cytokines. Thus, use of off surface matrix may provide the opportunity to electrochemically sense biomarkers sensitively to ng/ml range with negligible nonspecific binding and false signal in undiluted serum.

  13. Piezoelectric microcantilever serum protein detector (United States)

    Capobianco, Joseph A.

    The development of a serum protein detector will provide opportunities for better screening of at-risk cancer patients, tighter surveillance of disease recurrence and better monitoring of treatment. An integrated system that can process clinical samples for a number of different types of biomarkers would be a useful tool in the early detection of cancer. Also, screening biomarkers such as antibodies in serum would provide clinicians with information regarding the patient's response to treatment. Therefore, the goal of this study is to develop a sensor which can be used for rapid, all-electrical, real-time, label-fee, in-situ, specific quantification of cancer markers, e.g., human epidermal receptor 2 (Her2) or antibodies, in serum. To achieve this end, piezoelectric microcantilever sensors (PEMS) were constructed using an 8 mum thick lead magnesium niobate-lead titanate (PMN-PT) freestanding film as the piezoelectric layer. The desired limit of detection is on the order of pg/mL. In order to achieve this goal the higher frequency lateral extension modes were used. Also, as the driving and sensing of the PEMS is electrical, the PEMS must be insulated in a manner that allows it to function in aqueous solutions. The insulation layer must also be compatible with standardized bioconjugation techniques. Finally, detection of both cancer antigens and antibodies in serum was carried out, and the results were compared to a standard commercialized protocol. PEMS have demonstrated the capability of detecting Her2 at a concentration of 5 pg/mL in diluted human serum (1:40) in less than 1 hour. The approach can be easily translated into the clinical setting because the sensitivity is more than sufficient for monitoring prognosis of breast cancer patients. In addition to Her2 detection, antibodies in serum were assayed in order to demonstrate the feasibility of monitoring the immune response for antibody-dependent cellular cytotoxicity (ADCC) in patients on antibody therapies

  14. Disparate Proteome Responses of Pathogenic and Non-pathogenic Aspergilli to Human Serum Measured by Activity-Based Protein Profiling (ABPP)

    Energy Technology Data Exchange (ETDEWEB)

    Wiedner, Susan D.; Ansong, Charles; Webb-Robertson, Bobbie-Jo M.; Pederson, Leeanna M.; Fortuin, Suereta; Hofstad, Beth A.; Shukla, Anil K.; Panisko, Ellen A.; Smith, Richard D.; Wright, Aaron T.


    Aspergillus fumigatus is the primary pathogen causing the devastating pulmonary disease Invasive Aspergillosis in immunocompromised individuals. Genomic analysis shows high synteny between A. fumigatus and closely related rarely pathogenic Neosartorya fischeri and Aspergillus clavatus genomes. To investigate the presence of unique or highly inducible protein reactivity in the pathogen, we applied activity-based protein profiling to compare protein reactivity of all three fungi over time in minimal media growth and in response to human serum. We found 350 probe-reactive proteins exclusive to A. fumigatus, including known virulence associated proteins, and 13 proteins associated with stress response exclusive to A. fumigatus culture in serum. Though the fungi are highly orthologous, A. fumigatus has significantly more activity across varied biological process. Only 50% of expected orthologs of measured A. fumigatus reactive proteins were observed in N. fischeri and A. clavatus. Human serum induced processes uniquely or significantly represented in A. fumigatus include actin organization and assembly, transport, and fatty acid, cell membrane, and cell wall synthesis. Additionally, signaling proteins regulating vegetative growth, conidiation, and cell wall integrity, required for appropriate cellular response to external stimuli, had higher reactivity over time in A. fumigatus and N. fisheri, but not in A. clavatus. Together, we show that measured proteins and physiological processes identified solely or significantly over-represented in A. fumigatus reveal a unique adaptive response to human protein not found in closely related, but rarely aspergilli. These unique protein reactivity responses may reveal how A. fumigatus initiates pulmonary invasion leading to Invasive Aspergillosis.

  15. Serum Protein Profile Alterations in Hemodialysis Patients

    Energy Technology Data Exchange (ETDEWEB)

    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


    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.

  16. Protein expression and fucosylated glycans of the serum haptoglobin-β subunit in hepatitis B virus-based liver diseases

    Institute of Scientific and Technical Information of China (English)

    Hong Shu; Shu Zhang; Xiaonan Kang; Shan Li; Xue Qin; Chun Sun; Haojie Lu; Yinkun Liu


    Glycosylation, which regulates the configuration and function of glycoproteins, is the most important post-translational modification. The aim of this study was to observe the differential patterns in glycan and protein parts of the serum haptoglobin-β subunit (Hp-β) purified from patients with hepatitis B virus (HBV) infection, liver cirrhosis (LC), or hepatocellular carcinoma (HCC). 2-D gel electrophoresis and multiplexed proteomics staining technique were employed to investigate whether the Hp-β glycan level was proportional to the protein level. Multilectin blot, high-performance liquid chromatography (HPLC), and western blot analysis were carried out to identify the glycoform of Hp-β quantitatively. Our experiments showed that the ratio of total serum Hp-β to the glycosylated form of Hp-β varied among the patients with different liver diseases. The total Hp-β protein expression level was much higher in HCC than LC, while an incremental proportion of fucosylated Hp-β was also observed in LC and HCC patients compared with that in HBV and healthy controls. Differential fucosylation was firther identified as a Lewis X structure by HPLC and antihuman SialyI-Lewis X antibody. In conclusion, the aberrant alternation of Hp-β glycan and total protein expression may be a promising biomarker for early hepatocarcinogenesis.

  17. Proteomic evaluation of sheep serum proteins

    Directory of Open Access Journals (Sweden)

    Chiaradia Elisabetta


    Full Text Available Abstract Background The applications of proteomic strategies to ovine medicine remain limited. The definition of serum proteome may be a good tool to identify useful protein biomarkers for recognising sub-clinical conditions and overt disease in sheep. Findings from bovine species are often directly translated for use in ovine medicine. In order to characterize normal protein patterns and improve knowledge of molecular species-specific characteristics, we generated a two-dimensional reference map of sheep serum. The possible application of this approach was tested by analysing serum protein patterns in ewes with mild broncho-pulmonary disease, which is very common in sheep and in the peripartum period which is a stressful time, with a high incidence of infectious and parasitic diseases. Results This study generated the first reference 2-DE maps of sheep serum. Overall, 250 protein spots were analyzed, and 138 identified. Compared with healthy sheep, serum protein profiles of animals with rhino-tracheo-bronchitis showed a significant decrease in protein spots identified as transthyretin, apolipoprotein A1 and a significant increase in spots identified as haptoglobin, endopin 1b and alpha1B glycoprotein. In the peripartum period, haptoglobin, alpha-1-acid glycoprotein, apolipoprotein A1 levels rose, while transthyretin content dropped. Conclusions This study describes applications of proteomics in putative biomarker discovery for early diagnosis as well as for monitoring the physiological and metabolic situations critical for ovine welfare.

  18. Proteomic evaluation of sheep serum proteins


    Chiaradia Elisabetta; Avellini Luca; Tartaglia Micaela; Gaiti Alberto; Just Ingo; Scoppetta Fausto; Czentnar Zoltan; Pich Andreas


    Abstract Background The applications of proteomic strategies to ovine medicine remain limited. The definition of serum proteome may be a good tool to identify useful protein biomarkers for recognising sub-clinical conditions and overt disease in sheep. Findings from bovine species are often directly translated for use in ovine medicine. In order to characterize normal protein patterns and improve knowledge of molecular species-specific characteristics, we generated a two-dimensional reference...

  19. Multiplexed microfluidic quantification of proteins in serum (United States)

    Rajan, Nitin; Rajauria, Sukumar; Cleland, Andrew


    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.

  20. Development of a rapid and high-performance chemiluminescence immunoassay based on magnetic particles for protein S100B in human serum. (United States)

    Zhang, Huisheng; Qi, Suwen; Rao, Jie; Li, Qiaoliang; Yin, Li; Lu, Yuejun


    Protein S100B is a clinically useful non-invasive biomarker for brain cell damage. A rapid chemiluminescence immunoassay (CLIA) for S100B in human serum has been developed. Fluorescein isothiocyanate (FITC) and N-(aminobutyl)-N-(ethylisoluminol) (ABEI) are used to label two different monoclonal antibodies of anti-S100B. Protein S100B in serum combines with labeled antibodies and can form a sandwiched immunoreaction. A simplified separation procedure based on the use of magnetic particles (MPs) that were coated with anti-FITC antibody is performed to remove the unwanted materials. After adding the substrate solution, the relative light unit (RLU) of ABEI is measured and is found to be directly proportional to the concentration of S100B in serum. The relevant variables involved in the CLIA signals are optimized and the parameters of the proposed method are evaluated. The results demonstrate that the method is linear to 25 ng/mL S100B with a detection limit of 0.02 ng/mL. The coefficient of variation (CV) is < 5% and < 6% for intra- and interassay precision, respectively. The average recoveries are between 97 and 107%. The linearity-dilution effect produces a linear correlation coefficient of 0.9988. Compared with the commercial kit, the proposed method shows a correlation of 0.9897. The proposed method displays acceptable performance for quantification of S100B and is appropriate for use in clinical diagnosis.

  1. Identification of Differentially Expressed Serum Proteins in Infectious Purpura Fulminans

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    Ting He


    Full Text Available Purpura fulminans (PF is a life-threatening hemorrhagic condition. Because of the rarity and randomness of the disease, no improvement in treatment has been made for a long time. In this study, we assessed the serum proteome response to PF by comparing serum proteins between healthy controls and PF patient. Liquid chromatography with tandem mass spectrometry (LC-MS/MS approach was used after depleting 6 abundant proteins of serum. In total, 262 proteins were confidently identified with 2 unique peptides, and 38 proteins were identified significantly up- (≥2 or downregulated (≤0.5 based on spectral counting ratios (SpCPF/N. In the 38 proteins with significant abundance changes, 11 proteins were previously known to be associated with burn or sepsis response, but 27 potentially novel proteins may be specifically associated with PF process. Two differentially expressed proteins, alpha-1-antitrypsin (SERPINA1 and alpha-2 antiplasmin (SERPINF2, were validated by Western blot. This is the first study where PF patient and healthy controls are compared in a proteomic study to elucidate proteins involved in the response to PF. This study provides an initial basis for future studies of PF, and the differentially expressed proteins might provide new therapeutic targets to decrease the mortality of PF.

  2. Serotriflin, a CRISP family protein with binding affinity for small serum protein-2 in snake serum. (United States)

    Aoki, Narumi; Sakiyama, Akie; Kuroki, Kimiko; Maenaka, Katsumi; Kohda, Daisuke; Deshimaru, Masanobu; Terada, Shigeyuki


    Habu (Trimeresurus flavoviridis) serum contains 3 small serum proteins (SSP-1, SSP-2, and SSP-3) with molecular masses of 6.5 to 10 kDa. Gel filtration analysis showed that all the SSPs exist in high molecular mass forms of approximately 60 kDa in the serum. Ultrafiltration of Habu serum showed that SSPs dissociated from the complex below a pH of 4. An SSP-binding protein was purified from Habu serum by gel filtration, ion exchange, and reverse-phase HPLC. N-terminal sequencing yielded a 39-amino acid sequence, similar to the N-terminal region of triflin, which is a snake venom-derived Ca2+ channel blocker that suppresses smooth muscle contraction. The amino acid sequence of this protein, termed serotriflin, was established by peptide analysis and cDNA cloning. Serotriflin is a glycosylated protein and consists of 221 amino acids. Among the 3 SSPs, only SSP-2 formed a noncovalent complex with serotriflin. It was bound to triflin and serotriflin with high affinity, as evidenced by surface plasmon resonance. SSP-2 is considered to be a protein that prevents self injury by accidental leaking of venom into the blood.

  3. Influence of clotting time on the protein composition of serum samples based on LC-MS data

    NARCIS (Netherlands)

    Govorukhina, Natalia I.; de Vries, Marcel; Reijmers, Theo H.; Horvatovich, Peter; van der Zee, Ate G. J.; Bischoff, Rainer


    Many large, disease-related biobanks of serum samples have been established prior to the widespread use of proteomics in biomarker research. These biobanks may contain relevant information about the disease process, response to therapy or patient classifications especially with respect to long-term

  4. Reference intervals for acute phase protein and serum protein electrophoresis values in captive Asian elephants (Elephas maximus). (United States)

    Isaza, Ramiro; Wiedner, Ellen; Hiser, Sarah; Cray, Carolyn


    Acute phase protein (APP) immunoassays and serum protein electrophoresis (SPEP) are assays for evaluating the inflammatory response and have use as diagnostic tools in a variety of species. Acute phase proteins are markers of inflammation that are highly conserved across different species while SPEP separates and quantifies serum protein fractions based on their physical properties. In the current study, serum samples from 35 clinically healthy Asian elephants (Elephas maximus) were analyzed using automated assays for C-reactive protein, serum amyloid A, and haptoglobin and SPEP. Robust methods were used to generate reference intervals for the APPs: C-reactive protein (1.3-12.8 mg/l), serum amyloid A (0-47.5 mg/l), and haptoglobin (0-1.10 mg/ml). In addition, SPEP was performed on these samples to establish reference intervals for each protein fraction. A combination of APPs and SPEP measurements are valuable adjunctive diagnostic tools in elephant health care.

  5. Serum Protein Expression Profiling for Cancer Detection: Validation of a SELDI-Based Approach for Prostate Cancer

    Directory of Open Access Journals (Sweden)

    William E. Grizzle


    Full Text Available Multiple studies have reported that analysis of serum and other bodily fluids using surface enhanced laser desorption/ionization time of flight mass spectroscopy (SELDI-TOF-MS can identify a “fingerprint” or “signature” of spectral peaks that can separate patients with a specific disease from normal control patients. Ultimately, classification by SELDI-TOF-MS relies on spectral differences in position and amplitude of resolved peaks. Since the reproducibility of quantitation, resolution and mass accuracy of the SELDI-TOF-MS, or any high throughput mass spectrometric technique, has never been determined this method has come under some skepticism as to its clinical usefulness. This manuscript describes a detailed design of a three-phase study to validate the clinical usefulness of SELDI-TOF-MS in the identification of patients with prostatic adenocarcinoma (PCA. At the end of this validation study, the usefulness of the general SELDI-TOF-MS approach to identifying patients with PCA will be demonstrated and how it compares with PCA diagnosis by measuring prostate specific antigen.

  6. Protein Microarrays for Quantitative Detection of PAI-1 in Serum

    Institute of Scientific and Technical Information of China (English)

    Xu Ma; Qing-yun Zhang


    Objective:Plasminogen activator inhibitor-1 (PAl-1),one crucial component of the plasminogen activator system,is a major player in the pathogenesis of many vascular diseases as well as in cancer.High levels of PAI-1 in breast cancer tissue are associated with poor prognosis.The aim of this study is to evaluate rigorously the potential of serum PAl-1 concentration functioning as a general screening test in diagnostic or prognostic assays.Methods:A protein-microarray-based sandwich fluorescence immunoassay (FIA) was developed to detect PAl-1 in serum.Several conditions of this microarray-based FIA were optimized to establish an efficacious method.Serum specimens of 84 healthy women and 285 women with breast cancer were analyzed using the optimized FIA microarray.Results:The median serum PAl-1 level of breast cancer patients was higher than that of healthy women (109.7 ng/ml vs.63.4 ng/ml).Analysis of covariance revealed that PAl-1 levels of the two groups were significantly different (P<0.001) when controlling for an age effect on PAl-1 levels.However,PAl-1 values in TNM stage Ⅰ-Ⅳ patients respectively were not significantly different from each other.Conclusion:This microarray-based sandwich FIA holds potential for quantitative analysis of tumor markers such as PAl-1.

  7. iTRAQ-Based Proteomics Analysis of Serum Proteins in Wistar Rats Treated with Sodium Fluoride: Insight into the Potential Mechanism and Candidate Biomarkers of Fluorosis (United States)

    Wei, Yan; Zeng, Beibei; Zhang, Hua; Chen, Cheng; Wu, Yanli; Wang, Nanlan; Wu, Yanqiu; Shen, Liming


    Fluorosis induced by exposure to high level fluoride is quite widespread in the world. The manifestations of fluorosis include dental mottling, bone damage, and impaired malfunction of soft tissues. However, the molecular mechanism of fluorosis has not been clarified until now. To explore the underlying mechanisms of fluorosis and screen out serum biomarkers, we carried out a quantitative proteomics study to identify differentially expressed serum proteins in Wistar rats treated with sodium fluoride (NaF) by using a proteomics approach of isobaric tagging for relative and absolute quantitation (iTRAQ). We fed Wistar rats drinking water that had 50, 150, and 250 mg/L of dissolved NaF for 24 weeks. For the experimental duration, each rat was given an examination of the lower incisors to check for the condition of dental fluorosis (DF). By the end of the treatment, fluoride ion concentration in serum and lower incisors were detected. The results showed that NaF treatment can induce rat fluorosis. By iTRAQ analysis, a total of 37 differentially expressed serum proteins were identified between NaF-treated and control rats. These proteins were further analyzed by bioinformatics, out of which two proteins were validated by enzyme-linked immunoadsorbent assays (ELISA). The major proteins were involved in complement and coagulation cascade, inflammatory response, complement activation, defense response, and wound response, suggesting that inflammation and immune reactions may play a key role in fluorosis pathogenesis. These proteins may contribute to the understanding of the mechanism of fluoride toxicity, and may serve as potential biomarkers for fluorosis. PMID:27690006

  8. iTRAQ-Based Proteomics Analysis of Serum Proteins in Wistar Rats Treated with Sodium Fluoride: Insight into the Potential Mechanism and Candidate Biomarkers of Fluorosis

    Directory of Open Access Journals (Sweden)

    Yan Wei


    Full Text Available Fluorosis induced by exposure to high level fluoride is quite widespread in the world. The manifestations of fluorosis include dental mottling, bone damage, and impaired malfunction of soft tissues. However, the molecular mechanism of fluorosis has not been clarified until now. To explore the underlying mechanisms of fluorosis and screen out serum biomarkers, we carried out a quantitative proteomics study to identify differentially expressed serum proteins in Wistar rats treated with sodium fluoride (NaF by using a proteomics approach of isobaric tagging for relative and absolute quantitation (iTRAQ. We fed Wistar rats drinking water that had 50, 150, and 250 mg/L of dissolved NaF for 24 weeks. For the experimental duration, each rat was given an examination of the lower incisors to check for the condition of dental fluorosis (DF. By the end of the treatment, fluoride ion concentration in serum and lower incisors were detected. The results showed that NaF treatment can induce rat fluorosis. By iTRAQ analysis, a total of 37 differentially expressed serum proteins were identified between NaF-treated and control rats. These proteins were further analyzed by bioinformatics, out of which two proteins were validated by enzyme-linked immunoadsorbent assays (ELISA. The major proteins were involved in complement and coagulation cascade, inflammatory response, complement activation, defense response, and wound response, suggesting that inflammation and immune reactions may play a key role in fluorosis pathogenesis. These proteins may contribute to the understanding of the mechanism of fluoride toxicity, and may serve as potential biomarkers for fluorosis.

  9. Optimalisasi Pengikatan Tanin Daun Nangka dengan Protein Bovine Serum Albumin (Optimalisation Binding of Jackfruit Leaves Tannin with Bovine Serum Albumin Protein)


    Wahidin Teguh Sasongko; Lies Mira Yusiati; Zaenal Bachruddin; (Mugiono), Mugiono


    Tannins are high molecular weight polyphenol compounds with ability to bind proteins. Based on the structure, albumin are simple globular molecule protein. Optimalisation binding of jackfruit leave tannins to bovine serum (BSA) albumin was done in two stages. The first stage was to determine levels of tannins and condensed tannins in jackfruit leaves grown in mediterranean soil types. Second research was optimalisation binding of jackfruit leaf tannins with bovine serum albumin. In t...

  10. Evaluation of three high abundance protein depletion kits for umbilical cord serum proteomics

    Directory of Open Access Journals (Sweden)

    Nie Jing


    Full Text Available Abstract Background High abundance protein depletion is a major challenge in the study of serum/plasma proteomics. Prior to this study, most commercially available kits for depletion of highly abundant proteins had only been tested and evaluated in adult serum/plasma, while the depletion efficiency on umbilical cord serum/plasma had not been clarified. Structural differences between some adult and fetal proteins (such as albumin make it likely that depletion approaches for adult and umbilical cord serum/plasma will be variable. Therefore, the primary purposes of the present study are to investigate the efficiencies of several commonly-used commercial kits during high abundance protein depletion from umbilical cord serum and to determine which kit yields the most effective and reproducible results for further proteomics research on umbilical cord serum. Results The immunoaffinity based kits (PROTIA-Sigma and 5185-Agilent displayed higher depletion efficiency than the immobilized dye based kit (PROTBA-Sigma in umbilical cord serum samples. Both the PROTIA-Sigma and 5185-Agilent kit maintained high depletion efficiency when used three consecutive times. Depletion by the PROTIA-Sigma Kit improved 2DE gel quality by reducing smeared bands produced by the presence of high abundance proteins and increasing the intensity of other protein spots. During image analysis using the identical detection parameters, 411 ± 18 spots were detected in crude serum gels, while 757 ± 43 spots were detected in depleted serum gels. Eight spots unique to depleted serum gels were identified by MALDI- TOF/TOF MS, seven of which were low abundance proteins. Conclusions The immunoaffinity based kits exceeded the immobilized dye based kit in high abundance protein depletion of umbilical cord serum samples and dramatically improved 2DE gel quality for detection of trace biomarkers.

  11. Association of serum leptin with serum C-reactive protein in hemodialysis patients

    Directory of Open Access Journals (Sweden)

    Rastegari Ebrahim


    Full Text Available Introduction: Recent investigations have shown that leptin is cleared principally by the kidney. Objectives: To examine whether and how in patients on hemodialysis the level of C-reactive protein level correlate with serum leptin. Patients and Methods: The total patients were 36. The mean patients’ age were 46 (16 years. The median length of the time patients were on hemodialysis were 19 months. Results: The mean serum C-reactive protein was 8.7 (6.6 mg/l (median: 8 mg/l. The mean serum leptin was 9.4 (14 ng/ml (median: 5.75 ng/ml. In this study we found a significant inverse correlation of serum leptin with serum C-reactive protein (r= -0.57, p= 0.041 was seen. Conclusion: Our data supports, the positive effect of leptin on nutrition and support the theory of protective effects (reverse epidemiology of leptin in hemodialysis patients.

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

    Institute of Scientific and Technical Information of China (English)

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


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

  13. Interaction of Serum Proteins with Surface of Hemodialysis Fiber Membranes (United States)

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


    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.

  14. Comparative serum protein binding of anthracycline derivatives. (United States)

    Chassany, O; Urien, S; Claudepierre, P; Bastian, G; Tillement, J P


    The binding of doxorubicin, iododoxorubicin, daunorubicin, epirubicin, pirarubicin, zorubicin, aclarubicin, and mitoxantrone to 600 microM human serum albumin and 50 microM alpha 1-acid glycoprotein was studied by ultrafiltration at 37 degrees C and pH 7.4. Anthracycline concentrations (total and free) were determined by high-performance liquid chromatography (HPLC) with fluorometric detection. Binding to albumin (600 microM) varied from 61% (daunorubicin) to 94% (iododoxorubicin). The binding to alpha 1-acid glycoprotein (50 microM) was more variable, ranging from 31% (epirubicin) to 64% (zorubicin), and was essentially related to the hydrophobicity of the derivatives. Simulations showed that the total serum binding varied over a broad range from 71% (doxorubicin) to 96% (iododoxorubicin). We recently reported that the binding to lipoproteins of a series of eight anthracycline analogues could be ascribed to chemicophysical determinants of lipophilicity [2]. The present study was conducted to evaluate in vitro the contribution of albumin and alpha 1-acid glycoprotein to the total serum binding of these drugs.

  15. Grizzly bear corticosteroid binding globulin: Cloning and serum protein expression. (United States)

    Chow, Brian A; Hamilton, Jason; Alsop, Derek; Cattet, Marc R L; Stenhouse, Gordon; Vijayan, Mathilakath M


    Serum corticosteroid levels are routinely measured as markers of stress in wild animals. However, corticosteroid levels rise rapidly in response to the acute stress of capture and restraint for sampling, limiting its use as an indicator of chronic stress. We hypothesized that serum corticosteroid binding globulin (CBG), the primary transport protein for corticosteroids in circulation, may be a better marker of the stress status prior to capture in grizzly bears (Ursus arctos). To test this, a full-length CBG cDNA was cloned and sequenced from grizzly bear testis and polyclonal antibodies were generated for detection of this protein in bear sera. The deduced nucleotide and protein sequences were 1218 bp and 405 amino acids, respectively. Multiple sequence alignments showed that grizzly bear CBG (gbCBG) was 90% and 83% identical to the dog CBG nucleotide and amino acid sequences, respectively. The affinity purified rabbit gbCBG antiserum detected grizzly bear but not human CBG. There were no sex differences in serum total cortisol concentration, while CBG expression was significantly higher in adult females compared to males. Serum cortisol levels were significantly higher in bears captured by leg-hold snare compared to those captured by remote drug delivery from helicopter. However, serum CBG expression between these two groups did not differ significantly. Overall, serum CBG levels may be a better marker of chronic stress, especially because this protein is not modulated by the stress of capture and restraint in grizzly bears.

  16. Serum CRP protein as a differential marker in cancer. (United States)

    Juan, Hu; Qijun, Wang; Junlan, Liu


    The objective of this study was to measure the serum concentrations of C-reactive protein (CRP) in cancer patients and compare with those of immune disease patients and healthy individuals for discriminatory analysis. For this purpose, automatic systems for special protein analysis (Type: Drcon Diognostica Tarbox) was used to measure serum CRP concentrations in 276 cancer patients (Group A), 110 immune disease patients (Group B), 161 phlogistic patients (Group C), and 125 age-matched healthy individuals (Group D). Our data show that serum CRP concentrations in Group A were significantly higher than those in Groups B and D, whereas CRP concentrations in Group B were higher than those in Group D. The differences of serum CRP concentrations between Groups A and B as well as between Groups B and D were significant (P immune disease patients.

  17. QGQS Granule in SHR Serum Metabonomics Study Based on Tools of UPLC-Q-TOF and Renin-Angiotensin-Aldosterone System Form Protein Profilin-1 (United States)


    QGQS granule is effective for the therapeutic of hypertension in clinic. The aim of this research is to observe the antihypertension effect of QGQS granule on SHR and explain the mechanism of its lowering blood pressure. 30 SHR were selected as model group, captopril group, and QGQS group, 10 WKYr were used as control group, and RBP were measured on tail artery consciously. And all the serum sample analysis was carried out on UPLC-TOF-MS system to determine endogenous metabolites and to find the metabonomics pathways. Meanwhile, ELISA kits for the determination pharmacological indexes of PRA, AngI, AngII, and ALD were used for pathway confirmatory; WB for determination of profilin-1 protein expression was conducted for Ang II pathway analysis as well. It is demonstrated that QGQS granule has an excellent therapeutic effect on antihypertension, which exerts effect mainly on metabonomics pathway by regulating glycerophospholipid, sphingolipid, and arachidonic acid metabolism, and it could inhibit the overexpression of the profilin-1 protein. We can come to a conclusion that RAAS should be responsible mainly for the metabonomics pathway of QGQS granule on antihypertension, and it plays a very important role in protein of profilin-1 inhibition.

  18. Serum Copper and Plasma Protein Status in Normal Pregnancy

    Directory of Open Access Journals (Sweden)

    Nushrat Noor, Nasim Jahan, Nayma Sultana


    Full Text Available AbstractBackground: Gradual alteration of serum copper and some plasma protein levels may occur with advancement of pregnancy, which is associated with increased maternal and infant morbidity and mortality.Objective: To observe serum copper and plasma protein levels in normal pregnant women of different trimesters in order to find out their nutritional status.Methods: This cross sectional study was carried out in the Department of Physiology, Sir Salimullah Medical College (SSMC, Dhaka, between 1st January 2010 and December 2010. Ninety normal pregnant women of different trimesters with age 20-30 years were included in the study group. They were selected from Out Patient Department of Obstetrics and Gynaecology, SSMC. Age matched 30 non-pregnant women were taken as control. Serum copper level was measured by Spectrophotometric method, serum total protein and albumin levels were estimated by standard method. Statistical analysis was done by one way ANOVA, Bonferroni and Pearson’s correlation coefficient test as applicable.Results: Serum Cu levels were significantly higher in all trimesters of pregnant women compared to control. Again, this value was significantly higher in 3rd trimester than that of in 1st and 2nd trimester and also in 2nd trimester than that of in 1st trimester. In addition, mean serum total protein level was significantly lower in 3rd trimester than control but no statistically significant difference was observed among different trimesters. Again, mean serum albumin level was significantly lower in 2nd and 3rd trimester than 1st trimester and control. In addition, serum Cu concentration showed significant positive correlation with different trimesters of gestation.Conclusion: This study reveals that hypercupremia along with hypoproteinemia occur in pregnant women from 1st to 3rd trimester of gestation. This gradual alteration of micro and macronutrients become more profound with advancement of pregnancy.

  19. Serum Monocyte Chemoattractant Protein-1 in Pancreatic Cancer


    Jennifer Sullivan; Qiaoke Gong; Terry Hyslop; Harish Lavu; Galina Chipitsyna; Yeo, Charles J.; Arafat, Hwyda A


    Background/Aims. Pancreatic ductal adenocarcinoma (PDA) has etiological association with chronic inflammation. Elevated circulating levels of inflammatory mediators, such as monocyte chemoattractant protein-1 (MCP-1), are found in obese individuals. We hypothesized that serum MCP-1 levels are elevated in obese PDA patients. Methods. ELISA was used to analyze MCP-1 serum levels in PDA (n = 62) and intraductal papillary mucinous neoplasms (IPMN) (n = 27). Recursive partitioning statistical anal...

  20. 小麦饲粮中添加木聚糖酶对肉鹅血糖和血清总蛋白水平的影响%Effect of Wheat Based Diet Supplemented with Xylanase on Blood Sugar and Total Protein in Serum of Geese

    Institute of Scientific and Technical Information of China (English)

    王佳丽; 史东辉; 杨桂芹


    [Objective] The effects of wheat based diet supplemented with xylarmse on blood sugar and total protein in serum of geese were studied. [Method] By using the randomized design of single factor, the 1-day-old healthy goslings were divided into 6 groups and fed with corn based diet, wheat based diet and wheat based diet supplemented with xylanase at different concentrations respectively, the contents of blood sugar and total protein in serum were determined. [Result] The wheat based diet supplemented with xylanase could increase the blood sugar and total protein in serum of geese and wheat based diet supplemented with 0.2% xylanase generated the best effect, which was higher than those of corn based diet group. As for the concentration of protein in senun, wheat based diet supplemented with O. 2% xylarmse was significantly different from corn based diet and wheat based diet. [Conclusion] The wheat based diet supplemented with xylanase could enhance geese production.

  1. Differential serum protein markers and the clinical severity of asthma

    Directory of Open Access Journals (Sweden)

    Meyer N


    Full Text Available Norbert Meyer,1,2 Sarah Janine Nuss,1 Thomas Rothe,1 Alexander Siebenhüner,1 Cezmi A Akdis,2 Günter Menz11Hochgebirgsklinik Davos, Davos-Wolfgang, Switzerland; 2Swiss Institute of Allergy and Asthma Research (SIAF, Davos Platz, SwitzerlandBackground: Asthma is a heterogeneous disease characterized by different clinical phenotypes and the involvement of multiple inflammatory pathways. During airway inflammation, many cytokines and chemokines are released and some are detectable in the sera.Objective: Serum chemokines and cytokines, involved in airway inflammation in asthma patients, were investigated.Methods: A total of 191 asthma patients were classified by hierarchical cluster analysis, including the following parameters: forced expiratory volume in 1 second (FEV1, eosinophil cationic protein (ECP serum levels, blood eosinophils, Junipers asthma symptom score, and the change in FEV1, ECP serum levels, and blood eosinophils after 3 weeks of asthma therapy. Serum proteins were measured by multiplex analysis. Receiver operating characteristic (ROC curves were used to evaluate the validity of serum proteins for discriminating between asthma clusters.Results: Classification of asthma patients identified one cluster with high ECP serum levels, increased blood eosinophils, low FEV1 values, and good FEV1 improvement in response to asthma therapy (n=60 and one cluster with low ECP serum levels, low numbers of blood eosinophils, higher FEV1 values, and no FEV1 improvement in response to asthma therapy (n=131. Serum interleukin (IL-8, eotaxin, vascular endothelial growth factor (VEGF, cutaneous T-cell-attracting chemokine (CTACK, growth-related oncogene (GRO-α, and hepatocyte growth factor (HGF were significantly different between the two clusters of asthma patients. ROC analysis for serum proteins calculated a sensitivity of 55.9% and specificity of 75.8% for discriminating between them.Conclusion: Serum cytokine and chemokine levels might be

  2. Using competitive protein adsorption to measure fibrinogen in undiluted human serum (United States)

    Choi, Seokheun; Wang, Ran; Lajevardi-Khosh, Arad; Chae, Junseok


    We report a unique sensing mechanism based on competitive protein adsorption to measure fibrinogen, a cardiovascular biomarker, in undiluted human serum. The method uses physical adsorption of proteins to a surface rather than complex and time-consuming immobilization procedures. Two fibrinogen concentrations were differentiated in spiked in human serum [3.0 mg/ml (normal concentration) versus 3.2 mg/ml (abnormal concentration with heart disease)]. Real-time surface plasmon resonance signals were monitored as fibrinogen displaced a preadsorbed protein, IgM, on a hydrophobic gold surface. The relatively strong-affinity protein, IgM, was displaced primarily by fibrinogen and much less by other proteins in human serum.

  3. Serum monocyte chemoattractant protein-1 is a biomarker in patients with diabetes and periodontitis


    Preethi Radhakrishnan; Padma Srikanth; Seshadri, Krishna G.; Ramya Barani; Maitreya Samanta


    Introduction: The role of serum Monocyte Chemoattractant Protein-1 (MCP-1) as a biomarker of periodontitis is well documented; however, its role in diabetic patients with periodontitis is unknown. Aim : This study was conducted to determine the presence and concentration of serum MCP-1 in diabetic patients with and without periodontitis and correlate it glycemic status with periodontitis. Materials and Methods: Adult diabetic patients were enrolled and grouped into group I, II, and III based ...

  4. Competitive protein adsorption to polymer surface from human serum

    DEFF Research Database (Denmark)

    Holmberg, Maria; Jensen, Karin Bagger Stibius; Larsen, Niels Bent;


    Surface modification by "soft" plasma polymerisation to obtain a hydrophilic and non-fouling polymer surface has been validated using radioactive labelling. Adsorption to unmodified and modified polymer surfaces, from both single protein and human serum solutions, has been investigated. By using ...

  5. Genetic and environmental influences of surfactant protein D serum levels

    DEFF Research Database (Denmark)

    Sorensen, G.L.; Hjelmborg, J.V.; Kyvik, K.O.


    The collectin surfactant protein D (SP-D) is an important component of the pulmonary innate immune system, but SP-D is also present on extrapulmonary epithelial surfaces and in serum, where it has been used as a biomarker for pulmonary disease states. In this study, we investigate the mechanisms ...

  6. Blood serum components and serum protein test of Hybro-PG broilers of different ages

    Directory of Open Access Journals (Sweden)

    PRL Silva


    Full Text Available Blood serum samples of HYBRO PG broilers were analyzed, with 30 samples collected from 21-day-old broilers (G1, 30 from 35-day-old birds (G2, and 30 from 42-day-old birds (G3, with the aim of establishing normal values of some blood serum parameters. The activities of the enzymes gamma-glutamyl-transferase (GGT, aspartate aminotransferase (AST, creatine kinase (CK, alkaline phosphatase (ALP, and lactate dehydrogenase (LDH, serum levels of total calcium, calcium ion, phosphorus, sodium, potassium, magnesium, chlorides, creatinine, uric acid, triglycerides, cholesterol, total protein, albumin, total and indirect and direct bilirubin, and electrophoretic profile of serum proteins in acrylamide (SDS-PAGE and agarose gel were determined. There was no influence of age on total bilirubin and albumin levels. All the other evaluated parameters presented differences in at least one age group. Protein electrophoretic profile also changed as a function of age. The obtained results can be considered as normal for the studied ages, and therefore be used as references for the interpretation of laboratory exams of broilers of this genetic line in the evaluated ages.

  7. Beer consumption and changes in stability of human serum proteins. (United States)

    Gorinstein, S; Caspi, A; Goshev, I; Moncheva, S; Zemser, M; Weisz, M; Libman, I; Lerner, H T; Trakhtenberg, S; Martín-Belloso, O


    The aim of this study was to evaluate the influence of beer consumption (BC) on the functional and structural properties of human serum proteins (HSP). Thirty-eight volunteers (after coronary bypass) were divided into two groups: experimental (EG) and control (CG). Nineteen volunteers of the EG consumed 330 mL per day of beer (about 20 g of alcohol) for 30 consecutive days. The CG volunteers consumed mineral water instead of beer. Blood samples were collected from EG and CG patients before and after the experiment. Albumin (Alb), globulin (Glo), and methanol-precipitable proteins (MPP) from human serum were denatured with 8 M urea. Fluorescence and electrophoresis were employed in order to elucidate urea-induced conformational changes and structural behavior of proteins. The measured fluorescence emission spectra were used to estimate the stability of native and denatured protein fractions before and after BC. It was found that before BC the fractions most stable to urea denaturation were Glo, Alb, and MPP fractions. After BC in most of the beer-consuming patients (EG) some changes in native and denatured protein fractions were detected: a tendency to lower stability and minor structural deviations. These qualitative changes were more profound in MPP than in Alb and Glo. Thus, Glo is more resistible to alcohol influence than Alb, which in turn is more resistible than MPP. No serum protein changes were detected in patients of CG.

  8. [Electrophoretic studies of serum protein fractions in horses with laminitis]. (United States)

    Edinger, H; Miller, I; Stanek, C; Gemeiner, M


    The spectrum of serum proteins was evaluated in 46 horses affected with spontaneous laminitis and correlations between the severity of the disease and changes of the protein pattern were analyzed. The investigation was made in two groups; group A consisted of 21 horses of various breeds (warmblood, thoroughbred, standardbred) and group B of 25 ponys. Each group was subdivided according to the severity of the disease, using the OBEL-grade (OG) classification system. Serum proteins were separated by different one- and two-dimensional electrophoretic methods. Sera analysed by cellulose acetate electrophoresis showed a significant difference in the alpha 1-globulin fraction between OG II and OG IV affected horses. An increasing severity of the disease was correlated with a decrease of the alpha 1-globulins. The other protein fractions didn't show a uniform tendency. In group B there was a significant difference in the alpha 1-globulin fractions of OG II and OG III and in the beta 2-globulin fractions of OG I and OG II affected ponys. The acute phase proteins C3c, C4, Hp and fibronectin could be determined in a preliminary study in horse serum using the cross-reactivity of antibodies against the homologous human proteins.

  9. Antigenic specificity of serum antibodies in mice fed soy protein

    DEFF Research Database (Denmark)

    Christensen, Hanne Risager; Bruun, S.W.; Frøkiær, Hanne


    ingesting soy protein. Methods: Blood from mice fed a soy-containing diet was analyzed using ELISA and immunoblot for antibody reactivity towards various soy protein fractions and pure soy proteins/subunits. Mice bred on a soy-free diet were used as controls. Results: The detectable antigenic specificity...... of the serum antibodies of soy-consuming mice comprised glycinin and beta-conglycinin. Immunoblots with soy protein extract demonstrated antibody reactivity towards both the basic and the acidic chains of glycinin and the beta-conglycinin subunits with an individual response pattern among mice. Moreover......Background: Soybean protein is used in a number of food products but unfortunately is also a common cause of food allergy. Upon ingestion of soy protein, healthy mice like other animals and humans generate a soy-specific antibody response in the absence of signs of illness. Not much is known about...

  10. Discovery and identification of Serum Amyloid A protein elevated in lung cancer serum

    Institute of Scientific and Technical Information of China (English)


    Two hundred and eighteen serum samples from 175 lung cancer patients and 43 healthy individuals were analyzed by using Surface Enhaced Laser Desorption/Ionization Time of Flight Mass Spectrome- try (SELDI-TOF-MS). The data analyzed by both Biomarker Wizard? and Biomarker Patterns? software showed that a protein peak with the molecular weight of 11.6 kDa significantly increased in lung cancer. Meanwhile,the level of this biomarker was progressively increased with the clinical stages of lung cancer. The candidate biomarker was then obtained from tricine one-dimensional sodium dodecyl sul- fate-polyacrylamide gel electrophoresis by matching the molecular weight with peaks on WCX2 chips and was identified as Serum Amyloid A protein (SAA) by MALDI/MS-MS and database searching. It was further validated in the same serum samples by immunoprecipitation with commercial SAA antibody. To confirm the SAA differential expression in lung cancer patients, the same set of serum samples was measured by ELISA assay. The result showed that at the cutoff point 0.446(OD value)on the Receiver Operating Characteristic (ROC) curve, SAA could better discriminate lung cancer from healthy indi- viduals with sensitivity of 84.1% and specificity of 80%. These findings demonstrated that SAA could be characterized as a biomarker related to pathological stages of lung cancer.

  11. Serum protein profiling by solid phase extraction and mass spectrometry: A future diagnostics tool?

    DEFF Research Database (Denmark)

    Callesen, Anne K; Madsen, Jonna S; Vach, Werner;


    Serum protein profiling by MS is a promising method for early detection of disease. Important characteristics for serum protein profiling are preanalytical factors, analytical reproducibility and high throughput. Problems related to preanalytical factors can be overcome by using standardized...

  12. [Comparative immunochemical study of the serum proteins of Lacertilia (Reptilia)]. (United States)

    Blanc, F


    The immunochemical relationships of the serum proteins of Uromastix acanthinurus, Agama mutabilis, Oplurus cuvieri, O. quadrimaculatus, Chamaeleo chamaeleon, Chalcides ocellatus and Lacerta lepida were studied by means of immunoelectrophoresis with an antiserum anti-Uromastix successively absorbed. Homogeneity of the Iguania species was pointed out and they were found more closely related to the Scincid Chalcides ocellatus than to the Lacertid Lacerta lepida. Within the Iguania, the two Agamids are immunologically more closely related to the Chamaeleon than to the two Iguanids.

  13. Specific changes of serum proteins in Parkinson's disease patients.

    Directory of Open Access Journals (Sweden)

    Wenwen Lu

    Full Text Available The aim of this study is to identify and validate protein change in the serum from PD patients. We used serum samples from 21 PD patients and 20 age-matched normal people as control to conduct a comparative proteomic study. We performed 2-DE and analyzed the differentially expressed protein spots by LC-MS/MS. In PD group 13 spots were shown to be differentially expressed compared to control group. They were identified as 6 proteins. Among these, 3 proteins were confirmed by Western blot analysis. It showed that the frequency of fibrinogen γ-chain (FGG appeared 70% in PD, which could not be detected in control group. The protein of inter-alpha-trypsin inhibitor heavy chain H4 (ITI-H4 was found to exist two forms in serum. The full size (120 kDa of the protein was increased and the fragmented ITI-H4 (35 kDa was decreased in PD group. The ratio of full size ITI-H4 to fragmented ITI-H4 in PD patients was 3.85 ± 0.29-fold higher than in control group. Furthermore, fragmented Apo A-IV (∼ 26 kDa was mainly detected in control group, while it was rare to be found in PD group. Above findings might be useful for diagnosis of PD. When the expressions of FGG and 120 kDa ITI-H4 are increase, as well as ∼ 26 kDa Apo A-IV disappear would provide strong evidence for PD.

  14. An evaluation of multiplex bead-based analysis of cytokines and soluble proteins in archived lithium heparin plasma, EDTA plasma and serum samples

    DEFF Research Database (Denmark)

    Brøndum, Line; Sørensen, Brita Singers; Eriksen, Jesper Grau


    in plasma and serum from 86 head and neck cancer patients and 33 controls were evaluated: EGFR, leptin, OPN, VEGFR-1, VEGFR-2, IL-2, IL-13, PDGF-bb, TNF, PAI-1, SDF-1a, IL-4, IL-6, IL-8, eotaxin, G-CSF, VEGF, GRO-a, and HGF. RESULTS: The correlation between measurements of the same samples analyzed...

  15. Serum protein profiling by miniaturized solid-phase extraction and matrix-assisted laser desorption/ionization mass spectrometry

    DEFF Research Database (Denmark)

    Callesen, Anne K; Mohammed, Shabaz; Bunkenborg, Jakob;


    for translation of MALDI-MS based diagnostic methods to clinical applications. We have investigated a number of MALDI matrices and several miniaturized solid-phase extraction (SPE) methods for serum protein concentration and desalting with the aim of generating reproducible, high-quality protein profiles by MALDI...... mass spectra (m/z 1000-12,000) to be obtained from serum. In a proof-of-principle application, SPE with chelating material and MALDI-MS identified protein peaks in serum that had been previously reported for distinguishing a person diagnosed with breast cancer from a control. These preliminary results...

  16. The Acute-Phase Proteins Serum Amyloid A and C Reactive Protein in Transudates and Exudates (United States)

    Okino, Alessandra M.; Bürger, Cristiani; Cardoso, Jefferson R.; Lavado, Edson L.; Lotufo, Paulo A.; Campa, Ana


    The distinction between exudates and transudates is very important in the patient management. Here we evaluate whether the acute-phase protein serum amyloid A (SAA), in comparison with C reactive protein (CRP) and total protein (TP), can be useful in this discrimination. CRP, SAA, and TP were determined in 36 exudate samples (27 pleural and 9 ascitic) and in 12 transudates (9 pleural and 3 ascitic). CRP, SAA, and TP were measured. SAA present in the exudate corresponded to 10% of the amount found in serum, that is, the exudate/serum ratio (E/S) was 0.10 ± 0.13. For comparison, the exudate/serum ratio for CRP and TP was 0.39 ± 0.37 and 0.68 ± 0.15, respectively. There was a strong positive correlation between serum and exudate SAA concentration (r = 0.764;p < 0.0001). The concentration of SAA in transudates was low and did not overlap with that found in exudates (0.02-0.21 versus 0.8–360.5 g/mL). SAA in pleural and ascitic exudates results mainly from leakage of the serum protein via the inflamed membrane. A comparison of the E/S ratio of SAA and CRP points SAA as a very good marker in discriminating between exudates and transudates. PMID:16864904

  17. Protein Crystal Based Nanomaterials (United States)

    Bell, Jeffrey A.; VanRoey, Patrick


    This is the final report on a NASA Grant. It concerns a description of work done, which includes: (1) Protein crystals cross-linked to form fibers; (2) Engineering of protein to favor crystallization; (3) Better knowledge-based potentials for protein-protein contacts; (4) Simulation of protein crystallization.

  18. 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.

    Directory of Open Access Journals (Sweden)

    Chuan-Wei Yang

    Full Text Available 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-α. 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-α 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-α 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.

  19. Detection of H pylori antibody profile in serum by protein array

    Institute of Scientific and Technical Information of China (English)

    Feng-Chan Han; Xu-Jun Li; Hong Jiang; Li-Peng Qin; Ding Li; Yan-Hai Guo; Zhi-Guang Liu; Li Zhang; Xiao-Jun Yan


    AIM: To detect multiple H pylori antibodies in serum samples of individuals who carryH pyloriby protein array.METHODS: Recombinant H pyloriantigens, urease B subunit (UreB), vacuolating toxin A (VacA) and cytotoxin associated gene A protein (CagA), were prepared and immobilized in matrixes on nitrocellulose membrane by robotics to bind the specific immunoglobulin G (IgG) antibodies in serum. Staphylococcus protein A (SPA) labeled by colloid gold was used to integrate the immuno-complex and gave red color signal, The scanner based on charge-coupled device (CCD) could collect the image signal and convert it into digital signal.RESULTS: When human IgG was printed on the membrane in increasing concentrations and incubated with immunogold, a linear dose response curve was obtained and the detection limit for IgG was about 0.025 ng. The cutoff values, which were defined as the mean grey level plus 3 times of standard deviation, were 27.183, 28.546 and 27.402, for anti-UreB IgG, antiCagA IgG and anti-VacA IgG, respectively, as 400 human serum samples with negative H pylori antibodies were detected by the protein array. When 180 serum samples from patients in hospital were employed for detection of IgG against UreB, CagA and VacA, the sensitivity of the protein array was 93.4%, 95.4%, 96.0%, and the specificity was 94.8%, 94.4% and 97.5%, respectively,as compared with the results obtained by ELISA. The assay also showed high reproducibility, uniformity and stability, and the results were available within 30 min.CONCLUSION: The protein array is a very practical method for rapid detection of multiple antibodies in serum samples. It is especially useful for large scale epidemiological investigation of the infection of Hpylori.

  20. Development of gel-filter method for high enrichment of low-molecular weight proteins from serum.

    Directory of Open Access Journals (Sweden)

    Lingsheng Chen

    Full Text Available 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 μL 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.

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

    Energy Technology Data Exchange (ETDEWEB)

    Klein, N.W.


    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)

  2. Measures of bioavailable serum sex hormone levels in aging Chinese by protein chip

    Institute of Scientific and Technical Information of China (English)

    ZHOU Yong; DU Guanhua; CHANG Shuying; MENG Xiaoluo; YU Huafeng; WANG Luning; HE Jinggui; ZHANG Baohe; ZHANG Juntian; GENG Meiyu


    The purpose of this study was to develop a protein chip technique based on receptor binding assays to measure bioavailable serum sex hormone levels (BSSHL). 224 aging healthy Chinese were investigated to get the referenced values of BSSHL for the first time. In the assays recombined sex hormone receptor proteins were jointed to polysaccharide coated slides to make protein chip, and the dose-dependence curves of sex hormone on chip were prepared. The data showed that this method had good precision (CV<16%) and accuracy (Bias<10%), and the sensitivity could reach 1 pmol/L. From the results, BSSHL of men and women declined with aging, but no significant differences were observed. The BSSHL of aging men were higher than those of women. The bioavailable serum androgen level of men was 52―112 pmol/L, women's was 3―70 pmol/L and the whole group was 41.9―81.4 pmol/L. The bioavailable serum estrogen level of men was 0.8―3.0 pmol/L, women's was 1.2―2.5 pmol/L and the whole group was 0.6―2.64 pmol/L. Based on the assays, BSSHL measurement by protein chip can meet the needs of epidemiological studies in terms of speed, accuracy and sample volume required, and was helpful in quantitative assessment of aging people's health.

  3. The effect of serum magnesium levels and serum endothelin-1 levels on bone mineral density in protein energy malnutrition. (United States)

    Ozturk, C F; Karakelleoglu, C; Orbak, Z; Yildiz, L


    An inadequate and imbalanced intake of protein and energy results in protein-energy malnutrition (PEM). It is known that bone mineral density and serum magnesium levels are low in malnourished children. However, the roles of serum magnesium and endothelin-1 (ET-1) levels in the pathophysiology of bone mineralization are obscure. Thus, the relationships between serum magnesium and ET-1 levels and the changes in bone mineral density were investigated in this study. There was a total of 32 subjects, 25 of them had PEM and seven were controls. While mean serum ET-1 levels of the children with kwashiorkor and marasmus showed no statistically significant difference, mean serum ET-1 levels of both groups were significantly higher than that of the control group. Serum magnesium levels were lower than normal value in 9 (36%) of 25 malnourished children. Malnourished children included in this study were divided into two subgroups according to their serum magnesium levels. While mean serum ET-1 levels in the group with low magnesium levels were significantly higher than that of the group with normal magnesium levels (p malnutrition. Our study suggested that lower magnesium levels and higher ET-1 levels might be important factors in changes of bone mineral density in malnutrition. We recommend that the malnourished patients, especially with hypomagnesaemia, should be treated with magnesium early.

  4. Genetic analysis and serum level of cartilage oligomeric matrix protein in patients with pseudoachondroplasia

    Institute of Scientific and Technical Information of China (English)

    LIU Feng-xia; LI Zhi-ling; WEI Zhen-ji; MENG yan; REN Cui-ai; ZHANG Xu-de; YU Meng-xue; HUANG Shang-zhi


    Background Pseudoachondroplasia (PSACH) is an autosomal-dominant osteochondrodysplasia due to mutations in the gene encoding cartilage oligomeric matrix protein (COMP).Clinical diagnosis of PSACH is based primarily on family history, physical examination, and radiographic evaluation.There is evidence that decreased serum COMP concentration may serve as a diagnostic marker in PSACH.Here, we investigated the role of this gene and the serum COMP concentration in Chinese patients with PSACH.Methods A family with three patients and a sporadic case were recruited.Genomic and phenotypic data were recorded.The diagnosis of PSACH was made on the base of clinical evaluation.The genomic DNA was extracted from peripheral blood leukocytes.The 8-19 exons and flanking intron-exon boundary sequences of COMP were amplified by polymerase chain reaction (PCR) and screened for mutation by direct DNA sequencing.Serum COMP concentrations of 4 patients and age-compatible control group of 20 unrelated healthy subjects were analyzed on the basis of an ELISA Kit for human cartilage oligomeric matrix protein.Results A deletion (c.1447-1455del) was identified in exon 13 in the sporadic case.The mean serum COMP concentrations of four patients (3.12±2.28) were significantly lower than those of control group (10.86±2.21, P <0.05).There was no overlap in the distribution of serum COMP concentration between PSACH patients and controls.Conclusions Mutations in COMP gene are responsible for the PSACH.Serum COMP concentration may be suggested as an additional diagnostic marker to aid clinical findings in suspected cases of PSACH.

  5. Serum Monocyte Chemoattractant Protein-1 in Pancreatic Cancer

    Directory of Open Access Journals (Sweden)

    Jennifer Sullivan


    Full Text Available Background/Aims. Pancreatic ductal adenocarcinoma (PDA has etiological association with chronic inflammation. Elevated circulating levels of inflammatory mediators, such as monocyte chemoattractant protein-1 (MCP-1, are found in obese individuals. We hypothesized that serum MCP-1 levels are elevated in obese PDA patients. Methods. ELISA was used to analyze MCP-1 serum levels in PDA (n=62 and intraductal papillary mucinous neoplasms (IPMN (n=27. Recursive partitioning statistical analysis investigated the relationship between log MCP-1 and clinicopathological parameters. Results. Log MCP-1 values were significantly (P<0.05 elevated in patients with BMI ≥ 37.5. In patients with BMI < 37.5, average log MCP-1 values were significantly elevated in PDA patients when compared to IPMN patients. Within the IPMN group, higher log MCP-1 levels correlated with increased age. Recursive partitioning analysis of IPMN versus PDA revealed a strategy of predicting characteristics of patients who are more likely to have cancer. This strategy utilizes log MCP-1 as the primary factor and also utilizes smoking status, gender, and age. Conclusion. MCP-1 is a promising biomarker in pancreatic cancer. The potential of using MCP-1 to distinguish PDA from IPMN patients must be studied in larger populations to validate and demonstrate its eventual clinical utility.

  6. Changes in Serum Protein in Mice Infected with Hymenolepis nana Eggs


    SHIMODA, Kenji; OGO, Masanori; Sato, Shoichi; Ueda, Satoshi


    We investigated the changes in serum protein contents of male ddY mice following primary and secondary infections with Hymenolepis nana eggs. There was a marked decrease in the total serum protein contents throughout the course of both primary and secondary infections. The reduction of total serum protein contents occurred as early as 1 day after primary and secondary infections and was particularly prominent after secondary infection. A quiet similar pattern of reduction was observed in the ...

  7. Evaluation of steric exclusion chromatography on cryogel column for the separation of serum proteins. (United States)

    Wang, Chuan; Bai, Shu; Tao, Shi-Peng; Sun, Yan


    Steric exclusion chromatography (SXC) is a new mode of protein chromatography, in which large proteins are retained on hydrophilic stationary phase surface due to the steric exclusion of polyethylene glycol (PEG) in the mobile phase, and thereafter the retained proteins can be eluted by reducing PEG concentration. In this work, SXC was evaluated on a polyacrylamide cryogel monolith. Microscopic observation of γ-globulin precipitates on the gel surface in SXC was reported for the first time. Due to the compact packing of protein precipitates on the stationary phase surface, the dynamic retention capacity of the cryogel monolith for γ-globulin reached 20 mg/mL bed volume, much higher than those of cryogel beds in adsorption-based chromatography. The effect of molecular weight and concentration of PEG, solution pH and salt concentration on protein retention capacity was in agreement with the earlier work on SXC. Because the cryogel monoliths with interconnected macropores (10-100 μm) allow much easy flow-through of viscous PEG buffer, the SXC can be operated at low back pressure. Hence, the cryogel monoliths are more suitable for SXC than other monoliths of narrow pores reported previously. In the separation of bovine serum proteins, albumin was recovered in the breakthrough fraction with high purity, and globulin was over eight times concentrated in the elution pool. This work has, thus, demonstrated the rapid serum protein separation and concentration by SXC on the cryogel monolith columns.

  8. On the transfer of serum proteins to the rat intestinal juice

    DEFF Research Database (Denmark)

    Andersen, Vibeke; Hansen, G H; Olsen, J;


    The in vivo pattern of serum proteins in the rat small-intestinal juice was characterized by crossed immunoelectrophoresis. Immunoglobulins and albumin, alpha-1-antitrypsin, transferrin, and orosomucoid were present. Larger serum proteins were absent (ceruloplasmin, haptoglobin, alpha-1-macroglob......The in vivo pattern of serum proteins in the rat small-intestinal juice was characterized by crossed immunoelectrophoresis. Immunoglobulins and albumin, alpha-1-antitrypsin, transferrin, and orosomucoid were present. Larger serum proteins were absent (ceruloplasmin, haptoglobin, alpha-1...... was seen to leak out into the intestinal lumen from the opened interstitial spaces. A weak labelling was also found in the lysosomal/endosomal-like structures, especially in the crypt enterocytes, indicating pinocytosis of albumin. We conclude that the main reason for the occurrence of certain serum...... proteins in the intestinal juice is a selective passage through the capillary wall followed by passive intercellular transport via delivery of the serum in the interstitial space during disintegration of the enterocytes....

  9. Evaluation of serum biochemical marker concentrations and survival time in dogs with protein-losing enteropathy

    NARCIS (Netherlands)

    Equilino, Mirjam; Théodoloz, Vincent; Gorgas, Daniela; Doherr, Marcus G.; Heilmann, Romy M.; Suchodolski, Jan S.; Steiner, JöRg M.; Burgener, Iwan A.


    Results—Serum C-reactive protein concentration was high in 13 of 18 dogs with PLE and in 2 of 18 dogs with FRD. Serum concentration of canine pancreatic lipase immunoreactivity was high in 3 dogs with PLE but within the reference interval in all dogs with FRD. Serum a1-proteinase inhibitor concentra

  10. Maternal serum protein profile and immune response protein subunits as markers for non-invasive prenatal diagnosis of trisomy 21, 18, and 13

    KAUST Repository

    Narasimhan, Kothandaraman


    Objectives: To use proteomics to identify and characterize proteins in maternal serum from patients at high-risk for fetal trisomy 21, trisomy 18, and trisomy 13 on the basis of ultrasound and maternal serum triple tests. Methods: We performed a comprehensive proteomic analysis on 23 trisomy cases and 85 normal cases during the early second trimester of pregnancy. Protein profiling along with conventional sodium dodecyl sulfate polyacrylamide gel electrophoresis/Tandem mass spectrometry analysis was carried out to characterize proteins associated with each trisomy condition and later validated using Western blot. Results: Protein profiling approach using surface enhanced laser desorption/ionization time-of-flight mass (SELDI-TOF/MS) spectrometry resulted in the identification of 37 unique hydrophobic proteomic features for three trisomy conditions. Using sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by Matrix Assisted Laser Desorption Ionization - Time of Flight/Time of Flight (MALDI-TOF/TOF) and western blot, glyco proteins such as alpha-1-antitrypsin, apolipoprotein E, apolipoprotein H, and serum carrier protein transthyretin were identified as potential maternal serum markers for fetal trisomy condition. The identified proteins showed differential expression at the subunit level. Conclusions: Maternal serum protein profiling using proteomics may allow non-invasive diagnostic testing for the most common trisomies and may complement ultrasound-based methods to more accurately determine pregnancies with fetal aneuploidies. © 2013 John Wiley & Sons, Ltd.

  11. Spectroscopic study of drug-binding characteristics of unmodified and pNPA-based acetylated human serum albumin: Does esterase activity affect microenvironment of drug binding sites on the protein?

    Energy Technology Data Exchange (ETDEWEB)

    Moradi, Nastaran [Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah (Iran, Islamic Republic of); Faculty of Pharmaceutical Sciences, Kermanshah University of Medical Sciences, Kermanshah (Iran, Islamic Republic of); Ashrafi-Kooshk, Mohammad Reza [Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah (Iran, Islamic Republic of); Ghobadi, Sirous [Department of Biology, Faculty of Sciences, Razi University, Kermanshah (Iran, Islamic Republic of); Shahlaei, Mohsen [Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah (Iran, Islamic Republic of); Faculty of Pharmaceutical Sciences, Kermanshah University of Medical Sciences, Kermanshah (Iran, Islamic Republic of); Khodarahmi, Reza, E-mail: [Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah (Iran, Islamic Republic of); Faculty of Pharmaceutical Sciences, Kermanshah University of Medical Sciences, Kermanshah (Iran, Islamic Republic of)


    Human serum albumin (HSA) is the most prominent extracellular protein in blood plasma. There are several binding sites on the protein which provide accommodation for structurally-unrelated endogenous and exogenous ligands and a wide variety of drugs. “Esterase-like” activity (hydrolysis of p-nitrophenyl esters) by the protein has been also reported. In the current study, we set out to investigate the interaction of indomethacin and ibuprofen with the unmodified and modified HSA (pNPA-modified HSA) using various spectroscopic techniques. Fluorescence data showed that 1:1 binding of drug to HSA is associated with quenching of the protein intrinsic fluorescence. Decrease of protein surface hydrophobicity (PSH), alteration in drug binding affinity and change of the protein stability, after esterase-like activity and permanent acetylation of HSA, were also documented. Analysis of the quenching and thermodynamic parameters indicated that forces involved in drug–HSA interactions change upon the protein modification. - Highlights: • Binding propensity of indomethacin extremely decreased upon the protein acetylation. • There is no ibuprofen binding after protein acetylation. • Protein stability changes upon drug binding as well as protein acetylation. • Drug pharmacokinetics may be influenced under co-administration of HSA-modifier drugs.

  12. A preliminary screening study on the associated proteins in human psoriasis vulgaris by serum proteomics technologies

    Institute of Scientific and Technical Information of China (English)

    Zhankui Liu; Shengshun Tan; Chunshui Yu; Jinghua Fan; Zhuanli Bai; Junjie Li


    Objective:To investigate the optimum screening conditions of associated proteins in human psoriasis vulgaris by serum proteomics technique, and to screen the different expression proteins related with psoriasis vulgaris. Methods:Serum samples of peripheral blood were collected from newly diagnosed psoriasis vulgaris patients in the clinic, and 20 matched healthy persons.Serum albumin IgG was removed by filtering with ProteoExtract Albumin/IgG. After comparative proteomics analysis the different protein spots were identified using 2-DE and MS. Results :Electrophoresis figures with high resolution and reproducibility were obtained. Three different expression proteins were found only in the serum from psoriasis vulgaris patients,while nine other different proteins expressing from healthy volunteers. Conclusion:The protein expression was different in the serum between the psoriasis vulgaris patients and healthy volunteers. It was hoped that we could find the biomarkers related to psoriasis vulgaris by using proteomics.

  13. Identification and quantification of serum proteins secreted into the normal human jejunum

    DEFF Research Database (Denmark)

    Andersen, Vibeke; Hegnhøj, J H


    The in vivo transfer of serum proteins to the human intestinal lumen was characterized by crossed immunoelectrophoretic analyses of intestinal perfusates from four healthy volunteers. Serum proteins with molecular masses below 100 kDa and the immunoglobulins were found in human jejunal perfusates....... Larger serum proteins were either absent (alpha and beta lipoproteins) or present in small amounts (alpha 2-macroglobulin, haptoglobulin and ceruloplasmin). These results demonstrate the existence of a selective transfer of serum proteins to the intestinal lumen under physiological conditions....... The intestinal clearance rate was 0.1 ml serum per hour per 10 cm jejunum for albumin, prealbumin, alpha 1-antitrypsin, orosomucoid, transferrin and haemopexin. The rate of secretion of total protein to the jejunal lumen was 100 mg protein per hour per 10 cm jejunum. About 45% was due to immunoglobulins...

  14. Serum protein capillary electrophoresis and measurement of acute phase proteins in a captive cheetah (Acinonyx jubatus) population. (United States)

    Depauw, Sarah; Delanghe, Joris; Whitehouse-Tedd, Katherine; Kjelgaard-Hansen, Mads; Christensen, Michelle; Hesta, Myriam; Tugirimana, Pierrot; Budd, Jane; Dermauw, Veronique; Janssens, Geert P J


    Renal and gastrointestinal pathologies are widespread in the captive cheetah (Acinonyx jubatus) population but are often diagnosed at a late stage, because diagnostic tools are limited to the evaluation of clinical signs or general blood examination. Presently, no data are available on serum proteins and acute-phase proteins in cheetahs during health or disease, although they might be important to improve health monitoring. This study aimed to quantify serum proteins by capillary electrophoresis in 80 serum samples from captive cheetahs, categorized according to health status and disease type. Moreover, serum amyloid A concentrations were measured via a turbidimetric immunoassay validated in domestic cats, whereas haptoglobin and C-reactive protein were determined by non-species-specific functional tests. Cheetahs classified as healthy had serum protein and acute phase protein concentrations within reference ranges for healthy domestic cats. In contrast, unhealthy cheetahs had higher (P cheetahs suffering from chronic kidney disease were significantly greater compared to the reportedly healthy cheetahs. Our study indicates that serum proteins in the cheetah can be analyzed by routine capillary electrophoresis, whereas acute-phase proteins can be measured using available immunoassays or non-species-specific techniques, which are also likely to be applicable in other exotic felids. Moreover, results suggest that serum amyloid A and haptoglobin are important acute-phase proteins in the diseased cheetah and highlight the need to evaluate their role as early-onset markers for disease.

  15. Evaluation of the refractometric method for the determination of total protein in avian plasma or serum. (United States)

    Lumeij, J T; de Bruijne, J J


    Serum total protein concentrations in pigeon blood determined with the biuret method (TPB-se) were compared with total protein concentrations in plasma (TPR-pl) and serum (TPR-se) obtained by estimation from refractive index. The refractometric method consistently yielded higher values (Prefractometric method for determination of TP in pigeon blood is not recommended.


    Gumanova, N G; Klimushina, M V; Metelskaya, V A; Boitsov, S A


    The microchip technology represents convenient and relatively economic tool of analyzing specific biomarkers with the purpose to diagnose diseases, to evaluate effectiveness of therapy and to investigate signaling pathways. To analyze protein composition of blood serum certain types of finished microchips which were not applied previously on the territory of Russia. The detection from 2% to 5% out of matrix of chips depending on their variety was managed without preliminary depletion of serum (removal of proteins of major fractions). Hence, partial protein composition of blood serum can be analyzed with microchips even without preliminary removal of proteins of major fractions.

  17. Structural analysis and characterization of new small serum proteins from the serum of a venomous snake (Gloydius blomhoffii). (United States)

    Shioi, Narumi; Deshimaru, Masanobu; Terada, Shigeyuki


    Some snakes have several anti-toxic proteins in their sera that neutralize their own venom. Five new small serum proteins (SSPs) were isolated from Japanese mamushi (Gloydius blomhoffii) serum by gel-filtration and RP-HPLC, and their N-Terminal sequences were determined. The amino acid sequences of the precursor proteins were deduced from the nucleotide sequences of cDNAs encoding them. Due to the sequence similarity to those of SSPs in habu snake (Protobothrops flavoviridis) serum (>75% identity), these proteins were designated mSSP-1 to mSSP-5 as the homologs of habu proteins. mSSP-1 was stable at 100 °C and in the pH range of 1-10, and inhibited the proteolytic activity of a certain snake venom metalloproteinase. The inhibitory activity was extinguished by modifying the amino groups of mSSP-1. mSSP-1 is the first prostate secretory protein of the 94 amino acid-family protein with a carbohydrate chain in the Asn37 residue.

  18. Genetic and environmental influences of surfactant protein D serum levels

    DEFF Research Database (Denmark)

    Sørensen, Grith Lykke; Hjelmborg, Jacob v. B.; Kyvik, Kirsten Ohm


    defining the constitutional serum level of SP-D and determine the magnitude of the genetic contribution to serum SP-D in the adult population. Recent studies have demonstrated that serum SP-D concentrations in children are genetically determined and that a single nucleotide polymorphism (SNP) located...... correlation was significantly higher for monozygotic (MZ) twin pairs than for dizygotic (DZ) twin pairs. Serum SP-D variance was influenced by nonshared environmental effects and additive genetic effects. Multivariate analysis of MZ and DZ covariance matrixes showed significant genetic correlation among serum...... SP-D and metabolic variables. The Met11Thr variant explained a significant part of the heritability indicating that serum SP-D variance could be decomposed into non-shared environmental effects (e(2) = 0.19), additive genetic effects (h(2) = 0.42), and the effect of the Met11Thr variations (q(2) = 0.39)....

  19. Rabbit serum amyloid protein A: expression and primary structure deduced from cDNA sequences. (United States)

    Rygg, M; Marhaug, G; Husby, G; Dowton, S B


    Serum amyloid A protein (SAA), the precursor of amyloid protein A (AA) in deposits of secondary amyloidosis, is an acute phase plasma apolipoprotein produced by hepatocytes. The primary structure of SAA demonstrates high interspecies homology. Several isoforms exist in individual species, probably with different amyloidogenic potential. The nucleotide sequences of two different rabbit serum amyloid A cDNA clones have been analysed, one (corresponding to SAA1) 569 base pairs (bp) long and the other (corresponding to SAA2) 513 bp long. Their deduced amino acid sequences differ at five amino acid positions, four of which are located in the NH2-terminal region of the protein. The deduced amino acid sequence of SAA2 corresponds to rabbit protein AA previously described except for one amino acid in position 22. Eighteen hours after turpentine stimulation, rabbit SAA mRNA is abundant in liver, while lower levels are present in spleen. None of the other extrahepatic organs studied showed any SAA mRNA expression. A third mRNA species (1.9 kb) hybridizing with a single-stranded RNA probe transcribed from the rabbit SAA cDNA, was identified. SAA1 and SAA2 mRNA were found in approximately equal amounts in turpentine-stimulated rabbit liver, but seem to be coordinately decreased after repeated inflammatory stimulation.

  20. Serum Protein Profile Study of Clinical Samples Using High Performance Liquid Chromatography-Laser Induced Fluorescence

    DEFF Research Database (Denmark)

    Karemore, Gopal Raghunath; Ukendt, Sujatha; Rai, Lavanya


    The serum protein profiles of normal subjects, patients diagnosed with cervical cancer, and oral cancer were recorded using High Performance Liquid Chromatography combined with Laser Induced Fluorescence detection (HPLC-LIF). Serum protein profiles of the above three classes were tested for estab......The serum protein profiles of normal subjects, patients diagnosed with cervical cancer, and oral cancer were recorded using High Performance Liquid Chromatography combined with Laser Induced Fluorescence detection (HPLC-LIF). Serum protein profiles of the above three classes were tested...... for establishing the ability of HPLC-LIF protein profiling technique for discrimination, using hard clustering and Fuzzy clustering methods. The clustering algorithms have quite successfully classified the profiles as belonging to normal, cancer of cervix, and oral cancer conditions....

  1. Predicting brain metastases of breast cancer based on serum S100B and serum HER2

    DEFF Research Database (Denmark)

    Bechmann, Troels; Madsen, Jonna Skov; Brandslund, Ivan


    Brain metastases are a major cause of morbidity and mortality in breast cancer. The aim of the current study was to evaluate the prediction of brain metastases based on serum S100B and human epidermal growth factor receptor 2 (HER2). A total of 107 breast cancer patients were included in the curr......Brain metastases are a major cause of morbidity and mortality in breast cancer. The aim of the current study was to evaluate the prediction of brain metastases based on serum S100B and human epidermal growth factor receptor 2 (HER2). A total of 107 breast cancer patients were included...... in the current study from two prospective cohort studies with either elevated serum HER2 levels >15 ng/ml or brain metastases verified by magnetic resonance imaging (MRI) or computer tomography (CT). Following the exclusion of six patients, the remaining 101 patients were divided into two groups: Group 0 (n=55......), patients with normal MRI results; and group 1 (n=46), patients with brain metastases. The levels of serum S100B and HER2 in the two groups were analyzed prior to MRI or CT of the brain, and no significant differences were identified in the serum HER2 (P=0.060) or S100B levels (P=0.623) between the groups...

  2. Serum Proteins Stabilized Calcium Phosphate Nanoparticles and Its Effect on Bel-7402 Cells

    Institute of Scientific and Technical Information of China (English)


    Hydroxyapatite has a high affinity to biological macromolecules, especially to proteins. Bovine serum proteins were extracted to be used as stablizer to prepare calcium phosphate nanoparticles. 167.7 am and87.7 nm particles were respectively prepared by using bovine serum protein fractions at the concentration of 0.5mg/mL and 1.0 mg/mL. As the polysaccharide stabilized hydroxyapatite nanoparticles, the protein-stablized nanoparticles also inhibited the proliferation rate of Bel-7402 cells. It suggested that proteins could be applied to prepare calcium phosphate nanoparticles and it also has the anticancer effect.

  3. Blood glucose, serum insulin, serum growth hormone and serum glycosylated proteins during two years' oral contraception with low-estrogen combinations. (United States)

    Liukko, P; Erkkola, R; Lammintausta, R; Grönroos, M; Kloosterboer, H J


    Body weight, fasting blood glucose (GP) (BFG), serum immunoreactive insulin (IRI), serum growth hormone (GH) and serum glycosylated proteins were longitudinally followed in 2 groups of women during two years' oral contraception. One group (n = 10) received a combination of 0.030 mg ethinylestradiol and 0.150 mg levonorgestrel and the other (n = 10) a combination of 0.030 mg ethinylestradiol and 0.150 mg of desogestrel. There was a significant increase in BFG during the study and the values were still rising, when examined 2 months after discontinuation of the pill. Two subjects, reaching the level of 5.5 mmol/l showed normal pretreatment values, when investigated one year later. After 6 months' use of either preparation, GH significantly increased, remained on that level throughout the study and returned to the pretreatment level after discontinuation of the pill. Body weight, IRI and GPP did not change significantly during the study.

  4. Analysis of diferentially expressed protein from primary and recurrent ovarian cancer serum

    Institute of Scientific and Technical Information of China (English)

    Yuan Wang; Jin-Jin Yu; Ting Zhu; Ling Xu; Ming Xu; Yu-Zheng Huang; Hong Pu; Chun-Qing Yu


    ABSTRACT Objective:To study the value of the differentially expressed proteins from primary and recurrent ovarian cancer serum for early diagnosis of primary and recurrent ovarian cancer.Methods:WCX kit(BrukerDaltonicsGraBH) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS) technology were used to detect serum samples from49 patients with primary ovarian cancer and21 patients with recurrent disease.Results:In the mass range(Mr) from1000 to12000Da, eight differentially expressed protein peaks were screened from primary ovarian cancer serum.Among them, four protein peaks withMr1457,1857,2202, 7761 were lowly expressed and the others withMr2946,5333,5859,5901 were highly expressed. Ten diferentially expressed protein peaks were screened from recurrent ovarian cancer serum. Among them,1944,1980,2080,2661,2993,4450,4659,5359Da protein expressions were increased significantly, and1897,7868Da protein expressions were decreased significantly.The pattern of primary ovarian cancer was applied to8 early-stage ovarian cancer serum samples, and7 serum samples were successfully predicted with the accuracy of87.5%.The pattern of recurrent ovarian cancer was applied to9 without pelvic or abdominal mass recurrent ovarian cancer serum samples, and8 serum samples were successfully predicted with the accuracy of 88.9%.Conclusions:Combination ofMALDI-TOF-MS andWCX kit technology can directly screen the diferrential expressed protein from primary and recurrent ovarian cancer serum.They have clinical significance for enhancement of sensitivity and specificity of ovarian cancer diagnosis.


    Directory of Open Access Journals (Sweden)

    Nuzhat Sultana


    Full Text Available This study assessed the effect of oral administration of Aloe vera and was to evaluate total serum protein, albumin and globulin concentrations as well as albumin / globulin (A / G ratio. Twenty rabbits weighing 1000 – 1800 g were divided into 2 groups. Each group consisted of ten animals. One served as control and other group served as experimental group. Results show that animals after 07, 15 and 30 days dosing of Aloe vera showed highly significant decrease in total protein and globulin and highly significant decrease in Albumin after 15 and 30 days of dosing of Aloe vera in comparison to control animals group. It is concluded that the long-term use of Aloe vera may cause hypoglobinemia and hypoalbuminemia at 30 days of dosing and it could be due to the liver diseases, evidence of hepatotoxicity induced Aloe vera also reported in previous studies.

  6. On the transfer of serum proteins to the rat intestinal juice

    DEFF Research Database (Denmark)

    Andersen, Vibeke; Norén, Ove; Poulsen, Mona D;


    The in vivo pattern of serum proteins in the rat small-intestinal juice was characterized by crossed immunoelectrophoresis. Immunoglobulins and albumin, alpha-1-antitrypsin, transferrin, and orosomucoid were present. Larger serum proteins were absent (ceruloplasmin, haptoglobin, alpha-1......-macroglobulin, alpha and beta lipoproteins). Thus, apart from immunoglobulins, only serum proteins with a molecular mass less than approximately 100 kDa were demonstrated. The origin and epithelial transfer were further characterized, using albumin as a model. No sign of local synthesis of albumin...... proteins in the intestinal juice is a selective passage through the capillary wall followed by passive intercellular transport via delivery of the serum in the interstitial space during disintegration of the enterocytes....

  7. Differential expression profiling of serum proteins and metabolites for biomarker discovery (United States)

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


    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.

  8. Serum protein N-glycans profiling for the discovery of potential biomarkers for nonalcoholic steatohepatitis. (United States)

    Chen, Cuiying; Schmilovitz-Weiss, Hemda; Liu, Xue-en; Pappo, Orit; Halpern, Marisa; Sulkes, Jaqueline; Braun, Marius; Cohen, Maya; Barak, Nir; Tur-Kaspa, Ran; Vanhooren, Valerie; Van Vlierberghe, Hans; Libert, Claude; Contreras, Roland; Ben-Ari, Ziv


    The hepatic histology in nonalcoholic fatty liver disease can vary from isolated hepatic steatosis to steatohepatitis can progress to cirrhosis and liver-related death. The aim was to evaluate the use of blood serum N-glycan fingerprinting as a tool for differential diagnosis of nonalcoholic steatohepatitis from steatosis. A group of 47 patients with NAFLD was diagnosed by clinical laboratory analysis and ultrasonography, and was studied histologically using the Brunt's scoring system. The control group included 13 healthy individuals. N-glycan profiles of serum proteins were determined by DNA sequencer-based carbohydrate analytical profiling. We have found that the concentrations of two glycans (NGA2F and NA2) and their logarithm ratio of NGA2F versus NA2 (named GlycoNashTest) were associated with the degree of NASH-related fibrosis, but had no correlation with the grade of inflammation nor steatosis severity. When used to screen NAFLD patients, GlycoNashTest could identify advanced NASH-related fibrosis (F3-F4) with the diagnosis sensitivity of 89.5% and specificity of 71.4%. The serum N-glycan profile is a promising noninvasive method for detecting NASH or NASH-related fibrosis in NAFLD patients, which could be a valuable supplement to other markers currently used in diagnosis of NASH.

  9. Adsorption of human serum proteins onto TREN-agarose: purification of human IgG by negative chromatography. (United States)

    Bresolin, Igor Tadeu Lazzarotto; Borsoi-Ribeiro, Mariana; Caro, Juliana Rodrigues; dos Santos, Francine Petit; de Castro, Marina Polesi; Bueno, Sonia Maria Alves


    Tris(2-aminoethyl)amine (TREN) - a chelating agent used in IMAC - immobilized onto agarose gel was evaluated for the purification of IgG from human serum by negative chromatography. A one-step purification process allowed the recovery of 73.3% of the loaded IgG in the nonretained fractions with purity of 90-95% (based on total protein concentration and nephelometric analysis of albumin, transferrin, and immunoglobulins A, G, and M). The binding capacity was relatively high (66.63 mg of human serum protein/mL). These results suggest that this negative chromatography is a potential technique for purification of IgG from human serum.

  10. Occupational exposure levels of bioaerosol components are associated with serum levels of the acute phase protein Serum Amyloid A in greenhouse workers

    DEFF Research Database (Denmark)

    Madsen, Anne Mette; Thilsing, Trine; Bælum, Jesper;


    to elevated levels of bioaerosols. The objective of this study is to assess whether greenhouse workers personal exposure to bioaerosol components was associated with serum levels of the acute phase proteins Serum Amyloid A (SAA) and C-reactive protein (CRP). METHODS: SAA and CRP levels were determined...

  11. Serum Protein Fingerprint of Patients with Pancreatic Cancer by SELDI Technology

    Institute of Scientific and Technical Information of China (English)

    MA Ning; GE Chun-lin; LUAN Feng-ming; YAO Dian-bo; HU Chao-jun; LI Ning; LIU Yong-feng


    Objective:To study the serum protein fingerprint of patients with pancreatic cancer and to screen for protein molecules closely related to pancreatic cancer during the onset and progression of the disease using surface-enhanced laser desorption and ionization time of fight mass spectrometry(SELDI-TOF-MS).Methods:Serum samples from 20 pancreatic cancers,20 healthy volunteers and 18 patients with other pancreatic diseases.WCX magnetic beans and PBSII-C protein chips reader(Ciphergen Biosystems Ins.)were used.The protein fingerprint expression of all the Serum samples and the resulting profiles between cancer and normal were analyzed with Biomarker Wizard system.Results:A group of proteomic peaks were detected.Four differently expressed potential biomarkers were identified with the relative molecular weights of 5705 Da,4935 Da,5318 Da and 3243 Da.Among them,two proteins with m/z5705,5318Da down-regulated,and two proteins with m/z 4935,3243 Da were up-regulated in pancreatic cancers.Conclusion:SELDI technology can be used to screen significant proteins of differential expression in the serum of pancreatic cancer patients.These different proteins could be specific biomarkers of the patients with pancreatic cancer in the serum and have the potential value of further investigation.

  12. Serum C-reactive protein levels predict neurological outcome after aneurysmal subarachnoid hemorrhage

    Directory of Open Access Journals (Sweden)

    Flávio Ramalho Romero


    Full Text Available OBJECTIVES: Our aim was to evaluate the relationship between serum C-reactive protein (CRP levels and the neurological prognosis and development of vasospasm in patients with aneurysmal subarachnoid hemorrhage (aSAH. METHODS: Eighty-two adult patients with aSAH diagnoses were prospectively evaluated. Glasgow Coma Scale (GCS score, Hunt and Hess grade, Fisher grade, cranial CT scans, digital subtraction angiography studies and daily neurological examinations were recorded. Serial serum CRP measurements were obtained daily between admission and the tenth day. Glasgow Outcome Scale (GOS and the modified Rankin Scale (mRS were used to assess the prognosis. RESULTS: Serum CRP levels were related to severity of aSAH. Patients with lower GCS scores and higher Hunt and Hess and Fisher grades presented statistically significant higher serum CRP levels. Patients with higher serum CRP levels had a less favorable prognosis. CONCLUSIONS: Increased serum CRP levels were strongly associated with worse clinical prognosis in this study.

  13. Serum protein fractionation using supported molecular matrix electrophoresis. (United States)

    Dong, Weijie; Matsuno, Yu-ki; Kameyama, Akihiko


    Supported molecular matrix electrophoresis (SMME), in which a hydrophilic polymer such as PVA serves as a support within a porous PVDF membrane, was recently developed. This method is similar to cellulose acetate membrane electrophoresis but differs in the compatibility to glycan analysis of the separated bands. In this report, we describe the first instance of the application of SMME to human serum fractionation, and demonstrate the differences with serum fractionation by cellulose acetate membrane electrophoresis. The SMME membrane exhibited almost no EOF during electrophoresis, unlike the cellulose acetate membrane, but afforded comparative results for serum fractionation. The visualization of each fraction was achieved by conventional staining with dye such as Direct Blue-71, and objective quantification was obtained by densitometry after inducing membrane transparency with 1-nonene. Immunostaining was also achieved. Moreover, mass spectrometric analysis of both N-linked and O-linked glycans from the separated bands was demonstrated. Serum fractionation and glycan profiling of each fraction using SMME will enable novel insights into the relationships between various glycosylation profiles and disease states.

  14. Differential sensing using proteins: exploiting the cross-reactivity of serum albumin to pattern individual terpenes and terpenes in perfume. (United States)

    Adams, Michelle M; Anslyn, Eric V


    There has been a growing interest in the use of differential sensing for analyte classification. In an effort to mimic the mammalian senses of taste and smell, which utilize protein-based receptors, we have introduced serum albumins as nonselective receptors for recognition of small hydrophobic molecules. Herein, we employ a sensing ensemble consisting of serum albumins, a hydrophobic fluorescent indicator (PRODAN), and a hydrophobic additive (deoxycholate) to detect terpenes. With the aid of linear discriminant analysis, we successfully applied our system to differentiate five terpenes. We then extended our terpene analysis and utilized our sensing ensemble for terpene discrimination within the complex mixtures found in perfume.

  15. Drink fermented to the base of serum of milk and isolated soy protein / Bebida fermentada à base de soro de leite e isolado protéico de soja

    Directory of Open Access Journals (Sweden)

    Ivone Yurika Mizubuti


    Full Text Available The whey and soy protein were used as substrate to obtain a basic beverage, fermented by yogurt culture (Lactobacillus delbrueckii subsp. buigaricus / Streptococcus salivarius subsp. thermophilus YC-180 and by probiotic culture (Lactobacillus acidophilus La-5. The formulation ( whey 10g/100mL; soy protein 2,4 g/100mL; fiber 0,5 g/100mL and sucrose 15 g/100mL was fermented separately, and mixed (1:1 to obtain the final fermented product with 0.72% (lactic acid acidity, pH 4.3 and lactic acid bacteria count of 2,0 x 108 cfu/mL, adequate for production of the proposed beverage.O soro de leite e o isolado protéico de soja foram utilizados para obtenção de uma bebida base, fermentada com cultura de iogurte (Lactobacillus delbrueckii subsp. bulgaricus / Streptococcus salivarius subsp. thermophilus YC - 180 e cultura probiótica (Lactobacillus acidophilus La-5. A formulação (soro de leite 10 g/100mL; isolado protéico de soja 2,4 g/100mL; fibras 0,5 g/100mL e sacarose 15 g/100mL indicou que procedendo-se a fermentação seguida de mistura na proporção de 1:1, produziu-se um produto final com acidez de 0,72% de ácido láctico; pH 4,3 e contagem de bactérias lácticas de 2,0 x 108 ufc/mL adequado para a obtenção da bebida proposta.

  16. Effect of Spray Drying on Protein Content of Natural Rubber Serum (NRS

    Directory of Open Access Journals (Sweden)

    I. Aimi Izyana and M.N. Zairossani


    Full Text Available Natural rubber latex comprises approximately 70% natural rubber serum (NRS which is the aqueous part of the latex. The NRS is made up of mainly water and non-rubber components; sugar, protein and other lipids. The protein component of the NRS is separated using membrane separation and spray dried whereby the amino acid and protein content was monitored during the process. The optimum drying temperatures were first determined to obtain the maximum recovery of the protein powder, whereby the amino acid content was approximately 40 times that of the feed. The production of protein powder from NRS increases the rubber industry's competitiveness through value addition to the previously discarded NRS but also to the processing industry. NRS protein powder has a great opportunity to be developed as an alternative protein source.ABSTRAK: Susu getah asli mengandungi lebih kurang 70% serum getah asli (natural rubber serum (NRS iaitu bahagian susu getah yang berair. NRS sebahagian besarnya merupakan air dan komponen bukan getah; gula, protein dan lipid-lipid lain. Komponen protein NRS dipisahkan dengan menggunakan membran pemisah dan kaedah kering semburan dimana asid amino dan kandungan protein dipantau semasa proses tersebut. Pada mulanya, suhu pengeringan optimum ditentukan untuk mendapatkan hasil maksima serbuk protein, dimana kandungan asid amino adalah lebih kurang 40 kali suapan. Penghasilan serbuk protein daripada NRS meningkatkan daya saing industri getah dan industri pemprosesan menerusi pertambahan nilai terhadap NRS yang sebelum ini hanya dibuang. Serbuk protein NRS mempunyai peluang besar untuk dimajukan sebagai sumber protein alternatif.KEY WORDS:  protein, spray dry, natural rubber serum.

  17. Predictive value of brain-specific proteins in serum for neurodevelopmental outcome after birth asphyxia. (United States)

    Nagdyman, Nicole; Grimmer, Ingrid; Scholz, Tristess; Muller, Christian; Obladen, Michael


    Brain-specific proteins have been used to detect cerebral injury after birth asphyxia. Previous investigations suggest that serum protein S-100beta, brain-specific creatine kinase (CK-BB), and neuron-specific enolase (NSE) are capable of identifying patients with a risk of developing hypoxic-ischemic encephalopathy. Whether detection of elevated serum concentrations of these proteins reflects long-term neurodevelopmental impairment remains to be investigated. We examined serum protein S-100beta, NSE, and CK-BB at 2, 6, 12, and 24 h after birth in 29 asphyxiated infants and 20 control infants. Neurodevelopmental follow-up examinations were performed at 20 mo of age using the German revision of the Griffiths scales for developmental assessment. Elevated concentrations of serum protein S-100beta, NSE, and CK-BB within 24 h after asphyxia did not correlate with long-term neurodevelopmental delay. We conclude that serum protein S-100beta, NSE, and CK-BB, sampled on the first day of life, is of limited value in predicting severe brain damage after birth asphyxia.

  18. Label-free LC-MSe in tissue and serum reveals protein networks underlying differences between benign and malignant serous ovarian tumors.

    Directory of Open Access Journals (Sweden)

    Wouter Wegdam

    Full Text Available PURPOSE: To identify proteins and (molecular/biological pathways associated with differences between benign and malignant epithelial ovarian tumors. EXPERIMENTAL PROCEDURES: Serum of six patients with a serous adenocarcinoma of the ovary was collected before treatment, with a control group consisting of six matched patients with a serous cystadenoma. In addition to the serum, homogeneous regions of cells exhibiting uniform histology were isolated from benign and cancerous tissue by laser microdissection. We subsequently employed label-free liquid chromatography tandem mass spectrometry (LC-MSe to identify proteins in these serum and tissues samples. Analyses of differential expression between samples were performed using Bioconductor packages and in-house scripts in the statistical software package R. Hierarchical clustering and pathway enrichment analyses were performed, as well as network enrichment and interactome analysis using MetaCore. RESULTS: In total, we identified 20 and 71 proteins that were significantly differentially expressed between benign and malignant serum and tissue samples, respectively. The differentially expressed protein sets in serum and tissue largely differed with only 2 proteins in common. MetaCore network analysis, however inferred GCR-alpha and Sp1 as common transcriptional regulators. Interactome analysis highlighted 14-3-3 zeta/delta, 14-3-3 beta/alpha, Alpha-actinin 4, HSP60, and PCBP1 as critical proteins in the tumor proteome signature based on their relative overconnectivity. The data have been deposited to the ProteomeXchange with identifier PXD001084. DISCUSSION: Our analysis identified proteins with both novel and previously known associations to ovarian cancer biology. Despite the small overlap between differentially expressed protein sets in serum and tissue, APOA1 and Serotransferrin were significantly lower expressed in both serum and cancer tissue samples, suggesting a tissue-derived effect in serum

  19. Identification of serum biomarkers for lung cancer using magnetic bead-based SELDI-TOF-MS


    SONG, QI-BIN; Hu, Wei-Guo; Wang, Peng; Yao, Yi; Zeng, Hua-zong


    Aim: To identify novel serum biomarkers for lung cancer diagnosis using magnetic bead-based surface-enhanced laser desorption/ionization time-of-flight mass spectrum (SELDI-TOF-MS). Methods: The protein fractions of 121 serum specimens from 30 lung cancer patients, 30 pulmonary tuberculosis patients and 33 healthy controls were enriched using WCX magnetic beads and subjected to SELDI-TOF-MS. The spectra were analyzed using Bio-marker Wizard version 3.1.0 and Biomarker Patterns Software versio...

  20. A longitudinal study of serum cobalamins and its binding proteins in lactating women

    DEFF Research Database (Denmark)

    Mørkbak, A L; Ramlau-Hansen, C H; Møller, U K;


    were analysed. RESULTS: No significant differences were observed in serum cobalamins or its binding proteins related to supplementation with vitamin B12 or the duration of lactation. Serum cobalamins remained unchanged from 3 weeks to 9 months post-partum. Total TC (holoTC) (median+/-s.e. pmol...... in HC during a 9-month period post-partum. No differences were observed between the vitamin B12-supplemented and the unsupplemented groups. Thus, supplementation with vitamin B12 has no impact on the circulating level of serum cobalamins or its binding proteins in a Danish population of lactating......OBJECTIVE: To examine longitudinal changes in serum cobalamins, transcobalamin (TC) and haptocorrin (HC) during lactation and to investigate the influence of vitamin B12 supplementation on these parameters. DESIGN: A 9-month follow-up study. SUBJECTS AND METHODS: Lactating mothers (N=89) including...

  1. Relationship between serum adiopocyte fatty acid binding protein and atherosclerosis in chronic kidney disease

    Institute of Scientific and Technical Information of China (English)



    Objective To investigate the expression of serum adiopocyte fatty acid binding protein(A-FABP)in chronic kidney disease(CKD)and the role that A-FABP plays in CKD with atherosclerosis.Methods A total of 138 patients with CKD and 20 health control volunteers(HC)were involved in this study.The levels of serum AFABP,free fatty acid(FFA),interleukin-6(IL-6),

  2. Endometrial cancer risk prediction including serum-based biomarkers

    DEFF Research Database (Denmark)

    Fortner, Renée T; Hüsing, Anika; Kühn, Tilman;


    Endometrial cancer risk prediction models including lifestyle, anthropometric, and reproductive factors have limited discrimination. Adding biomarker data to these models may improve predictive capacity; to our knowledge, this has not been investigated for endometrial cancer. Using a nested case......-control study within the European Prospective Investigation into Cancer and Nutrition (EPIC) cohort, we investigated the improvement in discrimination gained by adding serum biomarker concentrations to risk estimates derived from an existing risk prediction model based on epidemiologic factors. Serum...... concentrations of sex steroid hormones, metabolic markers, growth factors, adipokines, and cytokines were evaluated in a step-wise backward selection process; biomarkers were retained at pdiscrimination was assessed using...

  3. Peptide Mimicrying Between SARS Coronavirus Spike Protein and Human Proteins Reacts with SARS Patient Serum

    Directory of Open Access Journals (Sweden)

    K.-Y. Hwa


    Full Text Available Molecular mimicry, defined as similar structures shared by molecules from dissimilar genes or proteins, is a general strategy used by pathogens to infect host cells. Severe acute respiratory syndrome (SARS is a new human respiratory infectious disease caused by SARS coronavirus (SARS-CoV. The spike (S protein of SARS-CoV plays an important role in the virus entry into a cell. In this study, eleven synthetic peptides from the S protein were selected based on its sequence homology with human proteins. Two of the peptides D07 (residues 927–937 and D08 (residues 942–951 were recognized by the sera of SARS patients. Murine hyperimmune sera against these peptides bound to proteins of human lung epithelial cells A549. Another peptide D10 (residues 490–502 stimulated A549 to proliferate and secrete IL-8. The present results suggest that the selected S protein regions, which share sequence homology with human proteins, may play important roles in SARS-CoV infection.

  4. Variation of Serum Protein Level in Xiang Piglets at Different Month of Age

    Institute of Scientific and Technical Information of China (English)

    ChiYujie; GaoXuejung; 等


    Usual clinieal methods were used to determine the concentrations of serum protein,albumin and globulin in Xiang piglets from the lst month to the 6th month of age.Thirteen piglets(eight males and five females)born is Spring were selected from the Livestock Experimental Station of NEAU.The results showed that the serum protein level was different at different month of age.The lowest level was found at the 1st month of age,and the highest at the 4th month of age.The difference in serum protein concentration was significant between different months of age.There was ,however,no significant difference between sexes at the same month of age.

  5. Abnormal serum thyroid hormones concentration with healthy functional gland: a review on the metabolic role of thyroid hormones transporter proteins. (United States)

    Azad, Reza Mansourian


    Laboratory findings can definitely help the patients not to enter into status, where the damage might be happen due to a miss-diagnosis based on clinical assessment alone. The secondary disease accompanied with thyroid patients should also carefully check out due to the interference which some diseases can cause in the amount of serum thyroid hormone, particularly the free thyroxin. The dilemma over thyroid clinical diagnosis occur due to variation on serum thyroid hormone which initiated by other non-thyroidal disorders which can play an important roles in metabolic disorders of thyroid hormone due to the alteration which occur on the serum level of thyroid hormone transporter proteins. The majority of serum thyroid hormones of up to 95-99% are bound to the carrier proteins mainly to Thyroxin-Binding Globulins (TBG), some transthyretin already known as pre-albumin and albumin which are all synthesis in the liver and any modification which alter their production may alter the status of thyroid hormones. It seems TBG, transthyretin and albumin carries 75, 20, 5% of thyroid hormones within blood circulation, respectively. The dilemma facing the thyroid hormones following disruption of thyroid hormone transporter protein synthesis originate from this fact that any alteration of these protein contribute to the alteration of total thyroid and free serum thyroid hormones which are in fact the biologically active form of thyroid hormones. The subsequent of latter implication result in miss-understanding and miss-diagnosis of thyroid function tests, with possible wrongly thyroid clinical care, followed by undesired therapy of otherwise healthy thyroid.

  6. High-protein diets in hyperlipidemia : effect of wheat gluten on serum lipids, uric acid, and renal function

    NARCIS (Netherlands)

    Jenkins, D.J.A.; Kendall, C.W.C.; Vidgen, E.; Augustin, L.S.A.; Erk, van M.; Geelen, A.; Parker, T.; Faulkner, D.; Vuksan, V.; Josse, R.G.; Leiter, L.A.; Connelly, P.W.


    BACKGROUND: The metabolic effects of diets high in vegetable protein have not been assessed despite much recent interest in the effect of soy proteins in reducing serum cholesterol. OBJECTIVE: We assessed the metabolic effects of diets high in vegetable protein (specifically, wheat gluten) on serum

  7. Serum proteins behavior in the urban population from a center with a lead-polluted atmospheric environment

    Energy Technology Data Exchange (ETDEWEB)

    Casin, I.; Ghelberg, N.W.


    Serum protein behavior was determined in an urban population from a center with a lead polluted atmospheric environment. The following increased amounts of serum proteins are found in 94 adult subjects: haptoglobin 75 percent; ceruloplasmin and transferrin 55 percent; IgA (immunoglobulin A) and IgG 40 percent; Igm and beta lc proteins 25 percent.

  8. Conjugation of biogenic and synthetic polyamines with serum proteins: A comprehensive review. (United States)

    Chanphai, P; Thomas, T J; Tajmir-Riahi, H A


    We have reviewed the conjugation of biogenic polyamines spermine (spm), spermidine (spmd) and synthetic polyamines 3,7,11,15-tetrazaheptadecane.4HCl (BE-333) and 3,7,11,15,19-pentazahenicosane.5HCl (BE-3333) with human serum albumin (HSA), bovine serum albumin (BSA) and milk beta-lactoglobulin (b-LG) in aqueous solution at physiological pH. The results of multiple spectroscopic methods and molecular modeling were analysed here and correlations between polyamine binding mode and protein structural changes were estabilished. Polyamine-protein bindings are mainly via hydrophilic and H-bonding contacts. BSA forms more stable conjugates than HSA and b-LG. Biogenic polyamines form more stable complexes than synthetic polyamines except in the case of b-LG, where the protein shows more hydrophobic character than HSA and BSA. The loading efficacies were 40-52%. Modeling showed the presence of several H-bonding systems, which stabilized polyamine-protein conjugates. Polyamine conjugation induced major alterations of serum protein conformations. The potential application of serum proteins in delivery of polyamines is evaluated here.

  9. Serum Profiling Using Protein Microarrays to Identify Disease Related Antigens



    Disease related antigens are of great importance in the clinic. They are used as markers to screen patients for various forms of cancer, to monitor response to therapy, or to serve as therapeutic targets (Chapman et al., Ann Oncol 18(5):868–873, 2007; Soussi et al., Cancer Res 60:1777–1788, 2000; Anderson and LaBaer, J Proteome Res 4:1123–1133, 2005; Levenson, Biochim Biophy Acta 1770:847–856, 2007). In cancer endogenous levels of protein expression may be disrupted or proteins may be express...

  10. The serum protein carbonyl content level in relation to exercise stress test

    Directory of Open Access Journals (Sweden)

    Titiporn Mekrungruangwong


    Full Text Available Background: Protein carbonyl (P is oxidatively-modified protein with diagnostic potential for acute myocardial infarction. However, many findings indicated the elevation of serum PC content level related to exercise, which could cause false positive results and limiting the specificity for acute coronary syndrome diagnosis. This study aims to evaluate the level of serum protein carbonyl content in healthy volunteers subjected to exercise stress test (EST. Materials and Methods: Serum from healthy volunteers was collected 5-10 min before performing EST and 1 hour after the EST was achieved. The serum was collected, and the serum PC content level was determined by spectrophotometric DNPH assay. Results: The serum PC content level after exercise stress test was significantly higher than that of before performing EST (0.373 ± 0.05 nM/mg vs. 0.275 ± 0.02 nM/mg, P < 0.0001. The results demonstrated that in both male and female, serum PC content level after EST was significantly higher than that of before performing EST (0.29 ± 0.03 nM/mg vs. 0.36 ± 0.05 nM/mg P < 0.0001 in male, 0.27 ± 0.02 nM/mg vs. 0.38 ± 0.06 nM/mg P < 0.0001 in female, respectively. Conclusions: This study demonstrated that exercise stress test could result in non-specificity and false positive increasing in serum PC content level in healthy subjects, which may cause misinterpretation when using PC as cardiac marker, especially in patients, who underwent exercise stress test or patients who performing heavy physical activities.

  11. Reproducibility of serum protein profiling by systematic assessment using solid-phase extraction and matrix-assisted laser desorption/ionization mass spectrometry

    DEFF Research Database (Denmark)

    Callesen, Anne K; Christensen, René Depont; Madsen, Jonna S


    for serum protein profiling we investigated a range of sample preparation techniques and developed a statistical method based on repeated analyses for evaluation of protein-profiling performance of MALDI MS. Two different solid-phase extraction (SPE) methods were investigated, namely custom......Protein profiling of human serum by matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) is potentially a new diagnostic tool for early detection of human diseases, including cancer. Sample preparation is a key issue in MALDI MS and the analysis of complex samples such as serum......-made microcolumns and commercially available magnetic beads. Using these two methods, nineteen different sample preparation methods for serum profiling by MALDI MS were systematically tested with regard to matrix selection, stationary phase, selectivity, and reproducibility. Microcolumns were tested with regard...

  12. Serum Advanced Oxidation Protein Products in Oral Squamous Cell Carcinoma: Possible Markers of Diagnostic Significance

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    Abhishek Singh Nayyar


    Full Text Available Background: The aim of this study was to measure the concentrations (levels ofserum total proteins and advanced oxidation protein products as markers of oxidantmediated protein damage in the sera of patients with oral cancers.Methods: The study consisted of the sera analyses of serum total protein andadvanced oxidation protein products’ levels in 30 age and sex matched controls, 60patients with reported pre-cancerous lesions and/or conditions and 60 patients withhistologically proven oral squamous cell carcinoma. One way analyses of variance wereused to test the difference between groups. To determine which of the two groups’ meanswere significantly different, the post-hoc test of Bonferroni was used. The results wereaveraged as mean ± standard deviation. In the above test, P values less than 0.05 weretaken to be statistically significant. The normality of data was checked before thestatistical analysis was performed.Results: The study revealed statistically significant variations in serum levels ofadvanced oxidation protein products (P<0.001. Serum levels of total protein showedextensive variations; therefore the results were largely inconclusive and statisticallyinsignificant.Conclusion: The results emphasize the need for more studies with larger samplesizes to be conducted before a conclusive role can be determined for sera levels of totalprotein and advanced oxidation protein products as markers both for diagnosticsignificance and the transition from the various oral pre-cancerous lesions and conditionsinto frank oral cancers.

  13. Serum immune-related proteins are differentially expressed during hibernation in the American black bear.

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    Brian A Chow

    Full Text Available 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 α2-macroglobulin, complement components C1s and C4, immunoglobulin μ and J chains, clusterin, haptoglobin, C4b binding protein, kininogen 1, α2-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.

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

    Institute of Scientific and Technical Information of China (English)


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

  15. Mass spectrometry-based serum proteome pattern analysis in molecular diagnostics of early stage breast cancer

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    Stobiecki Maciej


    .0003 increased level of osteopontin in blood of the group of cancer patients studied (however, the plasma level of osteopontin classified cancer samples with 88% sensitivity but only 28% specificity. Conclusion MALDI-ToF spectrometry of serum has an obvious potential to differentiate samples between early breast cancer patients and healthy controls. Importantly, a classifier built on MS-based serum proteome patterns outperforms available protein biomarkers analyzed in blood by immunoassays.

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

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    Rani Surekha


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

  17. Accelerated evolution of small serum proteins (SSPs)-The PSP94 family proteins in a Japanese viper. (United States)

    Aoki, Narumi; Matsuo, Hisashi; Deshimaru, Masanobu; Terada, Shigeyuki


    Five small serum proteins (SSPs) with molecular masses of 6.5-10 kDa were detected in Habu (Trimeresurus flavoviridis) serum; this included two novel proteins SSP-4 and SSP-5. The amino acid sequences of these proteins and of SSP-1, SSP-2, and SSP-3, which were reported previously, were determined on the basis of the nucleotide sequences of their cDNAs. Although these proteins exhibited only limited sequence identity to mammalian prostatic secretory protein of 94 amino acids (PSP94), the topological pattern of disulfide bonds in SSPs was identical to that of the mammalian proteins. SSP-3 and SSP-4 lacked approximately 30 residues at the C-terminal. Each of the full-length cDNAs encoded a mature protein of 62-90 residues and a highly conserved signal peptide. The evolutionary distances between SSPs estimated on the basis of the amino acid changes were significantly greater than those of the synonymous nucleotide substitutions; these finding, together with results from analyses of nonsynonymous to synonymous rates of change (dN/dS) suggest that snake SSPs have endured substantial accelerated adaptive protein evolution. Such accelerated positive selection in SSPs parallels other findings of similar molecular evolution in snake venom proteins and suggests that diversifying selection on both systems may be linked, and that snake SSP genes may have evolved by gene duplication and rapid diversification to facilitate the acquisition of various functions to block venom activity within venomous snakes.

  18. Identification of novel serum proteins in a Japanese viper: homologs of mammalian PSP94. (United States)

    Aoki, Narumi; Sakiyama, Akie; Deshimaru, Masanobu; Terada, Shigeyuki


    Three small serum proteins (SSP-1, -2, and -3), with molecular masses of 6.5-10kDa, were isolated from Habu (Trimeresurus flavoviridis) serum, and the amino acid sequences were determined by protein and cDNA analysis. Despite only limited sequence identity to any mammalian prostatic secretory protein of 94 amino acids (PSP94), all of the Cys residues in these SSPs were well conserved. SSPs are the first PSP94 family proteins to be identified in reptiles. SSP-1 and -3 weakly inhibited the proteolytic activity of a snake venom metalloproteinase. On the other hand, SSP-2 formed a tight complex with triflin, a snake venom-derived Ca(2+) channel blocker that suppresses the smooth muscle contraction. This suggests a role for SSP-2 in the self defense system of venomous snakes.

  19. Recombinant phosphoprotein based single serum dilution ELISA for rapid serological detection of Newcastle disease virus. (United States)

    Das, Moushumee; Kumar, Sachin


    Newcastle disease virus (NDV) is the causative agent of a highly contagious disease in avian species. All strains of NDV belong to avian paramyxovirus serotype-1. The disease is endemic in different parts of the world and vaccination is the only way to protect birds from NDV infection. The virus non-structural phosphoprotein (P) is the second most abundant protein and a major modulator of viral replication. Although P protein shows lesser evolutionary divergence among NDV isolates, it is known to be highly divergent among different avian paramyxovirus serotypes. In the present study, a recombinant P protein based single serum dilution ELISA was developed which showed better sensitivity, specificity and accuracy as compared to conventional methods for NDV detection. The recombinant P protein based ELISA could be an alternative to existing diagnostics against NDV infection in chickens.

  20. The role of serum β trace protein in the elderly diabetic patients with renal injury

    Institute of Scientific and Technical Information of China (English)



    Objective To study the role of serum β trace protein(βTP) in the evaluation of renal injury in elderly T2DM.Methods The 131 old patients with T2DM were classified into three groups: normal albuminuria group

  1. Protein and fat mobilization and associations with serum beta-hydroxybutyrate concentrations in dairy cows

    NARCIS (Netherlands)

    van der Drift, S. G. A.; Houweling, M.; Schonewille, J. T.; Tielens, A. G. M.; Jorritsma, R.


    The objective of this study was to obtain information on variation between dairy cows in muscle and fat tissue mobilization around parturition and to study the association between protein and fat mobilization and serum beta-hydroxybutyrate (BHBA) concentrations (hyperketonemia) in this period. Thirt

  2. Protein standardization IV: Value transfer procedure for the assignment of serum protein values from a reference preparation to a target material. (United States)

    Blirup-Jensen, S; Johnson, A M; Larsen, M


    A new approach for the assignment of values to serum proteins in a target material using a reference preparation has been developed. The procedure describes the general as well as the practical principles involved in the value assignment (with examples). Two models have been developed: 1) The direct value transfer between serum matrices and 2) the indirect value transfer from a pure protein preparation to a serum protein material. The necessary mathematical equations are developed and explained. The data reduction and statistical evaluation are discussed. The practical procedure (the transfer protocol) is based on six dilutions of the reference preparation assayed together with six dilutions of the target material. In this way imprecision is reduced and the proportionality of the two materials (i.e. the presence or absence of matrix effects) can be assessed directly by evaluating a single regression plot. If no matrix effects are found, the regression line will pass through zero with a slope equal to the ratio of the concentrations of the two materials. The transfer protocol is based on a multiple point value assignment obtained by several measurements a day repeated on several days, an important prerequisite being that all reconstitutions and dilutions are controlled by weighing.

  3. A novel protein isolated from the serum of rabbitfish (Siganus oramin) is lethal to Cryptocaryon irritans. (United States)

    Wang, Fang-Hua; Xie, Ming-Quan; Li, An-Xing


    The susceptibility of eight marine fish species cultured in South China were tested for infection by the parasitic ciliate, Cryptocaryon irritans, via a challenge examination and an immobilization assay. All species of fish (representing six different families) that we investigated were infected by C. irritans except the rabbitfish (Siganus oramin), which displayed resistance to C. irritans infection. The infection intensity of rabbitfish (0.92+/-0.97, prabbitfish serum were significantly higher (44.51+/-22.98, prabbitfish presented a strong killing effect to C. irritans in vitro. Light microscopy, scanning electron microscopy and fluorescence microscopy confirmed that rabbitfish serum could induce the theront cilia fall off, rupture of the cell membrane because of the swell and rupture of the macronucleus. Rabbitfish serum could also induce the rupture of the trophont membrane and content efflux. Herein a novel antiparasitic protein (APP) was isolated and purified from the serum of rabbitfish (S. oramin) by using a series of salting-out, cation exchange chromatography and two step of reversed phase high performance liquid chromatography (RP-HPLC). Analysis of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that APP was a homogenous polymeric protein with an N-terminal amino acid sequence of SSVEKNLAACLRDND. Its monomeric molecular mass, determined by matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometer (MALDI-TOF-TOF-MS), was found to be 61,739.87 Da. Results of homology analyses indicated that this protein was a newly discovered functional protein in the rabbitfish serum. Laser confocal fluorescence microscopy conformed that the action site of the APP was mainly on the cell membrane and nucleus of theront, which agreed with the results of light microscopy, fluorescence microscopy and scanning electron microscopy. These findings suggest that this protein may contribute considerably to the innate host

  4. A Multicenter Trial Defining a Serum Protein Signature Associated with Pancreatic Ductal Adenocarcinoma

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    Anna S. Gerdtsson


    Full Text Available Background. Pancreatic ductal adenocarcinoma (PDAC is an aggressive disease with rapid tumor progression and poor prognosis. This study was motivated by the lack of sensitive and specific PDAC biomarkers and aimed to identify a diagnostic, serum protein signature for PDAC. Methods. To mimic a real life test situation, a multicenter trial comprising a serum sample cohort, including 338 patients with either PDAC or other pancreatic diseases (OPD and controls with nonpancreatic conditions (NPC, was analyzed on 293-plex recombinant antibody microarrays targeting immunoregulatory and cancer-associated antigens. Results. Serum samples collected from different hospitals were analyzed and showed that (i sampling from five different hospitals could not be identified as a preanalytical variable and (ii a multiplexed biomarker signature could be identified, utilizing up to 10 serum markers that could discriminate PDAC from controls, with sensitivities and specificities in the 91–100% range. The first protein profiles associated with the location of the primary tumor in the pancreas could also be identified. Conclusions. The results demonstrate that robust enough serum signatures could be identified in a multicenter trial, potentially contributing to the development of a multiplexed biomarker immunoassay for improved PDAC diagnosis.


    Directory of Open Access Journals (Sweden)

    V. G. Levchenko


    Full Text Available Serum levels of myelin basic protein (MBP-bound immune complexes were studied in blood sera from women with gestosis, as compared with those in normal pregnancy and non-pregnant woman. The amounts of IgG-MBP complex in blood serum were determined by enzyme immunoassay using isolated anti-МBP-antibodies. The study has shown that about 0.05 mcg of IgG ml of blood serum are associated with myelin basic protein in unpregnant women or in normal pregnancy. Mild gestosis is accompanied by a 2-3-fold increase in MBP immunocomplex concentrations in blood serum. More severe stages of gestosis are characterized by its further rise, thus achieving maximal values of such MBP immune complexes (0.8 mcg/ml in patients with pre-eclampsia and eclampsia. Their amounts were reduced twice after the periods of eclampsia. Serum levels of MBP-bound IgGs may be used to determine severity of gestosis and to predict a risk of eclampsia in pregnant women.

  6. Seasonal trends in nesting leatherback turtle (Dermochelys coriacea) serum proteins further verify capital breeding hypothesis. (United States)

    Perrault, Justin R; Wyneken, Jeanette; Page-Karjian, Annie; Merrill, Anita; Miller, Debra L


    Serum protein concentrations provide insight into the nutritional and immune status of organisms. It has been suggested that some marine turtles are capital breeders that fast during the nesting season. In this study, we documented serum proteins in neophyte and remigrant nesting leatherback sea turtles (Dermochelys coriacea). This allowed us to establish trends across the nesting season to determine whether these physiological parameters indicate if leatherbacks forage or fast while on nesting grounds. Using the biuret method and agarose gel electrophoresis, total serum protein (median = 5.0 g/dl) and protein fractions were quantified and include pre-albumin (median = 0.0 g/dl), albumin (median = 1.81 g/dl), α1-globulin (median = 0.90 g/dl), α2-globulin (median = 0.74 g/dl), total α-globulin (median = 1.64 g/dl), β-globulin (median = 0.56 g/dl), γ-globulin (median = 0.81 g/dl) and total globulin (median = 3.12 g/dl). The albumin:globulin ratio (median = 0.59) was also calculated. Confidence intervals (90%) were used to establish reference intervals. Total protein, albumin and total globulin concentrations declined in successive nesting events. Protein fractions declined at less significant rates or remained relatively constant during the nesting season. Here, we show that leatherbacks are most likely fasting during the nesting season. A minimal threshold of total serum protein concentrations of around 3.5-4.5 g/dl may physiologically signal the end of the season's nesting for individual leatherbacks. The results presented here lend further insight into the interaction between reproduction, fasting and energy reserves and will potentially improve the conservation and management of this imperiled species.

  7. Comprehensive characterisation of pulmonary and serum surfactant protein D in COPD

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    Erpenbeck Veit J


    Full Text Available Abstract Background Pulmonary surfactant protein D (SP-D is considered as a candidate biomarker for the functional integrity of the lung and for disease progression, which can be detected in serum. The origin of SP-D in serum and how serum concentrations are related to pulmonary concentrations under inflammatory conditions is still unclear. Methods In a cross-sectional study comprising non-smokers (n = 10, young - (n = 10, elderly smokers (n = 20, and smokers with COPD (n = 20 we simultaneously analysed pulmonary and serum SP-D levels with regard to pulmonary function, exercise, repeatability and its quaternary structure by native gel electrophoresis. Statistical comparisons were conducted by ANOVA and post-hoc testing for multiple comparisons; repeatability was assessed by Bland-Altman analysis. Results In COPD, median (IQR pulmonary SP-D levels were lower (129(68 ng/ml compared to smokers (young: 299(190, elderly: 296(158 ng/ml; p Conclusions Pulmonary and serum SP-D levels are stable markers influenced by smoking and related to airflow obstruction and disease state. Smaller subunits of pulmonary SP-D and the rapid increase of serum SP-D levels in COPD due to exercise support the translocation hypothesis and its use as a COPD biomarker. Trial registration no interventional trial

  8. Serum Growth Arrest Specific Protein 6 (Gas-6 Levels in Patients with Schizophrenia

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    Fethullah Gerin


    Full Text Available Objective: We have investigated serum growth ar­rest-specific protein 6 (GAS-6 levels from patients with schizophrenia divided into acute phase remission phases as well as control group. Methods: This study was conducted in Psychiatry De­partment of Istanbul University Cerrahpasa Medical Fac­ulty. The patients who were diagnosed with schizophrenia after regular psychiatric examination according to DSM-IV criteria (n=22 as well as control subjects were includ­ed in the study. Schizophrenia patients with acute phase and remission phase were evaluated by Positive and Negative Syndrome Scale (PANSS and Clinical global Impression Scale (CGI-S. The serum GAS-6 levels of schizophrenia patients during acute phase and remission phase were compared with the serum GAS-6 levels of healthy controls. Serum GAS-6 levels were measured by commercial ELISA kits. Results: No difference was found in serum GAS-6 levels among the three groups; schizophrenia with acute phase, schizophrenia with remission phase, and controls. There were no correlations between serum GAS-6 levels and PANSS and CGI scores. Conclusion: To reach a definitive data and better in­terpretation about the relationship between GAS-6 and schizophrenia, future studies with larger groups of pa­tients with schizophrenia subdivided by drug naïve and treated with antipsychotics/other treatment modalities and controls are needed.

  9. Determination of ring-necked pheasant (Phasianus colchicus) serum protein concentrations by refractometry and the biuret method



    The purpose of this study was to evaluate the accuracy of hand-held refractometer in determining serum protein concentrations in ring-necked pheasants (Phasianus colchicus) as compared with the standard biuret method. The results indicated that serum protein values may be accurately determined in ring-necked pheasants with a hand-held refractometer. © Asian Network for Scientific Information, 2008.

  10. Interactions between DMPC liposomes and the serum blood proteins HSA and IgG. (United States)

    Sabín, Juan; Prieto, Gerardo; Ruso, Juan M; Messina, Paula V; Salgado, Francisco J; Nogueira, Montserrat; Costas, Miguel; Sarmiento, Félix


    The interaction between two serum blood proteins, namely human serum albumin (HSA) and human immunoglobulin G (IgG), with 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) liposomes has been studied in detail using dynamic light scattering, flow cytometry, enzyme-linked immunosorbent assay (ELISA), electrophoretic mobility, differential scanning calorimetry (DSC), and surface tension measurements. HSA and IgG interact with liposomes forming molecular aggregates that remain stable at protein concentrations beyond those of total liposome coverage. Both HSA and IgG penetrate into the liposome bilayer. An ELISA assay indicates that the Fc region of IgG is the one that is immersed in the DMPC membrane. The liposome-protein interaction is mainly of electrostatic nature, but an important hydrophobic contribution is also present.

  11. Immune Reactivity of Brucella Melitensis–Vaccinated Rabbit Serum with Recombinant Omp31 and Dnak Proteins

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    Mahmood Jeddi-Tehrani


    Full Text Available Background and objectives: Brucella melitensis infection is still a major health problem for human and cattle in developing countries and the Middle East.Materials and Methods: In this study, in order to screen immunogenic candidate antigens for the development of a Brucella subunit vaccine, a cytoplasmic protein (DnaK and an outer membrane protein (Omp31 of B. melitensis were cloned, expressed in E.coli BL21 and then purified using Ni-NTA agarose. Immunized serum was prepared from a rabbit inoculated with attenuated B. melitensis.Results and Conclusion: It was proved that immunized serum contains antibodies against recombinant Omp31 (rOmp31 and DnaK (rDnaK by Western blot and ELISA assays. The results may suggest the importance of these proteins as subunit vaccines against B. melitensis as well as targets for immunotherapy.

  12. Longitudinal determination of serum placental protein 13 during development of preeclampsia


    Huppertz, Berthold; Sammar, Marei; Chefetz, Ilana; Neumaier-Wagner, Peruka; Bartz, Clemens; Meiri, Hamutal


    Objective: To determine maternal serum placental protein 13 (PP13) in normal pregnancy and preeclampsia. Methods: A prospective, longitudinal study with 41 normal pregnant women, 18 cases with preterm delivery or cervix insufficiency and 4 with developing late-onset preeclampsia. Six hundred and sixty-six maternal blood samples were obtained every 2-4 weeks starting at 5-8 weeks gestation (10-12 samples/patient) and tested for serum PP13 by ELISA. Results: In normal pregnant women delivering ...

  13. Serum levels of C-reactive protein in adolescents with periodontitis

    DEFF Research Database (Denmark)

    López, Rodrigo; Baelum, Vibeke; Hedegaard, Chris Juul;


    Background: The results of several cross-sectional studies suggested a relationship between periodontitis and higher serum levels of C-reactive protein (CRP). Most of these studies were restricted to adult study groups with severe periodontal inflammation, and the potential effects of confounding...... ng/ml (31 to 183 ng/ml), respectively (P = 0.8). Conclusions: Serum levels of CRP were not significantly higher among subjects with periodontitis than among controls. However, a statistically significant positive association between percentages of sites with bleeding on probing and log...

  14. Competitive Protein Adsorption of Albumin and Immunoglobulin G from Human Serum onto Polymer Surfaces

    DEFF Research Database (Denmark)

    Holmberg, Maria; Hou, Xiaolin


    Competitive protein adsorption from human serum onto unmodified polyethylene terephthalate (PET) surfaces and plasma-polymerized PET surfaces, using the monomer diethylene glycol vinyl ether (DEGVE), has been investigated using radioactive labeling. Albumin and immunoglobulin G (IgG) labeled...... with two different iodine isotopes have been added to human serum solutions of different concentrations, and adsorption has been performed using adsorption times from approximately 5 s to 24 h. DEGVE surfaces showed indications of being nonfouling regarding albumin and IgG adsorption during competitive...

  15. Outer membrane protein A, peptidoglycan-associated lipoprotein, and murein lipoprotein are released by Escherichia coli bacteria into serum. (United States)

    Hellman, J; Loiselle, P M; Tehan, M M; Allaire, J E; Boyle, L A; Kurnick, J T; Andrews, D M; Sik Kim, K; Warren, H S


    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 OMP was identified as outer membrane protein A (OmpA) by immunoblotting studies using OmpA-deficient bacteria and recombinant OmpA protein. The 18-kDa OMP was identified as peptidoglycan-associated lipoprotein (PAL) based on peptide sequences from the purified protein and immunoblotting studies using PAL-deficient bacteria. The 5- to 9-kDa OMP was identified as murein lipoprotein (MLP) based on immunoblotting studies using MLP-deficient bacteria. The studies identify the OMPs released into human serum and into the circulation in an experimental model of sepsis as OmpA, PAL, and MLP.

  16. Calorimetric and spectroscopic properties of small globular proteins (bovine serum albumin, hemoglobin) after free radical generation

    Energy Technology Data Exchange (ETDEWEB)

    Farkas, N.; Belagyi, J.; Lorinczy, D


    Mild oxidation of -SH-containing proteins (serum albumin, hemoglobin) by Ce(IV)-ions in the presence of the spin trap phenyl-tert-butylnitrone (PBN) resulted in the appearance of strongly immobilized nitroxide free radicals which evidences the formation of thiyl radicals on the thiol site of the proteins. In hydroxyl free radical generating system a fraction of strongly immobilized nitroxide radicals was also detected in these proteins, which implies that the oxidation of a fraction of the thiol groups was also involved in the free radical reaction. According to the differential scanning calorimetry (DSC) experiments the melting processes of the proteins were calorimetrically irreversible, therefore the two-state kinetic model was used to evaluate the experiments. The results support the view that site-specific interaction of SH-containing proteins with hydroxyl and thiyl free radicals is able to modify the internal dynamics of proteins and affect the conformation of large molecules.

  17. Cellular Binding of Anionic Nanoparticles is Inhibited by Serum Proteins Independent of Nanoparticle Composition. (United States)

    Fleischer, Candace C; Kumar, Umesh; Payne, Christine K


    Nanoparticles used in biological applications encounter a complex mixture of extracellular proteins. Adsorption of these proteins on the nanoparticle surface results in the formation of a "protein corona," which can dominate the interaction of the nanoparticle with the cellular environment. The goal of this research was to determine how nanoparticle composition and surface modification affect the cellular binding of protein-nanoparticle complexes. We examined the cellular binding of a collection of commonly used anionic nanoparticles: quantum dots, colloidal gold nanoparticles, and low-density lipoprotein particles, in the presence and absence of extracellular proteins. These experiments have the advantage of comparing different nanoparticles under identical conditions. Using a combination of fluorescence and dark field microscopy, flow cytometry, and spectroscopy, we find that cellular binding of these anionic nanoparticles is inhibited by serum proteins independent of nanoparticle composition or surface modification. We expect these results will aid in the design of nanoparticles for in vivo applications.

  18. Effects of repeated psychological stress training on the spectrum of serum protein expression in special troops

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    Li ZHANG


    Full Text Available Objective To investigate the effects of repeated psychological stress training on the serum protein expression in soldiers under mental stress.Methods Ninety-six male commando soldiers were randomly assigned into the common psychological training group,the circulation psychological training group and the control group(32 each.After a 4-week training,all the soldiers were instructed to attend an one-day high-intensity simulated anti-riot exercise,and 3 days later attended another unannounced high-intensity simulated anti-riot exercise.Blood samples were collected from all the soldiers within 4 hours after each exercise,and the changes in serum protein expression were determined and statistically analyzed by using surface-enhanced laser desorption/ionization time of flight mass spectrometry(SELDI-TOF-MS combined with ProteinChip technology.Results The variance analysis showed that significant differences existed among the three groups(P < 0.05 in the relative contents of proteins with M/Z values of 6417.8,9134.2,15171.9 and 14972.7D after the first anti-riot exercise,and the relative contents of all the above mentioned proteins increased in the circulatory psychological training group;meanwhile,markedly increasing trends of the relative contents of all the proteins were observed in the three groups after the second anti-riot exercise(P < 0.05,and in control group the relative contents of the 4 above mentioned proteins were significantly higher than those after the first anti-riot exercise.Conclusion Psychological training may up-regulate the expression of serum proteins that are down-regulated after stress,and the repeated high-intensity mental training can rapidly improve the soldiers’ ability to counteract stress.

  19. Carvacrol attenuates serum levels of total protein, phospholipase A2 and histamine in asthmatic guinea pig

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    Mohammad Hossein Boskabady


    Full Text Available Objective: Pharmacological effects of carvacrol such as its anti-inflammatory activities have been shows. In this study the effects of carvacrol on serum levels of total protein (TP, phospholipase A2 (PLA2 and histamine in sensitized guinea pigs was evaluated. Materials and Methods: Sensitized guinea pigs were given drinking water alone (group S, drinking water containing three concentrations of carvacrol (40, 80 and 160 µg/ml or dexamethasone. Serum levels of TP, PLA2 and histamine were examined I all sensitized groups as well as a non-sensitized control group (n=6 for each group. Results: In sensitized animals, serum levels of TP, PLA2 and histamine were significantly increased compared to control animals (p

  20. Serum protein profiles predict coronary artery disease in symptomatic patients referred for coronary angiography

    Directory of Open Access Journals (Sweden)

    LaFramboise William A


    Full Text Available Abstract Background More than a million diagnostic cardiac catheterizations are performed annually in the US for evaluation of coronary artery anatomy and the presence of atherosclerosis. Nearly half of these patients have no significant coronary lesions or do not require mechanical or surgical revascularization. Consequently, the ability to rule out clinically significant coronary artery disease (CAD using low cost, low risk tests of serum biomarkers in even a small percentage of patients with normal coronary arteries could be highly beneficial. Methods Serum from 359 symptomatic subjects referred for catheterization was interrogated for proteins involved in atherogenesis, atherosclerosis, and plaque vulnerability. Coronary angiography classified 150 patients without flow-limiting CAD who did not require percutaneous intervention (PCI while 209 required coronary revascularization (stents, angioplasty, or coronary artery bypass graft surgery. Continuous variables were compared across the two patient groups for each analyte including calculation of false discovery rate (FDR ≤ 1% and Q value (P value for statistical significance adjusted to ≤ 0.01. Results Significant differences were detected in circulating proteins from patients requiring revascularization including increased apolipoprotein B100 (APO-B100, C-reactive protein (CRP, fibrinogen, vascular cell adhesion molecule 1 (VCAM-1, myeloperoxidase (MPO, resistin, osteopontin, interleukin (IL-1β, IL-6, IL-10 and N-terminal fragment protein precursor brain natriuretic peptide (NT-pBNP and decreased apolipoprotein A1 (APO-A1. Biomarker classification signatures comprising up to 5 analytes were identified using a tunable scoring function trained against 239 samples and validated with 120 additional samples. A total of 14 overlapping signatures classified patients without significant coronary disease (38% to 59% specificity while maintaining 95% sensitivity for patients requiring

  1. Characterisation of non-maternal serum proteins in amniotic fluid at weeks 16 to 18 of gestation

    DEFF Research Database (Denmark)

    Drøhse, H; Christensen, H; Myrhøj, Vibeke;


    Proteins found in amniotic fluid are mainly serum proteins, probably of maternal origin. About 5% of the total protein concentration has the potential of being fetal or decidual in origin. Only a few of these proteins have been isolated and characterised. In order to describe the foetal...... and decidual components in amniotic fluid more extensively, a polyspecific antiserum to amniotic fluid at weeks 16-18 of gestation was raised. Specificities in the antiserum to serum proteins were removed by adsorption. Several proteins of non-serum protein origin reacted with the antiserum. Three...... of these proteins were chosen for isolation and further characterisation. With the use of immunological methods, SDS-PAGE and N-terminal sequencing we identified two of the proteins as C-terminal propeptides of procollagen Type I and Type III, which have not hitherto been described in amniotic fluid. The third...

  2. Comparison of serum placental protein hormone levels in diabetic and normal pregnancy. (United States)

    Braunstein, G D; Mills, J L; Reed, G F; Jovanovic, L G; Holmes, L B; Aarons, J; Simpson, J L


    Conflicting data exist concerning maternal serum concentrations of placental hormones during pregnancy in women with diabetes mellitus. To resolve some of these discrepancies, women participating in the NICHD-Diabetes in Early Pregnancy Study were studied. In this collaborative study, pregnancy was identified within 21 days of conception by serum hCG measurements. We prospectively collected 185 blood samples from 35 insulin-dependent diabetic women and 166 blood samples from 31 control women, all between 5 and 37 weeks gestation. Serum concentrations of hCG, pregnancy-specific beta-1-glycoprotein, placental lactogen, and hCG alpha were measured serially. The relationship between serum hormone, fasting blood glucose, 1-h postprandial blood glucose, and glycosylated hemoglobin concentrations was compared. Serum hCG alpha levels were significantly lower in the diabetic women than in control women at multiple time points during the first and second trimesters, while no consistent differences in the serum concentrations of hCG or pregnancy-specific beta-1-glycoprotein were found between pregnant diabetic and control women. Serum placental lactogen levels were significantly lower in diabetic women at 9-10 weeks and 20 weeks gestation. There were no correlations between fasting blood glucose, 1-h postprandial blood glucose, or glycosylated hemoglobin and any of the placental protein levels in the diabetic women. These data are consistent with a defect in synthesis and/or secretion of hCG alpha by the cytotrophoblast during the first two trimesters of pregnancy in insulin-requiring diabetic women.

  3. Characterisation of non-maternal serum proteins in amniotic fluid at weeks 16 to 18 of gestation

    DEFF Research Database (Denmark)

    Drøhse, H; Christensen, H; Myrhøj, Vibeke;


    Proteins found in amniotic fluid are mainly serum proteins, probably of maternal origin. About 5% of the total protein concentration has the potential of being fetal or decidual in origin. Only a few of these proteins have been isolated and characterised. In order to describe the foetal and decid...

  4. Comparison Of Serum Calcium, Phosphorus And Total Protein Levels, In Pregnancy With Or Without Hypertensive Disorders

    Directory of Open Access Journals (Sweden)

    Rashid Pooraei M


    Full Text Available Background: Pre-eclampsia is characterized by hypertension development and proteinuria during pregnancy. Hypertension disorder is a leading cause of maternal and fetal morbidity and death in worldwide. Although the pathophysiology of hypertension during pregnancy is unclear, but there is consensus that early diagnosis and aggressive treatment is warranted to prevent complicated to both fetus and mother. The changes of serum trace elements during pregnancy are paramount important to predict and good understanding the situation of patients. The aim of this study was about this issue. Materials and Methods: In a case-control study we investigated the possible differences in the level of serum calcium, phosphorus and total protein in 55 healthy pregnant and 52 pregnant with hypertensive disease at 32-40 weeks of gestational age during the recent two years in Loghman hospital of Tehran. Some information such as age, blood group, parity number and blood pressure was taken from patients by a questionnaire. Results: The case population consisted of 22 sever preeclampsia, 15 mild preeclampsia, 8 eclampsia, and 7 chronic hypertensive. The mean serum calcium concentration (mg/dl was 9.180.74 in control group, 8.810.9 in mild preeclampsia, 7.850.38 in sever preeclampsia, 7.83 0.47 in eclampsia, 8.91 0.3 in chronic blood pressure. The mean serum phosphorus (mg/dl level observed, 4.27 in sever preeclampsia, 3.74 in eclampsia, 3.59 in mild eclampsia, 4.09 in chronic blood pressure, and 3.43 in control pregnant women. The mean serum total protein concentration level in sever preeclampsia and eclampsia was 5.46 and 5.04 mg/dl respectively. Conclusion: In conclusion, sever preeclampsia and eclampsia are associated with decreased level of calcium, total protein, and increased concentration of phosphorus.

  5. Effects of Egg White Protein Supplementation on Muscle Strength and Serum Free Amino Acid Concentrations

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    Yukari Kawano


    Full Text Available The aim of this study was to evaluate the effects of egg white protein compared to carbohydrate intake prior to exercise on fat free mass (FFM, one repetition maximum (1RM muscle strength and blood biochemistry in female athletes. Thirty healthy female collegiate athletes were recruited for this study and matched by sport type, body fat percentage and 1RM leg curl muscle strength. Participants were randomly divided into two groups: protein group (15.0 g egg white protein; 75 kcal and carbohydrate group (17.5 g maltodextrin, 78 kcal. Supplements were administered daily at the same time in a double-blind manner prior to training during an 8-week period. Measurements were performed before and after the 8-week regimen. The mean dietary energy intake did not change throughout the study period. FFM and 1RM assessments (i.e., leg curl, leg extension, squat, and bench press increased in both groups. Furthermore, serum urea and serum citrulline levels after the 8-week regimen increased significantly only in the protein group. Our findings indicated that compared to the carbohydrate supplement, the protein supplement was associated with some changes in protein metabolites but not with changes in body composition or muscle strength.

  6. Formation of long-lived reactive species of blood serum proteins by the action of heat. (United States)

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


    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)О2, (·)O2(-), (·)OН, 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.

  7. Serum Vascular Adhesion Protein-1 Predicts End-Stage Renal Disease in Patients with Type 2 Diabetes.

    Directory of Open Access Journals (Sweden)

    Hung-Yuan Li

    Full Text Available Diabetes is the leading cause of end-stage renal disease (ESRD worldwide. Vascular adhesion protein-1 (VAP-1 participates in inflammation and catalyzes the deamination of primary amines into aldehydes, hydrogen peroxide, and ammonia, both of which are involved in the pathogenesis of diabetic complications. We have shown that serum VAP-1 is higher in patients with diabetes and in patients with chronic kidney disease (CKD, and can predict cardiovascular mortality in subjects with diabetes. In this study, we investigated if serum VAP-1 can predict ESRD in diabetic subjects.In this prospective cohort study, a total of 604 type 2 diabetic subjects were enrolled between 1996 to 2003 at National Taiwan University Hospital, Taiwan, and were followed for a median of 12.36 years. The development of ESRD was ascertained by linking our database with the nationally comprehensive Taiwan Society Nephrology registry. Serum VAP-1 concentrations at enrollment were measured by time-resolved immunofluorometric assay.Subjects with serum VAP-1 in the highest tertile had the highest incidence of ESRD (p<0.001. Every 1-SD increase in serum VAP-1 was associated with a hazard ratio of 1.55 (95%CI 1.12-2.14, p<0.01 for the risk of ESRD, adjusted for smoking, history of cardiovascular disease, body mass index, hypertension, HbA1c, duration of diabetes, total cholesterol, use of statins, ankle-brachial index, estimated GFR, and proteinuria. We developed a risk score comprising serum VAP-1, HbA1c, estimated GFR, and proteinuria, which could predict ESRD with good performance (area under the ROC curve = 0.9406, 95%CI 0.8871-0.9941, sensitivity = 77.3%, and specificity = 92.8%. We also developed an algorithm based on the stage of CKD and a risk score including serum VAP-1, which can stratify these subjects into 3 categories with an ESRD risk of 0.101%/year, 0.131%/year, and 2.427%/year, respectively.In conclusion, serum VAP-1 can predict ESRD and is a useful biomarker to

  8. Structural characteristics of green tea catechins for formation of protein carbonyl in human serum albumin. (United States)

    Ishii, Takeshi; Mori, Taiki; Ichikawa, Tatsuya; Kaku, Maiko; Kusaka, Koji; Uekusa, Yoshinori; Akagawa, Mitsugu; Aihara, Yoshiyuki; Furuta, Takumi; Wakimoto, Toshiyuki; Kan, Toshiyuki; Nakayama, Tsutomu


    Catechins are polyphenolic antioxidants found in green tea leaves. Recent studies have reported that various polyphenolic compounds, including catechins, cause protein carbonyl formation in proteins via their pro-oxidant actions. In this study, we evaluate the formation of protein carbonyl in human serum albumin (HSA) by tea catechins and investigate the relationship between catechin chemical structure and its pro-oxidant property. To assess the formation of protein carbonyl in HSA, HSA was incubated with four individual catechins under physiological conditions to generate biotin-LC-hydrazide labeled protein carbonyls. Comparison of catechins using Western blotting revealed that the formation of protein carbonyl in HSA was higher for pyrogallol-type catechins than the corresponding catechol-type catechins. In addition, the formation of protein carbonyl was also found to be higher for the catechins having a galloyl group than the corresponding catechins lacking a galloyl group. The importance of the pyrogallol structural motif in the B-ring and the galloyl group was confirmed using methylated catechins and phenolic acids. These results indicate that the most important structural element contributing to the formation of protein carbonyl in HSA by tea catechins is the pyrogallol structural motif in the B-ring, followed by the galloyl group. The oxidation stability and binding affinity of tea catechins with proteins are responsible for the formation of protein carbonyl, and consequently the difference in these properties of each catechin may contribute to the magnitude of their biological activities.

  9. Effect of feed supplement on Milk Production, Fat % Total Serum Protein and Minerals in Lactating Buffalo

    Directory of Open Access Journals (Sweden)

    R.K. Verma


    Full Text Available A study was carried out to see the effect of feed supplement “Khurak” on milk yielding buffalo. The buffaloes were divided in two group. One group was offered “Khurak” as feed supplement for 7 days. Significant increase was observed in milk production, Total serum protein and calcium in khurak supplemented group (Treatment group. [Vet. World 2009; 2(5.000: 193-194

  10. Significant association between Helicobacter pylori infection and serum C-reactive protein



    Background: Helicobacter pylori (H. pylori) infection in gastric mucosa may cause systemic inflammatory reaction. This study aimed to examine the association between the infection and serum high sensitivity C-reactive protein (hsCRP). Methods: Subjects were comprised of three groups; 453 health checkup examinees from Yakumo town inhabitants in Hokkaido, Japan (YTI, 153 males and 300 females), 449 health checkup examinees (ENUH, 273 males and 176 females), and 255 female patients of an inferti...

  11. Effect of serum proteins on polystyrene nanoparticle uptake and intracellular trafficking in endothelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Guarnieri, Daniela; Guaccio, Angela; Fusco, Sabato; Netti, Paolo A., E-mail: [Istituto Italiano di Tecnologia, Center for Advanced Biomaterials for Health Care atCRIB (Italy)


    The physico-chemical properties of nanoparticles (NPs), such as small dimensions, surface charge and surface functionalization, control their capability to interact with cells and, in particular, with sub-cellular components. This interaction can be also influenced by the adsorption of molecules present in biological fluids, like blood, on NP surface. Here, we analysed the effect of serum proteins on 49 and 100 nm red fluorescent polystyrene NP uptake in porcine aortic endothelial (PAE) cells, as a model for vascular transport. To this aim, NP uptake kinetic, endocytic pathway and intracellular trafficking were studied by monitoring NPs inside cells through confocal microscopy and multiple particle tracking (MPT). We demonstrated that NPs are rapidly internalized by cells in serum-free (SF) medium, according to a saturation kinetic. Conversely, in 10% foetal bovine serum-enriched (SE) medium, NP uptake rate results drastically reduced. Moreover, NP internalization depends on an active endocytic mechanism that does not involve clathrin- and caveolae-mediated vesicular transport, in both SE and SF media. Furthermore, MPT data indicate that NP intracellular trafficking is unaffected by protein presence. Indeed, approximately 50-60% of internalized NPs is characterized by a sub-diffusive behaviour, whereas the remaining fraction shows an active motion. These findings demonstrate that the unspecific protein adsorption on NP surface can affect cellular uptake in terms of internalization kinetics, but it is not effective in controlling active and cellular-mediated uptake mechanisms of NPs and their intracellular routes.

  12. Detection and preliminary characterization of antibacterial protein(s in the serum of mud crab, Scylla serrata

    Directory of Open Access Journals (Sweden)

    V Meiyalagan


    Full Text Available Serum of mud crab, Scylla serrata has been found to possess significant antibacterial activity against some of the resident specific bacteria including Bacillus sp. N1, Bacillus flexus N3, Escherichia coli as well as crustacean pathogenic bacteria viz., Vibrio harveyi, V. alginolyticus and V. vulnificus. The physico-chemical characterization reveals the molecule responsible for antibacterial activity in the serum over 14 kDa, stable in the pH range of 6 to 8 and between the temperatures 20 to 40 ºC. Precipitation of respective molecule(s with 75 % ammonium sulphate or the supernatant obtained after precipitating the protein with 10 % TCA indicated that the molecule(s responsible for serum antibacterial activity appear to be proteinaceous in nature. Further studies demonstrated that antibacterial molecule(s against E. coli and V. harveyi appeared to be trypsin and pronase resistant and the molecule(s or domain responsible for antibacterial activity against Bacillus sp. N1 and B. flexus N3 appeared to be protease sensitive, thereby implicating possible involvement of multiple antibacterial factors in the serum of mud crab, S. serrata.

  13. Cellulose-based diagnostic devices for diagnosing serotype-2 dengue fever in human serum. (United States)

    Wang, Hsi-Kai; Tsai, Cheng-Han; Chen, Kuan-Hung; Tang, Chung-Tao; Leou, Jiun-Shyang; Li, Pi-Chun; Tang, Yin-Liang; Hsieh, Hsyue-Jen; Wu, Han-Chung; Cheng, Chao-Min


    Here, two types of cellulose-based in vitro diagnostic devices are demonstrated for the diagnosis of dengue virus infection in both buffer system and human serum: 1) paper-based ELISA for providing the semiquantitative information of the disease activity of serotype-2 dengue fever to healthcare persons (i.e., monitoring the disease activity with a specific serotype in single patients); 2) lateral flow immunoassays to screen for infection with serotype-2 dengue fever (i.e., rapid YES or NO diagnosis prepared for large populations, in terms of global public health). Paper-based ELISA (specific to serotype-2 dengue fever), which builds off of our previous studies and a revised previous ELISA procedure, owns multiple advantages: 1) high sensitivity (about 40 times higher than the current ELISA-based approaches, due to our therapeutic-based monoclonal antibody) and specificity (specific to dengue virus serotype-2 nonstructural protein-1 antigens); 2) tiny amount of sample and reagent used for single tests; 3) short operating duration (i.e., rapid diagnostic device); and, 4) inexpensiveness (appropriate for use in all developing and underdeveloped nations of the world). Due to the higher sensitivity and shorter operating duration of paper-based ELISA (compared with conventional ELISA, and lateral flow immunoassays also performed in this study), this study has not only been able to perform the diagnosis of dengue virus serotype-2 nonstructural protein-1 antigens in both buffer system and human serum but also to evaluate dengue virus serotype-2 envelope proteins in the buffer system, thus successfully achieving the first such use of these proteins as the target antigen for the development of diagnostic tools. These results provide a more comprehensive understanding for the genesis of dengue fever diagnostic tools (through antibody-antigen recognition).

  14. Effects of Regular Recreational Exercise Training on Serum ANGPTL3-Like Protein and Lipid Profile in Young Healthy Adults

    Directory of Open Access Journals (Sweden)

    Smol Ewelina


    Full Text Available Evidence of the role of ANGPTL3, a liver-secreted glycoprotein, in serum lipid turnover, led us to hypothesize that this protein may be involved in modification of the lipid profile induced by exercise-training. Given the lack of data regarding this issue, the main goal of the present study was to investigate the effects of regular participation in a recreational physical activity program on serum ANGPTL3 and selected lipid profile measures in young, apparently healthy female and male adults. We compared serum ANGPTL3, lipid profile measures, common lipid ratios, the Atherogenic Index of Plasma (AIP and glucose in fasting blood samples derived from 22 active physical education students including active females (AF, N=6 and males (AM, N=16 with samples from 28 relatively sedentary agematched peers, including female (SF, N=9 and male (SM, N=19 individuals not involved in any regular physical conditioning program. Despite high inter-individual variability of serum ANGPTL3, there was a general tendency toward higher serum ANGPTL3 and HDL-C in women compared to men, but without significant differences related to their physical activity status. Based on both routine lipid profile measures and lipid ratios, all participants had normal lipid profiles, normal glycemia, as well as favorable anthropometric indices not suggesting increased cardiometabolic risk. However, lower levels of the TG/HDL-C ratio and AIP in physically active compared to relatively sedentary participants, reflecting the predominance of large, buoyant LDL particles, strongly support the view of beneficial healthpromoting effects of regular participation in recreational sport activities.

  15. Post-transcriptional regulation of ribosomal protein genes during serum starvation in Entamoeba histolytica. (United States)

    Ahamad, Jamaluddin; Ojha, Sandeep; Srivastava, Ankita; Bhattacharya, Alok; Bhattacharya, Sudha


    Ribosome synthesis involves all three RNA polymerases which are co-ordinately regulated to produce equimolar amounts of rRNAs and ribosomal proteins (RPs). Unlike model organisms where transcription of rRNA and RP genes slows down during stress, in E. histolytica rDNA transcription continues but pre-rRNA processing slows down and unprocessed pre-rRNA accumulates during serum starvation. To investigate the regulation of RP genes under stress we measured transcription of six selected RP genes from the small- and large-ribosomal subunits (RPS6, RPS3, RPS19, RPL5, RPL26, RPL30) representing the early-, mid-, and late-stages of ribosomal assembly. Transcripts of these genes persisted in growth-stressed cells. Expression of luciferase reporter under the control of two RP genes (RPS19 and RPL30) was studied during serum starvation and upon serum replenishment. Although luciferase transcript levels remained unchanged during starvation, luciferase activity steadily declined to 7.8% and 15% of control cells, respectively. After serum replenishment the activity increased to normal levels, suggesting post-transcriptional regulation of these genes. Mutations in the sequence -2 to -9 upstream of AUG in the RPL30 gene resulted in the phenotype expected of post-transcriptional regulation. Transcription of luciferase reporter was unaffected in this mutant, and luciferase activity did not decline during serum starvation, showing that this sequence is required to repress translation of RPL30 mRNA, and mutations in this region relieve repression. Our data show that during serum starvation E. histolytica blocks ribosome biogenesis post-transcriptionally by inhibiting pre-rRNA processing on the one hand, and the translation of RP mRNAs on the other.

  16. Development of immunoturbidimetric assays for fourteen human serum proteins on the Hitachi 912. (United States)

    Ledue, Thomas B; Collins, Marilyn F; Ritchie, Robert F


    Many laboratories rely on dedicated nephelometers or turbidimeters and commercial reagent kits for the evaluation of serum proteins. However, with growing emphasis on cost containment, laboratories are forced to seek additional operational efficiencies by capitalizing on the use of existing analyzers whenever possible. In the present paper we describe the development of immunoturbidimetric assays for routine analysis of 14 human serum proteins (alpha1-antitrypsin, alpha2-macroglobulin, albumin, apolipoproteins Al and B, complement components 3 and 4, haptoglobin, immunoglobulins A, G, and M, orosomucoid, prealbumin, and transferrin) on the Hitachi 912, a general chemistry analyzer. With this system, we obtained excellent precision at levels corresponding to low, normal, and high physiologic concentrations of each protein (within-run imprecision CVs 0.97). We observed no significant interference from bilirubin (up to 718 micromol/l), hemoglobin (up to 8 g/l), triglyceride (up to 14.7 mmol/l) or rheumatoid factor (up to 4,140 IU/ml). Calibration for the 14 protein assays was stable for at least 7 days and onboard refrigerated reagents were stable for at least 3 months. The instrument's automated sample re-run feature minimized sample handling and helped to conserve specimens. In conclusion, the newly developed assays on the Hitachi 912 offer high throughput (>250 tests per hour) without the associated cost of a dedicated instrument for protein assays.

  17. Identification of (L)-fucose-binding proteins from the Nile tilapia (Oreochromis niloticus L.) serum. (United States)

    Argayosa, Anacleto M; Lee, Yuan C


    Lectins are carbohydrate-binding proteins with many biological functions including cellular recognition and innate immunity. In this study, a major l-fucose-binding lectin from the serum of Nile tilapia (Oreochromis niloticus L.), designated as TFBP, was isolated by l-fucose-BSA Sepharose CL6B affinity chromatography. The SDS-PAGE (10%) analysis of TFBP revealed a major band of approximately 23 kDa with an N-terminal amino acid sequence of DQTETAGQQSXPQDIHAVLREL which did not give significant similarities to the protein databases using BLASTp searches. Ruthenium red staining indicate positive calcium-binding property of TFBP. The purified TFBP agglutinated human type O erythrocytes but not the type A and B fresh erythrocytes. Live Aeromonas hydrophila and Enterococcus faecalis cells were also agglutinated by the lectin. The fucose-binding proteins were detected in the soluble protein extracts from the gills, gut, head kidneys, liver, serum and spleen using a fucose-binding protein probe (l-fucose-BSA-horseradish peroxidase). The binding of TFBP with the l-fucose-BSA probe was inhibited by l-fucose but not by alpha-methyl-d-mannose.

  18. A multiplex serum protein assay for determining the probability of colorectal cancer. (United States)

    Brock, Randall; Xiong, Bob; Li, Lily; Vanbogelen, Ruth A; Christman, Lori


    Our purpose is to develop a serum assay to determine an individual's probability of having colorectal cancer (CRC). We have discovered a protein panel yielding encouraging, clinically significant results. We evaluated 431 serum samples from donors screened for CRC by colonoscopy. We compared the concentration of seven proteins in individuals with CRC versus individuals found to be CRC free. The assay monitored a single peptide from each of seven proteins. Comparing CRC to normal samples in univariate two-sample t-tests, 6 of the 7 proteins yielded a p-value less than 0.01. Logistic regression was used to construct a model for determination of CRC probability. The model was fit on a randomly chosen training set of 321 samples. Using 6 of the 7 proteins (ORM1, GSN, C9, HABP2, SAA2, and C3) and a cut point of 0.4, an independent test set of 110 samples yielded a sensitivity of 93.75%, a specificity of 82.89% and a prevalence-adjusted negative predictive value (NPV) of 99.9775% for the assay. The results demonstrate that the assay has promise as a sensitive, non-invasive diagnostic test to provide individuals with an understanding of their own probability of having CRC.

  19. Hyperphosphatemia is a combined function of high serum PTH and high dietary protein intake in dialysis patients


    Streja, Elani; Lau, Wei Ling; Goldstein, Leanne; Sim, John J.; Molnar, Miklos Z.; Nissenson, Allen R; Csaba P Kovesdy; Kalantar-Zadeh, Kamyar


    Elevated serum phosphorus is associated with higher death risk in hemodialysis patients. Previous studies have suggested that both higher serum parathyroid hormone (PTH) level and higher dietary protein intake may contribute to higher serum phosphorus levels. However, it is not well known how these two factors simultaneously contribute to the combined risk of hyperphosphatemia in real patient-care scenarios. We hypothesized that the likelihood of hyperphosphatemia increases across higher seru...

  20. Structural modification of serum vitamin D3-binding protein and immunosuppression in AIDS patients. (United States)

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


    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.

  1. Milk-derived proteins and minerals alter serum osteocalcin in prepubertal boys after 7 days

    DEFF Research Database (Denmark)

    Mark, A.B.; Hoppe, Camilla; Michaelsen, K.F.


    We have previously shown that at equal protein content, milk, but not meat, decreased bone turnover in boys. This suggested that milk-derived components are important for bone metabolism. In the present study, we hypothesized that milk-derived proteins (whey and casein) affect bone turnover during...... growth depending on the content of milk minerals (calcium and phosphorus). This was a randomized, parallel, double-blind study. Eight-year-old boys (n = 57) received 1 of 4 milk drinks: whey protein with low or high content of minerals, or casein protein with low or high content of minerals. The amount...... of whey and casein was identical to their content in 1.5 L of milk. We measured serum osteocalcin (sOC), bone-specific alkaline phosphatase, and C-terminal telopeptides of type 1 collagen (immunoassay) and estimated dietary intake (3-day weighed food record) at baseline and after 7 days. Only s...

  2. Proteomic Analysis of Outer Membrane Proteins from Salmonella Enteritidis Strains with Different Sensitivity to Human Serum (United States)

    Dudek, Bartłomiej; Krzyżewska, Eva; Kapczyńska, Katarzyna; Rybka, Jacek; Pawlak, Aleksandra; Korzekwa, Kamila; Klausa, Elżbieta; Bugla-Płoskońska, Gabriela


    Differential analysis of outer membrane composition of S. Enteritidis strains, resistant to 50% normal human serum (NHS) was performed in order to find factors influencing the resistance to higher concentrations of NHS. Ten S. Enteritidis clinical strains, resistant to 50% NHS, all producing very long lipopolysaccharide, were subjected to the challenge of 75% NHS. Five extreme strains: two resistant and three sensitive to 75% NHS, were chosen for the further analysis of outer membrane proteins composition. Substantial differences were found in the levels of particular outer membrane proteins between resistant and sensitive strains, i.e. outer membrane protease E (PgtE) was present mainly in resistant strains, while sensitive strains possessed a high level of flagellar hook-associated protein 2 (FliD) and significantly higher levels of outer membrane protein A (OmpA). PMID:27695090

  3. Chromatographic Studies of Protein-Based Chiral Separations (United States)

    Bi, Cong; Zheng, Xiwei; Azaria, Shiden; Beeram, Sandya; Li, Zhao; Hage, David S.


    The development of separation methods for the analysis and resolution of chiral drugs and solutes has been an area of ongoing interest in pharmaceutical research. The use of proteins as chiral binding agents in high-performance liquid chromatography (HPLC) has been an approach that has received particular attention in such work. This report provides an overview of proteins that have been used as binding agents to create chiral stationary phases (CSPs) and in the use of chromatographic methods to study these materials and protein-based chiral separations. The supports and methods that have been employed to prepare protein-based CSPs will also be discussed and compared. Specific types of CSPs that are considered include those that employ serum transport proteins (e.g., human serum albumin, bovine serum albumin, and alpha1-acid glycoprotein), enzymes (e.g., penicillin G acylase, cellobiohydrolases, and α-chymotrypsin) or other types of proteins (e.g., ovomucoid, antibodies, and avidin or streptavidin). The properties and applications for each type of protein and CSP will also be discussed in terms of their use in chromatography and chiral separations.

  4. Adsorption and adhesion of common serum proteins to nanotextured gallium nitride. (United States)

    Bain, Lauren E; Hoffmann, Marc P; Bryan, Isaac; Collazo, Ramón; Ivanisevic, Albena


    As the broader effort towards device and material miniaturization progresses in all fields, it becomes increasingly important to understand the implications of working with functional structures that approach the size scale of molecules, particularly when considering biological systems. It is well known that thin films and nanostructures feature different optical, electrical, and mechanical properties from their bulk composites; however, interactions taking place at the interface between nanomaterials and their surroundings are less understood. Here, we explore interactions between common serum proteins - serum albumin, fibrinogen, and immunoglobulin G - and a nanotextured gallium nitride surface. Atomic force microscopy with a carboxyl-terminated colloid tip is used to probe the 'activity' of proteins adsorbed onto the surface, including both the accessibility of the terminal amine to the tip as well as the potential for protein extension. By evaluating the frequency of tip-protein interactions, we can establish differences in protein behaviour on the basis of both the surface roughness as well as morphology, providing an assessment of the role of surface texture in dictating protein-surface interactions. Unidirectional surface features - either the half-unit cell steppes of as-grown GaN or those produced by mechanical polishing - appear to promote protein accessibility, with a higher frequency of protein extension events taking place on these surfaces when compared with less ordered surface features. Development of a full understanding of the factors influencing surface-biomolecule interactions can pave the way for specific surface modification to tailor the bio-material interface, offering a new path for device optimization.

  5. Identification of serum monocyte chemoattractant protein-1 and prolactin as potential tumor markers in hepatocellular carcinoma.

    Directory of Open Access Journals (Sweden)

    Who-Whong Wang

    Full Text Available Early diagnosis of hepatocellullar carcinoma (HCC remains a challenge. The current practice of serum alpha-fetoprotein (AFP measurement is inadequate. Here we utilized a proteomic approach to identify novel serum biomarkers for distinguishing HCC patients from non-cancer controls. We profiled the serum proteins in a group of 58 resectable HCC patients and 11 non-HCC chronic hepatitis B (HBV carrier samples from the Singapore General Hospital (SGH using the RayBio® L-Series 507 Antibody Array and found 113 serum markers that were significantly modulated between HCC and control groups. Selected potential biomarkers from this list were quantified using a multiplex sandwich enzyme-linked immunosorbent assay (ELISA array in an expanded SGH cohort (126 resectable HCC patients and 115 non-HCC chronic HBV carriers (NC group, confirming that serum prolactin and monocyte chemoattractant protein-1 (MCP-1 were significantly upregulated in HCC patients. This finding of serum MCP-1 elevation in HCC patients was validated in a separate cohort of serum samples from the Mochtar Riady Institute for Nanotechnology, Indonesia (98 resectable HCC, 101 chronic hepatitis B patients and 100 asymptomatic HBV/HCV carriers by sandwich ELISA. MCP-1 and prolactin levels were found to correlate with AFP, while MCP-1 also correlated with disease stage. Subsequent receiver operating characteristic (ROC analysis of AFP, prolactin and MCP-1 in the SGH cohort and comparing their area under the ROC curve (AUC indicated that neither prolactin nor MCP-1 on their own performed better than AFP. However, the combination of AFP+MCP-1 (AUC, 0.974 had significantly superior discriminative ability than AFP alone (AUC, 0.942; p<0.001. In conclusion, prolactin and MCP-1 are overexpressed in HCC and are conveniently quantifiable in patients' sera by ELISA. MCP-1 appears to be a promising complementary biomarker for HCC diagnosis and this MCP-1+AFP model should be further evaluated as

  6. Sirtuin1: a promising serum protein marker for early detection of Alzheimer's disease.

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    Rahul Kumar

    Full Text Available Sirtuin (SIRT pathway has a crucial role in Alzheimer's disease (AD. The present study evaluated the alterations in serum sirtuin1 (SIRT1 concentration in healthy individuals (young and old and patients with AD and mild cognitive impairment (MCI. Blood samples were collected from 40 AD and 9 MCI patients as cases and 22 young healthy adults and 22 healthy elderly individuals as controls. Serum SIRT1 was estimated by Surface Plasmon Resonance (SPR, Western Blot and Enzyme Linked Immunosorbent Assay (ELISA. A significant (p<0.0001 decline in SIRT1 concentration was observed in patients with AD (2.27 ± 0.46 ng/µl and MCI (3.64 ± 0.15 ng/µl compared to healthy elderly individuals (4.82 ± 0.4 ng/µl. The serum SIRT1 concentration in healthy elderly was also significantly lower (p<0.0001 compared to young healthy controls (8.16 ± 0.87 ng/µl. This study, first of its kind, has demonstrated, decline in serum concentration of SIRT1 in healthy individuals as they age. In patients with AD and MCI the decline was even more pronounced, which provides an opportunity to develop this protein as a predictive marker of AD in early stages with suitable cut off values.

  7. Investigation of change of serum immunosuppressive acidic protein levels in gynecological tumors

    Institute of Scientific and Technical Information of China (English)


    Objective: To study the clinical significance for measuring serum immunosuppressive acidic protein(IAP) levels to diagnose and follow up survey patients with the gynecological tumor.Methods: Serum IAP levels were determined by IAP-single radial immunodiffusion test in 235 patients with the gynecological tumor,including 38 cases of benign tumor of ovary,41 cases of malignant tumor of ovary,66 cases of hysteromyoma,34 carcinomas of uterine cervix, 16 endometrial carcinomas,27 cases of chemotherapy,13 cases of recurrence, and the control group was 50 cases health women.Results: Serum IAP level was 889.4±207.8mg/L in malignant ovary tumors,which was significantly higher than that of health women and benign tumors of ovary (P<0.01).In patients with carcinoma of uterine cervix and endometrial carcinoma, their IAP levels were 741.4±212.6mg/L and 763.3±209.4mg/L,which were higher than those of the health women and benign tumor of ovary(P<0.01).After chemotherapy, serum IAP levels of malignant tumor of ovary were decreased;in patients with recurrence of tumor of ovary,IAP levels increased compared with the health women(P<0.01).Incidence of the abnormal value was 100%.Conclusion:Measuring IAP level of the gynecological tumor may be an auxiliary index for monitoring gynecological tumor and identifying benign and malignant tumor.

  8. Serum concentration and interaction properties of MBL/ficolin associated protein-1

    DEFF Research Database (Denmark)

    Skjoedt, Mikkel-Ole; Hummelshoj, Tina; Palarasah, Yaseelan;


    pathway (LCP) recognition molecules and MAP-1. We expressed recombinant MAP-1 in CHO DG44 cells, developed a quantitative ELISA assay based on a MAP-1 specific monoclonal capture antibody and measured the serum levels in 100 Danish blood donors. In addition we assessed the association properties between...

  9. Immunochemical quantitation of 3-(cystein-S-yl)acetaminophen adducts in serum and liver proteins of acetaminophen-treated mice. (United States)

    Pumford, N R; Hinson, J A; Potter, D W; Rowland, K L; Benson, R W; Roberts, D W


    Using a recently developed enzyme-linked immunosorbent assay specific for 3-(cystein-S-yl)acetaminophen adducts we have quantitated the formation of these specific adducts in liver and serum protein of B6C3F1 male mice dosed with acetaminophen. Administration of acetaminophen at doses of 50, 100, 200, 300, 400 and 500 mg/kg to mice resulted in evidence of hepatotoxicity (increase in serum levels of alanine aminotransferase and aspartate aminotransferase) at 4 hr in the 300, 400 and 500 mg/kg treatment groups only. The formation of 3-(cystein-S-yl)acetaminophen adducts in liver protein was not observed in the groups receiving 50, 100 and 200 mg/kg doses, but was observed in the groups receiving doses above 300 mg/kg of acetaminophen. Greater levels of adduct formation were observed at the higher doses. 3-(Cystein-S-yl)acetaminophen protein adducts were also observed in serum of mice receiving hepatotoxic doses of acetaminophen. After a 400 mg/kg dose of acetaminophen, 3-(cystein-S-yl)acetaminophen adducts in the liver protein reached peak levels 2 hr after dosing. By 12 hr the levels decreased to approximately 10% of the peak level. In contrast, 3-(cystein-S-yl)acetaminophen adducts in serum protein were delayed, reaching a sustained peak 6 to 12 hr after dosing. The dose-response correlation between the appearance of serum aminotransferases and 3-(cystein-S-yl)acetaminophen adducts in serum protein and the temporal correlation between the decrease in 3-(cystein-S-yl)acetaminophen adducts in liver protein and the appearance of adducts in serum protein are consistent with a hepatic origin of the adducts detected in serum protein.(ABSTRACT TRUNCATED AT 250 WORDS)

  10. Quantitative GSL-glycome analysis of human whole serum based on an EGCase digestion and glycoblotting method[S (United States)

    Furukawa, Jun-ichi; Sakai, Shota; Yokota, Ikuko; Okada, Kazue; Hanamatsu, Hisatoshi; Kobayashi, Takashi; Yoshida, Yasunobu; Higashino, Kenichi; Tamura, Tomohiro; Igarashi, Yasuyuki; Shinohara, Yasuro


    Glycosphingolipids (GSLs) are lipid molecules linked to carbohydrate units that form the plasma membrane lipid raft, which is clustered with sphingolipids, sterols, and specific proteins, and thereby contributes to membrane physical properties and specific recognition sites for various biological events. These bioactive GSL molecules consequently affect the pathophysiology and pathogenesis of various diseases. Thus, altered expression of GSLs in various diseases may be of importance for disease-related biomarker discovery. However, analysis of GSLs in blood is particularly challenging because GSLs are present at extremely low concentrations in serum/plasma. In this study, we established absolute GSL-glycan analysis of human serum based on endoglycoceramidase digestion and glycoblotting purification. We established two sample preparation protocols, one with and the other without GSL extraction using chloroform/methanol. Similar amounts of GSL-glycans were recovered with the two protocols. Both protocols permitted absolute quantitation of GSL-glycans using as little as 20 μl of serum. Using 10 healthy human serum samples, up to 42 signals corresponding to GSL-glycan compositions could be quantitatively detected, and the total serum GSL-glycan concentration was calculated to be 12.1–21.4 μM. We further applied this method to TLC-prefractionated serum samples. These findings will assist the discovery of disease-related biomarkers by serum GSL-glycomics. PMID:26420879

  11. Deglycosylation of serum vitamin D3-binding protein leads to immunosuppression in cancer patients. (United States)

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


    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.

  12. Biophysical analysis of the interaction of the serum protein human β2GPI with bacterial lipopolysaccharide

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    Anna Gries


    Full Text Available There are several human serum proteins for which no clear role is yet known. Among these is the abundant serum protein beta2-glycoprotein-I (β2GPI, 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 β2GPI 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, β2GPI 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 β2GPI 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.

  13. Proteomic analysis of changes in protein expression in serum from animals exposed to paraquat. (United States)

    Kim, Yong-Sik; Jung, Hana; Gil, Hyo-Wook; Hong, Sae-Yong; Song, Ho-Yeon


    Paraquat (PQ) poisoning remains a major public health concern in many countries. Extensive research has focused on finding early diagnostic biomarkers of acute PQ poisoning. In order to investigate the characterization of diagnostic biomarkers in PQ poisoning, we utilized proteomic analysis using serum from rats exposed to PQ, and we identified 8 differentially expressed proteins from over 500 protein spots. The expression of apolipoprotein E (ApoE), preprohaptoglobin (Pphg), a precursor of haptoglobin (Hp), and complement component 3 (C3) proteins was greatly induced by PQ exposure while the expression of fibrinogen γ-chain (FGG) and Ac-158 was dramatically reduced. To further investigate the possibility of ApoE, Pphg and FGG as useful diagnostic biomarkers of PQ poisoning, western blot and qRT-PCR analyses were conducted using cell lines as well as rat and human sera. The expression levels of ApoE, Hp and FGG were significantly altered in the presence of PQ in both rat and human serum suggesting that these proteins may be appropriate candidate molecular biomarkers for the early diagnosis of acute PQ intoxication.

  14. Localization of serum resistance-associated protein in Trypanosoma brucei rhodesiense and transgenic Trypanosoma brucei brucei. (United States)

    Bart, Jean-Mathieu; Cordon-Obras, Carlos; Vidal, Isabel; Reed, Jennifer; Perez-Pastrana, Esperanza; Cuevas, Laureano; Field, Mark C; Carrington, Mark; Navarro, Miguel


    African trypanosomes infect a broad range of mammals, but humans and some higher primates are protected by serum trypanosome lytic factors that contain apolipoprotein L1 (ApoL1). In the human-infective subspecies of Trypanosoma brucei, Trypanosoma brucei rhodesiense, a gene product derived from the variant surface glycoprotein gene family member, serum resistance-associated protein (SRA protein), protects against ApoL1-mediated lysis. Protection against trypanosome lytic factor requires the direct interaction between SRA protein and ApoL1 within the endocytic apparatus of the trypanosome, but some uncertainty remains as to the precise mechanism and location of this interaction. In order to provide more insight into the mechanism of SRA-mediated resistance to trypanosome lytic factor, we assessed the localization of SRA in T. b. rhodesiense EATRO3 using a novel monoclonal antibody raised against SRA together with a set of well-characterized endosomal markers. By three-dimensional deconvolved immunofluorescence single-cell analysis, combined with double-labelling immunoelectron microscopy, we found that ≈ 50% of SRA protein localized to the lysosome, with the remaining population being distributed through the endocytic pathway, but apparently absent from the flagellar pocket membrane. These data suggest that the SRA/trypanolytic factor interaction is intracellular, with the concentration within the endosomes potentially crucial for ensuring a high efficiency.

  15. Significance of serum troponin T and Creactive protein in the long-term prognosis of hemodialysis patients

    Institute of Scientific and Technical Information of China (English)



    Objective To investigate the long-term prognostic factors and the significance of serum cardiac troponin T(cTnT) and C-reactive protein(CRP) in maintenance hemodialysis(MHD) patients.Methods Clinical data of

  16. Computational modeling of serum-binding proteins and clearance in extrapolations across life stages and species for endocrine active compounds. (United States)

    Teeguarden, Justin G; Barton, Hugh A


    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.

  17. Tissue and serum samples of patients with papillary thyroid cancer with and without benign background demonstrate different altered expression of proteins

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    Mardiaty Iryani Abdullah


    Full Text Available Background Papillary thyroid cancer (PTC is mainly diagnosed using fine-needle aspiration biopsy. This most common form of well-differentiated thyroid cancer occurs with or without a background of benign thyroid goiter (BTG. Methods In the present study, a gel-based proteomics analysis was performed to analyse the expression of proteins in tissue and serum samples of PTC patients with (PTCb; n = 6 and without a history of BTG (PTCa; n = 8 relative to patients with BTG (n = 20. This was followed by confirmation of the levels of proteins which showed significant altered abundances of more than two-fold difference (p < 0.01 in the tissue and serum samples of the same subjects using ELISA. Results The data of our study showed that PTCa and PTCb distinguish themselves from BTG in the types of tissue and serum proteins of altered abundance. While higher levels of alpha-1 antitrypsin (A1AT and heat shock 70 kDa protein were associated with PTCa, lower levels of A1AT, protein disulfide isomerase and ubiquitin-conjugating enzyme E2 N seemed apparent in the PTCb. In case of the serum proteins, higher abundances of A1AT and alpha 1-beta glycoprotein were detected in PTCa, while PTCb was associated with enhanced apolipoprotein A-IV and alpha 2-HS glycoprotein (AHSG. The different altered expression of tissue and serum A1AT as well as serum AHSG between PTCa and PTCb patients were also validated by ELISA. Discussion The distinctive altered abundances of the tissue and serum proteins form preliminary indications that PTCa and PTCb are two distinct cancers of the thyroid that are etiologically and mechanistically different although it is currently not possible to rule out that they may also be due other reasons such as the different stages of the malignant disease. These proteins stand to have a potential use as tissue or serum biomarkers to discriminate the three different thyroid neoplasms although this requires further validation in clinically

  18. Detection of serum IgE antibody directed to aminothiazole using immobilized cephalosporins without protein conjugation

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    Akihito Yokoyama


    Full Text Available It is well known that allergic reactions may sometimes occur in patients under treatment with β-lactam antibiotics. For the detection of antidrug antibodies in vitro, conjugation with human serum albumin has been considered to be essential. In this study, we found that cefotiam, cefpirome, and ceftazidime could be immobilized without conjugation to carrier protein to construct a solid-phase enzyme-linked immunosorbent assay (ELISA system. We describe a patient (26-year-old female nurse with contact urticaria induced by antibiotics. Using the serum of this patient, we successfully detected IgE antibody directed to the aminothiazolyl group of cephalosporins, which has not previously been reported. Results suggest that the simple ELISA using unconjugated antibiotics could be applicable to patients with allergy to some cephalosporins and the aminothiazole side chain of the cephalosporins could cause an IgE-mediated allergic reaction.

  19. Fish protein intake induces fast-muscle hypertrophy and reduces liver lipids and serum glucose levels in rats. (United States)

    Kawabata, Fuminori; Mizushige, Takafumi; Uozumi, Keisuke; Hayamizu, Kohsuke; Han, Li; Tsuji, Tomoko; Kishida, Taro


    In our previous study, fish protein was proven to reduce serum lipids and body fat accumulation by skeletal muscle hypertrophy and enhancing basal energy expenditure in rats. In the present study, we examined the precise effects of fish protein intake on different skeletal muscle fiber types and metabolic gene expression of the muscle. Fish protein increased fast-twitch muscle weight, reduced liver triglycerides and serum glucose levels, compared with the casein diet after 6 or 8 weeks of feeding. Furthermore, fish protein upregulated the gene expressions of a fast-twitch muscle-type marker and a glucose transporter in the muscle. These results suggest that fish protein induces fast-muscle hypertrophy, and the enhancement of basal energy expenditure by muscle hypertrophy and the increase in muscle glucose uptake reduced liver lipids and serum glucose levels. The present results also imply that fish protein intake causes a slow-to-fast shift in muscle fiber type.

  20. A rapid lateral flow immunoassay for the detection of tyrosine phosphatase-like protein IA-2 autoantibodies in human serum.

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    Ingrid Kikkas

    Full Text Available 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.

  1. Serum Levels of Surfactant Proteins in Patients with Combined Pulmonary Fibrosis and Emphysema (CPFE.

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    Andriana I Papaioannou

    Full Text Available Emphysema and idiopathic pulmonary fibrosis (IPF present either per se or coexist in combined pulmonary fibrosis and emphysema (CPFE. Serum surfactant proteins (SPs A, B, C and D levels may reflect lung damage. We evaluated serum SP levels in healthy controls, emphysema, IPF, and CPFE patients and their associations to disease severity and survival.122 consecutive patients (31 emphysema, 62 IPF, and 29 CPFE and 25 healthy controls underwent PFTs, ABG-measurements, 6MWT and chest HRCT. Serum levels of SPs were measured. Patients were followed-up for 1-year.SP-A and SP-D levels differed between groups (p = 0.006 and p<0.001 respectively. In post-hoc analysis, SP-A levels differed only between controls and CPFE (p<0.05 and CPFE and emphysema (p<0.05. SP-D differed between controls and IPF or CPFE (p<0.001 for both comparisons. In IPF SP-B correlated to pulmonary function while SP-A, correlated to the Composite Physiological Index (CPI. Controls current smokers had higher SP-A and SP-D levels compared to non-smokers (p = 0.026 and p = 0.023 respectively. SP-D levels were higher in CPFE patients with extended emphysema (p = 0.042. In patients with IPF, SP-B levels at the upper quartile of its range (≥26 ng/mL presented a weak association with reduced survival (p = 0.05.In conclusion, serum SP-A and SP-D levels were higher where fibrosis exists or coexists and related to disease severity, suggesting that serum SPs relate to alveolar damage in fibrotic lungs and may reflect either local overproduction or overleakage. The weak association between high levels of SP-B and survival needs further validation in clinical trials.

  2. Interaction of bovine serum albumin protein with self assembled monolayer of mercaptoundecanoic acid (United States)

    Poonia, Monika; Agarwal, Hitesh; Manjuladevi, V.; Gupta, R. K.


    Detection of proteins and other biomolecules in liquid phase is the essence for the design of a biosensor. The sensitivity of a sensor can be enhanced by the appropriate functionalization of the sensing area so as to establish the molecular specific interaction. In the present work, we have studied the interaction of bovine serum albumin (BSA) protein with a chemically functionalized surface using a quartz crystal microbalance (QCM). The gold-coated quartz crystals (AT-cut/5 MHz) were functionalized by forming self-assembled monolayer (SAM) of 11-Mercaptoundecanoic acid (MUA). The adsorption characteristics of BSA onto SAM of MUA on quartz crystal are reported. BSA showed the highest affinity for SAM of MUA as compared to pure gold surface. The SAM of MUA provides carboxylated surface which enhances not only the adsorption of the BSA protein but also a very stable BSA-MUA complex in the liquid phase.

  3. Selective detection of protein homologues in serum using an OmpG nanopore (United States)

    Fahie, Monifa A.; Yang, Bib; Mullis, Martin; Holden, Matthew A.; Chen, Min


    Outer membrane protein G is a monomeric β-barrel porin that has seven flexible loops on its extracellular side. Conformational changes of these labile loops induce gating spikes in current recordings that we exploited as the prime sensing element for protein detection. The gating characteristics - open probability, frequency and current decrease - provide rich information for analyte identification. Here, we show that two antibiotin antibodies each induced a distinct gating pattern, which allowed them to be readily detected and simultaneously discriminated by a single OmpG nanopore in the presence of fetal bovine serum. Our results demonstrate the feasibility of directly profiling proteins in real-world samples with minimal or no sample pretreatment. PMID:26451707

  4. Modifications and oxidation of lipids and proteins in human serum detected by thermochemiluminescence. (United States)

    Shnizer, Sergei; Kagan, Tamara; Lanir, Amos; Maor, Irit; Reznick, Abraham Z


    Detection of electronically excited species (EES) in body fluids may constitute an important diagnostic tool in various pathologies. Examples of such products are triplet excited carbonyls (TEC), which can be a source for photon emission in the 400-550 nm range. The aim of the present study was to determine the actual contribution of lipid and protein components (protein carbonyls) to photon emission generated by thermochemiluminescence (TCL) during the heating of biological fluids. In this study, a new TCL Photometer device, designed by Lumitest Ltd, Israel, was used. Samples were heated to a constant temperature of 80 +/- 0.5 degrees C for 280 s and photon emission was measured at several time points. In order to compare the results of TCL measurements to conventional methods of detecting lipid and protein oxidation, each examined sample was also heated in a waterbath at 80 degrees C for 10-280 s. Lipid and protein oxidation were subsequently measured using conventional methods. The TCL of four polyunsaturated fatty acids (PUFA) with three to six double bonds was measured. The elevation of the PUFA TCL amplitude correlated with the increase in the number of double bonds of PUFA. A correlation between the increase in TCL intensity and protein carbonyl generation in bovine serum albumin (BSA) was also observed. In the venous blood serum, our study showed that an increase of TCL intensity during heating reflected the cleavage of TEC of lipid origin. Our study suggests that biological molecules such as proteins, lipids and other molecules, which may become unstable during heating, are capable of generating EES. We demonstrated that a TCL curve can be used as a kinetic model for measuring oxidative processes, which reflects modifications of different molecules involved in the oxidative stress phenomena.


    Directory of Open Access Journals (Sweden)

    L. S. Komarova


    Full Text Available Abstract. A role of intracellular proteins of eosinophils and mast cells remains unclear in the patients with hematological neoplasia. There is a substantial evidence that eosinophils possess some common mechanisms of cooperation with mast cells. Therapeutic interventions into key events controlling eosinophil migration may be a leading factor in treatment of hypereosinophylic states in onco-hematological disorders. Due to unknown functions of eosinophils in majority of eosinophilia-associated diseases, it would be useful to establish an algorithm of accurate diagnostics in the patients with eosinophilia, in order to choose more effective treatment in future.We studied serum levels of secretable eosinophil and mast cells proteins in oncohematological patients with increased eosinophil counts. The aim of our study was to test a significance of quantitative assay for tryptase and ECP in the patients with myelo- and lymphoproliferative diseases. The study group included thirty-eight patients with oncohematological diseases, accompanied by a marked eosinophilia (> 0.4 x 109/L. Eighteen patients with bronchial asthma (BA, and eight cases of solid tumors comprised a reference group for polyclonal eosinophilia. The levels of ECP and tryptase were measured in blood serum using a commercial fluoroimmunoenzyme assay («Pharmacia», Uppsala, Sweden. Total ECP levels were markedly increased in general group with hematological malignancies (p < 0.03, , and in cases of chronic GvHD (p < 0.03, and in a sub-group with lymphoproliferative disorders (р = 0.007 as compared to the group of non-hematological diseases.Serum levels of tryptase were significantly increased in the patients with chronic GvHD after allo-HSCT and lymphoproliferative diseases, as compared to the group of patients with solid tumors (р = 0.03, as well in GvHD compared with lymphoproliferative disorders (р < 0.05.A direct correlation was found between serum ECP levels and absolute

  6. The effects of GH and hormone replacement therapy on serum concentrations of mannan-binding lectin, surfactant protein D and vitamin D binding protein in Turner syndrome

    DEFF Research Database (Denmark)

    Gravholt, Claus Højbjerg; Leth-Larsen, Rikke; Lauridsen, Anna Lis


    function. In the present study we examined whether GH or hormone replacement therapy (HRT) in Turner syndrome (TS) influence the serum concentrations of MBL and two other proteins partaking in the innate immune defence, surfactant protein D (SP-D) and vitamin D binding protein (DBP). DESIGN: Study 1...

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

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    Jinghuan Li


    Full Text Available 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.

  8. Serum high-sensitivity C-reaction protein and heart fatty acid binding protein level and cardiac accidents in patients with unstable angina pectoris

    Institute of Scientific and Technical Information of China (English)



    Objective To investigate the relationship between serum high-sensitivity C-reaction protein (hs-CRP) and heart fatty acid binding protein (h-FABP) on cardiac accidents in patients with unstable angina pectoris (UAP). Methods Serum levels of hs-CRP, h-FABP, cardiac troponin-Ⅰ(cTn-Ⅰ) and creatine kinase MB isoenzyme (CK-MB) were measured and cardiac accidents within 2 weeks after the test were observed in 74 patients (male


    Hurwitz, S H; Meyer, K F


    The progress of an infection is usually associated with marked changes in the serum proteins. There may be an increase in the percentage of the total protein during some stage of the infection, and there is usually a change in the albumin-globulin ratio with an increase in the total globulins. This rise may antedate the development of any resistance by a considerable period of time. The non-protein constituents of the blood show fluctuations with a tendency to rise as the infection progresses. The process of immunization is in almost all instances associated with a definite increase in the globulins of the blood, and in some cases with a complete inversion of the normal albumin-globulin ratio. This may be produced both by living and dead organisms and by bacterial endotoxins. Massive doses usually result in an upset which shows no tendency to right itself during the period of observation. A rise in the globulins has been shown to occur long before the animal develops immune bodies in any appreciable concentration; and where the globulin curve and antibody curve appear to parallel one another, it can be shown by a careful analysis of both curves that there is a definite lack of correspondence at various periods of the experiment. Animals possessing a basic immunity show a more rapid rise in the globulin curve following inoculation. There is no parallelism between the leukocytic reaction and the globulin reaction. During periods of leukopenia the globulins may be as high as during the period of a leukocytosis. Bacterial endotoxins produce as striking an increase in the serum globulins as do living and killed bacteria. This would seem to indicate that a bacterial invasion of the organism is not absolutely essential for the globulin changes, and that the toxogenic factor in infection and immunity must play a part in the production of the changes noted. Inflammatory irritants injected intraperitoneally also result in a globulin increase. In this case the changes


    Directory of Open Access Journals (Sweden)

    Hristo HRISTEV


    Full Text Available The infl uence of lead (5mg/kg b.w and cadmium (2mg/kg b.w after chronic treatment of the rabbits on serum protein is investigated. Signifi cantly raised content of the cholesterol, ASAT and ALAT; hypo-albuminemia and hyperbetaglobulinemia of the background of one hypoproteinemia and low A/G coeffi cient are established. On basis of obtained result can to show degree of liver parenchyma damage and as trial for used the hyperbeta-globulinemia (at chronic treatment with cadmium is stronger markedly as indicator for delimitation of enteral from parenteral toxication, at that is noted hypergamma-globulinemia.

  11. Albumin-coated SPIONs: an experimental and theoretical evaluation of protein conformation, binding affinity and competition with serum proteins (United States)

    Yu, Siming; Perálvarez-Marín, Alex; Minelli, Caterina; Faraudo, Jordi; Roig, Anna; Laromaine, Anna


    The variety of nanoparticles (NPs) used in biological applications is increasing and the study of their interaction with biological media is becoming more important. Proteins are commonly the first biomolecules that NPs encounter when they interact with biological systems either in vitro or in vivo. Among NPs, super-paramagnetic iron oxide nanoparticles (SPIONs) show great promise for medicine. In this work, we study in detail the formation, composition, and structure of a monolayer of bovine serum albumin (BSA) on SPIONs. We determine, both by molecular simulations and experimentally, that ten molecules of BSA form a monolayer around the outside of the SPIONs and their binding strength to the SPIONs is about 3.5 × 10-4 M, ten times higher than the adsorption of fetal bovine serum (FBS) on the same SPIONs. We elucidate a strong electrostatic interaction between BSA and the SPIONs, although the secondary structure of the protein is not affected. We present data that supports the strong binding of the BSA monolayer on SPIONs and the properties of the BSA layer as a protein-resistant coating. We believe that a complete understanding of the behavior and morphology of BSA-SPIONs and how the protein interacts with SPIONs is crucial for improving NP surface design and expanding the potential applications of SPIONs in nanomedicine.The variety of nanoparticles (NPs) used in biological applications is increasing and the study of their interaction with biological media is becoming more important. Proteins are commonly the first biomolecules that NPs encounter when they interact with biological systems either in vitro or in vivo. Among NPs, super-paramagnetic iron oxide nanoparticles (SPIONs) show great promise for medicine. In this work, we study in detail the formation, composition, and structure of a monolayer of bovine serum albumin (BSA) on SPIONs. We determine, both by molecular simulations and experimentally, that ten molecules of BSA form a monolayer around the

  12. Serum protein response and renal failure in canine Babesia annae infection. (United States)

    Camacho, Angel Tomas; Guitian, Francisco Javier; Pallas, Estrella; Gestal, Juan Jesus; Olmeda, Sonia; Goethert, Heidi; Telford, Sam; Spielman, Andrew


    Babesia annae piroplasms have recently been recognised as a cause of infection and disease among dogs in Europe. The pathogenesis and clinical implications of this emerging disease remain poorly understood. We conducted this study to describe the electrophoretic profiles associated with the infection and to determine if B. annae associated azotaemia is caused by renal failure. We examined by microscopy 2,979 canine blood samples submitted to a diagnostic laboratory in NW Spain between September 2001 and April 2002. Small ring-shaped piroplasms were detected in blood smears of 87 samples and the identity of 58 of these presumptive cases were confirmed by PCR. This group of 58 infected dogs and a reference group of 15 healthy non-infected dogs were our study population. For all the dogs, serum protein response to -albumin, alpha-1 globulin, alpha-2 globulin, beta globulin and gamma globulin- was measured by capillary electrophoresis. The response of infected and non-infected dogs was compared and within infected dogs, the response of those with azotaemia (19) was compared with that of non-azotaemic dogs (39). Infected dogs presented a significant elevation of total proteins and all the different globulin fractions, and significantly lower levels of albumin compared to non-infected dogs. Among infected dogs, those presenting azotaemia had significantly lower concentrations of total proteins, albumin, beta and gamma globulins, and significantly higher values of alpha-2 globulin. Specific gravity was below the threshold of 1,025 for all dogs with azotaemia for which a urine sample was available (7) suggesting that azotaemia, in these dogs was of renal origin. Azotaemic dogs had higher concentrations of cholesterol and triglycerides, probably as a result of a liver compensatory response to the loss of proteins. We conclude that serum protein response in B. annae infected dogs corresponds to the pattern of a haemolytic syndrome with intense inflammatory reaction and that

  13. Metallomics for drug development: serum protein binding and analysis of an anticancer tris(8-quinolinolato)gallium(III) drug using inductively coupled plasma mass spectrometry. (United States)

    Ossipov, Konstantin; Foteeva, Lidia S; Seregina, Irina F; Perevalov, Sergei A; Timerbaev, Andrei R; Bolshov, Mikhail A


    The application of an inductively coupled plasma mass spectrometry (ICP-MS) assay for quantifying in vitro binding of a gallium-based anticancer drug, tris(8-quinolinolato)gallium(III), to serum albumin and transferrin and in human serum is described. The distribution of the drug between the protein-rich and protein-free fractions was assessed via ICP-MS measurement of total gallium in ultrafiltrates. Comparative kinetic studies revealed that the drug exhibits a different reactivity toward individual proteins. While the maximum possible binding to albumin (~10%) occurs practically immediately, interaction with transferrin has a step-like character and the equilibrium state (with more than 50% binding) is reached for about 48 h. Drug transformation into the bound form in serum, also very fast, results in almost quantitative binding (~95%). The relative affinity of protein-drug binding was characterized in terms of the association constants ranging from 10(3) to 10(4)M(-1). In order to further promote clinical testing of the gallium drug, the ICP-MS method was applied for direct quantification of gallium in human serum spiked with the drug. The detection limit for gallium was found to be as low as 20 ng L(-1). The repeatability was better than 8% (as RSD) and the achieved recoveries were in the range 99-103%.

  14. Unique expression of 35 KDa protein in serum and cystic fluid of women with malignant ovarian cysts substantiates its role in disease progression

    Institute of Scientific and Technical Information of China (English)

    Gulfam Ahmad; Muhammad Arslan


    Objective:The present study was undertaken as a part of a continuing search for proteomic based approaches for the diagnosis of ovarian cancers. Methods:The study comprised four groups of women with: i): malignant ovarian cysts, ii) benign ovarian cysts, iii): breast cancer (positive controls) and, iv) healthy females (negative controls). Serum and cystic fluids were processed for gel electrophoresis (SDS-PAGE). Serum and cystic follicular fluid estradiol and testosterone concentrations were measured by ELISA. Results:Proteomics analysis revealed the presence of a uniquely expressed protein with MW of 35 kDa in the serum and cystic fluid of patients with malignant ovarian cysts. On the other hand a protein of 100 kDa was not expressed in malignant ovarian patients whereas it was differentially expressed in the cystic fluid and serum of other three groups. Although mean values of estradiol levels were discernibly higher in patients with benign ovarian cysts as compared to those with malignant ovarian cysts and of positive and negative controls, difference was not significant. Mean concentrations of estradiol were significantly higher in cystic fluid aspirated from benign cysts as compared to that from malignant cysts. Conclusions:Taken together these data indicate a uniquely expressed protein of 35 kDa in patients with malignant ovarian cysts that may serve as a specific protein biomarker for the differential diagnosis of the ovarian cancer. However, a much larger sample of subjects is required to validate and confirm these findings.

  15. Effect of BCAA Supplementation on Serum C - Reactive Protein after Acute Resistance Exercise in Soccer Players

    Directory of Open Access Journals (Sweden)

    Mohammad Ghaderi


    Full Text Available Introduction: The present study was conducted to assess the effect of BCAA supplementation on serum C - reactive protein, after acute resistance exercise in soccer players.Methods: 20 soccer players in a randomized one-blind design completed in random 3 sets of 8-10 repetitions of 7 exercises. Subjects were randomized to two ten-member groups and consumed 200 mg. kg-1 BW of either BCAA or dextrin (placebo 30 minutes prior to exercise. To identify HS-CRP, venous blood samples were obtained 30 min prior to and immediately following exercise and at 1 hr, 2 hrs, 24hrs post exercise. Data were analyzed using 2-way repeated measure ANOVA and Bonferroni post hoc test. Results: Baseline serum values for Hs-CRP were not statistically different between groups in the 30 minutes before the exercise test (P>0.05. However there were significant increases (P<0.05 between the pre exercise and post exercise values for Hs-CRP from 24 hrs posttest, (P<0.05(. Importantly, the BCAA supplementation significantly reduced this Hs-CRP 24hrs post-test (P<0.05(.Conclusion: These results indicate that supplementary BCAA decreased serum concentrations of the inflammation biomarker Hs-CRP following resistance exercise. This observation suggests that BCAA supplementation may reduce the inflammation biomarker associated with resistance exercise.

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

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


    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 TFF3 correlated with extent of liver involvement in patient without pulmonary metastases and patients with higher TFF3 levels had significantly worse outcome (p 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.

  17. The importance of selecting a proper biological milieu for protein corona analysis in vitro: Human plasma versus human serum. (United States)

    Mirshafiee, Vahid; Kim, Raehyun; Mahmoudi, Morteza; Kraft, Mary L


    Nanoparticle (NP) exposure to biological fluids in the body results in protein binding to the NP surface, which forms a protein coating that is called the "protein corona". To simplify studies of protein-NP interactions and protein corona formation, NPs are incubated with biological solutions, such as human serum or human plasma, and the effects of this exposure are characterized in vitro. Yet, how NP exposure to these two different biological milieus affects protein corona composition and cell response has not been investigated. Here, we explore the differences between the protein coronas that form when NPs are incubated in human serum versus human plasma. NP characterization indicated that NPs that were exposed to human plasma had higher amounts of proteins bound to their surfaces, and were slightly larger in size than those exposed to human serum. In addition, significant differences in corona composition were also detected with gel electrophoresis and liquid chromatography-mass spectrometry/mass spectrometry, where a higher fraction of coagulation proteins and complement factors were found on the plasma-exposed NPs. Flow cytometry and confocal microscopy showed that the uptake of plasma-exposed NPs was higher than that of serum-exposed NPs by RAW 264.7 macrophage immune cells, but not by NIH 3T3 fibroblast cells. This difference is likely due to the elevated amounts of opsonins, such as fibrinogen, on the surfaces of the NPs exposed to plasma, but not serum, because these components trigger NP internalization by immune cells. As the human plasma better mimics the composition of the in vivo environment, namely blood, in vitro protein corona studies should employ human plasma, and not human serum, so the biological phenomena that is observed is more similar to that occurring in vivo.

  18. High Serum Lipopolysaccharide-Binding Protein Level in Chronic Hepatitis C Viral Infection Is Reduced by Anti-Viral Treatments (United States)

    Nien, Hsiao-Ching; Hsu, Shih-Jer; Su, Tung-Hung; Yang, Po-Jen; Sheu, Jin-Chuan; Wang, Jin-Town; Chow, Lu-Ping; Chen, Chi-Ling


    Background Lipopolysaccharide-binding protein (LBP) has been reported to associate with metabolic diseases, such as obesity, diabetes, and non-alcoholic fatty liver disease. Since chronic hepatitis C virus (HCV) infection is associated with metabolic derangements, the relationship between LBP and HCV deserves additional studies. This study aimed to determine the serum LBP level in subjects with or without HCV infection and investigate the change of its level after anti-viral treatments with or without interferon. Methods and Findings We recruited 120 non-HCV subjects, 42 and 17 HCV-infected subjects respectively treated with peginterferon α-2a/ribavirin and direct-acting antiviral drugs. Basic information, clinical data, serum LBP level and abdominal ultrasonography were collected. All the subjects provided written informed consent before being enrolled approved by the Research Ethics Committee of the National Taiwan University Hospital. Serum LBP level was significantly higher in HCV-infected subjects than non-HCV subjects (31.0 ± 8.8 versus 20.0 ± 6.4 μg/mL; p-value < 0.001). After multivariate analyses, LBP at baseline was independently associated with body mass index, hemoglobin A1c, alanine aminotransferase (ALT) and HCV infection. Moreover, the baseline LBP was only significantly positively associated with ALT and inversely with fatty liver in HCV-infected subjects. The LBP level significantly decreased at sustained virologic response (27.4 ± 6.6 versus 34.6 ± 7.3 μg/mL, p-value < 0.001; 15.9 ± 4.4 versus 22.2 ± 5.7 μg/mL, p-value = 0.001), regardless of interferon-based or -free therapy. Conclusions LBP, an endotoxemia associated protein might be used as an inflammatory biomarker of both infectious and non-infectious origins in HCV-infected subjects. PMID:28107471

  19. Serum Based Diagnosis of Asthma Using Raman Spectroscopy: An Early Phase Pilot Study (United States)

    Sahu, Aditi; Dalal, Krishna; Naglot, Sarla; Aggarwal, Parveen; Murali Krishna, C.


    The currently prescribed tests for asthma diagnosis require compulsory patient compliance, and are usually not sensitive to mild asthma. Development of an objective test using minimally invasive samples for diagnosing and monitoring of the response of asthma may help better management of the disease. Raman spectroscopy (RS) has previously shown potential in several biomedical applications, including pharmacology and forensics. In this study, we have explored the feasibility of detecting asthma and determining treatment response in asthma patients, through RS of serum. Serum samples from 44 asthma subjects of different grades (mild, moderate, treated severe and untreated severe) and from 15 reference subjects were subjected to Raman spectroscopic analysis and YKL-40 measurements. The force expiratory volume in 1 second (FEV1) values were used as gold standard and the serum YKL-40 levels were used as an additional parameter for diagnosing the different grades of asthma. For spectral acquisition, serum was placed on a calcium fluoride (CaF2) window and spectra were recorded using Raman microprobe. Mean and difference spectra comparisons indicated significant differences between asthma and reference spectra. Differences like changes in protein structure, increase in DNA specific bands and increased glycosaminoglycans-like features were more prominent with increase in asthma severity. Multivariate tools using Principal-component-analysis (PCA) and Principal-component based-linear-discriminant analysis (PC-LDA) followed by Leave-one-out-cross-validation (LOOCV), were employed for data analyses. PCA and PC-LDA results indicate separation of all asthma groups from the reference group, with minor overlap (19.4%) between reference and mild groups. No overlap was observed between the treated severe and untreated severe groups, indicating that patient response to treatment could be determined. Overall promising results were obtained, and a large scale validation study on

  20. An electrophoretic characterization of serum proteins of the collared peccary (Tayassu tajacu). (United States)

    Lochmiller, R L; Hellgren, E C; Varner, L W; Grant, W E


    Serum proteins of the collared peccary (Tayassu tajacu) were analyzed by agarose gel electrophoresis for wild adult males and females, nursing young, and reproductively-active females in captivity. Electrophoretic profiles of the adult peccary showed at least six distinct protein bands corresponding to the fractions: albumin, alpha-1, alpha-2, beta-1, beta-2, and gamma globulin. Globulin fractions of the peccary had different mobilities from the domestic swine. The only sexual dimorphisms were associated with the beta globulin:albumin ratio and the albumin:globulin ratio. Ingestion of colostrum in 1-day-old neonates was marked by a very large increase in gamma globulins. The only significant difference between pregnant and lactating females was in the alpha globulin:beta globulin ratio. Lactating females had higher concentrations of alpha-2 globulin than non-pregnant females.

  1. Protein and water dynamics in bovine serum albumin-water mixtures over wide ranges of composition. (United States)

    Panagopoulou, A; Kyritsis, A; Shinyashiki, N; Pissis, P


    Dielectric dynamic behavior of bovine serum albumin (BSA)-water mixtures over wide ranges of water fractions, from dry protein until 40 wt % in water, was studied through dielectric relaxation spectroscopy (DRS). The α relaxation associated with the glass transition of the hydrated system was identified. The evolution of the low temperature dielectric relaxation of small polar groups of the protein surface with hydration level results in the enhancement of dielectric response and the decrease of relaxation times, until a critical water fraction, which corresponds to the percolation threshold for protonic conductivity. For water fractions higher than the critical one, the position of the secondary ν relaxation of water saturates in the Arrhenius diagram, while contributions originating from water molecules in excess (uncrystallized water or ice) follow separate relaxation modes slower than the ν relaxation.

  2. Doping Human Serum Albumin with Retinoate Markedly Enhances Electron Transport Across the Protein

    CERN Document Server

    Amdursky, Nadav; Sheves, Mordechai; Cahen, David


    Electrons can migrate via proteins over distances that are considered long for non-conjugated systems. Proteins' nano-scale dimensions and the enormous flexibility of their structures and chemistry makes them fascinating subjects for investigating the mechanism of their electron transport (ETp) capacity. One particular attractive research direction is that of tuning their ETp efficiency by doping them with external small molecules. Here we report that solid-state ETp across human serum albumin (HSA) increases by more than two orders of magnitude upon retinoate (RA) binding to HSA. RA was chosen because optical spectroscopy has provided evidence for the non-covalent binding of at least three RA molecules to HSA and indications for their relative structural positions. The temperature dependence of ETp shows that both the activation energy and the distance-decay constant decrease with increasing RA binding to HSA. Furthermore, the observed transition from temperature-activated ETp above 190K to temperature-indep...

  3. Controlled release of bovine serum albumin from hydroxyapatite microspheres for protein delivery system

    Energy Technology Data Exchange (ETDEWEB)

    Boonsongrit, Yaowalak [Joining and Welding Research Institute, Osaka University, 11-1 Mihogaoka, Ibaraki, Osaka 567-0047 (Japan); Abe, Hiroya [Joining and Welding Research Institute, Osaka University, 11-1 Mihogaoka, Ibaraki, Osaka 567-0047 (Japan); Cooperative Research Center of Life Sciences, Kobe Gakuin University, Minatojima 1-1-3, Chuo-ku, Kobe 650-8586 (Japan)], E-mail:; Sato, Kazuyoshi [Joining and Welding Research Institute, Osaka University, 11-1 Mihogaoka, Ibaraki, Osaka 567-0047 (Japan); Naito, Makio [Joining and Welding Research Institute, Osaka University, 11-1 Mihogaoka, Ibaraki, Osaka 567-0047 (Japan); Cooperative Research Center of Life Sciences, Kobe Gakuin University, Minatojima 1-1-3, Chuo-ku, Kobe 650-8586 (Japan); Yoshimura, Masahiro [Materials and Structures Laboratory, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8503 (Japan); Ichikawa, Hideki; Fukumori, Yoshinobu [Faculty of Pharmaceutical Sciences, Kobe Gakuin University, Minatojima 1-1-3, Chuo-ku, Kobe 650-8586 (Japan); Cooperative Research Center of Life Sciences, Kobe Gakuin University, Minatojima 1-1-3, Chuo-ku, Kobe 650-8586 (Japan)


    Desorption behavior of a model protein (bovine serum albumin, BSA) on commercial hydroxyapatite (HAp) microspheres and its control were investigated for protein delivery system. The desorption behavior related strongly to the phosphate concentration in phosphate buffer solution: the amount of desorbed BSA increased when the phosphate concentration increased. In physiological buffer solution, which contains 10 mM phosphate, the initial burst release of BSA was observed: 70% of BSA was rapidly desorbed after 0.5 h, and 80% after 24 h. In contrast, the extremely low release profile of BSA was observed in distilled water. For the controlled release of BSA in physiological condition, the BSA-loaded HAp microspheres were encapsulated with a biodegradable polymer, poly(lactic acid-co-glycolic acid) (PLGA) by a solid-in oil-in water (S/O/W) emulsion solvent evaporation method. The initial burst was significantly reduced, and the BSA release was remarkably prolonged by the encapsulation.

  4. Serum acute phase proteins in cows with SARA (Subacute Ruminal Acidosis suspect

    Directory of Open Access Journals (Sweden)

    C. Cannizzo


    Full Text Available The aim of this study was to evaluate the variations of Acute Phase Proteins (APPs and other blood constituents during the onset of the sub-acute ruminal acidosis (SARA pathological status. A total of 108 cows from 12 dairy herds were randomly selected and divided into three Groups of 36 animals each. All animals were subjected to a rumenocentesis. Group A was composed by subjects with a rumen pH>5.8, Group B was composed by subjects with a rumen pH ≤5.5≤5.8 and Group C was composed by subjects with a rumen pH<5.5. Blood samples were collected by jugular venipuncture and Haptoglobin (Hp, Serum Amyloid A (SAA, Total Proteins, Albumin and White Blood Cells (WBC were determined. One-way ANOVA showed a statistical significance on Rumen pH, Hp, SAA. SARA seems not stimulate the APPs production from liver.

  5. Human serum albumin binding to silica nanoparticles--effect of protein fatty acid ligand. (United States)

    Ang, Joo Chuan; Henderson, Mark J; Campbell, Richard A; Lin, Jhih-Min; Yaron, Peter N; Nelson, Andrew; Faunce, Thomas; White, John W


    Neutron reflectivity shows that fatted (F-HSA) and defatted (DF-HSA) versions of human serum albumin behave differently in their interaction with silica nanoparticles premixed in buffer solutions although these proteins have close to the same surface excess when the silica is absent. In both cases a silica containing film is quickly established at the air-water interface. This film is stable for F-HSA at all relative protein-silica concentrations measured. This behaviour has been verified for two small silica nanoparticle radii (42 Å and 48 Å). Contrast variation and co-refinement have been used to find the film composition for the F-HSA-silica system. The film structure changes with protein concentration only for the DF-HSA-silica system. The different behaviour of the two proteins is interpreted as a combination of three factors: increased structural stability of F-HSA induced by the fatty acid ligand, differences in the electrostatic interactions, and the higher propensity of defatted albumin to self-aggregate. The interfacial structures of the proteins alone in buffer are also reported and discussed.

  6. On the mechanism of hydrogen evolution catalysis by proteins: A case study with bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Doneux, Th., E-mail: [Chimie Analytique et Chimie des Interfaces, Faculte des Sciences, Universite Libre de Bruxelles, Boulevard du Triomphe 2, CP 255, B-1050 Bruxelles (Belgium); Institute of Biophysics, Academy of Sciences of the Czech Republic, Kralovopolska 135, 612 65 Brno (Czech Republic); Ostatna, Veronika [Institute of Biophysics, Academy of Sciences of the Czech Republic, Kralovopolska 135, 612 65 Brno (Czech Republic); Palecek, Emil, E-mail: [Institute of Biophysics, Academy of Sciences of the Czech Republic, Kralovopolska 135, 612 65 Brno (Czech Republic)


    Highlights: > Proteins catalyse hydrogen evolution at mercury electrodes. > The adsorbed protein is the mediator and the buffer proton donor is the substrate. > The characteristics of the catalytic peak are connected to the protein properties. - Abstract: The catalysis of the hydrogen evolution reaction (HER) by proteins has been known for decades but was only recently found to be useful for electroanalytical purposes. The mechanism of the catalytic process is investigated at hanging mercury drop electrodes by cyclic voltammetry, with bovine serum albumin as a model system. It is shown that the catalyst is the protein in the adsorbed state. The influence of various parameters such as the accumulation time, scan rate or buffer concentration is studied, and interpreted in the framework of a surface catalytic mechanism. Under the experimental conditions used in the work, a 'total catalysis' phenomenon takes place, the rate of HER being limited by the diffusion of the proton donor. The adequacy of the existing models is discussed, leading to a call for the development of more refined models.

  7. Localization of the immunoglobulins G, A and M, beta-trace protein and gamma-trace protein on isoelectric focusing of serum and cerebrospinal fluid by immunofixation. (United States)

    Laurenzi, M A; Link, H


    The mobility of the immunoglobulins G, A and M, beta-trace protein and gramma-trace protein on isoleectric focusing of serum and CSF was determined by immunofixation using specific antisera. Polyclonal IgG migrated as multiple bands between pH 4.7--8.6, polyclonal IgA as multiple bands between pH 4.9--6.1 in CSF and serum. IgM could not be identified in normal CSF or serum. beta-trace protein gave three bands at pH 8.0, 8.4 and 7.4--7.5, respectively, while gamma-trace protein gave one single band at pH 9.5--greater than 9.5. Oligoclonal IgG in CSF in multiple sclerosis and neurosyphilis migrated between pH 8.6--greater than 9.5 and was easily discriminated from other proteins.

  8. Clorazepate dipotassium and diazepam in renal insufficiency: serum concentrations and protein binding of diazepam and desmethyldiazepam. (United States)

    Ochs, H R; Rauh, H W; Greenblatt, D J; Kaschell, H J


    5 patients with chronic renal failure on maintenance hemodialysis and 5 healthy matched controls received single 20-mg intravenous doses of clorazepate dipotassium. Clearance of pharmacologically active unbound desmethyldiazepam was reduced in renal failure patients as opposed to controls, and free fraction in serum was greater. Since desmethyldiazepam distribution was reduced in renal patients, elimination half-life was actually shorter than in controls (36 vs. 57 h). In 10 dialysis patients receiving chronic diazepam treatment (5-15 mg/day), steady-state concentrations of diazepam (56 ng/ml) and desmethyldiazepam (77 ng/ml) were significantly lower than in age- and weight-matched controls receiving similar doses (189 and 216 ng/ml, respectively). However after correction for the higher free fractions of both compounds in renal patients as opposed to controls, steady-state concentrations of unbound drug were found to be similar between groups. Interpretation of kinetic variables and steady-state serum concentrations of extensively protein-bound drugs requires consideration of alterations in protein binding that may occur in disease states.

  9. [Refractometric measurement of total serum protein, comparison of refractometry and biuret test (author's transl)]. (United States)

    Liappis, N; Jäkel, A


    Study on the Refractometric Determination of Total Protein in Serum, Comparison of the Refractometric and Biuret Method. Total protein concentration in serum was determined by the aid of the Abbé-refractometer and the biuret method. Both methods showed a good precision and accuracy. The investigation was carried out in 241 sera with normal bilirubin (up to 1 mg/100 ml), cholesterol (up to 200 mg/100 ml) and urea (up to 23,0 mg/100 ml) concentration, in 43 sera with increased (10,6-26,6 mg/100 ml) bilirubin concentration, in 129 sera with increased (200-520 mg/100 ml) cholesterol concentration and in 43 sera with increased (23,0-155,3 mg/100 ml) urea concentration. The comparison of the refractometric values with the values obtained by the biuret method in the 241 sera with normal bilirubin, cholesterol and urea concentration (correlation coefficient = 0,971) showed a close correlation and in the 43 sera with increased bilirubin concentration (correlation coefficient = 0,958) an acceptable correlation. However no close correlations were observed in the 129 sera with increased cholesterol concentration and in the 43 sera with increased urea concentration. The correlation lines diverged proportional with the increase of cholesterol and urea concentration from the expected correlation lines.

  10. Active fragments of the antihemorrhagic protein HSF from serum of habu (Trimeresurus flavoviridis). (United States)

    Aoki, Narumi; Deshimaru, Masanobu; Terada, Shigeyuki


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

  11. AGR3 in breast cancer: prognostic impact and suitable serum-based biomarker for early cancer detection. (United States)

    Garczyk, Stefan; von Stillfried, Saskia; Antonopoulos, Wiebke; Hartmann, Arndt; Schrauder, Michael G; Fasching, Peter A; Anzeneder, Tobias; Tannapfel, Andrea; Ergönenc, Yavuz; Knüchel, Ruth; Rose, Michael; Dahl, Edgar


    Blood-based early detection of breast cancer has recently gained novel momentum, as liquid biopsy diagnostics is a fast emerging field. In this study, we aimed to identify secreted proteins which are up-regulated both in tumour tissue and serum samples of breast cancer patients compared to normal tissue and sera. Based on two independent tissue cohorts (n = 75 and n = 229) and one serum cohort (n = 80) of human breast cancer and healthy serum samples, we characterised AGR3 as a novel potential biomarker both for breast cancer prognosis and early breast cancer detection from blood. AGR3 expression in breast tumours is significantly associated with oestrogen receptor α (Pbreast tumours (multivariate hazard ratio: 2.186, 95% CI: 1.008-4.740, Pbreast cancer patients (n = 40, mainly low stage tumours) compared to healthy controls (n = 40). To develop a suitable biomarker panel for early breast cancer detection, we measured AGR2 protein in human serum samples in parallel. The combined AGR3/AGR2 biomarker panel achieved a sensitivity of 64.5% and a specificity of 89.5% as shown by receiver operating characteristic (ROC) curve statistics. Thus our data clearly show the potential usability of AGR3 and AGR2 as biomarkers for blood-based early detection of human breast cancer.

  12. Serum peptide/protein profiling by mass spectrometry provides diagnostic information independently of CA125 in women with an ovarian tumor

    DEFF Research Database (Denmark)

    Callesen, Anne; Madsen, Jonna S; Iachina, Maria;


    In the present study, the use of a robust and sensitive mass spectrometry based protein profiling analysis was tested as diagnostic tools for women with an ovarian tumor. The potential additional diagnostic value of serum protein profiles independent of the information provided by CA125 were also...... investigated. Protein profiles of 113 serum samples from women with an ovarian tumor (54 malign and 59 benign) were generated using MALDI-TOF MS. A total of 98 peaks with a significant difference (pwomen with benign tumors/cysts and malignant ovarian tumors were identified. After...... average linkage clustering, a profile of 46 statistical significant mass peaks was identified to distinguish malignant tumors and benign tumors/cysts. In the subgroup of women with normal CA125 values (

  13. Total sialic acid, total protein and total sugar levels in serum and saliva of oral squamous cell carcinoma patients: A case control study

    Directory of Open Access Journals (Sweden)

    Nidhi Dhakar


    Conclusion: The present study showed a significant increase in serum sialic acid, salivary sialic acid and serum protein from control to OSCC and suggests that these markers may be reliable in diagnosis and predicting prognosis.

  14. Marsupial and monotreme serum immunoglobulin binding by proteins A, G and L and anti-kangaroo antibody. (United States)

    Vaz, Paola K; Hartley, Carol A; Browning, Glenn F; Devlin, Joanne M


    Serological studies are often conducted to examine exposure to infectious agents in wildlife populations. However, specific immunological reagents for wildlife species are seldom available and can limit the study of infectious diseases in these animals. This study examined the ability of four commercially available immunoglobulin-binding reagents to bind serum immunoglobulins from 17 species within the Marsupialia and Monotremata. Serum samples were assessed for binding, using immunoblots and ELISAs (Enzyme-linked immunosorbent assays), to three microbially-derived proteins - staphylococcal protein A, streptococcal protein G and peptostreptococcal protein L. Additionally, an anti-kangaroo antibody was included for comparison. The inter- and intra-familial binding patterns of the reagents to serum immunoglobulins varied and evolutionary distance between animal species was not an accurate predictor of the ability of reagents to bind immunoglobulins. Results from this study can be used to inform the selection of appropriate immunological reagents in future serological studies in these clades.

  15. Protein-bound carbohydrates in breast cancer. Liquid-chromatographic analysis for mannose, galactose, fucose, and sialic acid in serum. (United States)

    Mrochek, J E; Dinsmore, S R; Tormey, D C; Waalkes, T P


    We describr high-resolution chromatographic analysis for protein-bound sialic acid in serum, with use of a cerate oxidimetric detector. Values for sera from normal women averaged 680.5 mg/liter, with a coefficient of variation of 23%. Including data obtained by previously developed chromatographic procedures for protein-bound mannose, galactose, and fucsoe, we assessed sera from breast-cancer patients whose malignancy had been categorized as either stable, responsive, or progressive (based on clinical observations spaced from two to five months apart). All of 12 responsive patients had decreases of protein-bound fucose averaging 34.5% (SD, 16.1) and all of 10 patients with progressive disease had increases averaging 38.3% (SD 21.5). Changes in fucose averaged less than 6.7% (SD, 4.9) for eight patients with clinically stable breast cancer. Changes in protein-bound mannose, galactose, and sialic acid did not correlate as well as did fucose with the clinical disease status of the patients.

  16. Protein-bound carbohydrates in breast cancer. Liquid-chromatographic analysis for mannose, galactose, fucose, and sialic acid in serum

    Energy Technology Data Exchange (ETDEWEB)

    Mrochek, J.E. (Oak Ridge National Lab., TN); Dinsmore, S.R.; Tormey, D.C.; Waalkes, T.P.


    High-resolution chromatographic analysis for protein-bound sialic acid in serum, with use of a cerate oxidimetric detector is described. Values for sera from normal women averaged 680.5 mg/liter, with a coefficient of variation of 23%. Including data obtained by previously developed chromatographic procedures for protein-bound mannose, galactose, and fucose, sera from breast-cancer patients whose malignancy had been categorized as either stable, responsive, or progressive (based on clinical observations spaced from two to five months apart) were assessed. All of 12 responsive patients had decreases of protein-bound fucose averaging 34.5% (SD, 16.1) and all of 10 patients with progressive disease had increases averaging 38.3% (SD 21.5). Changes in fucose averaged less than 6.7% (SD, 4.9) for eight patients with clinically stable breast cancer. Changes in protein-bound mannose, galactose, and sialic acid did not correlate as well as did fucose with the clinical disease status of the patients.

  17. Protein-based composite materials

    Directory of Open Access Journals (Sweden)

    Xiao Hu


    Full Text Available Protein-based composite biomaterials have been actively pursued as they can encompass a range of physical properties to accommodate a broader spectrum of functional requirements, such as elasticity to support diverse tissues. By optimizing molecular interfaces between structural proteins, useful composite materials can be fabricated as films, gels, particles, and fibers, as well as for electrical and optical devices. Such systems provide analogies to more traditional synthetic polymers yet with expanded utility due to the material's tunability, mechanical properties, degradability, biocompatibility, and functionalization, such as for drug delivery, biosensors, and tissue regeneration.

  18. Serum concentration and increased temperature alter Mayaro virus RNA and protein synthesis in Aedes albopictus (mosquito)-infected cells. (United States)

    Motta, M C; Fournier, M V; Carvalho, M G


    We have previously shown the inhibition of Mayaro virus multiplication in Aedes albopictus-infected cells maintained at a supraoptimal temperature for growth (37 degrees C) and a stimulation of virus production in response to high serum concentrations in the incubation medium. In the present study, we addressed the question of how the effect of continuous heat stress and high serum concentration soon after infection interfere with virus macromolecule synthesis. Cells maintained at 28 degrees C in the presence of 2% serum synthesized a viral genomic RNA of 12 kb and a subgenomic RNA of 5.2 kb 6 h postinfection. Analysis of the protein profile showed the presence of the viral nucleocapsid protein of 34 kDa (P34). However, if infected cells were maintained at 37 degrees C, a smear starting immediately below the 5.2-kb RNA was noticed and the viral P34 was not detected by SDS-PAGE. Addition of 10% serum to the growth medium of infected cells maintained at 37 degrees C results in a viral RNA profile and protein synthesis similar to those observed in cultures kept at 28 degrees C, i.e., the smear was not observed and the P34 protein was detected. The results suggest that the inhibition of virus multiplication by temperature may be related to the inhibition of viral nonstructural protein synthesis early during infection. The presence of high serum levels in the incubation medium protects macromolecule synthesis against heat stress.

  19. Leptin Parameters are Associated with Inflammation and Both Anthropometric and Serum Markers of Protein-Energy Wasting in Hemodialysis Patients

    Directory of Open Access Journals (Sweden)

    Abdullah ÖZKÖK


    Full Text Available OBJECTIVE: Protein-energy wasting (PEW and inflammation are common and related to increased cardiovascular mortality in hemodialysis (HD patients. Herein, we aimed to investigate the relationships between adipocytokines, inflammation and serum and anthropometric markers of PEW syndrome in HD patients. MATERIAL and METHODS: Seventy-four HD patients (mean age=62±13 years, male/female=44/30 were enrolled. Serum leptin, adiponectin and IL-6 levels were measured by ELISA. Anthropometric measurements such as triceps skin fold thickness (TSFT, mid-arm circumference (MAC were performed. Bioelectrical impedance analysis was also done. The degree of malnutrition was evaluated with subjective global assessment (SGA. RESULTS: The leptin/fat ratio was associated with serum IL-6 (r=0.32, p=0.008, serum albumin (r= -0.37, p=0.002 and TSFT (r= -0.41, p=0.001. Serum adiponectin levels were positively correlated with serum albumin (r=0.29,p=0.01 and negatively associated with the leptin/fat ratio (r= -0.30, p=0.01. Serum CRP levels were significantly lower in the high adiponectin group (1.20 ±0.92 vs 2.02 ±2.18 μg/mL, p=0.042. On linear regression analysis for predicting the leptin/fat ratio, gender and serum albumin were retained as significant in the model. CONCLUSION: Serum leptin parameters are associated with inflammation and both anthropometric and serum markers of protein-energy wasting. Adipocytokines may be the link between PEW and inflammation in uremia.

  20. The relationship between serum adipocytokines and Graves' ophthalmopathy: A hospital-based study. (United States)

    Kim, Bo-Yeon; Mok, Ji-Oh; Kang, Sung-Koo; Jang, Sun-Young; Jung, Chan-Hee; Kim, Chul-Hee


    Adipocytokines are thought to be associated with inflammatory disorders and autoimmune diseases. However, limited information is available on the relationship between serum adipocytokine levels, Graves' disease (GD), and Graves' ophthalmopathy (GO). The present study examined the relationship between serum adipocytokine levels and GD and GO. A total of 80 patients with GD participated in this study. The medical records of patients were reviewed retrospectively. GO activity was assessed using the clinical activity score (CAS). GO severity was assessed by the modified NOSPECS classification and included soft tissue involvement, proptosis, and extraocular muscle involvement. Serum adiponectin, leptin, resistin, and retinol-binding protein 4 (RBP-4) levels were measured using commercially available enzyme-linked immunosorbent assays. The prevalence of GO was 36.3%. Serum adiponectin, leptin, and resistin levels were significantly higher in patients with GO than in those without GO. The CAS was positively correlated with serum adiponectin and leptin levels. The total eye score was positively correlated with serum adiponectin, leptin, resistin, and RBP-4 levels. A multivariate analysis revealed that serum leptin and resistin levels were associated with the presence of GO after adjusting for clinical factors. Free thyroxine was negatively correlated with serum leptin level. These results suggest that adipocytokines, such as leptin and resistin, may play a role in inflammatory and autoimmune processes of GD and GO. Future studies with larger numbers of patients are required to establish relationships between serum adipocytokines levels and GO and ascertain the role of adipocytokines in GD and GO.

  1. C-reactive protein serum level in patients with psoriasis before and after treatment with narrow-band ultraviolet B* (United States)

    Farshchian, Mahmoud; Ansar, Akram; Sobhan, Mohammadreza; Hoseinpoor, Valiollah


    Background C-reactive protein is an inflammatory biomarker and its level increases in the serum of psoriatic patients. Its level is also associated with Psoriasis Area and Severity Index score. Objective The aim of this study was to assess the decrement of serum C-reactive protein level with narrow-band ultraviolet B (NB-UVB) therapy. Methods C-reactive protein serum levels in psoriasis patients were measured before and after treatment with NB-UVB and the data were analyzed in relation to the Psoriasis Area and Severity Index score improvement. Results Baseline C-reactive protein levels among psoriatic patients were higher than normal. These levels decreased significantly after treatment (P<0.001). At the beginning of the study, patients with higher levels of C-reactive protein also had more extensive and severe skin involvement. The highest decrease in C-reactive protein was observed in patients who responded better to the treatment and achieved a higher Psoriasis Area and Severity Index 75%. There was an association between baseline Psoriasis Area and Severity Index scores and C-reactive protein levels. Conclusion Patients with moderate to severe plaque-type psoriasis had active systemic inflammation, which was demonstrated by increased levels of C-reactive protein. Furthermore, skin disease severity was correlated with C-reactive protein levels. Phototherapy healed the psoriatic skin lesions and reduced inflammation, while decreasing C-reactive protein levels. PMID:27828628

  2. Preparation of protein imprinted materials by hierarchical imprinting techniques and application in selective depletion of albumin from human serum (United States)

    Liu, Jinxiang; Deng, Qiliang; Tao, Dingyin; Yang, Kaiguang; Zhang, Lihua; Liang, Zhen; Zhang, Yukui


    Hierarchical imprinting was developed to prepare the protein imprinted materials, as the artificial antibody, for the selective depletion of HSA from the human serum proteome. Porcine serum albumin (PSA) was employed as the dummy template for the fabrication of the recognition sites. To demonstrate the advantages of the hierarchical imprinting, molecularly imprinted polymers prepared by hierarchical imprinting technique (h-MIPs) were compared with those obtained by bulk imprinting (b-MIPs), in terms of the binding capacity, adsorption kinetics, selectivity and synthesis reproducibility. The binding capacity of h-MIPs could reach 12 mg g-1. And saturation binding could be reached in less than 20 min for the h-MIPs. In the protein mixture, h-MIPs exhibit excellent selectivity for PSA, with imprinting factors as about 3.6, much higher than those for non-template proteins. For the proteomic application, the identified protein group number in serum treated by h-MIPs was increased to 422, which is 21% higher than that obtained from the original serum, meanwhile the identified protein group number for the Albumin Removal kit was only 376. The results demonstrate that protein imprinted polymers prepared by hierarchical imprinting technique, might become the artificial antibodies for the selective depletion of high abundance proteins in proteome study.

  3. An integrated bioanalytical platform for supporting high-throughput serum protein binding screening. (United States)

    Zhang, Jun; Shou, Wilson Z; Vath, Marianne; Kieltyka, Kasia; Maloney, Jennifer; Elvebak, Larry; Stewart, Jeremy; Herbst, John; Weller, Harold N


    Quantification of small molecules using liquid chromatography/tandem mass spectrometry (LC/MS/MS) on a triple quadrupole mass spectrometer has become a common practice in bioanalytical support of in vitro adsorption, distribution, metabolism and excretion (ADME) screening. The bioanalysis process involves primarily three indispensable steps: MS/MS optimization for a large number of new chemical compounds undergoing various screening assays in early drug discovery, high-throughput sample analysis with LC/MS/MS for those chemically diverse compounds using the optimized MS/MS conditions, and post-acquisition data review and reporting. To improve overall efficiency of ADME bioanalysis, an integrated system was proposed featuring an automated and unattended MS/MS optimization, a staggered parallel LC/MS/MS for high-throughput sample analysis, and a sophisticated software tool for LC/MS/MS raw data review as well as biological data calculation and reporting. The integrated platform has been used in bioanalytical support of a serum protein binding screening assay with high speed, high capacity, and good robustness. In this new platform, a unique sample dilution scheme was also introduced. With this dilution design, the total number of analytical samples was reduced; therefore, the total operation time was reduced and the overall throughput was further improved. The performance of the protein binding screening assay was monitored with two controls representing high and low binding properties and an acceptable inter-assay consistency was achieved. This platform has been successfully used for the determination of serum protein binding in multiple species for more than 4000 compounds.

  4. Evaluation of the use of serum C-reactive protein concentration to predict outcome in puppies infected with canine parvovirus

    DEFF Research Database (Denmark)

    McClure, Vanessa; van Schoor, Mirinda; Thompson, Peter N.;


    Objective-To evaluate associations of serum C-reactive protein (CRP) concentration with duration of hospitalization and with outcome in puppies with canine parvoviral enteritis. Design-Prospective observational study. Animals-79 client-owned puppies with naturally acquired canine parvovirus.......7%, respectively (considered moderately accurate). Conclusions and Clinical Relevance-Although serum CRP concentration was associated with outcome in puppies with canine parvovirus enteritis, it did not prove to be a good predictor of outcome when used alone....

  5. The role of serum cholinesterase activity and S100B protein in the evaluation of organophosphate poisoning. (United States)

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


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

  6. Brix refractometry in serum as a measure of failure of passive transfer compared to measured immunoglobulin G and total protein by refractometry in serum from dairy calves. (United States)

    Hernandez, D; Nydam, D V; Godden, S M; Bristol, L S; Kryzer, A; Ranum, J; Schaefer, D


    A series of trials were conducted to evaluate Brix refractometry (Brix %) for the assessment of failure of passive transfer (FPT) in dairy calves compared to: (1) serum IgG (reference standard) when measured by radial immunodiffusion (RID) or a turbidometric immunoassay (TIA), and (2) serum total protein refractometry (STP). For the serum samples tested with TIA, STP, and Brix % (n = 310; Holstein calves), the median concentrations were 21.3 g/L IgG, 58 g/L STP, and 9.2%, respectively. For the serum samples tested with RID, STP and Brix % (n = 112; Jersey calves), the mean concentrations were 38 g/L IgG, 68 g/L STP, and 10.2%, respectively. For samples tested with only Brix % and STP (n = 265; Holstein calves), median STP and Brix % were 50 g/L STP and 8.5%, respectively. Correlations between Brix % and RID, and between Brix % and TIA were equal (r = 0.79, respectively). Brix % and STP were positively correlated (r = 0.99). Brix % estimated serum IgG concentrations determined by TIA and RID (r(2) = 0.63, 0.62, respectively). When FPT was defined as serum IgG Brix % ≤ 8.5% showed optimal sensitivity (100%) and specificity (89.2%) to predict FPT. At the same IgG cut-point, an STP value of ≤ 52 g/L showed a similar sensitivity (100%) and specificity (80.4%) to predict FPT. Brix refractometry predicted successful transfer of passive immunity in dairy calves, but further evaluation as a diagnostic tool for the diagnosis of FPT is warranted.

  7. Increased serum C-reactive protein concentrations in dogs with congestive heart failure due to myxomatous mitral valve disease

    DEFF Research Database (Denmark)

    Reimann, M. J.; Ljungvall, I.; Hillstrom, A.;


    Cardiovascular disease in humans and dogs is associated with mildly increased circulating concentrations of C-reactive protein (CRP). Few studies have evaluated associations between circulating CRP and canine myxomatous mitral valve disease (MMVD) and the results reported have been divergent....... The aim of this study was to investigate whether serum concentrations of CRP, determined using a novel automated canine-specific high -sensitivity CRP assay (Gentian hsCRP), were associated with severity of MMVD and selected clinical variables in dogs. The study included 188 client-owned dogs...... with different severities of MMVD. Dogs were classified based on ACVIM consensus statement guidelines (group A, n = 58; group BI, n = 56; group B2, n = 38; group C, n = 36). Data were analysed using descriptive statistics and multiple regression analysis. Dogs with congestive heart failure (CHF; group C) had...

  8. [Evaluation of the relations between serum proteins electrophoresis and other laboratory tests in monoclonal gammopathies (author's transl)]. (United States)

    Ramacciotti, P G; Lazzari, L; Minardi, P


    We have considered interesting to determine monoclonal gammopathies incidence, in 2191 serum proteins electrophoresis performed in our laboratory from January to December 1974. We have found 15 cases of monoclonal gammopathies, some cases combined with Mieloma (3 cases), some other with other with non specific diseases. We have considered the relations between type of gammopathy and other laboratory tests useful for any other diagnose: they are: immunochemical analysis, E.S.R., red and white count, total proteins, Bence Jones protein.

  9. Day/night changes in serum S100B protein concentrations in acute paranoid schizophrenia. (United States)

    Morera-Fumero, Armando L; Díaz-Mesa, Estefanía; Abreu-Gonzalez, Pedro; Fernandez-Lopez, Lourdes; Cejas-Mendez, Maria Del Rosario


    There are day/night and seasonal changes in biological markers such as melatonin and cortisol. Controversial changes in serum S100B protein levels have been described in schizophrenia. We aim studying whether serum S100B levels present day/night variations in schizophrenia patients and whether S100B levels are related to psychopathology. Sixty-five paranoid schizophrenic inpatients participated in the study. Psychopathology was assessed with the Positive and Negative Syndrome Scale (PANSS) at admission and discharge. Blood was drawn at 12:00 (midday) and 00:00 (midnight) hours at admission and discharge. Sixty-five healthy subjects matched by age, gender and season acted as control group. At admission and discharge patients had significantly higher serum S100B concentrations at midday and midnight than healthy subjects. At admission, patients showed a day/night variation of S100B levels, with higher S100B levels at 12:00 than at 00:00h (143.7±26.3pg/ml vs. 96.9±16.6pg/ml). This day/night difference was not present in the control group. Midday and midnight S100B at admission decreased when compared to S100B at discharge (midday, 143.7±26.3 vs. 83.0±12, midnight 96.9±16.6 vs. 68.6±14.5). There was a positive correlation between the PANSS positive subscale and S100B concentrations at admission. This correlation was not present at discharge.

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

    Directory of Open Access Journals (Sweden)

    Anckarsäter Henrik


    Full Text Available Abstract 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 CRP were determined by ELISA in a population of 270 42-year-old women recruited from the population registry who had been assessed using the Temperament and Character Inventory. Self-reported previous or ongoing depression was also recorded. Unpaired two-tailed t-tests were used for comparison between two groups and correlations were evaluated by the calculation of Pearson's r-coefficient. Results The temperament trait harm avoidance was positively (r = 0.227, p r = -0.261, p p-values corrected for multiple comparisons. The correlations between the personality traits and CRP were observed also after exclusion of subjects reporting ongoing depression (n = 26. Whereas women reporting ongoing depression showed significantly increased levels of CRP as compared to non-depressed women (n = 155, women reporting a history of depression displayed no significant difference in CRP levels as compared to women that reported that they had never been depressed. Conclusion Serum levels of CRP in women was found to be associated with the personality traits harm avoidance and self-directedness. In addition, moderately elevated levels may be a state dependent marker of depression.

  11. Significant association between Helicobacter pylori infection and serum C-reactive protein

    Directory of Open Access Journals (Sweden)

    Yoshiko Ishida, Koji Suzuki, Kentaro Taki, Toshimitsu Niwa, Shozo Kurotsuchi, Hisao Ando, Akira Iwase, Kazuko Nishio, Kenji Wakai, Yoshinori Ito, Nobuyuki Hamajima


    Full Text Available Background: Helicobacter pylori (H. pylori infection in gastric mucosa may cause systemic inflammatory reaction. This study aimed to examine the association between the infection and serum high sensitivity C-reactive protein (hsCRP. Methods: Subjects were comprised of three groups; 453 health checkup examinees from Yakumo town inhabitants in Hokkaido, Japan (YTI, 153 males and 300 females, 449 health checkup examinees (ENUH, 273 males and 176 females, and 255 female patients of an infertility clinic (PIC, Nagoya University Hospital. Twenty participants with hsCRP more than 1 mg/dl were excluded from the analysis. Those with hsCRP more than 0.1mg/dl were defined as high hsCRP individuals. H. pylori infection status was examined with a serum IgG antibody test. Results: When the three groups were combined, the geometric mean of hsCRP concentration was significantly higher among the seropositives (0.047mg/dl than among the seronegatives (0.035mg/dl; p<0.0001 by a t-test. The percentage of high hsCRP individuals was also higher in the seropositives than in the seronegatives among any group; 23.3% and 20.1% in YTI, 22.0% and 16.0% in ENUH, and 32.7% and 18.7% in PIC, respectively, although the difference was significant only in ENUH. The summary odds ratio of the high hsCRP for the seropositives relative to the seronegatives was 1.38 (95% confidence interval, 1.01-1.89, when age, sex, body mass index, smoking, and subject group were adjusted by a logistic model. Conclusions: In three groups, hsCRP was higher among the infected individuals. The summary odd ratio indicated that H. pylori infection could influence the serum hsCRP level.

  12. Changes of serum high sensitive C-reactive protein in patients with acute cerebral infarction

    Institute of Scientific and Technical Information of China (English)

    Yan Du; Yan Ren; Ying Li


    BACKGROUND: Serum high sensitive C-reactive protein (hs-CRP), which regards as a high sensitive mark of systemic inflammatory response syndrome, can provide a lot of valuable information for the treatment and prognosis of cerebrovascular disease.OBJECTIVE: To observe the differences of blood glucose, lipid, homocysteine and previous disease history among patients with acute cerebral infarction at various levels of hs-CRP and compare changes of hs-CRP of patients with various degrees ofneurologic impairment.DESIGN: Contrast observation.SETTING: Department of Neurology, Shenzhou Hospital, Shenyang Medical College.PARTICIPANTS: A total of 102 patients with acute cerebral infarction were selected from Department of Neurology, Shenzhou Hospital of Shenyang Medical College from February 2005 to September 2006,including 55 males and 47 females aged from 55 to 86 years. All accepted patients met the diagnostic criteria of cerebral infarction established by the Fourth National Cerebrovascular Disease Academic Meeting and were diagnosed with CT or MRI examination. All patients provided the confirmed consent. Based on clinical criteria of neurologic impairment established by the Fourth National Cerebrovascular Disease Academic Meeting, patients were randomly divided into mild group (0- 15 points, n =46), moderate group (16- 30points, n =38) and severe group (31 - 45 points, n =18). In addition, based on hs-CRP level within 72 hours,patients were divided into normal group (hs-CRP ≤ 3 mg/L, n =53) and increasing group (hs-CRP > 3 mg/L,n =49).METHODS: ① 2 mL venous blood was selected from hospitalized patients in the next morning to separate serum. Quantitative measurement of hs-CRP was dealt with Latex Enhnced Turbidimetric Immunoassay (LETIA). ② Fasting venous blood was colleted from hospitalized patients in the next morning to measure numeration of white blood cells, fibrinogen, blood glucose, total cholesterol (TC), triacylglycerol (TG), high density lipoprotein

  13. Detection of the Inflammation Biomarker C-Reactive Protein in Serum Samples: Towards an Optimal Biosensor Formula

    Directory of Open Access Journals (Sweden)

    Wellington M. Fakanya


    Full Text Available 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.

  14. Effect of protein, probiotic, and symbiotic supplementation on serum biological health markers of molted layers. (United States)

    Anwar, H; Rahman, Z U; Javed, I; Muhammad, F


    Dietary zinc was used to induce molt in 200 White Leghorn birds in caged housing at the age of 70 wk. The birds were equally and randomly allocated to 4 groups each of 50 birds as G1 (control; CP 16%, no supplement), G2 (CP 18%, no other supplement), G3 (CP 16%, symbiotic Perfectin: 85 mg•L(-1) in drinking water daily), and G4 (CP 16%, probiotic Protexin: 85 mg•L(-1) in drinking water daily) after the completion of molt. The sampling was conducted 3 times at 5% production, peak production, and end of production with 15 birds being killed at each sampling from each group to collect the blood for harvesting of serum. Serum health markers including total oxidant status, total antioxidant capacity, homocysteine concentration, and paraoxonase, arylesterase, and ceruloplasmin activity were determined by the prescribed assays. The overall total antioxidant capacity was increased, whereas total oxidant status and homocysteine concentrations were reduced significantly (P ≤ 0.01) in all the supplemented groups compared with the control. The paraoxonase and ceruloplasmin activity were enhanced (P ≤ 0.01) in the supplemented groups compared with the control, and arylesterase activity was increased (P ≤ 0.01) in only G2 compared with the other groups. Although protein supplementation significantly reduced the oxidative stress, supplementation with symbiotic and probiotic also improved the health status by decreasing the oxidative stress in the birds.

  15. The Effect of Cinnamon Extract on Serum Proteins Levels of Male Balb/c Mice

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    M Modaresi


    Full Text Available Background & Aim: Cinnamon is a plant with many pharmaceutical effects. The present research evaluated the effects of Cinnamon bark extract on serum proteins level in male Balb/c mice. Methods: In this experimental study, 40 small Balb/c mice were chosen and divided into 5 groups: a control group, a case group, and three treatment groups. Normal saline was administered as placebo to the case group while the control group received no injections.. Cinnamon extract in doses of 50, 100 and 200 mg/Kg/48hr were injected intraperitoneally for 20 days to treatment groups. The levels of pre albumin, albumin, alpha-1, alpha-2, beta and gamma globulins were separated electrophoretically and calculated from the pattern of electrophoretogram. Results: The result indicated that the levels of pre albumin decreased significantly in two experimental groups (doses of 50 and 100 mg/kg, the levels of alpha-1 and beta in 200 group, alpha-2 level in 100 and gamma in 100 and 200 groups increased significantly .The injection of 100 mg/Kg/48h extract of Cinnamon decreased (p<0.05 the albumin level in plasma in treatment group as compared to the control group. Conclusion: Results of this study indicated that the serum level of globulins has not changed dramatically by the extract of Cinnamon. Since albumin synthesis occurs in the liver cells‚ thus administration of Cinnamon may affect the function of liver cells.


    Directory of Open Access Journals (Sweden)

    Iván Darío BRAVO-TOBAR


    Full Text Available SUMMARY Chagas disease is a public health problem worldwide. The availability of diagnostic tools to predict the development of chronic Chagas cardiomyopathy is crucial to reduce morbidity and mortality. Here we analyze the prognostic value of adenosine deaminase serum activity (ADA and C-reactive protein serum levels (CRP in chagasic individuals. One hundred and ten individuals, 28 healthy and 82 chagasic patients were divided according to disease severity in phase I (n = 35, II (n = 29, and III (n = 18. A complete medical history, 12-lead electrocardiogram, chest X-ray, and M-mode echocardiogram were performed on each individual. Diagnosis of Chagas disease was confirmed by ELISA and MABA using recombinant antigens; ADA was determined spectrophotometrically and CRP by ELISA. The results have shown that CRP and ADA increased linearly in relation to disease phase, CRP being significantly higher in phase III and ADA at all phases. Also, CRP and ADA were positively correlated with echocardiographic parameters of cardiac remodeling and with electrocardiographic abnormalities, and negatively with ejection fraction. CRP and ADA were higher in patients with cardiothoracic index ≥ 50%, while ADA was higher in patients with ventricular repolarization disturbances. Finally, CRP was positively correlated with ADA. In conclusion, ADA and CRP are prognostic markers of cardiac dysfunction and remodeling in Chagas disease.




    SUMMARY Chagas disease is a public health problem worldwide. The availability of diagnostic tools to predict the development of chronic Chagas cardiomyopathy is crucial to reduce morbidity and mortality. Here we analyze the prognostic value of adenosine deaminase serum activity (ADA) and C-reactive protein serum levels (CRP) in chagasic individuals. One hundred and ten individuals, 28 healthy and 82 chagasic patients were divided according to disease severity in phase I (n = 35), II (n = 29), and III (n = 18). A complete medical history, 12-lead electrocardiogram, chest X-ray, and M-mode echocardiogram were performed on each individual. Diagnosis of Chagas disease was confirmed by ELISA and MABA using recombinant antigens; ADA was determined spectrophotometrically and CRP by ELISA. The results have shown that CRP and ADA increased linearly in relation to disease phase, CRP being significantly higher in phase III and ADA at all phases. Also, CRP and ADA were positively correlated with echocardiographic parameters of cardiac remodeling and with electrocardiographic abnormalities, and negatively with ejection fraction. CRP and ADA were higher in patients with cardiothoracic index ≥ 50%, while ADA was higher in patients with ventricular repolarization disturbances. Finally, CRP was positively correlated with ADA. In conclusion, ADA and CRP are prognostic markers of cardiac dysfunction and remodeling in Chagas disease. PMID:26603224

  18. 基于亲和素-生物素双夹心体系测定外源性蛋白血清动态浓度的非抗体依赖新方法%A Novel Method for Assessment of Dynamic Concentration of Exogenous Protein in Serum: an Antibody-independent Sandwich System Based on Avidin-biotin Interaction

    Institute of Scientific and Technical Information of China (English)

    余颖欣; 刘艳君; 焦德龙; 文李艳; 徐江平; 富宁


    For the assessment of the dynamic concentration of exogenous protein or peptide in serum, an antibody- independent sandwich system based on the avidin-biotin interaction was established and validated. The exogenous protein or polypeptide labeled with biotin was added to the microplate coated with streptavidin, and followed by adding HRP-streptavidin to complete the sandwich system. This sandwich system was validated with respect to specificity, sensitivity, accuracy (recovery) and reproducibility. While the sensitivity of detection could reach to 0.3125 μg/L, the sensitivity and the range of detection can also be adjusted by changing coating concentration of streptavidin. Recoveries ranged from 97.82% to 107.29%, and the intra- and inter-assay variation was < 5.76% and < 8.42%, respectively. Thus, the well-validated streptavidin sandwich system was successfully applied to determine dynamic concentrations of Biotin-HAS (human serum albumin) and Biotin-OVM (Ovomucoid) in mouse serum, respectively. Most importantly, this novel method, where generation of antibody or radionuclide is not needed, may provide a new choice for assessment or monitoring of exogenous protein or peptide in pharmacokinetics study.%建立一种非抗体依赖的检测外源性蛋白多肽分子代谢动力学血清浓度及动态变化的新方法.链亲和素为捕获分子,加入待测生物素标记蛋白或合成肽,辣根过氧化物酶标记链亲和素为检测分子,构成链亲和素-生物素标记大分子-酶链亲和素的双夹心体系,并进行特异性、敏感性、准确性及稳定性评价.本检测体系灵敏度高,可达0.3125 μg/L,且可通过改变亲和素包被浓度调整检出敏感度与检测范围.准确性回收率为97.82%~107.92%,批内、批间变异系数分别<5.76%和<8.42%,并成功应用在生物素标记人血清白蛋白(biotin-HSA)与生物素标记鸡卵黏蛋白(biotin-OVM)的小鼠血清浓度动态检测.本方法不依赖抗体

  19. The proform of eosinophil major basic protein: a new maternal serum marker for adverse pregnancy outcome

    DEFF Research Database (Denmark)

    Pihl, Kasper; Larsen, Torben; Rasmussen, Steen;


    MBP median was significantly reduced in pregnancies with SGA (0.81 MoM), spontaneous preterm delivery (0.83 MoM), preeclampsia (0.88 MoM) and gestational hypertension (0.60 MoM). The best screening performance was found for preeclampsia including the covariates proMBP and nulliparity yielding an area under......OBJECTIVE: To establish the first trimester serum levels of the proform of eosinophil major basic protein (proMBP) in pregnancies with adverse outcome. Furthermore, to determine the screening performance using proMBP alone and in combination with other first trimester markers. METHODS: A case......-control study was conducted in a primary hospital setting. The proMBP concentration was measured in cases with small-for-gestational age (SGA) (n = 150), spontaneous preterm delivery (n = 88), preeclampsia (n = 40), gestational hypertension (n = 10) and in controls (n = 500). Concentrations were converted...

  20. Blood group and serum protein polymorphisms in turpu kapu population of vizianagaram district, Andhra Pradesh

    Directory of Open Access Journals (Sweden)

    V Komal Madhavi


    Full Text Available Data on two blood group and three serum protein polymorphisms of the Turpu Kapu, an endogamous population of Vizianagaram District, Andhra Pradesh (AP is presented. The gene frequencies for the blood group systems ABO and Rh are within the ranges of distribution reported earlier among the caste populations of Andhra Pradesh. The study population shows highest frequency of Hp1 allele and the lowest frequency of Hp2 allele compared to the other populations of AP. The Cp system is monomorphic, all individuals being the BB type. The GC system exhibits polymorphism with the gene frequencies of GC1 and GC2 alleles showing the highest and lowest frequencies, respectively, as compared to the caste populations reported earlier. The c2 test suggest that this population is in Hardy-Weinberg Equilibrium.

  1. Serum amyloid A and protein AA: molecular mechanisms of a transmissible amyloidosis. (United States)

    Westermark, Gunilla T; Westermark, Per


    Systemic AA-amyloidosis is a complication of chronic inflammatory diseases and the fibril protein AA derives from the acute phase reactant serum AA. AA-amyloidosis can be induced in mice by an inflammatory challenge. The lag phase before amyloid develops can be dramatically shortened by administration of a small amount of amyloid fibrils. Systemic AA-amyloidosis is transmissible in mice and may be so in humans. Since transmission can cross species barriers it is possible that AA-amyloidosis can be induced by amyloid in food, e.g. foie gras. In mice, development of AA-amyloidosis can also be accelerated by other components with amyloid-like properties. A new possible risk factor may appear with synthetically made fibrils from short peptides, constructed for tissue repair.

  2. Measurement of neonatal equine immunoglobulins for assessment of colostral immunoglobulin transfer: comparison of single radial immunodiffusion with the zinc sulfate turbidity test, serum electrophoresis, refractometry for total serum protein, and the sodium sulfite precipitation test. (United States)

    Rumbaugh, G E; Ardans, A A; Ginno, D; Trommershausen-Smith, A


    Four procedures for assessment of adequacy of colostral immunoglobulin (Ig) transfer in foals were evaluated. Results of zinc sulfate turbidity test, serum electrophoresis, total serum protein refractometry, and sodium sulfite precipitation test were compared with immunoglobulin G content determined by single radial immunodiffusion. The zinc sulfate turbidity test gave acceptable results for IgG, except that hemolyzed serum samples gave higher than expected values. A correction factor for hemolyzed serum was found to be useful. Serum electrophoresis was a satisfactory method of estimating IgG content. Total serum protein values may not be a valid basis for estimating IgG content, inasmuch as postsuckling total protein values were found to decrease in some foals in which passive transfer of IgG had been adequate. Sodium sulfite precipitation reactions were too unpredictable to be of value for determination of neonatal IgG concentration.

  3. Serum Angiopoietin-Like Protein 2 Concentrations Are Independently Associated with Heart Failure.

    Directory of Open Access Journals (Sweden)

    Chi-Lun Huang

    Full Text Available Angiopoietin-like protein 2 (ANGPTL2, which is mainly expressed from adipose tissue, is demonstrated to be involved in obesity, metabolic syndrome, and atherosclerosis. Because several adipocytokines are known to be associated with heart failure (HF, here we investigated the association of ANGPTL2 and HF in Taiwanese subjects.A total of 170 symptomatic HF patients and 130 age- and sex-matched controls were enrolled from clinic. The echocardiography was analyzed in each patient, and stress myocardial perfusion study was performed for clinical suspicion of coronary artery disease. Detailed demographic information, medications, and biochemical data were recorded. Circulating adipocytokines, including tumor necrosis factor-alpha (TNF-α, adiponectin, adipocyte fatty acid-binding protein (A-FABP and ANGPTL2, were analyzed. Compared with the control group subjects, serum ANGPTL2 concentrations were significantly higher in HF group patients. In correlation analyses, ANGPTL2 level was positively correlated to creatinine, fasting glucose, triglyceride, hsCRP, TNF-α, NT-proBNP and A-FABP levels, and negatively correlated with HDL-C and left ventricular ejection fraction. In multiple regression analysis, A-FABP, hsCRP, and HDL-C levels remained as independent predictors for ANGPTL2 level. To determine the association between serum ANGPTL2 concentrations and HF, multivariate logistic regression analyses were performed with subjects divided into tertiles by ANGPTL2 levels. For the subjects with ANGPTL2 levels in the highest tertile, their risk of HF was about 2.97 fold (95% CI = 1.24-7.08, P = 0.01 higher than those in the lowest tertile.Our results demonstrate a higher circulating ANGPTL2 level in patients with HF, and the upregulating ANGPTL2 levels might be associated with metabolic derangements and inflammation.

  4. Size-dependent interaction of silica nanoparticles with lysozyme and bovine serum albumin proteins (United States)

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


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

  5. Detection of kappa and lambda light chain monoclonal proteins in human serum: automated immunoassay versus immunofixation electrophoresis. (United States)

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


    Recently, turbidimetric immunoassays for detecting and quantifying kappa and lambda 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 kappa and lambda light chains compared to standard immunofixation electrophoresis. Sera demonstrating a restricted band of protein migration (other than a definite M spike) by serum protein electrophoresis (SPE), which may represent early monoclonal proteins, were also examined. When compared to IFE, percent agreement, sensitivity, and specificity for the kappa-FLC and lambda-FLC were 94.6, 72.9, and 99.5% and 98.5, 91.4, and 99.7%, respectively, in detecting monoclonal proteins involving free and heavy-chain-bound light chains. The majority of sera (73.7%) demonstrating a restricted band of protein migration on SPE demonstrated abnormal IFE patterns suggestive of multiple myeloma or monoclonal gammopathy of unknown significance, but gave normal kappa/lambda FLC ratios using the turbidimetric immunoassays. In conclusion, the kappa and lambda FLC assays are significantly less sensitive (72.9 to 91.4%) than IFE, but specific in detecting serum monoclonal proteins. Moreover, the kappa/lambda ratio has little value in routine screening since the majority of sera with abnormal IFE patterns had normal kappa/lambda FLC ratios.

  6. Comparative evaluation of serum and salivary immunoglobulin G and A levels with total serum protein in oral submucous fibrosis patients: A case control study

    Directory of Open Access Journals (Sweden)

    M Kandasamy


    Full Text Available Aim and Objective: The objective of this study is to estimate and compare the serum and salivary immunoglobulin G and A (IgG, IgA levels in various stages of oral submucous fibrosis (OSMF patients and relate it to total serum protein (TSP and hemoglobin (Hb levels. Materials and Methods: The sample for the present study comprised a total of 20 healthy controls, 20 OSMF patients. About 5 ml 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 cyanmethemoglobin methods, respectively. Results: Serum and salivary IgA and IgG levels were statistically significantly increased (P < 0.001 in OSMF patients when compared to controls. Also serum and salivary IgG and IgA levels showed significantly increased (P < 0.01 in all the three staging of OSMF when compared to control group. Hb levels and TSP levels were significantly decreased (P < 0.001 in OSMF patients when compared to controls. One-way ANOVA, Pearson's correlation, and unpaired t -test were used for statistical analysis. Conclusion: The elevated levels of IgG and IgA are also in favor of polygammapathy, which are nonspecific and nondiagnostic objective reflections of an underlying disease. Decreased TSP is a result of host response and Hb, acts as an indicator of nutritional status plays an important role. It is also observed from the present study that the severity of OSMF was directly proportional to the estimated elevated levels of the major IgG and IgA. A need is also felt for the knowledge of immunoprofile estimation in etiology and pathogenesis that would prove a great asset in the proper assessment of this condition.

  7. Prokaryotic expression, purification, and production of polyclonal antibody against novel human serum inhibited related protein I (SI1). (United States)

    Ma, Mingxing; Ma, Jie; Shi, Yinghui; Wu, Hong; Zhao, Wenxiu; Huang, Weiwei; Jiao, Yang; Tan, Deyong


    A novel serum inhibited related gene (SI1) has been cloned in our lab by using mRNA differential display analysis of U251 cells in the presence or absence of serum, the expression of SI1 was dramatically inhibited by the addition of serum to serum starved cells. Previous reports suggested the potential significance of SI1 in regulating the cell cycle. In this study, the plasmid construction, protein expression and purification, as well as the generation of anti-SI1 polyclonal antibody are described. A full-length cDNA of Si1 was inserted in a prokaryotic expression plasmid pET28-b(+) and efficiently expressed in E. coli Rosetta (DE3) strain after induction by isopropyl-b-D: -thiogalactoside. The expressed 6His-tagged SI1 fusion protein was purified by Ni(+) affinity column and then used to immunize Balb/C mice, and the anti-SI1 polyclonal antibody was purified by protein A column. To determine the sensitivity and specificity of the antibody against SI1, a cell lysate of pEGFP-N2-SI1 plasmid transiently transfected Hela cell was identified by anti-GFP monoclonal antibody and anti-SI1 polyclonal antibody. Both the GFP-SI1 fusion protein and endogenous SI1 protein in Hela cell can be recognized by the anti-SI1 polyclonal antibody. The anti-SI1 polyclonal antibody will provide a useful tool for further characterization of SI1.

  8. Comparing serum responses to acute feedings of an extensively hydrolyzed whey protein concentrate versus a native whey protein concentrate in rats: a metabolomics approach. (United States)

    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


    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 protein sources led to a dramatic increase in free fatty acids compared with control (up to 6-fold increases, q protein sources.

  9. Detection of Colorectal Cancer by Serum and Tissue Protein Profiling: A Prospective Study in a Population at Risk

    Directory of Open Access Journals (Sweden)

    Judith Y.M.N. Engwegen


    Full Text Available Colorectal cancer (CRC is the second most common cause of cancer-related death in Europe and its prognosis is largely dependent on stage at diagnosis. Currently, there are no suitable tumour markers for early detection of CRC. In a retrospective study we previously found discriminative CRC serum protein profiles with surface enhanced laser desorption ionisation—time of flight mass spectrometry (SELDI-TOF MS. We now aimed at prospective validation of these profiles. Additionally, we assessed their applicability for follow-up after surgery and investigated tissue protein profiles of patients with CRC and adenomatous polyps (AP. Serum and tissue samples were collected from patients without known malignancy with an indication for colonoscopy and patients with AP and CRC during colonoscopy. Serum samples of controls (CON; n = 359, patients with AP (n = 177 and CRC (n = 73, as well as tissue samples from AP (n = 52 and CRC (n = 47 were analysed as described previously. Peak intensities were compared by non-parametric testing. Discriminative power of differentially expressed proteins was assessed with support vector machines (SVM. We confirmed the decreased serum levels of apolipoprotein C-1 in CRC in the current population. No differences were observed between CON and AP. Apolipoprotein C-I levels did not change significantly within 1 month post-surgery, although a gradual return to normal levels was observed. Several proteins differed between AP and CRC tissue, among which a peak with similar mass as apolipoprotein C-1. This peak was increased in CRC compared to AP. Although we prospectively validated the serum decrease of apolipoprotein C-1 in CRC, serum protein profiles did not yield SVM classifiers with suitable sensitivity and specificity for classification of our patient groups.


    Institute of Scientific and Technical Information of China (English)

    覃舒文; 史轶蘩; 邓洁英


    Objective. To study the value of serum insulin-like growth factor binding protein-3 (IGFBP-3) levels in differential diagnosis of growth hormone deficiency (GHD). Methods. To measure serum IGFBP-3 levels by RIA in normal children and adolescents, GHD children and short-stature children without GHD. Results. Serum level of IGFBP-3 in 129 children with untreated GHD and with no pubertal development was 1.6± 0.9 mg/L, which was less than that in normal group of the same age, but overlapped with the normal children in Tanner stage I. After six -month treatment with recombinant human growth hormone (rhGH), serum level of IGFBP-3 in 59 GHD significantly increased from 1.3± 0.7 mg/L to 2.7± 0.9 mg/L, accompanied by an increase of body heights, growth velocities and serum level of IGF-1. Serum level of IGFBP-3 in 55 short-stature children without GHD was 3.3± 2.2 mg/L, which was not significantly different from that in normal group. Conclusion. Serum IGFBP-3 level can reflect the status of GH secretion in children with GHD and is a useful marker for differential diagnosis of GHD.

  11. Expression of Serum Retinol Binding Protein and Transthyretin within Mouse Gastric Ghrelin Cells.

    Directory of Open Access Journals (Sweden)

    Angela K Walker

    Full Text Available Ghrelin is an orexigenic peptide hormone produced mainly by a distinct group of dispersed endocrine cells located within the gastric oxyntic mucosa. Besides secreted gene products derived from the preproghrelin gene, which include acyl-ghrelin, desacyl-ghrelin and obestatin, ghrelin cells also synthesize the secreted protein nesfatin-1. The main goal of the current study was to identify other proteins secreted from ghrelin cells. An initial gene chip screen using mRNAs derived from highly enriched pools of mouse gastric ghrelin cells demonstrated high levels of serum retinol-binding protein (RBP4 and transthyretin (TTR, both of which are known to circulate in the bloodstream bound to each other. This high expression was confirmed by quantitative RT-PCR using as template mRNA derived from the enriched gastric ghrelin cell pools and from two ghrelin-producing cell lines (SG-1 and PG-1. RBP4 protein also was shown to be secreted into the culture medium of ghrelin cell lines. Neither acute nor chronic caloric restriction had a significant effect on RBP4 mRNA levels within stomachs of C57BL/6J mice, although both manipulations significantly decreased stomach TTR mRNA levels. In vitro studies using PG-1 cells showed no effect on RBP4 release of octanoic acid, epinephrine or norepinephrine, all of which are known to act directly on ghrelin cells to stimulate ghrelin secretion. These data provide new insights into ghrelin cell physiology, and given the known functions of RBP4 and TTR, support an emerging role for the ghrelin cell in blood glucose handling and metabolism.

  12. Polymorphism in SFTPD gene affects assembly and constitutional serum levels of surfactant protein D in a Lebanese population

    DEFF Research Database (Denmark)

    Fakih, Dalia; Chamat, Soulaima; Medlej-Hashim, Myrna;


    the distribution profile of serum constitutional SP-D in the Lebanese population and to investigate the genetic influence of single nucleotide polymorphism (SNP) rs721917 within SP-D gene (SFTPD) on its multimerization and its serum basal level. The C/T alleles at this SNP level lead respectively either...... to a threonine or methionine at position 11 of the SP-D protein. SP-D was measured by ELISA in serum taken from 97 healthy subjects. The p.Met11Thr SNP was genotyped. Different forms of SP-D were separated by gel filtration chromatography. Basal serum SP-D levels varied extensively, over a range of 108.3 ng...

  13. Expression of serum human epididymal secretory protein E4 at low grade and high grade serous carcinomas

    Institute of Scientific and Technical Information of China (English)

    Ya-Fei Zhu; Lin-Sheng He; Zhen-Dong Zhang; Qing-Shui Huang


    Objective: To investigate the value of serum human epididymis protein 4 (HE4) in differential diagnosis of patients with low-grade serous (LGSC) and high-grade serous carcinoma (HGSC) serous ovarian cancer. Methods: LGSC and HGSC serous ovarian cancer were diagnosed by the two-tier grade system, serum levels of HE4 and carbohydrate antigen 125 (CA125) were measured by ELISA and radioisotope method, respectively in 60 serous ovarian cancer patients. HE4 and TP53 protein in cancer tissue were measured by immunohistochemical method. Results: The difference in density of HE4 and TP53 protein was significant between LGSC and HGSC tissue, while serum CA125 did not show significant difference between different serum samples. There was significant difference in serum HE4 levels between LGSC and HGSC, and the result was different within FIGO (I+II) stage, suggesting HE4 was not a reliable biomarker for the discrimination between LGSC and HGSC. HE4 had potential as a biomarker for the discrimination between LGSC and HGSC but the role in early diagnosis was limited. Conclusions: HE4 may be a reliable marker for differential diagnosis of LGSC and HGSC. But its role in early diagnosis of LGSC and HGSC need to be confirmed from the perspective of two-tier grade system.

  14. Assessment of lectin and HILIC based enrichment protocols for characterization of serum glycoproteins by mass spectrometry

    DEFF Research Database (Denmark)

    Calvano, Cosima D; Zambonin, Carlo G; Jensen, Ole Nørregaard


    glycosylation profiles are associated with certain human ailments. Glycoprotein analysis by mass spectrometry of biological samples, such as blood serum, is hampered by sample complexity and the low concentration of the potentially informative glycopeptides and -proteins. We assessed the utility of lectin...... of 63 glycosylation sites in 38 proteins were identified by both methods, demonstrating distinct differences and complementarity. Serial application of custom-made microcolumns of mixed, immobilized lectins proved efficient for recovery and analysis of glycopeptides from serum samples of breast cancer...

  15. Yersinia enterocolitica serum resistance proteins YadA and ail bind the complement regulator C4b-binding protein.

    Directory of Open Access Journals (Sweden)

    Vesa Kirjavainen

    Full Text Available Many pathogens are equipped with factors providing resistance against the bactericidal action of complement. Yersinia enterocolitica, a Gram-negative enteric pathogen with invasive properties, efficiently resists the deleterious action of human complement. The major Y. enterocolitica serum resistance determinants include outer membrane proteins YadA and Ail. Lipopolysaccharide (LPS O-antigen (O-ag and outer core (OC do not contribute directly to complement resistance. The aim of this study was to analyze a possible mechanism whereby Y. enterocolitica could inhibit the antibody-mediated classical pathway of complement activation. We show that Y. enterocolitica serotypes O:3, O:8, and O:9 bind C4b-binding protein (C4bp, an inhibitor of both the classical and lectin pathways of complement. To identify the C4bp receptors on Y. enterocolitica serotype O:3 surface, a set of mutants expressing YadA, Ail, O-ag, and OC in different combinations was tested for the ability to bind C4bp. The studies showed that both YadA and Ail acted as C4bp receptors. Ail-mediated C4bp binding, however, was blocked by the O-ag and OC, and could be observed only with mutants lacking these LPS structures. C4bp bound to Y. enterocolitica was functionally active and participated in the factor I-mediated degradation of C4b. These findings show that Y. enterocolitica uses two proteins, YadA and Ail, to bind C4bp. Binding of C4bp could help Y. enterocolitica to evade complement-mediated clearance in the human host.

  16. Serum levels of growth hormone binding protein in children with normal and precocious puberty

    DEFF Research Database (Denmark)

    Juul, A; Fisker, Sidse; Scheike, Thomas Harder


    To study the regulation of GHBP serum levels by gonadal steroids in normal and precocious puberty.......To study the regulation of GHBP serum levels by gonadal steroids in normal and precocious puberty....

  17. Towards structure-based protein drug design. (United States)

    Zhang, Changsheng; Lai, Luhua


    Structure-based drug design for chemical molecules has been widely used in drug discovery in the last 30 years. Many successful applications have been reported, especially in the field of virtual screening based on molecular docking. Recently, there has been much progress in fragment-based as well as de novo drug discovery. As many protein-protein interactions can be used as key targets for drug design, one of the solutions is to design protein drugs based directly on the protein complexes or the target structure. Compared with protein-ligand interactions, protein-protein interactions are more complicated and present more challenges for design. Over the last decade, both sampling efficiency and scoring accuracy of protein-protein docking have increased significantly. We have developed several strategies for structure-based protein drug design. A grafting strategy for key interaction residues has been developed and successfully applied in designing erythropoietin receptor-binding proteins. Similarly to small-molecule design, we also tested de novo protein-binder design and a virtual screen of protein binders using protein-protein docking calculations. In comparison with the development of structure-based small-molecule drug design, we believe that structure-based protein drug design has come of age.

  18. Variation of Serum Levels of Glucose and Protein During the Day in Sheep of Different Sex

    Directory of Open Access Journals (Sweden)

    Carmen Galván Doria


    Full Text Available In order to establish the effect of sampling hour and sex on blood glucose levels and total protein, there were monitored six Colombian native sheep for 3 days. The blood samples were taken by jugular venipuncture, after asepsis and stored at -20°C to its subsequent processing through Spectrophotometry. The results were tabulated in Excel and analyzed using statistical program SAS 9.1. The mean values found were: glucose 75.57 ± 27.5 mg/dL for females and 83.70 ± 37.7 mg/dL for males; total protein 8.47 ± 2.76 g/dL for females and 7.14 ± 3.26 g/dL for males; albumin 4.29 ± 1.41 g/dL for females and 3.83 ± 1.46 g/dL for males; globulin 4.62 ± 2.35 g/dL for females and 3.92 ± 2.45 g/dL for males. The day and the hour of sampling affect serum concentrations (p < 0.05 of evaluated metabolites, also there was variation of themself regarding sex (p < 0.05.

  19. Developmental regulation of expression of C-reactive protein and serum amyloid A in Syrian hamsters. (United States)

    Dowton, S B; Waggoner, D J; Mandl, K D


    The fetal and maternal concentration of various plasma proteins alters during pregnancy. Cells in the livers of fetal hamsters accumulate serum amyloid A (SAA) and C-reactive protein (CRP) mRNA, major acute phase reactants, when lipopolysaccharide is administered to the fetal circulation. No fetal SAA or CRP mRNA response is seen when the mother is stimulated at a remote site by endotoxin or a nonspecific inflammatory agent. In addition, cells of the fetal hamster liver do not respond by accumulating SAA mRNA when exposed to the specific cytokines, tumor necrosis factor, IL-1, and IL-6. CRP mRNA levels increased in fetal livers after administration of tumor necrosis factor and IL-1. These data suggest that cells contained in the fetal liver can respond during an acute phase reaction but that the capacity of some acute phase reactant genes to respond to cytokines may be developmentally regulated. Studies of immature hamsters after birth show that the responses of CRP and SAA genes to lipopolysaccharide, tumor necrosis factor, IL-1, and IL-6 are reduced when compared with induction of mRNA accumulation for these acute phase reactants in adult animals.

  20. Evaluation of a type 1 diabetes serum cohort by SELDI-TOF MS protein profiling

    DEFF Research Database (Denmark)

    Albrethsen, J.; Kaas, A.; Schonle, E.;


    Proteomics analysis of serum from patients with type 1 diabetes (T1D) may lead to novel biomarkers for prediction of disease and for patient monitoring. However, the serum proteome is highly sensitive to sample processing and before proteomics biomarker research serum cohorts should preferably be...... at different times after diagnosis. Three members of the apolipoprotein family increased with time in patient serum collected 1, 6, and 12 months after diagnosis (ANOVA, p...

  1. Benefits and Limitations of Protein Hydrolysates as Components of Serum-Free Media for Animal Cell Culture Applications (United States)

    Lobo-Alfonso, Juliet; Price, Paul; Jayme, David

    Increased understanding of influential factors for the cultivation of animal cells, combined with heightened regulatory concern over potential transmission of adventitious contaminants associated with serum and other animal-derived components, has elevated interest in using protein hydrolysates as serum replacements or nutrient supplements. This paper reviews the chemistry and biology of various hydrolysates derived from animal, plant and microbial sources. It provides specific examples of a beneficial selection of plant and yeast hydrolysates as ingredients of serum-free nutrient formulations for bioproduction applications of cultured mammalian and insect cells. Strategies for customizing and optimizing nutrients for specialized applications and general benefits and limitations of protein hydrolysates for biopharmaceutical production are also discussed.

  2. Grafting of bovine serum albumin proteins on plasma-modified polymers for potential application in tissue engineering. (United States)

    Kasálková, Nikola Slepičková; Slepička, Petr; Kolská, Zdeňka; Hodačová, Petra; Kučková, Stěpánka; Svorčík, Václav


    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.

  3. The mechanisms of complement activation in normal bovine serum and normal horse serum against Yersinia enterocolitica O:9 strains with different outer membrane proteins content. (United States)

    Miętka, K; Brzostek, K; Guz-Regner, K; Bugla-Płoskońska, G


    Yersinia enterocolitica is a common zoonotic pathogen and facultative intracellular bacterium which can survive within blood cells. Cattle and horses are considered a reservoir of Y. enterocolitica which often causes several serious syndromes associated with yersiniosis such as abortions, premature births or infertility. The aim of our investigation was to determine the vitality of Y. enterocolitica O:9 strains (Ye9) in bovine and horse sera (NBS and NHrS) and explain the role of outer membrane proteins (OMPs) in serum resistance of these bacteria. Our previous studies demonstrated moderate human serum (NHS) resistance of the wild type Ye9 strain, whereas mutants lacking YadA, Ail or OmpC remained sensitive to the bactericidal activity of NHS. The present study showed that the wild type of Ye9 strain was resistant to the bactericidal activity of both NHrS and NBS, while Ye9 mutants lacking the YadA, Ail and OmpC proteins were sensitive to NHrS and NBS as well as to NHS. The mechanisms of complement activation against Ye9 strains lacking Ail and YadA were distinguished, i.e. activation of the classical/lectin pathways decisive in the bactericidal mechanism of complement activation of NBS, parallel activation of the classical/lectin and alternative pathways of NHrS. In this research the mechanism of independent activation of the classical/lectin or the alternative pathway of NBS and NHrS against Ye9 lacking OmpC porin was also established. The results indicate that serum resistance of Ye9 is multifactorial, in which extracellular structures, i.e. outer membrane proteins (OMPs) such as Ail, OmpC or YadA, play the main role.

  4. The interaction of platinum-based drugs with native biologically relevant proteins

    NARCIS (Netherlands)

    Brauckmann, Christine; Wehe, Christoph A.; Kieshauer, Michael; Lanvers-Kaminsky, Claudia; Sperling, Michael; Karst, Uwe


    This study focuses on the identification of the products that are formed upon binding of therapeutically relevant platinum complexes to proteins like beta-lactoglobulin A (LGA), human serum albumin (HSA), or human hemoglobin (HB). The respective proteins were incubated with the platinum-based antica

  5. Myelin protein zero and its antibody in serum as biomarkers of n-hexane-induced peripheral neuropathy and neurotoxicity effects

    Institute of Scientific and Technical Information of China (English)

    Jia Xiaowei; Liu Qingjun; Zhang Yanshu; Dai Yufei; Duan Huawei; Bin Ping; Niu Yong


    Background Chronic exposure to n-hexane can lead to peripheral neuropathy that no effective treatment regimen could be applied presently.This study investigated whether myelin protein zero (P0) protein and its antibody could be used to distinguish n-hexane intoxication and protect workers from peripheral neuropathy.Methods We compared P0 protein and its antibody among three levels of n-hexane-exposed groups,which included 18 patients with n-hexane-induced peripheral neuropathy as case group,120 n-hexane-exposed workers as n-hexaneexposed control group,and 147 non-hexane-exposed participants used as control group.ELISA method was applied to detect P0 protein and its antibody.Results P0 protein in serum was significantly higher in the case group and n-hexane-exposed control group in comparison with the control group (P<0.01).Compared with the n-hexane-exposed control group,the case group also had significant increase of P0 protein (P<0.01).After 6 months therapy,P0 protein was observed to decrease significantly in the case group (P<0.01).The P0 antibody in serum was significantly higher in the n-hexane-exposed control group than in the control group (P<0.01),but not significantly different between cases and controls.Conclusions P0 antibodies in serum may be a short-term effect biomarker for n-hexane exposure.P0 protein in serum may be an early effective biomarker for peripheral nerve neuropathy and its biological limit value needs investigation in the future study.

  6. Soy proteins and isoflavones reduce interleukin-6 but not serum lipids in older women: a randomized controlled trial. (United States)

    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


    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

  7. Assessment of 7.5% NaCl /6% Dextran-70 (HSD) Effects on Serum or Plasma Protein Determinations (United States)


    determined by modified Lowry, dye-binding, and an automated biuret method, as well as by refractometry , before, and at various times following HSD...protein concentrations determined by the biuret assay or refractometry when dextran serum concentrations exceeded 1.2 g/dl. The in vivo studies... refractometry , before, and at various times following HSD infusion in both euvolemic and hemorrhaged animals. Other studies analyzed plasma protein

  8. Serum leptin, C-reactive protein, and cancer mortality in the NHANES III. (United States)

    Wulaningsih, Wahyu; Holmberg, Lars; Ng, Tony; Rohrmann, Sabine; Van Hemelrijck, Mieke


    Adipokines, such as leptin, may affect cancer through its link with inflammation and obesity. We investigated the association between leptin, C-reactive protein, and risk of cancer death while accounting general and abdominal obesity. From the Third National Health and Examination Survey (NHANES III), we selected 5957 adult men and women with baseline measurements of serum leptin and CRP. Multivariable Cox regression was used to assess leptin and CRP levels (low, moderate, high) in relation to risk of cancer death. Stratification analyses were performed for obesity as defined by body mass index (BMI) and waist circumference. Fine and Gray regression was performed to account for death from cardiovascular disease and other causes as competing events. A total of 385 participants died of cancer during a mean follow-up of 18 years. After adjusting for BMI and waist circumference, an inverse association with log-transformed leptin was found for women, with a hazard ratio (HR) of 0.81 (95% confidence interval [CI]: 0.51-1.30) and 0.40 (95% CI: 0.24-0.68) for moderate and high compared to low levels of leptin, respectively; P(trend) = 0.0007). No association for leptin was observed in men, but higher CRP corresponded to increased risk of dying from cancer (HR: 2.98; 95% CI: 1.57-5.64 for the highest vs. lowest categories of CRP). Similar associations were observed with competing risk analysis also adjusted for BMI and waist circumference. Contrasting associations of serum leptin and CRP with cancer mortality may indicate sex-specific biological or environmental pathways linking obesity and cancer in men and women which warrant mechanistic investigations.

  9. Studies on the role of serum proteins in the ovarian function of the Tsetse fly, Glossina palpalis palpalis

    NARCIS (Netherlands)

    Takken, W.


    Nutritional factors in the diet of the obligatory haematophagous tsetse fly Glossina p. palpalis have been investigated, as an initial step towards the development of an artificial diet for this insect. Emphasis was laid on the role of serum proteins in the reproductive physiology, particularly with

  10. Acromegaly: the significance of serum total and free IGF-I and IGF-binding protein-3 in diagnosis

    NARCIS (Netherlands)

    A-J. van der Lely (Aart-Jan); W.W. de Herder (Wouter); J.A.M.J.L. Janssen (Joop); S.W.J. Lamberts (Steven)


    textabstractWe have studied the physiological and clinical relevance of measurements of serum total and free IGF-I and IGF-binding protein-3 (IGFBP-3) in 57 previously untreated patients with active acromegaly (32 males, 25 females; mean age 47 years) as compared with sex- and age-

  11. Evidence that kidney function but not type 2 diabetes determines retinol-binding protein 4 serum levels

    DEFF Research Database (Denmark)

    Henze, Andrea; Frey, Simone K; Raila, Jens


    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...

  12. High level of serum cholesteryl ester transfer protein inactive hepatitis C virus infection

    Institute of Scientific and Technical Information of China (English)


    AIM To determine the significance of cholesteryl estertransfer protein (CETP) in lipoprotein abnormalities inchronic hepatitis C virus (HCV) infection.METHODS: We evaluated the significance of the serumconcentration of CETP in 110 Japanese patients withchronic HCV infection. Fifty-five patients had active HCVinfection, and HCV eradication had been achieved in55. The role of CETP in serum lipoprotein abnormalities,specifically, in triglyceride (TG) concentrations in thefour major classes of lipoproteins, was investigatedusing Pearson correlations in conjunction with multipleregression analysis and compared them between thosewith active HCV infection and those in whom eradicationhad been achieved.RESULTS: The serum CETP levels of patients withactive HCV infection were significantly higher than thoseof patients in whom HCV eradication was achieved(mean ± SD, 2.84 ± 0.69 μg/mL vs 2.40 ± 1.00 μg/mL, P = 0.008). In multiple regression analysis, HCVinfection status (active or eradicated) was an independentfactor significantly associated with the serumCETP level. TG concentrations in low-density lipoprotein(mean ± SD, 36.25 ± 15.28 μg/mL vs 28.14 ± 9.94μg/mL, P = 0.001) and high-density lipoprotein (HDL)(mean ± SD, 25.9 ± 7.34 μg/mL vs 17.17 ± 4.82μg/mL, P 〈 0.001) were significantly higher in patients with active HCV infection than in those in whom HCVeradication was achieved. The CETP level was stronglycorrelated with HDL-TG in patients with active HCVinfection (R = 0.557, P 〈 0.001), whereas CETP wasnot correlated with HDL-TG in patients in whom HCVeradication was achieved (R = -0.079, P = 0.56).CONCLUSION: Our results indicate that CETP playsa role in abnormalities of lipoprotein metabolism inpatients with chronic HCV infection.

  13. The interaction of human serum albumin with selected lanthanide and actinide ions: Binding affinities, protein unfolding and conformational changes. (United States)

    Ali, Manjoor; Kumar, Amit; Kumar, Mukesh; Pandey, Badri N


    Human serum albumin (HSA), the most abundant soluble protein in blood plays critical roles in transportation of biomolecules and maintenance of osmotic pressure. In view of increasing applications of lanthanides- and actinides-based materials in nuclear energy, space, industries and medical applications, the risk of exposure with these metal ions is a growing concern for human health. In present study, binding interaction of actinides/lanthanides [thorium: Th(IV), uranium: U(VI), lanthanum: La(III), cerium: Ce(III) and (IV)] with HSA and its structural consequences have been investigated. Ultraviolet-visible, Fourier transform-infrared, Raman, Fluorescence and Circular dichroism spectroscopic techniques were applied to study the site of metal ions interaction, binding affinity determination and the effect of metal ions on protein unfolding and HSA conformation. Results showed that these metal ions interacted with carbonyl (CO..:)/amide(N..-H) groups and induced exposure of aromatic residues of HSA. The fluorescence analysis indicated that the actinide binding altered the microenvironment around Trp214 in the subdomain IIA. Binding affinity of U(VI) to HSA was slightly higher than that of Th(IV). Actinides and Ce(IV) altered the secondary conformation of HSA with a significant decrease of α-helix and an increase of β-sheet, turn and random coil structures, indicating a partial unfolding of HSA. A correlation was observed between metal ion's ability to alter HSA conformation and protein unfolding. Both cationic effects and coordination ability of metal ions seemed to determine the consequences of their interaction with HSA. Present study improves our understanding about the protein interaction of these heavy ions and their impact on its secondary structure. In addition, binding characteristics may have important implications for the development of rational antidote for the medical management of health effects of actinides and lanthanides.

  14. Predicting residual kidney function in hemodialysis patients using serum β-trace protein and β2-microglobulin. (United States)

    Wong, Jonathan; Sridharan, Sivakumar; Berdeprado, Jocelyn; Vilar, Enric; Viljoen, Adie; Wellsted, David; Farrington, Ken


    Residual kidney function (RKF) contributes significant solute clearance in hemodialysis patients. Kidney Diseases Outcomes Quality Initiative (KDOQI) guidelines suggest that hemodialysis dose can be safely reduced in those with residual urea clearance (KRU) of 2 ml/min/1.73 m(2) or more. However, serial measurement of RKF is cumbersome and requires regular interdialytic urine collections. Simpler methods for assessing RKF are needed. β-trace protein (βTP) and β2-microglobulin (β2M) have been proposed as alternative markers of RKF. We derived predictive equations to estimate glomerular filtration rate (GFR) and KRU based on serum βTP and β2M from 191 hemodialysis patients based on standard measurements of KRU and GFR (mean of urea and creatinine clearances) using interdialytic urine collections. These modeled equations were tested in a separate validation cohort of 40 patients. A prediction equation for GFR that includes both βTP and β2M provided a better estimate than either alone and contained the terms 1/βTP, 1/β2M, 1/serum creatinine, and a factor for gender. The equation for KRU contained the terms 1/βTP, 1/β2M, and a factor for ethnicity. Mean bias between predicted and measured GFR was 0.63 ml/min and 0.50 ml/min for KRU. There was substantial agreement between predicted and measured KRU at a cut-off level of 2 ml/min/1.73 m(2). Thus, equations involving βTP and β2M provide reasonable estimates of RKF and could potentially be used to identify those with KRU of 2 ml/min/1.73 m(2) or more to follow the KDOQI incremental hemodialysis algorithm.

  15. Detection of African Swine Fever Virus Antibodies in Serum and Oral Fluid Specimens Using a Recombinant Protein 30 (p30) Dual Matrix Indirect ELISA (United States)

    Giménez-Lirola, Luis G.; Mur, Lina; Rivera, Belen; Mogler, Mark; Sun, Yaxuan; Lizano, Sergio; Goodell, Christa; Harris, D. L. Hank; Rowland, Raymond R. R.; Gallardo, Carmina; Sánchez-Vizcaíno, José Manuel; Zimmerman, Jeff


    In the absence of effective vaccine(s), control of African swine fever caused by African swine fever virus (ASFV) must be based on early, efficient, cost-effective detection and strict control and elimination strategies. For this purpose, we developed an indirect ELISA capable of detecting ASFV antibodies in either serum or oral fluid specimens. The recombinant protein used in the ELISA was selected by comparing the early serum antibody response of ASFV-infected pigs (NHV-p68 isolate) to three major recombinant polypeptides (p30, p54, p72) using a multiplex fluorescent microbead-based immunoassay (FMIA). Non-hazardous (non-infectious) antibody-positive serum for use as plate positive controls and for the calculation of sample-to-positive (S:P) ratios was produced by inoculating pigs with a replicon particle (RP) vaccine expressing the ASFV p30 gene. The optimized ELISA detected anti-p30 antibodies in serum and/or oral fluid samples from pigs inoculated with ASFV under experimental conditions beginning 8 to 12 days post inoculation. Tests on serum (n = 200) and oral fluid (n = 200) field samples from an ASFV-free population demonstrated that the assay was highly diagnostically specific. The convenience and diagnostic utility of oral fluid sampling combined with the flexibility to test either serum or oral fluid on the same platform suggests that this assay will be highly useful under the conditions for which OIE recommends ASFV antibody surveillance, i.e., in ASFV-endemic areas and for the detection of infections with ASFV isolates of low virulence. PMID:27611939

  16. An Eco-Friendly Direct Injection HPLC Method for Methyldopa Determination in Serum by Mixed-Mode Chromatography Using a Single Protein-Coated Column. (United States)

    Emara, Samy; Masujima, Tsutomu; Zarad, Walaa; Kamal, Maha; Fouad, Marwa; El-Bagary, Ramzia


    A simple, rapid and environment-friendly direct injection HPLC method for the determination of methyldopa (MTD) in human serum has been developed and validated. The method was based on cleanup and separation of MTD from serum by mixed-mode liquid chromatography using a single protein-coated TSK gel ODS-80 TM analytical column (50 × 4.0 mm i.d., 5 µm). The protein-coated column exhibited excellent resolution, selectivity and functioned in two chromatographic modes: size-exclusion chromatography [i.e., solid-phase extraction (SPE) for serum proteins] and reversed-phase chromatography for the final separation of MTD. SPE and HPLC separation were carried out simultaneously with a green mobile phase consisting of acetate buffer (0.1 M, pH 2.4) at a flow rate of 1 mL/min and at room temperature (23 ± 1°C). The eluent was monitored at emission and excitation wavelengths of 320 and 270 nm, respectively. A calibration curve was linear over the range of 0.1-30 µg/mL with a detection limit of 0.027 µg/mL. This online SPE method was successfully applied to real samples obtained from patients receiving MTD therapy.

  17. Effect of Different Protein Levels On Nutrient Digestion Metabolism and Serum Biochemical Parameters in Calves

    Institute of Scientific and Technical Information of China (English)

    LI Hui; DIAO Qi-yu; ZHANG Nai-feng; TU Yan; WANG Ji-feng


    The current study has been performed to examine the effects of different dietary protein levels on the growth,nutrient digestion and absorption,as well as biochemical parameters in calves.Nine healthy newborn calves were selected,randomly divided into 3 groups and fed 3 milk replacers with different protein levels(18,22,and 26%),respectively.Five period-digestion-metabolism trials were taken between 12-20,22-30,32-40,42-50,and 52-60 days after birth.All 3 groups showed a similar growth curve during 11 to 61 experimental days,however,the growth rate of the 22%crude protein(CP) group was 8.89%higher than that of the 18%CP group and 19.48%higher than that of the 26%CP group.respectively. The apparent digestibility of dry material(DM)declined gradually with age,whereas,the apparent digestibility of N,extract ether(EE)rose slightly.Compared to the 22%CP and 26%CP group,calves fed with 18%CP apparently had a lower digestibility than DM,EE,and nitrogen(N).The average apparent digestibilities of N were 69.39,75.36,and 74.55%, respectively.Both the apparent digestibility and retention of calcium(Ca)and phosphorus(P)were steady throughout the experiment,but the average apparent digestibility of P in the 26%CP group was only 63.83%,which was markedly lower than that of the 18%CP group(70.40%)and 22%CP group(69.73%).In addition,the sernm concentrations of total protein(TP),albumin(ALB),and globulin(GLOB)of the 22%CP group were higher than those in the 18%CP and 26%CP groups.The urea N(BUN)content in the 18%CP group,on the other hand,was significantly lower than that of the other two groups.The highest glucose(GLU)concentration was found in the 22%CP group(5.38 mmol L-1),at the end of the trials.The protein levels in the milk replacer affected the digestion metabolism of nutrition and the serum biochemical parameters of calves at different physiological phases.Calves fed with 22%CP milk replacer had a better growth performance than the other groups.

  18. Impact of Elevated Hemoglobin and Serum Protein on Vasovagal Reaction from Blood Donation.

    Directory of Open Access Journals (Sweden)

    Takeshi Odajima

    Full Text Available We conducted a cross-sectional study to elucidate factors contributing to vasovagal reaction (VVR, the most frequent side effect following whole blood and apheresis donations. Complications recorded at the collection sites after voluntary donations by the Japanese Red Cross Tokyo Blood Center (JRC, in the 2006 and 2007 fiscal years, were analyzed by both univariate analysis and the multivariate conditional logistic regression model. Of 1,119,716 blood donations over the full two years, complications were recorded for 13,320 donations (1.18%, among which 67% were VVR. There were 4,303 VVR cases which had sufficient information and could be used for this study. For each VVR case, two sex- and age-matched controls (n = 8,606 were randomly selected from the donors without complications. Age, sex, body mass index (BMI, predonation blood pressure, pulse and blood test results, including total protein, albumin, and hemoglobin, were compared between the VVR group and the control group. In univariate analysis, the VVR group was significantly younger, with a lower BMI, higher blood pressure and higher blood protein and hemoglobin levels than the control group (p<0.001. Furthermore, blood protein and hemoglobin levels showed dose-dependent relationships with VVR incidences by the Cochran-Armitage trend test (p<0.01. For both sexes, after adjusting for confounders with the multivariate conditional logistic regression model, the higher than median groups for total protein (male: OR 1.97; 95%CI 1.76,-2.21; female: OR 2.29; 95%CI 2.05-2.56, albumin (male: 1.75; 1.55-1.96; female: 1.76; 1.57-1.97 and hemoglobin (male: 1.98; 1.76-2.22; female: 1.62; 1.45-1.81 had statistically significant higher risk of VVR compared to the lower than median groups. These elevated serum protein and hemoglobin levels might offer new indicators to help understand VVR occurrence.

  19. Impact of Elevated Hemoglobin and Serum Protein on Vasovagal Reaction from Blood Donation (United States)

    Tanba, Taiko; Yoshinaga, Kentaro; Motoji, Toshiko; Munakata, Masaya; Nakajima, Kazunori; Minami, Mutsuhiko


    We conducted a cross-sectional study to elucidate factors contributing to vasovagal reaction (VVR), the most frequent side effect following whole blood and apheresis donations. Complications recorded at the collection sites after voluntary donations by the Japanese Red Cross Tokyo Blood Center (JRC), in the 2006 and 2007 fiscal years, were analyzed by both univariate analysis and the multivariate conditional logistic regression model. Of 1,119,716 blood donations over the full two years, complications were recorded for 13,320 donations (1.18%), among which 67% were VVR. There were 4,303 VVR cases which had sufficient information and could be used for this study. For each VVR case, two sex- and age-matched controls (n = 8,606) were randomly selected from the donors without complications. Age, sex, body mass index (BMI), predonation blood pressure, pulse and blood test results, including total protein, albumin, and hemoglobin, were compared between the VVR group and the control group. In univariate analysis, the VVR group was significantly younger, with a lower BMI, higher blood pressure and higher blood protein and hemoglobin levels than the control group (p<0.001). Furthermore, blood protein and hemoglobin levels showed dose-dependent relationships with VVR incidences by the Cochran-Armitage trend test (p<0.01). For both sexes, after adjusting for confounders with the multivariate conditional logistic regression model, the higher than median groups for total protein (male: OR 1.97; 95%CI 1.76,-2.21; female: OR 2.29; 95%CI 2.05–2.56), albumin (male: 1.75; 1.55–1.96; female: 1.76; 1.57–1.97) and hemoglobin (male: 1.98; 1.76–2.22; female: 1.62; 1.45–1.81) had statistically significant higher risk of VVR compared to the lower than median groups. These elevated serum protein and hemoglobin levels might offer new indicators to help understand VVR occurrence. PMID:26894814

  20. Assessing the Association between Serum Ferritin, Transferrin Saturation, and C-Reactive Protein in Northern Territory Indigenous Australian Patients with High Serum Ferritin on Maintenance Haemodialysis

    Directory of Open Access Journals (Sweden)

    Sandawana William Majoni


    Full Text Available Objective. To determine the significance of high serum ferritin observed in Indigenous Australian patients on maintenance haemodialysis in the Northern Territory, we assessed the relationship between ferritin and transferrin saturation (TSAT as measures of iron status and ferritin and C-reactive protein (CRP as markers of inflammation. Methods. We performed a retrospective cohort analysis of data from adult patients (≥18 years on maintenance haemodialysis (>3 months from 2004 to 2011. Results. There were 1568 patients. The mean age was 53.9 (11.9 years. 1244 (79.3% were Indigenous. 44.2% (n=693 were male. Indigenous patients were younger (mean age [52.3 (11.1 versus 57.4 (15.2, p<0.001] and had higher CRP [14.7 mg/l (7–35 versus 5.9 mg/l (1.9–17.5, p<0.001], higher median serum ferritin [1069 µg/l (668–1522 versus 794.9 µg/l (558.5–1252.0, p<0.001], but similar transferrin saturation [26% (19–37 versus 28% (20–38, p=0.516]. We observed a small positive correlation between ferritin and TSAT (r2=0.11, p<0.001, no correlation between ferritin and CRP (r2 = 0.001, p<0.001, and positive association between high serum ferritin and TSAT (p<0.001, Indigenous ethnicity (p<0.001, urea reduction ratio (p=0.001, and gender (p<0.001 after adjustment in mixed regression analysis. Conclusion. Serum ferritin and TSAT may inadequately reflect iron status in this population. The high ferritin was poorly explained by inflammation.

  1. Assessing the Association between Serum Ferritin, Transferrin Saturation, and C-Reactive Protein in Northern Territory Indigenous Australian Patients with High Serum Ferritin on Maintenance Haemodialysis (United States)

    Lawton, Paul D.; Barzi, Federica; Cass, Alan; Hughes, Jaquelyne T.


    Objective. To determine the significance of high serum ferritin observed in Indigenous Australian patients on maintenance haemodialysis in the Northern Territory, we assessed the relationship between ferritin and transferrin saturation (TSAT) as measures of iron status and ferritin and C-reactive protein (CRP) as markers of inflammation. Methods. We performed a retrospective cohort analysis of data from adult patients (≥18 years) on maintenance haemodialysis (>3 months) from 2004 to 2011. Results. There were 1568 patients. The mean age was 53.9 (11.9) years. 1244 (79.3%) were Indigenous. 44.2% (n = 693) were male. Indigenous patients were younger (mean age [52.3 (11.1) versus 57.4 (15.2), p < 0.001]) and had higher CRP [14.7 mg/l (7–35) versus 5.9 mg/l (1.9–17.5), p < 0.001], higher median serum ferritin [1069 µg/l (668–1522) versus 794.9 µg/l (558.5–1252.0), p < 0.001], but similar transferrin saturation [26% (19–37) versus 28% (20–38), p = 0.516]. We observed a small positive correlation between ferritin and TSAT (r2 = 0.11, p < 0.001), no correlation between ferritin and CRP (r2 = 0.001, p < 0.001), and positive association between high serum ferritin and TSAT (p < 0.001), Indigenous ethnicity (p < 0.001), urea reduction ratio (p = 0.001), and gender (p < 0.001) after adjustment in mixed regression analysis. Conclusion. Serum ferritin and TSAT may inadequately reflect iron status in this population. The high ferritin was poorly explained by inflammation. PMID:28243472

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

    Directory of Open Access Journals (Sweden)

    Dan He

    Full Text Available To correlate serum level of monocyte chemoattractant protein-1 (MCP-1 with postoperative recurrence of spinal tuberculosis in the Chinese Han population.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.A total of 169 patients with spinal tuberculosis were included in the study and followed up for an average of 2.2 ± 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.Postoperative recurrence of spinal tuberculosis is likely to increase the serum level of MCP-1.

  3. Increased serum C-reactive protein level in Japanese patients of psoriasis with cardio- and cerebrovascular disease. (United States)

    Takahashi, Hidetoshi; Iinuma, Shin; Honma, Masaru; Iizuka, Hajime


    Psoriasis is a chronic inflammatory skin disease, which may be associated with metabolic syndrome accompanied by cardio- and cerebrovascular diseases. We investigated the relation between serum C-reactive protein (CRP) and cardio- and cerebrovascular diseases in Japanese psoriasis vulgaris patients. Ninety-seven psoriasis vulgaris patients and 79 healthy controls were assessed for serum CRP levels by immunoturbidimetry. The data were analyzed in terms of Psoriasis Area and Severity Index (PASI) scores, and comorbidity of cardio- and cerebrovascular disease and metabolic syndrome. Serum CRP levels in psoriasis vulgaris patients were significantly higher than those of healthy controls. There was no significant difference between male and female CRP levels in either psoriasis or healthy controls. No correlation was detected between PASI scores and serum CRP levels, either. Psoriasis with cardio- and cerebrovascular disease showed significantly higher CRP levels compared with those without the diseases. Furthermore, psoriasis with metabolic syndrome showed significantly higher serum CRP levels than those without the metabolic syndrome. In conclusion, serum CRP level is increased in psoriasis, and may be a useful marker for the prediction of the future risk of cardio- and cerebrovascular disease.

  4. Quantification of intermediate-abundance proteins in serum by multiple reaction monitoring mass spectrometry in a single-quadrupole ion trap. (United States)

    Lin, Shanhua; Shaler, Thomas A; Becker, Christopher H


    A method is presented to quantify intermediate-abundance proteins in human serum using a single-quadrupole linear ion trap mass spectrometer-in contrast, for example, to a triple-quadrupole mass spectrometer. Stable-isotope-labeled (tryptic) peptides are spiked into digested protein samples as internal standards, aligned with the traditional isotope dilution approach. As a proof-of-concept experiment, four proteins of intermediate abundance were selected, coagulation factor V, adiponectin, C-reactive protein (CRP), and thyroxine binding globulin. Stable-isotope-labeled peptides were synthesized with one tryptic sequence from each of these proteins. The normal human serum concentration ranges of these proteins are from 1 to 30 microg/mL (or 20 to 650 pmol/mL). These labeled peptides and their endogenous counterparts were analyzed by LC-MS/MS using multiple reaction monitoring, a multiplexed form of the selected reaction monitoring technique. For these experiments, only one chromatographic dimension (on-line reversed-phase capillary column) was used. Improved limits of detection will result with multidimensional chromatographic methods utilizing more material per sample. Standard curves of the spiked calibrants were generated with concentrations ranging from 3 to 700 pmol/mL using both neat solutions and peptides spiked into the complex matrix of digested serum protein solution where ion suppression effects and interferences are common. Endogenous protein concentrations were determined by comparing MS/MS peak areas of the endogenous peptides to the isotopically labeled internal calibrants. The derived concentrations from a normal human serum pool (neglecting loss of material during sample processing) were 9.2, 110, 120, and 246 pmol/mL for coagulation factor V, adiponectin, CRP, and thyroxine binding globulin, respectively. These concentrations generally agree with the reported normal ranges for these proteins. As a measure of analytical reproducibility of this

  5. Acute phase protein concentrations in serum and milk from healthy cows, cows with clinical mastitis and cows with extramammary inflammatory conditions

    NARCIS (Netherlands)

    Nielsen, B.H.; Jacobsen, S.; Andersen, P.H.; Niewold, T.A.; Heegaard, P.M.H.


    The concentrations of the two acute phase proteins, serum amyloid A and haptoglobin, in serum and milk were compared in 10 cows with clinical mastitis, 11 cows with extramammary inflammatory conditions and 10 clinically healthy control cows. The concentrations of both acute phase proteins were highe

  6. 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

    DEFF Research Database (Denmark)

    Pekkinen, Minna; Saarnio, Elisa; Viljakainen, Heli T.


    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 frac...

  7. A novel and cost effective method of removing excess albumin from plasma/serum samples and its impacts on LC-MS/MS bioanalysis of therapeutic proteins. (United States)

    Liu, Guowen; Zhao, Yue; Angeles, Aida; Hamuro, Lora L; Arnold, Mark E; Shen, Jim X


    We have developed an innovative method to remove albumin from plasma/serum samples for the LC-MS/MS quantitation of therapeutic proteins. Different combinations of organic solvents and acids were screened for their ability to remove albumin from plasma and serum samples. Removal efficiency was monitored by two signature peptides (QTALVELVK and LVNEVTEFAK) from albumin. Isopropanol with 1.0% trichloroacetic acid was found to be the most effective combination to remove albumin while retaining the protein of interest. Our approach was compared with a commercial albumin depletion kit on both efficiency of albumin removal and recovery of target proteins. We have demonstrated that our approach can remove 95% of the total albumin in human plasma samples while retaining close to 100% for two of three therapeutic proteins tested, with the third one at 60-80%. The commercial kit removed 98% of albumin but suffered at least 50% recovery loss for all therapeutic proteins when compared to our approach. Using BMS-C as a probe compound, the incorporation of the albumin removal approach has improved both assay sensitivity and ruggedness, compared to the whole plasma protein digestion approach alone. An LC-MS/MS method was developed and validated based on this new approach for the analysis of BMS-C in monkey serum. This assay was successfully applied to a toxicological study. When the albumin removal method was used in another clinical LC-MS/MS method, the sensitivity improved 10-fold to 50 ng/mL LLOQ comparing to a typical pellet digestion method.

  8. Protein standardization V: value transfer. A practical protocol for the assignment of serum protein values from a Reference Material to a Target Material. (United States)

    Blirup-Jensen, Søren; Johnson, A Myron; Larsen, Marianne


    We present a practical protocol for the assignment of values to serum proteins in a Target Material using a Reference Material. This protocol is based on the model of Direct Value Transfer between serum matrices and is intended to improve the value assignment of commercial calibrators using the Reference Material CRM 470 (now labeled ERM-DA 470) or similar reference materials. The procedure describes the general as well as the practical principles involved in the value assignment (with examples). The practical transfer protocol is based on multiple assays of 6 dilutions of the Reference Material and 6 dilutions of the Target Material. The transfer protocol requires several measurements a day repeated on several days, an important prerequisite being that all reconstitutions and dilutions are controlled by weighing thus reducing uncertainty in the transfer. In open systems that allow the use of the Reference Material as calibrator and the Target Material as samples, the proportionality of the two materials (the presence or absence of matrix effects) can now be directly assessed by evaluating a single regression plot. If no matrix effects are found, the regression line will pass through zero with a slope equal to the ratio of the concentrations of the two materials. In closed systems, the dedicated commercial calibrator has to be used as such; the Reference Material and the Target Material are now assayed as samples against this calibrator. Two regression plots are therefore obtained; if no matrix effects are present among the two materials and the calibrator, both the Reference and Target Materials will show zero intercepts, and the ratio of the two slopes will equal the ratio of the concentrations.

  9. The associations of diet with serum insulin-like growth factor I and its main binding proteins in 292 women meat-eaters, vegetarians, and vegans. (United States)

    Allen, Naomi E; Appleby, Paul N; Davey, Gwyneth K; Kaaks, Rudolf; Rinaldi, Sabina; Key, Timothy J


    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 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.

  10. Serum protein profile study of normal and cervical cancer subjects by high performance liquid chromatography with laser-induced fluorescence. (United States)

    Sujatha; Rai, Lavanya; Kumar, Pratap; Mahato, Krishna K; Kartha, Vasudevan B; Santhosh, Chidangil


    High performance liquid chromatography with high sensitivity laser-induced fluorescence detection is used to study the protein profiles of serum samples from healthy volunteers and cervical cancer subjects. The protein profiles are subjected to principal component analysis (PCA). PCA shows that the large number of chromatograms of a given class of serum samples--say normal/malignant--can be expressed in terms of a small number of factors (principal components). Three parameters--scores of the factors, squared residuals, and Mahalanobis distance--are derived from PCA. The parameters are observed to have a narrow range for protein profiles of standard calibration sets formed from groups of clinically confirmed normal/malignant classes. Limit tests using match/no match of the parameters of any test sample with parameters derived for the standard calibration sets give very good discrimination between malignant and normal samples with high sensitivity (approximately 100%) aand specificity (approximately 94%).

  11. Semi-automated competitive protein binding analysis of serum thyroxine on reusable Sephadex columns and its advantages over radioimmunoassay. (United States)

    Alexander, N M


    Competitive protein-binding analysis of serum thyroxine on small, reusable, Sephadex columns has been further studied and improved. The improved, semi-automated procedure results in reduced working time and costs. It has also been established that triiodothyronine crossreacts only 1/6 to 1/9 as well as thyroxine, and can be ignored because it represents only about 1/80 of the total serum iodothyronine content. The economic and methodological advantages of the improved method over radioammunoassay and other displacement assays are discussed.

  12. Insight into the Interaction of Graphene Oxide with Serum Proteins and the Impact of the Degree of Reduction and Concentration. (United States)

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


    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.

  13. Increased serum C-reactive protein level is associated with increased storage lower urinary tract symptoms in men with benign prostatic hyperplasia.

    Directory of Open Access Journals (Sweden)

    Shun-Fa Hung

    Full Text Available OBJECTIVE: Chronic inflammation is considered as one of the contributing mechanisms of lower urinary tract symptoms (LUTS. Serum C-reactive protein (CRP level is the widely used biomarker of inflammatory status. This study investigated the association between serum CRP level in men with benign prostatic hyperplasia (BPH and lower urinary tract symptoms (LUTS before and after medical treatment. METHODS: A total of 853 men with BPH and LUTS were enrolled. All patients completed the International Prostate Symptoms Score (IPSS questionnaire and urological examinations. The parameters of uroflowmetry (maximum flow rate, Qmax; voided volume, VV, post-void residual (PVR, total prostate volume (TPV and transition zone index (TZI, serum prostate specific antigen (PSA, and serum CRP levels were obtained. All patients were treated with alpha-blocker or antimuscarinic agent based on the IPSS voiding to storage subscore ratio (IPSS-V/S. Correlation analyses were performed between serum CRP levels with age, IPSS, TPV, TZI, Qmax, PVR, VV, PSA and between baseline and post treatment. RESULTS: The mean age was 66.9 ± 11.6 years old and the mean serum CRP levels were 0.31 ± 0.43 mg/dL. Univariate analyses revealed serum CRP levels were significantly associated with age (p<0.001, PSA levels (p = 0.005 and VV (p = 0.017, but not significantly associated with TPV (p = 0.854 or PVR (p = 0.068. CRP levels were positively associated with urgency (p<0.001 and nocturia (p<0.001 subscore of IPSS, total IPSS (p = 0.008 and storage IPSS (p<0.001 and negatively associated with IPSS- V/S ratio (p = 0.014. Multivariate analyses revealed that serum CRP levels were significantly associated with age (p = 0.004 and storage IPSS subscore p<0.001. Patients with IPSS-V/S<1 and treated with tolterodine for 3 months had significant decrease of CRP levels after treatment. CONCLUSION: Serum CRP levels are associated with storage LUTS and sensory bladder disorders, suggesting chronic

  14. Serum heat shock protein 60 in acute heart failure: a new biomarker? (United States)

    Bonanad, Clara; Núñez, Julio; Sanchis, Juan; Bodi, Vicente; Chaustre, Fabian; Chillet, Marifina; Miñana, Gema; Forteza, Maria José; Palau, Patricia; Núñez, Eduardo; Navarro, David; Llàcer, Angel; Chorro, Francisco J


    Heat shock protein 60 (HSP60) is a mitochondrial protein constitutively expressed in the majority of cells, and its expression is up-regulated by a variety of stressors. In heart failure, HSP60 is released from cardiomyocytes. The authors speculate that increased serum HSP60 (sHSP60) may be related to the severity of heart failure. This investigation sought to assess the association between sHSP60 and the composite end point of death/readmission in patients with acute heart failure (AHF). A total of 132 consecutive patients were admitted for AHF. The independent association between sHSP60 and the end point was assessed with Cox regression. During a median follow-up of 7 months (interquartile range, 3-14), 35 (26.5%) deaths, 40 (30.3%) readmissions, and 65 (49.2%) deaths/readmission were identified. Patients who exhibited the outcome showed higher median sHSP60 values (6.15 ng/mL [8.49] vs 4.71 ng/mL [7.55] P=.010). A monotonic increase in the incidence of the composite end point was observed when moving from lower to higher tertile (4.74, 4.76, and 6.98 per 10 patients-years of follow-up, P for trend <.001). After adjusting for established risk factors, only patients in the upper tertile showed an increased risk of death/readmission (hazard ratio, 2.63; 95% confidence interval, 1.29-5.37; P=.008). In patients with AHF, high sHSP60 was related to a higher risk for subsequent death/readmission for AHF.

  15. Serum levels of hypersensitive-C-reactive protein in moderate and severe acne

    Directory of Open Access Journals (Sweden)

    M R Namazi


    Full Text Available Background: Elevation of C-reactive protein (CRP has been reported to occur in psoriasis, urticaria, acne, rosacea and many other dermatological and nondermatological conditions. Chronic systemic inflammation has been implicated in the development of neuropsychiatric/degenerative disorders, atherosclerosis, coronary artery disease, diabetes mellitus and even carcinogenesis. The present study is designed to determine whether the level of inflammation created by acne vulgaris could be high enough to raise the serum levels of high-sensitive CRP. Materials and Methods: Forty-two patients with moderate and severe acne vulgaris were enrolled, along with 44 age and sex matched healthy blood donors as controls. Hypersensitive-CRP (Hs-CRP was measured in both groups. Results: Hypersensitive-C-reactive protein levels in the case group varied between 0 and 28.1 μg/ml with an average of 2.24 ± 4.87 μg/ml (mean ± standard deviation and a median of 0.6 μg/ml (interquartile range [IQR] =0.3, 1.4 μg/ml. Hs-CRP levels of the control group varied between 0 and 14 μg/ml with an average of 3.12 ± 3.67 μg/ml and a median of 1.5 μg/ml (IQR = 0.55, 5.0 μg/ml. No significant difference of Hs-CRP level between the two groups was seen (t = -0.961, 95% confidence interval: Lower = -2.6942, upper = 0.9377; P = 0.339. Additionally, no significant difference in the level of Hs-CRP was noted between the moderate and severe acne groups (95% confidence interval: Lower = -5.2495, upper = 1.6711; P = 0.165. Conclusion: Acne vulgaris, even in its severe grades (excluding acne fulminans and acne conglobata, does not induce significant inflammation at the systemic level.

  16. Serum amyloid P (female protein) of the Syrian hamster. Gene structure and expression. (United States)

    Rudnick, C M; Dowton, S B


    The structure and expression of the gene encoding serum amyloid P (SAP) component of the Syrian hamster have been studied by isolation of cosmid clones, nucleotide sequence analyses, and quantitation of nuclear run-on transcripts, nuclear RNA, mRNA, and protein levels. Hamster SAP, originally identified as female protein (FP), is a unique pentraxin because pretranslational expression of this gene is modulated by mediators of inflammation and sex steroids. SAP(FP) levels are high in sera from female hamsters and low in males. The response to inflammation is divergent; SAP(FP) levels decrease in females and increase in males during an acute phase response. The SAP(FP) gene encodes a 211 amino acid residue mature polypeptide as well as a 22-residue signal peptide. The intron/exon organization is similar to that of other pentraxins, but additional transcripts are generated from alternate polyadenylation sites in the 3' region. Circulating levels of SAP(FP) and the corresponding hepatic transcript levels are augmented by estrogen, while testosterone, dexamethasone, and progesterone cause a decrease in these levels. In addition the cytokines interleukin-1, -6, and tumor necrosis factor mediate a decrease in hepatic SAP(FP) transcript levels in female hamsters but did not cause a significant elevation of SAP(FP) mRNA in livers of male hamsters. The differences in expression of the SAP(FP) gene between male and female hamsters and between unstimulated male hamsters and male hamsters stimulated with an injection of lipopolysaccharide are due, at least in part, to alterations in transcription.

  17. Serum cartilage oligomeric matrix protein: is there a repeated bout effect?

    Directory of Open Access Journals (Sweden)

    Michael Behringer


    Full Text Available The primary aim of the present study was to investigate if there is a repeated bout effect for cartilage tissue, evident in the marker serum cartilage oligomeric matrix protein (sCOMP. Ten healthy male subjects (26.4±3.14 years performed two high impact interventions (100 drop jumps with a 30 second interval carried out at a 3 week interval. After each intervention, sCOMP and muscle soreness were assessed on 8 and 6 occasions respectively. Muscle soreness was determined via a visual analog scale with a maximum pain score of 10. sComp levels did not show a blunted response after the second bout (Bout 1: 12.2±3.3 U/L−1; Bout 2: 13.1±4.0 U/L−1; P>0.05. Remarkably, sCOMP increased from baseline levels by 16% after bout 1 and 15% after bout 2. Muscle soreness was blunted following the second intervention (Bout 1: 5.0±1.8; Bout 2: 1.6±0.8. Unlike the known repeated bout effect for muscle damage markers, sCOMP levels do not show a blunted response after two similar loading interventions. This information on biomarker behavior is essential to clinicians attempting to use this marker as an indicator of cartilage damage associated with the development or progression of osteoarthritis.

  18. Detection of pregnancy by radioimmunoassay of a pregnancy serum protein (PSP60) in cattle

    Energy Technology Data Exchange (ETDEWEB)

    Mialon, M.M.; Renand, G.; Camous, S.; Martal, J.; Menissier, F. (Institut National de Recherches Agronomiques (INRA), 78 - Jouy-en-Josas (France))


    The accuracy and efficiency of pregnancy diagnoses in cattle by pregnancy serum protein (PSP60) radioimmunoassay, a progesterone radioimmunoassay or oestrus detection were compared. Blood samples were taken from 349 suckling heifers and cows ( 1 191 inseminations) at 28, 35, 50 and 90 d post-insemination for PSP60 determination and at 22-23 d for progesterone. Females were declared nonpregnant when plasma PSP60 concentration was lower than 0.2 ng/ml at 28, 35 and 50 d and 0.5 ng/ml at 90 d. When compared with rectal palpation at 90 d, the accuracy of positive (negative) diagnoses by progesterone assay was 80% (100%) in heifers and 75% (99%) in cows. The accuracy of positive diagnoses by PSP60 assay increased with gestation stage from 90% on d 28 in heifers (74% in cows) to 100% (99% in cows) at the time of rectal palpation. This accuracy was 84% on d 28 in cows when the interval from calving to blood sampling was higher than 115 d. Whatever the stage, the accuracy of negative diagnoses was higher than 90%. Efficiency in detecting pregnant or nonpregnant females on d 28 was equivalent to the progesterone assay. The method for detecting oestrus applied in this experiment was as efficient as the PSP60 or progesterone test at any stage of gestation. The PSP60 test is very flexible, which makes its use particularly interesting in naturally mated suckling herds because of the uncertainty regarding the date of fertilization. (authors).

  19. Haemophilus influenzae P4 Interacts With Extracellular Matrix Proteins Promoting Adhesion and Serum Resistance. (United States)

    Su, Yu-Ching; Mukherjee, Oindrilla; Singh, Birendra; Hallgren, Oskar; Westergren-Thorsson, Gunilla; Hood, Derek; Riesbeck, Kristian


    Interaction with the extracellular matrix (ECM) is one of the successful colonization strategies employed by nontypeable Haemophilus influenzae (NTHi). Here we identified Haemophilus lipoprotein e (P4) as a receptor for ECM proteins. Purified recombinant P4 displayed a high binding affinity for laminin (Kd = 9.26 nM) and fibronectin (Kd = 10.19 nM), but slightly less to vitronectin (Kd = 16.51 nM). A P4-deficient NTHi mutant showed a significantly decreased binding to these ECM components. Vitronectin acquisition conferred serum resistance to both P4-expressing NTHi and Escherichia coli transformants. P4-mediated bacterial adherence to pharynx, type II alveolar, and bronchial epithelial cells was mainly attributed to fibronectin. Importantly, a significantly reduced bacterial infection was observed in the middle ear of the Junbo mouse model when NTHi was devoid of P4. In conclusion, our data provide new insight into the role of P4 as an important factor for Haemophilus colonization and subsequent respiratory tract infection.

  20. Functional analysis of the larval serum protein gene promoter from silkworm,Bombyx mori.

    Institute of Scientific and Technical Information of China (English)

    TANG Shunming; YI Yongzhu; SHEN Xingjia; ZHANG Zhifang; LI Yiren; HE Jialu


    The regulation region of larval serum protein gene, Bombyx mori. (BmLSP), consisting of the first intron, the first exon, the central promoter region and 5′-upstream region, is cloned from genomic DNA from the silkworm variety of Suju×Minghu. Using PCR and restriction endonuclease methods, a series of luciferase reporter plasmids, driven by different length of BmLSP promoters, are constructed. Via the transient expression system in BmN cells, the effects of the regulation elements and foreign insect hormones on the BmLSP promoter activity are investigated. The results demonstrate that the promoter activity of BmLSP is 5.8- or 4.4-fold higher than that of BmLSPs whose first intron or the element in 5′-upstream region harboring the homologous sequence with the first intron of light-chain fibroin gene (EHIF) is deleted, respectively, suggesting that both the first intron and EHIF contain the main positive cis-acting elements. However, the inactive mariner transposable element (MTE) in 5′-upstream region presents a negative effect. Furthermore, the effects of juvenile hormone analogue (JHA) on the BmLSP promoter activity show a typical dose-dependent manner, that is, low concentration treatments increase the BmLSP promoter activity and high concentration treatments decrease it. Meanwhile, insect ecdysone (MH) treatments present no significant effect.

  1. Knowledge Transposition from Tropical Fish Serum Proteins to Fundamental Education Students Through Biochemical Models

    Directory of Open Access Journals (Sweden)

    E.V.M. Maciel de Carvalho


    Full Text Available The subject was represented and discussed at The National Week of Science and Technology, UFPE, an initiative from The Ministry of Science and Technology to encourage children and people in science and technology activities. The work aimed to renew the importance to transmit knowledge from simple, imaginative, biochemical models and interactive teaching. The stand tool contained an aquarium with fishes, five scale models showing peptide bond, carbohydrate inhibited lectin molecule, hemagglutination reaction, lectin-bacterium surface interaction and enzyme-substract-inhibitor. Posters described tropical fish importance and methods applied to obtain fish serum and organs to purify lectins and protein inhibitors as well as to extract tissue DNA; notions were transmitted on fish immunology and diseases. The students were attracted and impressed with the exotic fishes most cultivated in Brazil; they asked if it is necessary to kill the fish to extract lectin and about lectin importance. Students were also interested to know if all fish enzyme/inhibitors are favorable to the own fish organism. The work succeeded to inform and stimulate future scientists in the field and to awake their scientific curiosity.

  2. The elevation of serum napsin A in idiopathic pulmonary fibrosis, compared with KL-6, surfactant protein-A and surfactant protein-D

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    Samukawa Takuya


    Full Text Available Abstract Background Napsin A, an aspartic protease, is mainly expressed in alveolar type-II cells and renal proximal tubules and is a putative immunohistochemical marker for pulmonary adenocarcinomas. This study sought to determine whether napsin A could be measured in the serum to evaluate its relationship to idiopathic pulmonary fibrosis (IPF and determine whether renal dysfunction might affect serum napsin A levels. Methods Serum levels of napsin A were measured in 20 patients with IPF, 34 patients with lung primary adenocarcinoma, 12 patients with kidney diseases, and 20 healthy volunteers. Surfactant protein (SP-A, SP-D, and Krebs von den Lungen-6 (KL-6 levels in serum and pulmonary function tests were also evaluated in IPF patients. Results Circulating levels of napsin A were increased in patients with IPF, as compared with healthy controls, and they correlated with the severity of disease. Moreover, the serum napsin A levels were not elevated in patients with pulmonary adenocarcinoma or renal dysfunction. The distinguishing point between IPF and the controls was that the area under the receiver operating characteristic curve (ROC of napsin A was larger than that of KL-6, SP-A, or SP-D. Conclusion These findings suggest that serum napsin A may be a candidate biomarker for IPF.

  3. The associations between serum adiponectin, leptin, C-reactive protein, insulin, and serum long-chain omega-3 fatty acids in Labrador Retrievers

    Directory of Open Access Journals (Sweden)

    Streeter RM


    Full Text Available Renee M Streeter,1 Angela M Struble,1 Sabine Mann,2 Daryl V Nydam,2 John E Bauer,3 Marta G Castelhano,1 Rory J Todhunter,1 Bethany P Cummings,4 Joseph J Wakshlag11Department of Clinical Sciences, 2Department of Population Medicine, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA; 3Department of Clinical Sciences, Texas A&M University, College Station, TX, USA; 4Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, USAAbstract: Obesity has been associated with an increased inflammatory response and insulin resistance due to adipose tissue–derived adipokines and increases in C-reactive protein (CRP. Dogs appear to be similar to other species with the exception of adiponectin, which might not be affected by obesity status. Serum long-chain polyunsaturated fatty acid concentrations have been positively and negatively associated with serum adipokines. The aim of the study was to examine the relationship between leptin, CRP, adiponectin, and insulin to body condition score (BCS and to the long-chain omega-3 fatty acids in serum lipoproteins, including alpha-linolenic acid, eicosapentaenoic acid (EPA, docosapentanenoic acid (DPA, and docosahexaenoic acid (DHA as a reflection of dietary omega-3 status in the Labrador Retriever. Seventy-seven Labrador Retrievers were evaluated for BCS, percent fasting serum lipoprotein fatty acid concentrations, as well as serum leptin, adiponectin, insulin, and CRP. A multivariable general linear regression model was constructed to examine the association between the dependent variables leptin, CRP, adiponectin, and insulin and the predictor variables of BCS, age, and sex, as well as concentrations of alpha-linolenic acid, EPA, DHA, and DPA. Adiponectin concentration was positively associated with age (P<0.0008, EPA (P=0.027 and negatively associated with DHA (P=0.008. Leptin concentration was positively associated with an increased DHA (P=0.009, BCS (P

  4. Acute phase protein concentrations in serum and milk from healthy cows, cows with clinical mastitis and cows with extramammary inflammatory conditions

    DEFF Research Database (Denmark)

    Nielsen, B.H.; Jacobsen, S.; Andersen, P.H.


    The concentrations of the two acute phase proteins, serum amyloid A and haptoglobin, in serum and milk were compared in 10 cows with clinical mastitis, 11 cows with extramammary inflammatory conditions and 10 clinically healthy control cows. The concentrations of both acute phase proteins were...... higher in the serum and milk of the cows with mastitis than in the cows in the other two groups. Four of the cows with extramammary inflammatory conditions had serum amyloid A concentrations in serum above 100 mug/ml, but negligible concentrations in milk, indicating that a pathogen must be present...... in the mammary gland for serum amyloid A to accumulate in milk. The acute phase protein concentrations in milk increased significantly with increasing somatic cell count, suggesting that they may be indicators of the severity of an infection....

  5. In vitro and in vivo interactions of selected nanoparticles with rodent serum proteins and their consequences in biokinetics

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    Wolfgang G. Kreyling


    Full Text Available When particles incorporated within a mammalian organism come into contact with body fluids they will bind to soluble proteins or those within cellular membranes forming what is called a protein corona. This binding process is very complex and highly dynamic due to the plethora of proteins with different affinities and fractions in different body fluids and the large variation of compounds and structures of the particle surface. Interestingly, in the case of nanoparticles (NP this protein corona is well suited to provide a guiding vehicle of translocation within body fluids and across membranes. This NP translocation may subsequently lead to accumulation in various organs and tissues and their respective cell types that are not expected to accumulate such tiny foreign bodies. Because of this unprecedented NP accumulation, potentially adverse biological responses in tissues and cells cannot be neglected a priori but require thorough investigations. Therefore, we studied the interactions and protein binding kinetics of blood serum proteins with a number of engineered NP as a function of their physicochemical properties. Here we show by in vitro incubation tests that the binding capacity of different engineered NP (polystyrene, elemental carbon for selected serum proteins depends strongly on the NP size and the properties of engineered surface modifications. In the following attempt, we studied systematically the effect of the size (5, 15, 80 nm of gold spheres (AuNP, surface-modified with the same ionic ligand; as well as 5 nm AuNP with five different surface modifications on the binding to serum proteins by using proteomics analyses. We found that the binding of numerous serum proteins depended strongly on the physicochemical properties of the AuNP. These in vitro results helped us substantially in the interpretation of our numerous in vivo biokinetics studies performed in rodents using the same NP. These had shown that not only the

  6. Human serum albumin adsorption on TiO2 from single protein solutions and from plasma. (United States)

    Sousa, S R; Moradas-Ferreira, P; Saramago, B; Melo, L Viseu; Barbosa, M A


    In the present work, the adsorption of human serum albumin (HSA) on commercially pure titanium with a titanium oxide layer formed in a H(2)O(2) solution (TiO(2) cp) and on TiO(2) sputtered on Si (TiO(2) sp) was analyzed. Adsorption isotherms, kinetic studies, and work of adhesion determinations were carried out. HSA exchangeability was also evaluated. Surface characterization was performed by atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), and wettability studies. The two TiO(2) surfaces have very distinct roughnesses, the TiO(2) sp having a mean R(a) value 14 times smaller than the one of TiO(2) cp. XPS analysis revealed consistent peaks representative of TiO(2) on sputtered samples as well as on Ti cp substrate after 48 h of H(2)O(2) immersion. Nitrogen was observed as soon as protein was present, while sulfur, present in disulfide bonds in HSA, was observed for concentrations of protein higher than 0.30 mg/mL. The work of adhesion was determined from contact angle measurements. As expected from the surface free energy values, the work of adhesion of HSA solution is higher for the TiO(2) cp substrate, the more hydrophilic one, and lower for the TiO(2) sp substrate, the more hydrophobic one. The work of adhesion between plasma and the substrates assumed even higher values for the TiO(2) cp surface, indicating a greater interaction between the surface and the complex protein solutions. Adsorption studies by radiolabeling of albumin ((125)I-HSA) suggest that rapid HSA adsorption takes place on both surfaces, reaching a maximum value after approximately 60 min of incubation. For the higher HSA concentrations in solution, a multilayer coverage was observed on both substrates. After the adsorption step from single HSA solutions, the exchangeability of adsorbed HSA molecules by HSA in solution was evaluated. The HSA molecules adsorbed on TiO(2) sp seem to be more easily exchanged by HSA itself than those adsorbed on TiO(2) cp after 24 h. In

  7. Serum sex hormone and growth arrest-specific protein 6 levels in male patients with coronary heart disease

    Directory of Open Access Journals (Sweden)

    Rui Zhao


    Full Text Available Epidemiological studies have shown a high prevalence of low serum testosterone levels in men with cardiovascular disease. Moreover, the tyrosine kinase receptor Axl, the ligand of which is growth arrest-specific protein 6 (GAS6, is expressed in the vasculature, and serum GAS6 levels are associated with endothelial dysfunction and cardiovascular events. Testosterone regulates GAS6 gene transcription directly, which inhibits calcification of vascular smooth muscle cells and provides a mechanistic insight into the cardioprotective action of androgens. This study was designed to determine the correlation between serum GAS6 and testosterone levels in male patients with coronary heart disease (CHD. We recruited 225 patients with CHD and 102 apparently healthy controls. Serum concentrations of GAS6 and soluble Axl were quantified by an enzyme-linked immunosorbent assay. Levels of high-sensitivity C-reactive protein, testosterone, estradiol, and other routine biochemical markers were also measured. Testosterone decreased from 432.69 ± 14.40 to 300.76 ± 6.23 ng dl−1 (P < 0.001 and GAS6 decreased from 16.20 ± 0.31 to 12.51 ± 0.19 ng ml−1 (P < 0.001 in patients with CHD, compared with control subjects. Multiple linear regression analysis showed that serum testosterone and GAS6 levels were positively associated in male patients with CHD. Alterations in GAS6 levels may influence the development of CHD. Downregulation of GAS6/Axl signaling in the presence of low sex hormone levels during disease progression is a potential mechanism by which GAS6 affects CHD. This study provides novel results regarding the influence of sex hormones on serum GAS6 levels in patients with CHD.

  8. Protein-protein interaction based on pairwise similarity

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    Zaki Nazar


    Full Text Available Abstract Background Protein-protein interaction (PPI is essential to most biological processes. Abnormal interactions may have implications in a number of neurological syndromes. Given that the association and dissociation of protein molecules is crucial, computational tools capable of effectively identifying PPI are desirable. In this paper, we propose a simple yet effective method to detect PPI based on pairwise similarity and using only the primary structure of the protein. The PPI based on Pairwise Similarity (PPI-PS method consists of a representation of each protein sequence by a vector of pairwise similarities against large subsequences of amino acids created by a shifting window which passes over concatenated protein training sequences. Each coordinate of this vector is typically the E-value of the Smith-Waterman score. These vectors are then used to compute the kernel matrix which will be exploited in conjunction with support vector machines. Results To assess the ability of the proposed method to recognize the difference between "interacted" and "non-interacted" proteins pairs, we applied it on different datasets from the available yeast saccharomyces cerevisiae protein interaction. The proposed method achieved reasonable improvement over the existing state-of-the-art methods for PPI prediction. Conclusion Pairwise similarity score provides a relevant measure of similarity between protein sequences. This similarity incorporates biological knowledge about proteins and it is extremely powerful when combined with support vector machine to predict PPI.

  9. Serum Amyloid A Protein Concentration in Blood is Influenced by Genetic Differences in the Cheetah (Acinonyx jubatus). (United States)

    Franklin, Ashley D; Schmidt-Küntzel, Anne; Terio, Karen A; Marker, Laurie L; Crosier, Adrienne E


    Systemic amyloid A (AA) amyloidosis is a major cause of morbidity and mortality among captive cheetahs. The self-aggregating AA protein responsible for this disease is a byproduct of serum amyloid A (SAA) protein degradation. Transcriptional induction of the SAA1 gene is dependent on both C/EBPβ and NF-κB cis-acting elements within the promoter region. In cheetahs, 2 alleles exist for a single guanine nucleotide deletion in the putative NF-κB binding site. In this study, a novel genotyping assay was developed to screen for the alleles. The results show that the SAA1A (-97delG) allele is associated with decreased SAA protein concentrations in the serum of captive cheetahs (n = 58), suggesting genetic differences at this locus may be affecting AA amyloidosis prevalence. However, there was no significant difference in the frequency of the SAA1A (-97delG) allele between individuals confirmed AA amyloidosis positive versus AA amyloidosis negative at the time of necropsy (n = 48). Thus, even though there is evidence that having more copies of the SAA1A (-97delG) allele results in a potentially protective decrease in serum concentrations of SAA protein in captive cheetahs, genotype is not associated with this disease within the North American population. These results suggest that other factors are playing a more significant role in the pathogenesis of AA amyloidosis among captive cheetahs.

  10. Determinants of serum concentrations of lipopolysaccharide-binding protein (LBP in the adult population: the role of obesity.

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    Arturo Gonzalez-Quintela

    Full Text Available BACKGROUND AND AIM: Assessment of serum concentration of lipopolysaccharide (LPS-binding protein (LBP has been suggested as a useful biomarker to indicate activation of innate immune responses to microbial products. We investigated LBP concentrations and associations with demographics, lifestyle factors, and common metabolic abnormalities in adults. We also examined if LBP concentrations were associated with common polymorphisms in genes coding for LBP (rs2232618, CD14 (rs2569190, and TLR4 (rs4986790, the molecules responsible for the innate immune response to LPS, or serum levels of soluble CD14 (sCD14 and proinflammatory cytokines. METHODS: Serum LBP was measured with a commercial immunoassay in a random sample of the adult population (n = 420, 45% males, age 18-92 years from a single municipality. RESULTS: Serum LBP concentrations increased with age (P<0.001 and were higher in individuals who were overweight or obese than in normal-weight individuals (P<0.001. Similarly, LBP concentrations were higher in individuals with metabolic syndrome than in individuals without it (P<0.001. Among metabolic syndrome components, LBP concentrations were independently associated with abdominal obesity (P = 0.002 and low concentrations of HDL-cholesterol (P<0.001. Serum LBP concentrations tended to be independently associated with smoking (P = 0.05, but not with alcohol consumption. Likewise, there was not significant association between LBP concentrations and gene polymorphisms. Concentrations of LBP significantly correlated with serum levels of proinflammatory cytokines (IL-6 and IL-8, sCD14, and with liver enzymes. CONCLUSIONS: Serum LBP concentrations increased with age. Overweight, obesity, and having metabolic syndrome (particularly, low HDL cholesterol levels were associated with higher LBP concentrations. These findings are consistent with microbial exposure playing a role in these inflammatory, metabolic abnormalities.

  11. Identification of Serum Monocyte Chemoattractant Protein-1 and Prolactin as Potential Tumor Markers in Hepatocellular Carcinoma

    NARCIS (Netherlands)

    Wang, Who-Whong; Ang, Soo Fan; Kumar, Rajneesh; Heah, Charmain; Utama, Andi; Tania, Navessa Padma; Li, Huihua; Tan, Sze Huey; Poo, Desmond; Choo, Su Pin; Chow, Wan Cheng; Tan, Chee Kiat; Toh, Han Chong


    Early diagnosis of hepatocellullar carcinoma (HCC) remains a challenge. The current practice of serum alpha-fetoprotein (AFP) measurement is inadequate. Here we utilized a proteomic approach to identify novel serum biomarkers for distinguishing HCC patients from non-cancer controls. We profiled the

  12. Proteinuria: The diagnostic strategy based on urine proteins differentiation

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    Stojimirović Biljana B.


    Full Text Available Basal glomerular membrane represents mechanical and electrical barrier for passing of the plasma proteins. Mechanical barrier is composed of cylindrical pores and filtration fissure, and negative layer charge in exterior and interior side of basal glomerular membrane, made of heparan sulphate and sialoglicoproteine, provides certain electrical barrier. Diagnostic strategy based on different serum and urine proteins enables the differentiation of various types of proteinuria. Depending on etiology of proteinuria it can be prerenal, renal and postrenal. By analyzing albumin, armicroglobulin, immunoglobulin G and armacroglobulin, together with total protein in urine, it is possible to detect and differentiate causes of prerenal, renal (glomerular, tubular, glomerulo-tubular and postrenal proteinuria. The adequate and early differentiation of proteinuria type is of an immense diagnostic and therapeutic importance.

  13. Bovine serum albumin surface imprinted polymer fabricated by surface grafting copolymerization on zinc oxide rods and its application for protein recognition. (United States)

    Li, Xiangjie; Zhou, Jingjing; Tian, Lei; Li, Wei; Zhang, Baoliang; Zhang, Hepeng; Zhang, Qiuyu


    A novel bovine serum albumin (BSA) surface imprinted polymer based on ZnO rods was synthesized by surface grafting copolymerization. It exhibited an excellent recognition performance to bovine serum albumin. The adsorption capacity and imprinting factor of bovine serum albumin could reach 89.27 mg/g and 2.35, respectively. Furthermore, the fluorescence property of ZnO was used for tracing the process of protein imprinting and it implied the excellent optical sensing property of this material. More importantly, the hypothesis that the surface charge of carrier could affect the imprinting process was confirmed. That is, ZnO with positive surface charge could not only improve the recognition specificity of binding sites to template proteins (pI 7). It was also important that the reusability of ZnO@BSA molecularly imprinted polymers was satisfactory. This implied that the poor mechanical/chemical stability of traditional zinc oxide sensors could be solved by the introduction of surface grafting copolymerization. These results revealed that the ZnO@BSA molecularly imprinted polymers are a promising optical/electrochemical sensor element.

  14. Isoprenaline increases serum levels of monocyte chemoattractant protein-1 and tissue inhibitor of matrix metalloproteinases type Ⅰ in Wistar rats

    Institute of Scientific and Technical Information of China (English)

    LEI He-ping; ZHANG Meng-zhen; YANG Xiang-yu; HOU Xing-hua; LIN Qiu-xiong; YANG Min; ZHONG Shi-long


    Background Treatment of rats with the beta-adrenergic agonist Isoprenaline (ISO) results in cardiac hypertrophy and myocardial fibrosis.In the present work,we aimed to study the in vivo effects of ISO on serum levels of monocyte chemoattractant protein-1 and tissue inhibitor of matrix metalloproteinases type Ⅰ in Wistar rats.Methods ISO (5 mg· kg-1) or Saline were injected subcutaneously into Wistar rats once a day for 3 or 7 consecutive days.Ventricular remodeling and cardiac function were evaluated by echocardiography.Sections of heart were stained with hematoxylin-eosin (HE) for histopathology or with Masson's trichrome for collagen visualization.In addition,heart tissue immunohistochemistry for α-SMA was also analyzed.The serum levels of tissue inhibitor of matrix metalloproteinases type Ⅰ (TIMP-1) and monocyte chemoattractant protein-1 (MCP-1) were determined by Luminex multiplex technology.Results ISO induced cardiac dysfunction in rats after 3 or 7 days of treatment.ISO caused significant increase of myocardial disorder and fibrosis withincreased α-SMA expression.ISO treated aats showed a significant increase in the serum levels of TIMP-1 and MCP-1.Conclusions Our study suggests that ISO induces profound cardiac remodeling accompanied with increase of serum TIMP-1 and MCP-1.

  15. Bilirubin scavenges chloramines and inhibits myeloperoxidase-induced protein/lipid oxidation in physiologically relevant hyperbilirubinemic serum. (United States)

    Boon, A C; Hawkins, C L; Coombes, J S; Wagner, K H; Bulmer, A C


    Hypochlorous acid (HOCl), an oxidant produced by myeloperoxidase (MPO), induces protein and lipid oxidation, which is implicated in the pathogenesis of atherosclerosis. Individuals with mildly elevated bilirubin concentrations (i.e., Gilbert syndrome; GS) are protected from atherosclerosis, cardiovascular disease, and related mortality. We aimed to investigate whether exogenous/endogenous unconjugated bilirubin (UCB), at physiological concentrations, can protect proteins/lipids from oxidation induced by reagent and enzymatically generated HOCl. Serum/plasma samples supplemented with exogenous UCB (≤250µM) were assessed for their susceptibility to HOCl and MPO/H2O2/Cl(-) oxidation, by measuring chloramine, protein carbonyl, and malondialdehyde (MDA) formation. Serum/plasma samples from hyperbilirubinemic Gunn rats and humans with GS were also exposed to MPO/H2O2/Cl(-) to: (1) validate in vitro data and (2) determine the relevance of endogenously elevated UCB in preventing protein and lipid oxidation. Exogenous UCB dose-dependently (P<0.05) inhibited HOCl and MPO/H2O2/Cl(-)-induced chloramine formation. Albumin-bound UCB efficiently and specifically (3.9-125µM; P<0.05) scavenged taurine, glycine, and N-α-acetyllysine chloramines. These results were translated into Gunn rat and GS serum/plasma, which showed significantly (P<0.01) reduced chloramine formation after MPO-induced oxidation. Protein carbonyl and MDA formation was also reduced after MPO oxidation in plasma supplemented with UCB (P<0.05; 25 and 50µM, respectively). Significant inhibition of protein and lipid oxidation was demonstrated within the physiological range of UCB, providing a hypothetical link to protection from atherosclerosis in hyperbilirubinemic individuals. These data demonstrate a novel and physiologically relevant mechanism whereby UCB could inhibit protein and lipid modification by quenching chloramines induced by MPO-induced HOCl.

  16. Prediction of lung cancer based on serum biomarkers by gene expression programming methods. (United States)

    Yu, Zhuang; Chen, Xiao-Zheng; Cui, Lian-Hua; Si, Hong-Zong; Lu, Hai-Jiao; Liu, Shi-Hai


    In diagnosis of lung cancer, rapid distinction between small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) tumors is very important. Serum markers, including lactate dehydrogenase (LDH), C-reactive protein (CRP), carcino-embryonic antigen (CEA), neurone specific enolase (NSE) and Cyfra21-1, are reported to reflect lung cancer characteristics. In this study classification of lung tumors was made based on biomarkers (measured in 120 NSCLC and 60 SCLC patients) by setting up optimal biomarker joint models with a powerful computerized tool - gene expression programming (GEP). GEP is a learning algorithm that combines the advantages of genetic programming (GP) and genetic algorithms (GA). It specifically focuses on relationships between variables in sets of data and then builds models to explain these relationships, and has been successfully used in formula finding and function mining. As a basis for defining a GEP environment for SCLC and NSCLC prediction, three explicit predictive models were constructed. CEA and NSE are frequently- used lung cancer markers in clinical trials, CRP, LDH and Cyfra21-1 have significant meaning in lung cancer, basis on CEA and NSE we set up three GEP models-GEP 1(CEA, NSE, Cyfra21-1), GEP2 (CEA, NSE, LDH), GEP3 (CEA, NSE, CRP). The best classification result of GEP gained when CEA, NSE and Cyfra21-1 were combined: 128 of 135 subjects in the training set and 40 of 45 subjects in the test set were classified correctly, the accuracy rate is 94.8% in training set; on collection of samples for testing, the accuracy rate is 88.9%. With GEP2, the accuracy was significantly decreased by 1.5% and 6.6% in training set and test set, in GEP3 was 0.82% and 4.45% respectively. Serum Cyfra21-1 is a useful and sensitive serum biomarker in discriminating between NSCLC and SCLC. GEP modeling is a promising and excellent tool in diagnosis of lung cancer.

  17. Serum protein profile study of clinical samples using high performance liquid chromatography-laser induced fluorescence: case of cervical and oral cancers (United States)

    Karemore, Gopal; Sujatha, .; Rai, Lavanya; Pai, Keerthilatha M.; Kartha, V. B.; Santhosh C., .


    The serum protein profiles of normal subjects, patients diagnosed with cervical cancer, and oral cancer were recorded using High Performance Liquid Chromatography combined with Laser Induced Fluorescence detection (HPLC-LIF). Serum protein profiles of the above three classes were tested for establishing the ability of HPLC-LIF protein profiling technique for discrimination, using hard clustering and Fuzzy clustering methods. The clustering algorithms have quite successfully classified the profiles as belonging to normal, cancer of cervix, and oral cancer conditions.

  18. Rapid Detection of Ricin in Serum Based on Cu-Chelated Magnetic Beads Using Mass Spectrometry (United States)

    Zhao, Yong-Qiang; Song, Jian; Wang, Hong-Li; Xu, Bin; Liu, Feng; He, Kun; Wang, Na


    The protein toxin ricin obtained from castor bean plant (Ricinus communis) seeds is a potent biological warfare agent due to its ease of availability and acute toxicity. In this study, we demonstrated a rapid and simple method to detect ricin in serum in vitro. The ricin was mixed with serum and digested by trypsin, then all the peptides were efficiently extracted using Cu-chelated magnetic beads and were detected with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The specific ricin peptides were identified by Nanoscale Ultra Performance liquid chromatography coupled to tandem mass spectrometry according to their sequences. The assay required 2.5 hours, and a characteristic peptide could be detected down to 4 ng/μl and used as a biomarker to detect ricin in serum. The high sensitivity and simplicity of the procedure makes it valuable in clinical practice.

  19. Chromatography of carbon nanotubes separated albumin from other serum proteins: a method for direct analysis of their interactions. (United States)

    Kuboki, Yoshinori; Koshikawa, Takamitu; Takita, Hiroko; Fujisawa, Ryuichi; Lee, Min-ho; Abe, Shigeaki; Akasaka, Tsukasa; Uo, Motohiro; Watari, Fumio; Sammons, Rachel


    Chromatography technology was employed to clarify the mechanism of interaction between multi-wall carbon nanotubes (MWCNT) and proteins. A column (16x100 mm) was packed with purified MWCNT, and various proteins were eluted with phosphate buffered saline (PBS) with and without gradient systems. It was found that albumin in bovine serum was eluted immediately from the column without any adsorption to MWCNT. Conversely, the non-albumin proteins, including a protein of 85 kDa molecular mass and a group of proteins with molecular masses higher than 115 kDa, exhibited considerably high affinity towards MWCNT. A sample of pure bovine serum albumin was also eluted immediately from the column, while lysozyme did not elute as a peak with PBS, but eluted with 0.6 M NaCl. Fundamentally, carbon nanotubes are devoid of any electrical charge. Therefore, other forces including the hydrogen bonds, hydrophilic interactions, and van der Waals forces were most probably responsible for the differential elution behaviors. In conclusion, this chromatographic method provided a simple and direct analysis of the interactions between carbon nanotubes and the various proteins.

  20. Fetal bovine serum influences the stability and bioactivity of resveratrol analogues: A polyphenol-protein interaction approach. (United States)

    Tang, Fen; Xie, Yixi; Cao, Hui; Yang, Hua; Chen, Xiaoqing; Xiao, Jianbo


    Fetal bovine serum (FBS) is a universal growth supplement of cell and tissue culture media. Herein, the influences of FBS on the stability and antioxidant activity of 21 resveratrol analogues were investigated using a polyphenol-protein interaction approach. The structure-stability relationships of resveratrol analogues in FBS showed a clear decrease in the stability of hydroxylated resveratrol analogues in the order: resorcinol-type>pyrogallol-type>catechol-type. The glycosylation and methoxylation of resveratrol analogues enhanced their stability. A linear relationship between the stability of resveratrol analogues in FBS and the affinity of resveratrol analogues-FBS interaction was found. The oxidation process is not the only factor governing the stability of resveratrol analogues in FBS. These results facilitated the insightful investigation of the role of polyphenol-protein interactions in serum, thereby providing some fundamental clues for future clinical research and pharmacological studies on natural small molecules.

  1. Selection of symptomatic patients with Crohn's disease for abdominopelvic computed tomography: role of serum C-reactive protein.

    LENUS (Irish Health Repository)

    Desmond, Alan N


    Results of previous studies have shown that repeated abdominopelvic computed tomography (CT) examinations can lead to substantial cumulative diagnostic radiation exposure in patients with Crohn\\'s disease (CD). Improved selection of patients referred for CT will reduce unnecessary radiation exposure. This study examines if serum C-reactive protein (CRP) concentration predicts which symptomatic patients with CD are likely to have significant disease activity or disease complications (such as abscess) detected on abdominopelvic CT.

  2. Maternal protein and folic acid intake during gestation does not program leptin transcription or serum concentration in rat progeny. (United States)

    Chmurzynska, Agata; Stachowiak, Monika; Pruszynska-Oszmalek, Ewa


    Maternal nutrition during gestation influences the development of the fetus, thereby determining its phenotype, including nutrient metabolism, appetite, and feeding behavior. The control of appetite is a very complex process and can be modulated by orexigenic and anorexigenic mediators such as leptin, which is involved in the regulation of energy homeostasis by controlling food intake and energy expenditure. Leptin transcription and secretion are regulated by numerous factors, nutrition being one of them. The present study was designed to test whether maternal nutrition can permanently affect leptin gene transcription and leptin serum concentration in rat progeny. Moreover, we analyzed whether leptin expression and secretion in response to high-fat postweaning feeding depends on the maternal diet during gestation. Pregnant rats were fed either a normal protein, normal folic acid diet (the AIN-93 diet); a protein-restricted, normal folic acid diet; a protein-restricted, folic acid-supplemented diet; or a normal protein, folic acid-supplemented diet. After weaning, the progeny was fed either the AIN-93 diet or a high-fat diet. Neither maternal nutrition nor the postweaning diet significantly affected Lep transcription. High-fat feeding after weaning was associated with higher serum leptin concentration, but the reaction of an organism to the fat content of the diet was not determined by maternal nutrition during gestation. There was no correlation between Lep mRNA level and serum leptin concentration. Global DNA methylation in adipose tissue was about 30% higher in rats fed postnatally the high-fat diet (P folic acid content in the maternal diet had no significant programming effect on Lep transcription and serum leptin concentration in the rats.

  3. Increased serum cartilage oligomeric matrix protein levels and decreased patellar bone mineral density in patients with chondromalacia patellae.



    BACKGROUND: Chondromalacia patellae is a potentially disabling disorder characterised by features of patellar cartilage degradation. OBJECTIVE: To evaluate markers of cartilage and bone turnover in patients with chondromalacia patellae. METHODS: 18 patients with chondromalacia patellae were studied. Serum cartilage oligomeric matrix protein (s-COMP) and bone sialoprotein (s-BSP) levels were measured by enzyme linked immunosorbent assay (ELISA) and compared with those of age and sex matched he...

  4. Pioglitazone Lowers Serum Retinol Binding Protein 4 by Suppressing its Expression in Adipose Tissue of Obese Rats

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    Chaoyu Zhu


    Full Text Available Background/Aims: Pioglitazone, a peroxisome proliferator-activated receptor γ activator, is clinically used to treat insulin resistance. However, the underlying mechanism of pioglitazone's action remains unclear. We investigated whether, and how, pioglitazone modulates serum level of retinol binding protein 4 (RBP4, an adipocytokine associated with obesity and insulin resistance. Methods: Insulin sensitivity was determined by oral glucose tolerance test, and RBP4 expression was detected by RT-PCR and Western blotting. Results: Pioglitazone treatment significantly decreased serum RBP4 levels in obese rats, which was correlated with reduced body weight and increased insulin sensitivity. Moreover, pioglitazone greatly decreased RBP4 mRNA and protein levels in adipose tissue but not in the liver. Consistently, pioglitazone treatment significantly reduced RBP4 protein expression in 3T3-L1 adipocytes but not in HepG2 cells. Conclusion: These results demonstrate that pioglitazone inhibits the level of serum RPB4 by suppressing RBP4 expression in adipose tissue of obese rats, suggesting that inhibiting RBP4 expression in adipocytes may provide a mechanism by which pioglitazone improves insulin sensitivity in insulin-resistant subjects.

  5. Integrative proteomics and tissue microarray profiling indicate the association between overexpressed serum proteins and non-small cell lung cancer.

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    Yansheng Liu

    Full Text Available 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.

  6. Differences in urine cadmium associations with kidney outcomes based on serum creatinine and cystatin C

    Energy Technology Data Exchange (ETDEWEB)

    Weaver, Virginia M., E-mail: [Division of Occupational and Environmental Health, Department of Environmental Health Sciences, Johns Hopkins University Bloomberg School of Public Health, 615N. Wolfe St., Rm. 7041, Baltimore, MD 21205 (United States); Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD (United States); Welch Center for Prevention, Epidemiology and Clinical Research, Johns Hopkins Medical Institutions, Baltimore, MD (United States); Kim, Nam-Soo; Lee, Byung-Kook [Institute of Industrial Medicine, SoonChunHyang University, Asan (Korea, Republic of); Parsons, Patrick J. [Laboratory of Inorganic and Nuclear Chemistry, Wadsworth Center, New York State Department of Health, Albany, NY (United States); Department of Environmental Health Sciences, School of Public Health, University at Albany, Albany, NY (United States); Spector, June [Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA (United States); Fadrowski, Jeffrey [Welch Center for Prevention, Epidemiology and Clinical Research, Johns Hopkins Medical Institutions, Baltimore, MD (United States); Department of Pediatrics, Johns Hopkins School of Medicine, Baltimore, MD (United States); Jaar, Bernard G. [Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD (United States); Welch Center for Prevention, Epidemiology and Clinical Research, Johns Hopkins Medical Institutions, Baltimore, MD (United States); Department of Epidemiology, Johns Hopkins University Bloomberg School of Public Health, Baltimore, MD (United States); Steuerwald, Amy J. [Laboratory of Inorganic and Nuclear Chemistry, Wadsworth Center, New York State Department of Health, Albany, NY (United States); Department of Environmental Health Sciences, School of Public Health, University at Albany, Albany, NY (United States); Todd, Andrew C. [Department of Preventive Medicine, Mount Sinai School of Medicine, New York, NY (United States); and others


    Cadmium is a well-known nephrotoxicant; chronic exposure increases risk for chronic kidney disease. Recently, however, associations between urine cadmium and higher creatinine-based estimated glomerular filtration rate (eGFR) have been reported. Analyses utilizing alternate biomarkers of kidney function allow evaluation of potential mechanisms for these observations. We compared associations of urine cadmium with kidney function measures based on serum cystatin C to those with serum creatinine in 712 lead workers. Mean (standard deviation) molybdenum-corrected urine cadmium, Modification of Diet in Renal Disease (MDRD) eGFR and multi-variable cystatin C eGFR were 1.02 (0.65) {mu}g/g creatinine, and 97.4 (19.2) and 112.0 (17.7) mL/min/1.73 m{sup 2}, respectively. The eGFR measures were moderately correlated (r{sub s}=0.5; p<0.001). After adjustment, ln (urine cadmium) was not associated with serum cystatin-C-based measures. However, higher ln (urine cadmium) was associated with higher creatinine-based eGFRs including the MDRD and an equation incorporating serum cystatin C and creatinine (beta-coefficient=4.1 mL/min/1.73 m{sup 2}; 95% confidence interval=1.6, 6.6). Urine creatinine was associated with serum creatinine-based but not cystatin-C-based eGFRs. These results support a biomarker-specific, rather than a kidney function, effect underlying the associations observed between higher urine cadmium and creatinine-based kidney function measures. Given the routine use of serum and urine creatinine in kidney and biomarker research, additional research to elucidate the mechanism(s) for these associations is essential.

  7. Bone morphogenic protein-2 regulates the myogenic differentiation of PMVECs in CBDL rat serum-induced pulmonary microvascular remodeling

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Chang; Chen, Lin; Zeng, Jing; Cui, Jian; Ning, Jiao-nin [Department of Anesthesia, Southwest Hospital, The Third Military Medical University, Chongqing 400038 (China); Wang, Guan-song [Institute of Respiratory Disease, Xinqiao Hospital, The Third Military Medical University, Chongqing 400037 (China); Belguise, Karine; Wang, Xiaobo [Université P. Sabatier Toulouse III and CNRS, LBCMCP, 31062 Toulouse Cedex 9 (France); Qian, Gui-sheng [Institute of Respiratory Disease, Xinqiao Hospital, The Third Military Medical University, Chongqing 400037 (China); Lu, Kai-zhi [Department of Anesthesia, Southwest Hospital, The Third Military Medical University, Chongqing 400038 (China); Yi, Bin, E-mail: [Department of Anesthesia, Southwest Hospital, The Third Military Medical University, Chongqing 400038 (China)


    Hepatopulmonary syndrome (HPS) is characterized by an arterial oxygenation defect induced by intrapulmonary vasodilation (IPVD) that increases morbidity and mortality. In our previous study, it was determined that both the proliferation and the myogenic differentiation of pulmonary microvascular endothelial cells (PMVECs) play a key role in the development of IPVD. However, the molecular mechanism underlying the relationship between IPVD and the myogenic differentiation of PMVECs remains unknown. Additionally, it has been shown that bone morphogenic protein-2 (BMP2), via the control of protein expression, may regulate cell differentiation including cardiomyocyte differentiation, neuronal differentiation and odontoblastic differentiation. In this study, we observed that common bile duct ligation (CBDL)-rat serum induced the upregulation of the expression of several myogenic proteins (SM-α-actin, calponin, SM-MHC) and enhanced the expression levels of BMP2 mRNA and protein in PMVECs. We also observed that both the expression levels of Smad1/5 and the activation of phosphorylated Smad1/5 were significantly elevated in PMVECs following exposure to CBDL-rat serum, which was accompanied by the down-regulation of Smurf1. The blockage of the BMP2/Smad signaling pathway with Noggin inhibited the myogenic differentiation of PMVECs, a process that was associated with relatively low expression levels of both SM-α-actin and calponin in the setting of CBDL-rat serum exposure, although SM-MHC expression was not affected. These findings suggested that the BMP2/Smad signaling pathway is involved in the myogenic differentiation of the PMVECs. In conclusion, our data highlight the pivotal role of BMP2 in the CBDL-rat serum-induced myogenic differentiation of PMVECs via the activation of both Smad1 and Smad5 and the down-regulation of Smurf1, which may represent a potential therapy for HPS-induced pulmonary vascular remodeling. - Highlights: • CBDL-rat serum promotes the myogenic

  8. [Relationship between serum levels of C-reactive protein and alpha1-antitrypsin and insulin resistance in obese women]. (United States)

    Ramírez Alvarado, María Matilde; Sánchez Roitz, César


    Adipose tissue produces cytokines involved in insulin resistance (IR) such as IL-6, IL-8, TNF-alpha and proinflammatory molecules such as C reactive protein (CRP). alpha1-antitrypsin is an inflammation-sensitive plasma protein. The objective of this study is to determine the correlation between serum CRP high-sensitivity (CRPhs) and alpha1-antitrypsin levels with IR indices in obese Venezuelan women. The study population consisted of 15 normal weight women (BMI 21.8 +/- 1.9 kg/m2) and 15 obese women (BMI 35.3 +/- 5.3 kg/m2). Obese and lean women underwent a 2 h-75 g oral glucose tolerance test and the following indices were calculated: homeostatic model assessment of insulin resistance (HOMA-IR), homeostatic model assessment of beta cell function (HOMA-beta), Matsuda Index and Insulinogenic Index. The relationship between serum CRPhs and alpha1-antitrypsin levels and these indices were determined. Obese women had higher CRPhs levels (p = 0.001) compared with normal weight women. In obese women, serum CRPhs levels were positively correlated with HOMA-IR (r = 0.73, p = 0.0021), HOMA-beta (r = 0.53, p = 0.031) and negatively correlated with the Matsuda Index (r = -0.60, p = 0.017). No correlation between serum levels of alpha1-antitrypsin and IR indices in the obese group and the lean group was observed. There was a relation between serum CRPhs levels and insulin resistance, suggesting a role of subclinical inflammation in IR.

  9. Serum liver fatty acid binding protein levels correlate positively with obesity and insulin resistance in Chinese young adults.

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    Juan Shi

    Full Text Available BACKGROUND: Liver fatty acid-binding protein (FABP1 plays an inconclusive role in adiposity. We investigated the association of serum FABP1 levels with obesity and insulin resistance in Chinese young people under 30 years old. METHODOLOGY AND PRINCIPAL FINDINGS: Cross-sectional analysis including 200 obese and 172 normal-weight subjects matched for age and sex, anthropometric measurements were performed and serum FABP1 and biochemical characteristics were measured. Insulin resistance was determined by homeostasis model assessment of insulin resistance (HOMA-IR and by the insulin sensitivity index (S(i derived from Bergman's minimal model. FABP1 levels in obese subjects were significantly higher than those in normal-weight subjects (p<0.001 and the significance remained after adjustment for age, gender, alanine and aspartate aminotransferases (p<0.001. Serum FABP1 levels were significantly correlated with many metabolic-related parameters, with BMI and triglycerides as the independent determinants. FABP1 levels remained an independent risk factor of insulin resistance assessed by binary S(i (OR = 1.868 per SD unit, 95% CI [1.035-3.373], p = 0.038 after adjustment for age, sex, BMI, waist circumference, systolic blood pressure, serum triacylglycerol, total cholesterol, HDL- and LDL-cholesterol,. FABP1 levels were also elevated with an increasing number of components of the metabolic syndrome (p for trend <0.001. Multiple regression modeling for the MetS and its components demonstrated that hypertriglyceridemia and low HDL-cholesterol were significantly correlated to serum FABP1 levels. CONCLUSIONS AND SIGNIFICANCE: Serum FABP1 correlates positively with obesity and insulin resistance in Chinese young adults. Our data supports the fact that FABP1 might be an important mediator participating in fatty acid metabolism and energy balance.


    Directory of Open Access Journals (Sweden)

    F. Jadali MD,


    Full Text Available ObjectiveBacterial meningitis is still a life threatening epidemiological problem especiallyin many developing countries; considering its dire consequences, its promptand accurate diagnosis has become a priority for clinicians. Because of thevarious limitations of conventionally used laboratory techniques, we evaluatedand compared the diagnostic utility of C-reactive protein(CRP and lactatedehydrogenase (LDHin serum and cerebrospinal fluid (CSFin the diagnosisof bacterial meningitis and its effectivity in distinguishing it from asepticmeningitis (AP.Material and MethodsA total of 125 pediatric cases, aged between 1 month and 12 years, includingpatients with bacterial meningitis (n=45, aseptic meningitis (n=42 and acontrol group (n=38, were retrospectively analyzed on the basis of datafrom the initial clinical examinations. Cultures, smears and other commonserum and CSF indices were compared with serum and CSF CRP levels andLDH activity.ResultsCompared with each of the other variables, there were significant differencesin the mean values of serum-CRP, CSF-glucose, CSF-LDH and CSF/serumLDH ratio between the bacterial and aseptic meningitis groups (p<0.001.Of all the tests applied, the highest sensitivity (95% and negative predictivevalue (95% belonged to CSF-LDH activity and the most specific (100% testwith the highest positive predictive value (100% was CSF-CRP titration aswell as smear and culture. Combination of CSF-CRP serum-CRP, and CSFLDHyielded the highest sensitivity (100% and negative predictive value butthe combined application of CSF-LDH and CSF-CRP proved to be the mostspecific and efficient.ConclusionIn the presence of a normal CRP titration and low glucose level in CSF,bacterial meningitis is excluded, whereas elevated level of CSF-LDH activityis a valid confirmatory predictor of BM. In addition, combination of thesethree tests with serum CRP is far more effective than the separate determinationof any of these parameters.

  11. A blood-based screening tool for Alzheimer's disease that spans serum and plasma: findings from TARC and ADNI.

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    Sid E O'Bryant

    Full Text Available CONTEXT: There is no rapid and cost effective tool that can be implemented as a front-line screening tool for Alzheimer's disease (AD at the population level. OBJECTIVE: To generate and cross-validate a blood-based screener for AD that yields acceptable accuracy across both serum and plasma. DESIGN, SETTING, PARTICIPANTS: Analysis of serum biomarker proteins were conducted on 197 Alzheimer's disease (AD participants and 199 control participants from the Texas Alzheimer's Research Consortium (TARC with further analysis conducted on plasma proteins from 112 AD and 52 control participants from the Alzheimer's Disease Neuroimaging Initiative (ADNI. The full algorithm was derived from a biomarker risk score, clinical lab (glucose, triglycerides, total cholesterol, homocysteine, and demographic (age, gender, education, APOE*E4 status data. MAJOR OUTCOME MEASURES: Alzheimer's disease. RESULTS: 11 proteins met our criteria and were utilized for the biomarker risk score. The random forest (RF biomarker risk score from the TARC serum samples (training set yielded adequate accuracy in the ADNI plasma sample (training set (AUC = 0.70, sensitivity (SN = 0.54 and specificity (SP = 0.78, which was below that obtained from ADNI cerebral spinal fluid (CSF analyses (t-tau/Aβ ratio AUC = 0.92. However, the full algorithm yielded excellent accuracy (AUC = 0.88, SN = 0.75, and SP = 0.91. The likelihood ratio of having AD based on a positive test finding (LR+ = 7.03 (SE = 1.17; 95% CI = 4.49-14.47, the likelihood ratio of not having AD based on the algorithm (LR- = 3.55 (SE = 1.15; 2.22-5.71, and the odds ratio of AD were calculated in the ADNI cohort (OR = 28.70 (1.55; 95% CI = 11.86-69.47. CONCLUSIONS: It is possible to create a blood-based screening algorithm that works across both serum and plasma that provides a comparable screening accuracy to that obtained from CSF analyses.

  12. Serum levels of S100B and NSE proteins in Alzheimer's disease patients

    Directory of Open Access Journals (Sweden)

    Souza Diogo O


    Full Text Available Abstract Background Alzheimer's disease is the most common dementia in the elderly, and the potential of peripheral biochemical markers as complementary tools in the neuropsychiatric evaluation of these patients has claimed further attention. Methods We evaluated serum levels of S100B and neuron-specific enolase (NSE in 54 mild, moderate and severe Alzheimer's disease (AD patients and in 66 community-dwelling elderly. AD patients met the probable NINCDS-ADRDA criteria. Severity of dementia was ascertained by the Clinical Dementia Rating (CDR scale, cognitive function by the Mini Mental State Examination (MMSE, and neuroimage findings with magnetic resonance imaging. Serum was obtained from all individuals and frozen at -70°C until analysis. Results By comparing both groups, serum S100B levels were lower in AD group, while serum NSE levels were the same both groups. In AD patients, S100B levels were positively correlated with CDR scores (rho = 0.269; p = 0.049 and negatively correlated with MMSE scores (rho = -0.33; P = 0.048. NSE levels decreased in AD patients with higher levels of brain atrophy. Conclusions The findings suggest that serum levels of S100B may be a marker for brain functional condition and serum NSE levels may be a marker for morphological status in AD.

  13. Quantitation of a PEGylated protein in monkey serum by UHPLC-HRMS using a surrogate disulfide-containing peptide: A new approach to bioanalysis and in vivo stability evaluation of disulfide-rich protein therapeutics. (United States)

    Zheng, Naiyu; Zeng, Jianing; Manney, Amy; Williams, Lakenya; Aubry, Anne-Françoise; Voronin, Kimberly; Buzescu, Adela; Zhang, Yan J; Allentoff, Alban; Xu, Carrie; Shen, Hongwu; Warner, William; Arnold, Mark E


    To quantify a therapeutic PEGylated protein in monkey serum as well as to monitor its potential in vivo instability and methionine oxidation, a novel ultra high performance liquid chromatography-high resolution mass spectrometric (UHPLC-HRMS) assay was developed using a surrogate disulfide-containing peptide, DCP(SS), and a confirmatory peptide, CP, a disulfide-free peptide. DCP(SS) was obtained by eliminating the step of reduction/alkylation before trypsin digestion. It contains an intact disulfide linkage between two peptide sequences that are essential for drug function but susceptible to potential in vivo cleavages. HRMS-based single ion monitoring (SIM) on a Q Exactive™ mass spectrometer was employed to improve assay specificity and sensitivity for DCP(SS) due to its poor fragmentation and low sensitivity with SRM detection. The assay has been validated for the protein drug in monkey serum using both surrogate peptides with excellent accuracy (within ±4.4%Dev) and precision (within 7.5%CV) with a lower limit of quantitation (LLOQ) at 10 ng mL(-1). The protein concentrations in monkey serum obtained from the DCP(SS)-based assay not only provided important pharmacokinetic parameters, but also confirmed in vivo stability of the peptide regions of interest by comparing drug concentrations with those obtained from the CP-based assay or from a ligand-binding assay (LBA). Furthermore, UHPLC-HRMS allowed simultaneous monitoring of the oxidized forms of both surrogate peptides to evaluate potential ex vivo/in vivo oxidation of one methionine present in each of both surrogate peptides. To the best of our knowledge, this is the first report of using a surrogate disulfide-containing peptide for LC-MS bioanalysis of a therapeutic protein.

  14. Comparison of serum amyloid A and C-reactive protein as diagnostic markers of systemic inflammation in dogs

    DEFF Research Database (Denmark)

    Christensen, Michelle Brønniche; Langhorn, Rebecca; Goddard, Amelia


    The diagnostic performance of canine serum amyloid A (SAA) was compared with that of C-reactive protein (CRP) in the detection of systemic inflammation in dogs. Sera from 500 dogs were retrospectively included in the study. C-reactive protein and SAA were measured using validated automated assays....... The overlap performance, clinical decision limits, overall diagnostic performance, correlations, and agreement in the clinical classification between these 2 diagnostic markers were compared. Significantly higher concentrations of both proteins were detected in dogs with systemic inflammation (SAA range: 48.......75 to > 2700 mg/L; CRP range: 0.4 to 907.4 mg/L) compared to dogs without systemic inflammation (SAA range: 1.06 to 56.4 mg/L; CRP range: 0.07 to 24.7 mg/L). Both proteins were shown to be sensitive and specific markers of systemic inflammation in dogs. Significant correlations and excellent diagnostic...

  15. Stromal cells from human long-term marrow cultures, but not cultured marrow fibroblasts, phagocytose horse serum constituents: studies with a monoclonal antibody that reacts with a species-specific epitope common to multiple horse serum proteins. (United States)

    Charbord, P; Tippens, D; Wight, T S; Gown, A M; Singer, J W


    This report describes an IgG1 mouse monoclonal antibody derived after immunization of mice with washed stromal cells from human, long-term bone marrow cultures. The antigen recognized by the antibody (BMS-1) is a carbohydrate-containing prosthetic group that is common to and specific for multiple horse serum proteins. These proteins are avidly ingested by stromal cells and concentrated in endocytic vesicles. Cultured smooth muscle cells took up the horse proteins in a similar manner to marrow stromal cells while cultured marrow fibroblasts, endothelial cells, and hepatoma cells did not. These data indicate that marrow stromal cells specifically accumulate horse serum proteins which might partially explain the horse serum requirement for long-term marrow culture maintenance. The data also suggest further similarities between marrow stromal and smooth muscle cells and additional differences between marrow fibroblasts and marrow stromal cells.

  16. A process efficiency assessment of serum protein removal from milk using ceramic graded permeability microfiltration membrane. (United States)

    Tremblay-Marchand, D; Doyen, A; Britten, M; Pouliot, Y


    Microfiltration (MF) is a well-known process that can be used in the dairy industry to separate caseins from serum proteins (SP) in skim milk using membranes with a pore diameter of 0.1μm. Graded permeability ceramic membranes have been studied widely as means of improving milk fractionation by overcoming problems encountered with other MF membranes. The ideal operating parameters for process efficiency in terms of membrane selectivity, permeate flux, casein loss, SP transmission, energy consumption, and dilution with water remain to be determined for this membrane. Our objective was to evaluate the effects of transmembrane pressure (TMP), volumetric concentration factor (VCF), and diafiltration on overall process efficiency. Skim milk was processed using a pilot-scale MF system equipped with 0.72-m(2) graded permeability membranes with a pore size of 0.1μm. In the first experiment, in full recycle mode, TMP was set at 124, 152, 179, or 207 kPa by adjusting the permeate pressure at the outlet. Whereas TMP had no significant effect on permeate and retentate composition, 152 kPa was found to be optimal for SP removal during concentration and concentration or diafiltration experiments. When VCF was increased to 3×, SP rejection coefficient increased along with energy consumption and total casein loss, whereas SP removal rate decreased. Diafiltering twice allowed an increase in total SP removal but resulted in a substantial increase in energy consumption and casein loss. It also reduced the SP removal rate by diluting permeate. The membrane surface area required for producing cheese milk by blending whole milk, cream, and MF retentate (at different VCF) was estimated for different cheese milk casein concentrations. For a given casein concentration, the same quantity of permeate and SP would be produced, but less membrane surface area would be needed at a lower retentate VCF. Microfiltration has great potential as a process of adding value to conventional

  17. Effect of extracranial electric stimulation at cerebellar fastigial nucleus on serum C-reactive protein of patients with acute cerebral infarction

    Institute of Scientific and Technical Information of China (English)


    BACKGROUND: Some reports indicate that electric and/or chemical stimulation at various brain sites of experimental animals can raise regional cerebral blood flow and improve cerebral circulation; however, its mechanism is still unclear.OBJECTIVE: To observe the effects of electric stimulation at cerebellar fastigial nucleus on serum C-reactive protein of patients with acute cerebral infarction.DESIGN: Non-randomized synchronized contrast study.SETTING: The Second People's Hospital of Xinxiang City.PARTICIPANTS: A total of 54 patients with acute cerebral infarction were selected from the Department of Neurology, the Second People's Hospital of Xinxiang from December 2005 to December 2006. There were 31 males and 23 females, and their ages ranged from 56 to 80 years. All patients met the diagnostic criteria of the Fourth National Cerebrovascular Academic Meeting, were finally diagnosed by using CT examination,and provided the confirmed consent. Based on therapeutic demands, patients were divided into electric stimulation group and routine treatment group with 27 cases in each group. In addition, 21 healthy subjects,including 11 males and 10 females and aging 53 - 78 years, were selected as the control group. All the subjects in the control group did not have any histories of cerebrovascular diseases and severe body diseases.METHODS: Based on routine drug therapy, patients in the electric stimulation group were also treated by using CVFT-010M cerebral circulation function therapeutic device (made in Shanghai). Electrode was fixed at bilateral mastoid in the first group and at extensible sides of upper limbs in the second group. Electric stimulation was given twice a day and lasted for 30 minutes each time. Ten days were regarded as a course.Parameters of device: mode Ⅲ, frequency 198%, and intensity 90% - 110% (bionic current). Patients in the routine treatment group received the routine drug treatment. Content of serum C-reactive protein was measured in both

  18. A supercritical fluid-based coating technology. 3: preparation and characterization of bovine serum albumin particles coated with lipids. (United States)

    Ribeiro Dos Santos, I; Richard, J; Thies, C; Pech, B; Benoit, J-P


    Solvent-free microparticles, loaded with bovine serum albumin as a model protein, were produced using a novel supercritical (SC) fluid-based coating technology. Coating material consists either of trimyristin (Dynasan 114) or of Gelucire 50-02. Microparticles obtained were characterized as regards their morphology, protein content and in vitro release profile. A discontinuous coating made of micro-needles of trimyristin led to an initial burst release of approximately 70% in 30 min. However, a prolonged release of the BSA could be achieved in a phosphate buffer solution at 37 degrees C over a 24 h period from particles coated with Gelucire 50-02. Furthermore, it was shown that BSA does not undergo any degradation after SC CO(2) treatment under the conditions used in the coating process.

  19. Improved corn protein based articles (United States)

    Developing higher value uses for zein (corn protein), a potential major co-product of the bio-ethanol industry, will improve the economics of this business. Historically, zein was predominantly used in the textile fiber industry. Unfortunately the techniques used at that time to modify the zein cann...

  20. Serum insulin-like growth factor-I, iron, C-reactive protein, and serum amyloid A for prediction of outcome in dogs with pyometra. (United States)

    Jitpean, Supranee; Holst, Bodil Ström; Höglund, Odd V; Pettersson, Ann; Olsson, Ulf; Strage, Emma; Södersten, Fredrik; Hagman, Ragnvi


    Pyometra, accumulation of pus in the uterus, is a bacterial infection that frequently initiates systemic inflammation. The disease may have lethal consequences when the systemic effects are severe or complications occur. Markers for identifying high-risk patients and predicting outcome are therefore in high demand. The objective of this study was to measure serum concentrations of insulin-like growth factor-I (IGF-I), iron, C-reactive protein (CRP), and serum amyloid A (SAA) in bitches with pyometra and to explore the possible value of these variables for detection of increased morbidity. In total, 31 bitches were diagnosed with pyometra and destined for surgical treatment (ovariohysterectomy) and 17 healthy bitches were included in the study. Concentrations of IGF-I and iron were lower in the pyometra group (mean concentration 221.2 ± 22.5 ng/mL and 16.9 ± 1.6 μmol/L, respectively) compared with the healthy control group (mean concentration 366.7 ± 46.2 ng/mL and 38.1 ± 2.7 μmol/L, respectively). In contrast, concentrations of CRP and SAA were significantly higher in bitches with pyometra (mean concentrations 212.9 ± 17.3 mg/L and 119.9 ± 8.5 mg/L, respectively) compared with the control group (<5 mg/L and <10 mg/L, respectively). None of the explored variables were associated with morbidity as measured by duration of postoperative hospitalization. In conclusion, IGF-I and iron concentrations were decreased in pyometra, whereas SAA and CRP concentrations were increased in the disease. Although unspecific, measurement of these variables may be valuable as adjunctive markers for prognosis in cases of pyometra.

  1. Serum acute phase proteins as biomarkers of pleuritis and cranio-ventral pulmonary consolidation in slaughter-aged pigs. (United States)

    Saco, Yolanda; Fraile, Lorenzo; Giménez, Mercè; Alegre, Ana; López-Jimenez, Rosa; Cortey, Martí; Segalés, Joaquim; Bassols, Anna


    The purpose of this study was to investigate the relationship between the existence of lung lesions in pigs at slaughter and the concentration of the serum acute phase proteins (APP), haptoglobin (Hp), pig-major acute protein (Pig-MAP) and C-reactive protein (CRP). A total of 24 pig farms were selected out of a larger farm database previously screened to study risk factors associated with pleuritis and cranio-ventral pulmonary consolidation (CVPC) lesions at slaughter-aged pigs in Spain. The farms were classified as "pleuritis negative (P-) or positive (P+)" and as "CVPC negative (M-) or positive (M+)" and divided into four groups according to a 2X2 factorial design (P-M-, P-M+, P+M-, P+M+). Also at slaughter, blood from 20 randomly selected pigs from each farm was collected. Obtained serum samples were used to measure acute phase proteins. All APP concentrations were significantly higher for M+ farms than for M- ones. However, only Hp and Pig-MAP showed significantly higher concentrations for P+ farms than for P- ones. Pig-MAP was the most sensitive biomarker since it was able to clearly discriminate between P-/P+ and M-/M+ groups (ppleuritis and good for CVPC lesions. CRP was able to discriminate for CVPC lesions but not for pleuritis. The present results indicate that Pig-MAP and, possibly Hp, may be used as potential markers to characterise and discriminate respiratory lesions in swine herds at slaughter.

  2. Changes in Holstein cow milk and serum proteins during intramammary infection with three different strains of Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Robert Claude


    Full Text Available Abstract Background Staphylococcus aureus is one of the most prevalent pathogens to cause mastitis in dairy cattle. Intramammary infection of dairy cows with S. aureus is often subclinical, due to the pathogen's ability to evade the innate defense mechanisms, but this can lead to chronic infection. A sub-population of S. aureus, known as small colony variant (SCV, displays atypical phenotypic characteristics, causes persistent infections, and is more resistant to antibiotics than parent strains. Therefore, it was hypothesized that the host immune response will be different for SCV than its parental or typical strains of S. aureus. In this study, the local and systemic immune protein responses to intramammary infection with three strains of S. aureus, including a naturally occurring bovine SCV strain (SCV Heba3231, were characterized. Serum and casein-depleted milk cytokine levels (interleukin-8, interferon-γ, and transforming growth factor-β1, as well as serum haptoglobin concentrations were monitored over time after intramammary infection with each of the three S. aureus strains. Furthermore, comparative proteomics was used to evaluate milk proteome profiles during acute and chronic phases of S. aureus intramammary infection. Results Serum IL-8, IFN-γ, and TGF-β1 responses differed in dairy cows challenged with different strains of S. aureus. Changes in overall serum haptoglobin concentrations were observed for each S. aureus challenge group, but there were no significant differences observed between groups. In casein-depleted milk, strain-specific differences in the host IFN-γ response were observed, but inducible IL-8 and TGF-β1 concentrations were not different between groups. Proteomic analysis of the milk following intramammary infection revealed unique host protein expression profiles that were dependent on the infecting strain as well as phase of infection. Notably, the protein, component-3 of the proteose peptone (CPP3, was

  3. Localization of the domains of the Haemophilus ducreyi trimeric autotransporter DsrA involved in serum resistance and binding to the extracellular matrix proteins fibronectin and vitronectin. (United States)

    Leduc, Isabelle; Olsen, Bonnie; Elkins, Christopher


    Resisting the bactericidal activity of naturally occurring antibodies and complement of normal human serum is an important element in the evasion of innate immunity by bacteria. In the gram-negative mucosal pathogen Haemophilus ducreyi, serum resistance is mediated primarily by the trimeric autotransporter DsrA. DsrA also functions as an adhesin for the extracellular matrix proteins fibronectin and vitronectin and mediates attachment of H. ducreyi to keratinocytes. We sought to determine the domain(s) of the 236-residue DsrA protein required for serum resistance and extracellular matrix protein binding. A 140-amino-acid truncated protein containing only the C-terminal portion of the passenger domain and the entire translocator domain of DsrA exhibited binding to fibronectin and vitronectin and conferred serum resistance to an H. ducreyi serum-sensitive strain. A shorter DsrA construct consisting of only 128 amino acids was unable to bind to extracellular matrix proteins but was serum resistant. We concluded that neither fibronectin binding nor vitronectin binding is required for high-level serum resistance in H. ducreyi.

  4. Correlation between the serum level of advanced oxidation protein products and the cognitive function in patients with obstructive sleep apnea hypopnea syndrome

    Institute of Scientific and Technical Information of China (English)



    Objective To observe the change of the cognitive function and the serum level of advanced oxidation protein products(AOPP) in patients with obstructive sleep apnea hypopnea syndrome(OSAHS),and then to investigate

  5. Serum protein capillary electrophoresis and measurement of acute phase proteins in a captive cheetah (Acinonyx jubatus) population

    DEFF Research Database (Denmark)

    Depauw, Sarah; Delanghe, Joris; Whitehouse-Tedd, Katherine


    Renal and gastrointestinal pathologies are widespread in the captive cheetah (Acinonyx jubatus) population but are often diagnosed at a late stage, because diagnostic tools are limited to the evaluation of clinical signs or general blood examination. Presently, no data are available on serum...

  6. The Development and Characterization of Protein-Based Stationary Phases for Studying Drug-Protein and Protein-Protein Interactions


    Sanghvi, Mitesh; Moaddel, Ruin; Wainer, Irving W.


    Protein-based liquid chromatography stationary phases are used in bioaffinity chromatography for studying drug-protein interactions, the determination of binding affinities, competitive and allosteric interactions, as well as for studying protein-protein interactions. This review addresses the development and characterization of protein-based stationary phase, and the application of these phases using frontal and zonal chromatography techniques. The approach will be illustrated using immobili...

  7. Evaluation of Sialic Acid and Acute Phase Proteins (Haptoglobin and Serum Amyloid A in Clinical and Subclinical Bovine Mastitis

    Directory of Open Access Journals (Sweden)

    S. Nazifi*, M. Haghkhah1, Z. Asadi, M. Ansari-Lari2, M. R. Tabandeh3, Z. Esmailnezhad and M. Aghamiri


    Full Text Available The present study was conducted to evaluate the concentrations of sialic acids (total, lipid bound and protein bound and their correlation with acute phase proteins (haptoglobin and serum amyloid A in clinical and subclinical mastitis of cattle. Thirty subclinical mastitic cows with positive California mastitis test (CMT test and no clinical signs of mastitis, 10 clinical mastitic cows and 10 healthy cows with negative CMT test and normal somatic cell count were selected. Milk and blood samples were collected after confirmation of clinical and subclinical mastitis by somatic cell count and bacterial identification. Serum haptoglobin (Hp, serum amyloid A (SAA, total sialic acid (TSA, lipid bound sialic acid (LBSA and protein bound sialic acid (PBSA were measured by validated standard methods. Haptoglobin and SAA increased significantly in both types of mastitis compared with control group (P<0.001. However, the ratio of HP/SAA was significantly different from the control group only in clinical mastitis. The results showed that TSA and LBSA were significantly different in control group compared with clinical and subclinical mastitis (P<0.001. Protein bound sialic acid did not change in subclinical mastitis in comparison with control group (P=0.86. There was positive correlation between LBSA and PBSA in clinical mastitis (r=0.72, P=0.02 whereas significant negative correlation was observed between LBSA and PBSA in subclinical mastitis (r=-0.62, P<0.001. Results also showed no correlation between Hp and SAA with each other or with any other parameters in study groups.

  8. Serum Lipid Transfer Proteins in Hypothyreotic Patients Are Inversely Correlated with Thyroid-Stimulating Hormone (TSH) Levels (United States)

    Skoczyńska, Anna; Wojakowska, Anna; Turczyn, Barbara; Zatońska, Katarzyna; Wołyniec, Maria; Rogala, Natalia; Szuba, Andrzej; Bednarek-Tupikowska, Grażyna


    Background Plasma cholesteryl ester transfer protein (CETP) activity is often decreased in patients with hypothyroidism, whereas less is known about the phospholipid transfer protein (PLTP). We aimed to evaluate simultaneously serum CETP and PLTP activity in patients diagnosed with hypothyroidism. Material/Methods The selection criteria for control group members (without thyroid dysfunction) in this case to case study were levels of total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol (HDL-C), and triglycerides similar to those in study group patients (101 patients diagnosed with hypothyroidism). Serum CETP and PLTP activities were measured by homogenous fluorometric assays using synthetic donor particle substrates. Results Serum CETP and PLTP activities in hypothyreotic patients were lower (p<0.001) compared with those in healthy subjects. This lowering was associated with significant changes in HDL-C subclasses: decrease in HDL2- and increase in HDL3 cholesterol levels. Multiple linear regression analyses adjusted for age, sex, body mass index, smoking habits, and alcohol drinking showed a strong association between hypothyroidism and activity of lipid transfer proteins. A linear inverse relationship between thyroid-stimulating hormone (TSH) and CETP (r=−0.21; p<0.01) and between TSH and PLTP (r=−0.24; p<0.001) was shown. There also was a positive correlation (p<0.001) between CETP and HDL2 cholesterol (r=0.27) and between PLTP and HDL2 cholesterol (r=0.37). A negative correlation between CETP and HDL3 cholesterol (r=−0.22: p<0.01) and between PLTP and HDL3 cholesterol (r=−0.24; p<0.001) has been demonstrated as well. Conclusions The decreased HDL2 and increased HDL3 cholesterol levels in subjects with hypothyroidism are consequences of decreased activity of lipid transfer proteins. These changes are early symptoms of lipid disturbances in hypothyroidism. PMID:27899788

  9. Placental lactogen administration reverses the effect of low-protein diet on maternal and fetal serum somatomedin levels in the pregnant rat.



    Female rats were studied on day 20 of pregnancy after being fed either a 5% lactalbumin (low protein) diet or a 20% lactalbumin (adequate) diet for the last 2 weeks of pregnancy. Rats on the lower intake of protein showed decreased serum levels of rat placental lactogen and reduced numbers of lactogenic receptors in the maternal liver. These changes were accompanied by much reduced serum levels of somatomedins IGF I(insulin-like growth factor) and II (multiplication-stimulating activity, MSA)...

  10. New multi protein patterns differentiate liver fibrosis stages and hepatocellular carcinoma in chronic hepatitis C serum samples

    Institute of Scientific and Technical Information of China (English)

    Thomas G(o)bel; Sonja Vorderwülbecke; Katarzyna Hauck; Holger Fey; Dieter H(a)ussinger; Andreas Erhardt


    AIM: To identify a multi serum protein pattern as well as single protein markers using surface-enhanced laser desorption/ionisation time-of-flight mass spectrometry (SELDI-TOF-MS) for detection and differentiation of liver fibrosis (F1-F2), liver cirrhosis (F4) and hepatocellular carcinoma (HCC) in patients with chronic hepatitis C virus (HCV).METHODS: Serum samples of 39 patients with F1/F2 fibrosis, 44 patients with F4 fibrosis, 34 patients with HCC were applied to CM10 arrays and analyzed using the SELDI-TOF ProteinChip System (PBS-Ⅱ c; Ciphergen Biosystems) after anion-exchange fractionation. All patients had chronic hepatitis C and histologically confirmed fibrosis stage/HCC. Data were analyzed for protein patterns by multivariate statistical techniques and artificial neural networks.RESULTS: A 4 peptide/protein multimarker panel (7486,12843, 44293 and 53598 Da) correctly identified HCCs with a sensitivity of 100% and specificity of 85% in a two way-comparison of HCV-cirrhosis versus HCV-HCC training samples (AUROC 0.943). Sensitivity and specificity for identification of HCC were 68% and 80% for random test samples. Cirrhotic patients could be discriminated against patients with F1 or F2 fibrosisusing a 5 peptide/protein multimarker pattern (2873,6646, 7775, 10525 and 67867 Da) with a specificity of 100% and a sensitivity of 85% in training samples (AUROC 0.976) and a sensitivity and specificity of 80% and 67% for random test samples. Combination of the biomarker classifiers with APRI score and alfa-fetopotein (AFP) improved the diagnostic performance. The 6646 Da marker protein for liver fibrosis was identified as apolipoprotein C-Ⅰ.CONCLJSION: SELDI-TOF-MS technology combined with protein pattern analysis seems a valuable approach for the identification of liver cirrhosis and hepatocellular carcinoma in patients with chronic hepatitis C. Most probably a combination of different serum markers will help to identify liver cirrhosis and early

  11. Predicting disease-related proteins based on clique backbone in protein-protein interaction network. (United States)

    Yang, Lei; Zhao, Xudong; Tang, Xianglong


    Network biology integrates different kinds of data, including physical or functional networks and disease gene sets, to interpret human disease. A clique (maximal complete subgraph) in a protein-protein interaction network is a topological module and possesses inherently biological significance. A disease-related clique possibly associates with complex diseases. Fully identifying disease components in a clique is conductive to uncovering disease mechanisms. This paper proposes an approach of predicting disease proteins based on cliques in a protein-protein interaction network. To tolerate false positive and negative interactions in protein networks, extending cliques and scoring predicted disease proteins with gene ontology terms are introduced to the clique-based method. Precisions of predicted disease proteins are verified by disease phenotypes and steadily keep to more than 95%. The predicted disease proteins associated with cliques can partly complement mapping between genotype and phenotype, and provide clues for understanding the pathogenesis of serious diseases.

  12. Interleukin-28B polymorphisms and interferon gamma inducible protein-10 serum levels in seronegative occult hepatitis C virus infection. (United States)

    Bartolomé, Javier; Castillo, Inmaculada; Quiroga, Juan Antonio; Carreño, Vicente


    Polymorphisms upstream interleukin (IL)-28B gene and serum levels of interferon gamma inducible protein-10 (IP-10) are associated with spontaneous and treatment-induced hepatitis C virus (HCV) clearance. Patients with seronegative occult HCV infection are anti-HCV and serum HCV-RNA negative but have viral RNA in liver and abnormal values of liver enzymes. We examined if the rs12979860 polymorphism of IL-28B and serum IP-10 levels differ between chronic and seronegative occult CV infection. IL-28B polymorphism was determined with allele specific TaqMan probes in total DNA isolated from peripheral blood mononuclear cells and IP-10 by an enzyme-linked immunosorbent assay in serum from 99 patients with seronegative occult HCV infection and 130 untreated patients with chronic hepatitis C. IL-28B genotypes were also determined in 54 healthy volunteers. Prevalence of the IL-28B CC genotype was significantly higher in seronegative occult HCV infection (52/99; 52.5%) than in chronic hepatitis C (32/130; 24.6%, P occult HCV infection, HCV-RNA load in liver was significantly lower in those with the IL-28B CC genotype than in those with CT + TT genotypes (2.8 × 10(5)  ± 5.8 × 10(4) vs. 4.1 × 10(5)  ± 5.9 × 10(4)  copies/μg of total RNA respectively; P = 0.023). Mean serum IP-10 levels were significantly lower in patients with seronegative occult HCV infection than in patients with chronic hepatitis C (160.8 ± 17.9 vs. 288.7 ± 13.3 pg/ml respectively; P occult HCV infection in comparison with chronic hepatitis C.

  13. A novel multiplex-protein array for serum diagnostics of colon cancer: a case–control study

    Directory of Open Access Journals (Sweden)

    Bünger Stefanie


    Full Text Available Abstract Background More than 1.2 million new cases of colorectal cancer are reported each year worldwide. Despite actual screening programs, about 50% of the patients are diagnosed at advanced tumor stages presenting poor prognosis. Innovative screening tools could aid the detection at early stages and allow curative treatment interventions. Methods A nine target multiplex serum protein biochip was generated and evaluated using a training- and validation-set of 317 highly standardized, liquid nitrogen preserved serum samples comprising controls, adenomas, and colon cancers. Results Serum levels of CEA, IL-8, VEGF, S100A11, MCSF, C3adesArg, CD26, and CRP showed significant differences between cases and controls. The largest areas under the receiver operating characteristics curve were observed for CEA, IL-8, and CRP. At threshold levels yielding 90% specificity, sensitivities for CEA, IL-8 and CRP were 26%, 22%, and 17%, respectively. The most promising marker combinations were CEA + IL-8 reaching 37% sensitivity at 83% specificity and CEA + CRP with 35% sensitivity at 81% specificity. In an independent validation set CEA + IL-8 reached 47% sensitivity at 86% specificity while CEA + CRP obtained 39% sensitivity at 86% specificity. Early carcinomas were detected with 33% sensitivity for CEA + IL-8 and 28% for CEA + CRP. Conclusions Apart from CEA, IL-8, and CRP, the screening value of additional blood markers and the potential advantage of combining serum biochip testing with fecal occult blood testing needs to be studied. Multiplex biochip array technology utilizing serum samples offers an innovative approach to colorectal cancer screening.

  14. Serum lipopolysaccharide binding protein levels predict severity of lung injury and mortality in patients with severe sepsis.

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    Jesús Villar

    Full Text Available BACKGROUND: There 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 FINDINGS: LBP serum levels at study entry, at 48 hours and at day-7 were measured in 180 patients with severe sepsis. Data regarding the nature of infections, disease severity, development of acute lung injury (ALI and acute respiratory distress syndrome (ARDS, and intensive care unit (ICU outcome were recorded. LBP serum levels were similar in survivors and non-survivors at study entry (117.4+/-75.7 microg/mL vs. 129.8+/-71.3 microg/mL, P = 0.249 but there were significant differences at 48 hours (77.2+/-57.0 vs. 121.2+/-73.4 microg/mL, P<0.0001 and at day-7 (64.7+/-45.8 vs. 89.7+/-61.1 microg/ml, p = 0.017. At 48 hours, LBP levels were significantly higher in ARDS patients than in ALI patients (112.5+/-71.8 microg/ml vs. 76.6+/-55.9 microg/ml, P = 0.0001. An increase of LBP levels at 48 hours was associated with higher mortality (odds ratio 3.97; 95%CI: 1.84-8.56; P<0.001. CONCLUSIONS/SIGNIFICANCE: Serial LBP serum measurements may offer a clinically useful biomarker for identification of patients with severe sepsis having the worst outcomes and the highest probability of developing sepsis-induced ARDS.

  15. Sensitive Procedure for Rapid Detection of Human Brucellosis, Based on PCR Method in Contaminated Serum Samples

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    Eslam Ghezelsofla


    Full Text Available AbstractBackground and objective: Brucellosis is a zoonosis transmittable to humans poses a significant public health problem in many developing countries and requires rapid and accurate diagnostic methods. Here, our aim was to develop a diagnostic polymerase chain reaction (PCR assay in artificially contaminated serum samples as a model for rapid and accurate laboratory confirmation of human brucellosis. Material and methods: In this study, initially the standard Brucella abortus strain (2308 were cultured on Brucella agar medium and then colonies were inactivated by formalin 10 %. Genomic DNA was extracted from inactivated bacterial colonies. Serial dilutions of bacterial-DNA were prepared in fetal bovine serum (FBS and water and subsequently DNA extraction were repeated on these artificially contaminated samples. The two pairs of primers amplified two different fragments included in: a gene encoding an outer membrane protein (omp-2 (primers JPF/JPR and a sequence 16S rRNA of B. abortus (primers F4/R2. Results: The two primers assayed showed a difference in sensitivity for detecting Brucella DNA, ranging between 5 pg and 50 pg for artificially contaminated serum samples and 50Fg and 5 pg for contaminated control samples. Therefore, the sensitivity of PCR using F4/R2 primers was greater than the PCR using JPF/JPR primers.Conclusion: Although the sensitivity of PCR using these primers was affected by serum inhibitors, they are still the most sensitive and they could provide a useful tool for the diagnosis of human brucellosis.

  16. Moringa oleifera aqueous leaf extract inhibits reducing monosaccharide-induced protein glycation and oxidation of bovine serum albumin. (United States)

    Nunthanawanich, Pornpimon; Sompong, Weerachat; Sirikwanpong, Sukrit; Mäkynen, Kittana; Adisakwattana, Sirichai; Dahlan, Winai; Ngamukote, Sathaporn


    Advanced glycation end products (AGEs) play an important factor for pathophysiology of diabetes and its complications. Moringa oleifera is one of the medicinal plants that have anti-hyperglycemic activity. However, anti-glycation property of Moringa oleifera leaf extract on the different types of reducing monosaccharides-induced protein glycation has not been investigated. Therefore, the aim of this study was to examine the protective effect of Moringa oleifera aqueous leaf extract (MOE) on reducing sugars-induced protein glycation and protein oxidation. Total phenolic content of MOE was measured using the Folin-Ciocalteu method. Bovine serum albumin was incubated with 0.5 M of reducing sugars (glucose or fructose) with or without MOE (0.5-2.0 mg/mL) for 1, 2, 3 and 4 weeks. The results found that total phenolic content was 38.56 ± 1.50 mg gallic acid equivalents/g dry extract. The formation of fluorescent and non-fluorescent AGEs [N (ε)-(carboxymethyl) lysine (CML)] and the level of fructosamine were determined to indicate protein glycation, whereas the level of protein carbonyl content and thiol group were examined for protein oxidation. MOE (0.5-2.0 mg/mL) significantly inhibited the formation of fluorescent, N (ε)-CML and markedly decreased fructosamine level (P < 0.05). Moreover, MOE significantly prevented protein oxidation manifested by reducing protein carbonyl and the depletion of protein thiol in a dose-dependent manner (P < 0.05). Thus, the findings indicated that polyphenols containing in MOE have high potential for decreasing protein glycation and protein oxidation that may delay or prevent AGE-related diabetic complications.

  17. Targeted non-covalent self-assembled nanoparticles based on human serum albumin

    NARCIS (Netherlands)

    Bunschoten, Anton; Buckle, Tessa; Kuil, Joeri; Luker, Gary D.; Luker, Kathryn E.; Nieweg, Omgo; van Leeuwen, Fijs W. B.


    Human serum albumin (HSA) is a biological nanocarrier that forms non-covalent complexes with a number of synthetic and biomolecules. Previously we demonstrated radiolabeled HSA-based nanoparticles can form non-covalent complexes with fluorescent cyanine dyes yielding imaging agents for surgical guid

  18. Redox chemistry of the molecular interactions between tea catechins and human serum proteins under simulated hyperglycemic conditions. (United States)

    Özyurt, Hazal; Luna, Carolina; Estévez, Mario


    Carbonylation is an irreversible modification in oxidized proteins that has been directly related to a number of health disorders including Type 2 diabetes. Dietary antioxidants have been proposed to counteract the oxidative stress occurring under hyperglycemic conditions. An understanding of the nature and consequences of the molecular interactions between phytochemicals and human plasma proteins is of utmost scientific interest. Three tea catechins namely epicatechin (EC), epigallocatechin (EGC) and epigallocatechin-3-gallate (EGCG) were tested for (i) their affinity to bind to human serum albumin (HSA) and human hemoglobin (HH) and (ii) their ability to inhibit tryptophan (Trp) depletion and for the formation of specific protein carbonyls and pentosidine in the aforementioned proteins. Both proteins (20 mg mL(-1)) were allowed to react with postprandial plasmatic concentrations of the catechins (EC: 0.7 μM, EGC: 1.8 μM, and EGCG: 0.7 μM) under simulated hyperglycemic conditions (12 mM glucose/0.2 mM Fe(3+)/37 °C/10 days). The three catechins were able to inhibit Trp oxidation and protein carbonylation in both plasma proteins. Some anti-glycation properties were linked to their binding affinities. The molecular interactions reported in the present study may explain the alleged beneficial effects of tea catechins against the redox impairment linked to hyperglycemic conditions.

  19. Hydrogel based drug carriers for controlled release of hydrophobic drugs and proteins

    NARCIS (Netherlands)

    Ke Peng,


    The aim of this study is to prepare in situ forming hydrogels based on biocompatible polymers for the controlled release of hydrophobic drug and proteins. In order to load hydrophobic drug to the hydrophilic hydrogel matrix, beta-cyclodextrin and human serum albumin was introduced to the hydrogel ne

  20. Effect of certified reference material 470 (CRM 470) on national quality assurance programs for serum proteins in Europe. (United States)

    Johnson, A M; Whicher, J T


    The introduction of the international reference material for serum proteins, CRM 470, has resulted in significant reduction of the among-laboratory variance for most proteins assayed in European national quality assurance programs. In general, the CVs have decreased by 5 to 65%. However, both among- and within-manufacturer variances in many cases remain unacceptably high. In addition, concentration-dependent differences in variance and bias are present for some proteins. Although some variance will persist, reducing variance and bias to levels required for the institution of universal reference ranges will necessitate more accurate transfer of values to calibrants and controls and improved calibration curve fitting by manufacturers, as well as better quality control within laboratories.

  1. Comparison of serum leptin, glucose, total cholesterol and total protein levels in fertile and repeat breeder cows

    Directory of Open Access Journals (Sweden)

    Saime Guzel


    Full Text Available In the present study we measured serum glucose, leptin, total cholesterol and total protein concentrations in repeat breeder cows and compared them with fertile cows. For this aim, 20 repeat breeder cows and 20 fertile cows were used as material. Repeat breeder cows were found to have lower levels of leptin and glucose as compared with fertile ones. No significant differences in total cholesterol and total protein levels were observed between the two groups. No significant correlation of leptin with glucose, total cholesterol and total protein was observed in fertile and repeat breeder cows. Low concentrations of glucose and leptin can have some effects on reproductive problems as repeat breeder and help to understand potential mechanisms impairing fertility in repeat breeder cows.

  2. Activity-Based Protein Profiling of Microbes

    Energy Technology Data Exchange (ETDEWEB)

    Sadler, Natalie C.; Wright, Aaron T.


    Activity-Based Protein Profiling (ABPP) in conjunction with multimodal characterization techniques has yielded impactful findings in microbiology, particularly in pathogen, bioenergy, drug discovery, and environmental research. Using small molecule chemical probes that react irreversibly with specific proteins or protein families in complex systems has provided insights in enzyme functions in central metabolic pathways, drug-protein interactions, and regulatory protein redox, for systems ranging from photoautotrophic cyanobacteria to mycobacteria, and combining live cell or cell extract ABPP with proteomics, molecular biology, modeling, and other techniques has greatly expanded our understanding of these systems. New opportunities for application of ABPP to microbial systems include: enhancing protein annotation, characterizing protein activities in myriad environments, and reveal signal transduction and regulatory mechanisms in microbial systems.

  3. The use of serum glial fibrillary acidic protein measurements in the diagnosis of neuromyelitis optica spectrum optic neuritis.

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    Mithu Storoni

    Full Text Available BACKGROUND: Glial fibrillary acidic protein (GFAP is a specific intermediate filament of the cytoskeleton of the astrocyte and may be used as a specific marker for astrocytic damage. It is detectable in the cerebrospinal fluid following a relapse caused by Multiple Sclerosis (MS and Neuromyelitis Optica (NMO spectrum disease. Higher levels are found following an NMO-related relapse. It is not known if GFAP is also detectable in the serum following such relapses. In particular, it is not known if lesions limited to the optic nerve release GFAP in sufficient quantities to be detectable within the serum. The aim of this study was to ascertain the extent to which serum GFAP levels can distinguish between an episode of optic neuritis (ON related to NMO spectrum disease and ON from other causes. METHODOLOGY/PRINCIPAL FINDINGS: Out of 150 patients consecutively presenting to our eye hospital over the period March 2009 until July 2010, we were able to collect a serum sample from 12 patients who had presented with MS-related ON and from 10 patients who had presented with NMO spectrum disease-related ON. We also identified 8 patients with recurrent isolated ON and 8 patients with a corticosteroid-dependent optic neuropathy in the absence of any identified aetiology. GFAP was detectable in the serum of all but three patients (two patients with MS-related ON and one with recurrent optic neuritis. The median serum GFAP level in the patient group with NMO spectrum disease was 4.63 pg/mL whereas in all other cases combined together, this was 2.14 pg/mL. The difference was statistically significant (P = 0.01. A similar statistically significant difference was found when cases with pathology limited to the optic nerve were compared (P = 0.03. CONCLUSIONS: Glial pathology in NMO related optic neuritis is reflected in elevated serum GFAP levels independently of whether or not there is extra-optic nerve disease.

  4. [Pesticide detection in Costarican vegetables based on the inhibition of serum and erythrocytic human cholinesterases]. (United States)

    Nevermann, Karl Schosinsky; Guzmán, Eugenia Quintana


    A simple and low cost method able to detect the presence of pesticides, organophosphates and carbamates based on the inhibition of serum and erythrocytic cholinesterases, was used in lettuce (Lactuca sativa), cilantro (Coriandum santivum) and celery (Apium graveolens) obtained from the Ferias del Agricultor from Valle Central of Costa Rica. The percentage inhibition of cholinesterases is related to the presence of plaguicide in the vegetable. Thirteen percent of the analyzed samples were positive for plaguicides using serum cholinesterase and 33% for erythrocytic cholinesterase. Washing and cooking the vegetables does not eliminate the presence of plaguicides but they lower slightly the concentration. Statistical evidence (p = 0.0001) indicates that erythrocytic cholinesterase has higher analytical sensitivity than serum cholinesterase. It is very important to establish the degree of contamination with pesticides in these agricultural products because they are exposed to direct contamination by fumigation, soil contamination and irrigation water, and are products that are often consumed without adequate cooking and washing.

  5. G-protein beta3 subunit gene variant is unlikely to have a significant influence on serum uric acid level in Japanese workers. (United States)

    Suwazono, Yasushi; Kobayashi, Etsuko; Uetani, Mirei; Miura, Katsuyuki; Morikawa, Yuko; Ishizaki, Masao; Kido, Teruhiko; Nakagawa, Hideaki; Nogawa, Koji


    The C825T variant of the G-protein beta3 subunit (GNB3) gene has attracted renewed attention as a candidate gene for obesity, hypertension and hyperuricemia. The main role of G-protein is to translate signals from the cell surface into a cellular response. The 825T allele is associated with a splice variant of GNB3 protein and enhanced G-protein activation. We examined the relationship between this variant and the risk of hyperuricemia in Japanese workers. The study subjects were 1,452 men and 1,169 women selected from 3,834 men and 2,591 women in 1997. On the basis of common clinical criteria, hyperuricemia I was defined as serum uric acid >or= 7.0 mg/dl in men and 6.0 mg/dl in women or taking antihyperuricemic medication. The hyperuricemia I group consisted of 186 men and 20 women and its control of 1,266 men and 1,149 women. Hyperuricemia II was defined as serum uric acid > 5.7 mg/dl (median) in men and 3.9 mg/dl (median) in women or taking antihyperuricemic medication. The hyperuricemic II group consisted of 684 men and 570 women and its control of 768 men and 599 women. To replicate previous significant results in young Caucasian men, we selected these criteria because the authors of the study in young Caucasian men adopted the median in their subjects as a cut-off. The statistical power was estimated as 99% based on the significant results in Caucasians. Genotype and allele distributions in men and women with hyperuricemia I and II were not significantly different from those in the corresponding control groups. Logistic regression analysis on hyperuricemia I and II, and multiple regression on serum uric acid level demonstrated no significant effect of the C825T genotype. Despite the sufficient statistical power, this study could not demonstrate the significant influence of C825T on hyperuricemia or serum uric acid. The targeting of this polymorphism is unlikely to be beneficial in the prevention of hyperuricemia in the general Japanese population.

  6. Studies on the effect of long-term exposure to nitrogen dioxide on serum and liver proteins level and enzyme activity in guinea pigs. (United States)

    Drozdz, M; Kucharz, E; Ludyga, K; Molska-Drozda, T


    Forty male guinea pigs were exposed to nitrogen dioxide in a concentration of 2 mg/m3, 8 hours daily for a period of 180 days. Forty male animals were used as a control group. The following changes were found in intoxicated animals: the decrease of total protein and seromucoid concentration in blood serum and the decrease of total protein, perchloric acid-soluble proteins, protein-bound hexosamines and sialic acids content, in liver tissue. Electrophoretic examination of the serum proteins showed the increase of alpha 1- and beta 2-globulins and the decrease of albumin concentration. Changes in the level of glycoproteins fractions and protein-bound carbohydrates in blood serum were described also. Estimation of enzymes activity showed the decrease of alanine and aspartate transaminase activity in blood serum caused by the strong decrease of the cytoplasmic fraction of these enzymes. However the simultaneous increase of the mitochondrial fraction of transaminases activity was observed. The decrease of the activity of choline esterase was found also. Similar changes of enzymes activity were found in liver tissue. Histopathological studies were done for the further clearing the influenze of nitrogen dioxide on serum and liver proteins concentration and enzymes activity. It was found that after long-term exposure to nitrogen dioxide the destruction processes may be observed in the liver. The possible mechanism of the nitrogen dioxide-induced damage of protein metabolism is discussed.

  7. Profiling serum antibodies to Mycobacterium tuberculosis proteins in rhesus monkeys with nontuberculous Mycobacteria. (United States)

    Min, Fangui; Pan, Jinchun; Wu, Ruike; Chen, Meiling; Kuang, Huiwen; Zhao, Weibo


    Recent evidence indicates that the prevalence of diseases caused by nontuberculous mycobacteria (NTM) has been increasing in both human and animals. In this study, antibody profiles of NTM in rhesus monkeys (Macaca mulatta) were determined and compared with those of monkeys infected with Mycobacterium tuberculosis complex (MTBC). Antibodies against 10 M. tuberculosis proteins, purified protein derivative (PPD), and mammalian old tuberculin (MOT) were detected in 14 monkeys naturally infected with NTM by indirect ELISA. Sera from 10 monkeys infected with MTBC and 10 healthy monkeys were set as controls. All antigens showed high serological reactivities to MTBC infections and low reactivities in healthy monkeys. NTM infections showed strong antibody responses to MOT and PPD; moderate antibody responses to 16kDa, U1, MPT64L, 14kDa, and TB16.3; and low antibody responses to 38kDa, Ag85b, CFP10, ESAT-6, and CFP10-ESAT-6. According to the criteria of MTBC, only CFP10, ESAT-6, and CFP10-ESAT-6 showed negative antibody responses in all NTM infections. Taken together, these results suggest that positive results of a PPD/MOT-based ELISA in combination with results of antibodies to M. tuberculosis-specific antigens, such as CFP10 and ESAT-6, could discriminate NTM and MTBC infections. Two positive results indicate an MTBC infection, and a negative result for an M. tuberculosis-specific antigen may preliminarily predict an NTM infection.

  8. Intentional formation of a protein corona on nanoparticles: Serum concentration affects protein corona mass, surface charge, and nanoparticle-cell interaction. (United States)

    Gräfe, Christine; Weidner, Andreas; Lühe, Moritz V D; Bergemann, Christian; Schacher, Felix H; Clement, Joachim H; Dutz, Silvio


    The protein corona, which immediately is formed after contact of nanoparticles and biological systems, plays a crucial role for the biological fate of nanoparticles. In the here presented study we describe a strategy to control the amount of corona proteins which bind on particle surface and the impact of such a protein corona on particle-cell interactions. For corona formation, polyethyleneimine (PEI) coated magnetic nanoparticles (MNP) were incubated in a medium consisting of fetal calf serum (FCS) and cell culture medium. To modulate the amount of proteins bind to particles, the composition of the incubation medium was varied with regard to the FCS content. The protein corona mass was estimated and the size distribution of the participating proteins was determined by means of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Additionally, the zeta potential of incubated particles was measured. Human blood-brain barrier-representing cell line HBMEC was used for in vitro incubation experiments. To investigate the consequences of the FCS dependent protein corona formation on the interaction of MNP and cells flow cytometry and laser scanning microscopy were used. Zeta potential as well as SDS-PAGE clearly reveal an increase in the amount of corona proteins on MNP with increasing amount of FCS in incubation medium. For MNP incubated with lower FCS concentrations especially medium-sized proteins of molecular weights between 30kDa and 100kDa could be found within the protein corona, whereas for MNP incubated within higher FCS concentrations the fraction of corona proteins of 30kDa and less increased. The presence of the protein corona reduces the interaction of PEI-coated MNP with HBMEC cells within a 30min-incubation.

  9. Serum Proteins and Alkaline Phosphatase Levels in Patients with Tuberous Sclerosis (United States)

    Fischer, M. H.; And Others


    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)

  10. Surfactant protein D (SP-D) serum levels in patients with community-acquired pneumonia

    DEFF Research Database (Denmark)

    Leth-Larsen, Rikke; Nordenbaek, Claudia; Tornoe, Ida;


    SP-D is a lectin involved in the first line of defense against microorganisms. It is primarily found in the lung but also at extrapulmonary sites and in the circulation. An immunoassay for the quantification of SP-D in serum was established and the SP-D concentration was measured in consecutive b...

  11. Independent Candidate Serum Protein Biomarkers of Response to Adalimumab and to Infliximab in Rheumatoid Arthritis: An Exploratory Study. (United States)

    Ortea, Ignacio; Roschitzki, Bernd; López-Rodríguez, Rosario; Tomero, Eva G; Ovalles, Juan G; López-Longo, Javier; de la Torre, Inmaculada; González-Alvaro, Isidoro; Gómez-Reino, Juan J; González, Antonio


    Response to treatment of rheumatoid arthritis shows large inter-individual variability. This heterogeneity is observed with all the anti-rheumatic drugs, including the commonly used TNF inhibitors. It seems that drug-specific and target-specific factors lead individual patients to respond or not to a given drug, although this point has been challenged. The search of biomarkers distinguishing responders from non-responders has included shotgun proteomics of serum, as a previous study of response to infliximab, an anti-TNF antibody. Here, we have used the same study design and technology to search biomarkers of response to a different anti-TNF antibody, adalimumab, and we have compared the results obtained for the two anti-TNF drugs. Search of biomarkers of response to adalimumab included depletion of the most abundant serum proteins, 8-plex isobaric tag for relative and absolute quantitation (iTRAQ) labeling, two-dimensional liquid chromatography fractionation and relative quantification with a hybrid Orbitrap mass spectrometer. With this approach, 264 proteins were identified in all the samples with at least 2 peptides and 95% confidence. Nine proteins showed differences between non-responders and responders (P treatment of rheumatoid arthritis and indicate that they are notably drug-specific.

  12. Immunohistochemical Study of Serum Proteins in Normal and Cadmium-Treated Mouse Testis by in vivo Cryotechnique

    Institute of Scientific and Technical Information of China (English)

    Xiao-gang LIAO; Nobuo TERADA; Zi-long LI; Nobuhiko OHNO; Yasuhisa FUJII; Shinichi OHNO


    Objective To investigate time-dependent changes of serum proteins permeability inthe normal and cadmium(Cd)-treated mouse testis, reflecting tight junctional (TJ) barriersof Sertoli cellsMethods The serum proteins, albumin and immunoglobulin-G(IgG), in the seminiferoustubules were firstly immobilized by the "in vivo cryotechnique", in which the dynamicblood circulation was always kept. The cryofixed testicular tissues were then processedfor the freeze-substitution method, and embedded in the paraffin wax. Serial sectionsof 5μm thickness were immunostained by anti-mouse albumin or IgG antibody withperoxidase immunostaining, and also stained with hematoxylin-eosine (HE)formorphological observation.Results In normal seminiferous tubules, albumin immunoreaction products werelocalized around peritubular myoid cells and among Leydig cells, as well as in bloodvessels. They were also localized as arch-like patterns around some spermatogonia inbasal compartments. The number of the immunopositive arch structures was differentaccording to developmental stages of the seminiferous cycle, judging from thearrangement of germ cells by HE-staining. The patterns of localization of IgGimmunostaining in normal mouse testis were similar to that of albumin. In 24 h afterCd-treatment, some enlarged spaces and vesicular formation in the seminiferousepithelium were observed on the paraffin sections by HE-staining. The albumin or IgG immunolocalization was seen not only in the basal compartments, but also in theadluminal compartments between Sertolicells and germ cells.Conclusion The structural changes of inter-Sertoli TJ barriers in vivo, such as differentimmunostaining patterns of serum proteins between the normal and Cd-treated mouseseminiferous tubules, could be clearly detected by the "in vivo cryotechnique" withalbumin or IgG immunohistochemistry.

  13. Evaluation of Fucosylated Haptoglobin and Mac-2 Binding Protein as Serum Biomarkers to Estimate Liver Fibrosis in Patients with Chronic Hepatitis C. (United States)

    Tawara, Seiichi; Tatsumi, Tomohide; Iio, Sadaharu; Kobayashi, Ichizou; Shigekawa, Minoru; Hikita, Hayato; Sakamori, Ryotaro; Hiramatsu, Naoki; Miyoshi, Eiji; Takehara, Tetsuo


    Fucosylated haptoglobin (Fuc-Hpt) and Mac-2 binding protein (Mac-2 bp) are identified as cancer biomarkers, based on the results from a glyco-proteomic analysis. Recently, we reported that these glyco-biomarkers were associated with liver fibrosis and/or ballooning hepatocytes in patients with nonalcoholic fatty liver disease (NAFLD). We evaluated the ability of these glycoproteins to estimate liver fibrosis in 317 patients with chronic hepatitis C. We measured the serum Fuc-Hpt and Mac-2 bp levels using a lectin-antibody ELISA and ELISA, respectively. The serum levels of both Fuc-Hpt and Mac-2 bp increased with the progression of liver fibrosis. The multivariate analysis revealed that Mac-2 bp was an independent factor associated with moderate liver fibrosis (F ≥ 2). In contrast, Fuc-Hpt was an independent factor associated with advanced liver fibrosis (F ≥ 3). In terms of evaluating liver fibrosis, the serum levels of these glycomarkers were correlated with well-known liver fibrosis indexes, such as the aspartate aminotransferase to platelet ratio index (APRI) and Fibrosis-4 (FIB4) index. An assay that combined the APRI or FIB4 index and the Fuc-Hpt or Mac-2 bp levels increased the AUC value for diagnosing hepatic fibrosis. Interestingly, the cumulative incidence of hepatocellular carcinoma (HCC) was significantly higher in the patients with elevated serum levels of Fuc-Hpt and Mac-2 bp. In conclusion, both Fuc-Hpt and Mac-2 bp could be useful glyco-biomarkers of liver fibrosis and predictors of HCC in patients with chronic hepatitis C.

  14. Quantitative Proteomics Identifies Serum Response Factor Binding Protein 1 as a Host Factor for Hepatitis C Virus Entry

    Directory of Open Access Journals (Sweden)

    Gisa Gerold


    Full Text Available Hepatitis C virus (HCV enters human hepatocytes through a multistep mechanism involving, among other host proteins, the virus receptor CD81. How CD81 governs HCV entry is poorly characterized, and CD81 protein interactions after virus binding remain elusive. We have developed a quantitative proteomics protocol to identify HCV-triggered CD81 interactions and found 26 dynamic binding partners. At least six of these proteins promote HCV infection, as indicated by RNAi. We further characterized serum response factor binding protein 1 (SRFBP1, which is recruited to CD81 during HCV uptake and supports HCV infection in hepatoma cells and primary human hepatocytes. SRFBP1 facilitates host cell penetration by all seven HCV genotypes, but not of vesicular stomatitis virus and human coronavirus. Thus, SRFBP1 is an HCV-specific, pan-genotypic host entry factor. These results demonstrate the use of quantitative proteomics to elucidate pathogen entry and underscore the importance of host protein-protein interactions during HCV invasion.

  15. Neomycin-phenolic conjugates: polycationic amphiphiles with broad-spectrum antibacterial activity, low hemolytic activity and weak serum protein binding. (United States)

    Findlay, Brandon; Zhanel, George G; Schweizer, Frank


    Here we present a proof-of-concept study, combining two known antimicrobial agents into a hybrid structure in order to develop an emergent cationic detergent-like interaction with the bacterial membrane. Six amphiphilic conjugates were prepared by copper (I)-catalyzed 1,3-dipolar cycloaddition between a neomycin B-derived azide and three alkyne-modified phenolic disinfectants. Three conjugates displayed good activity against a variety of clinically relevant Gram positive and Gram negative bacteria, including MRSA, without the high level of hemolysis or strong binding to serum proteins commonly observed with other cationic antimicrobial peptides and detergents.

  16. Independent Candidate Serum Protein Biomarkers of Response to Adalimumab and to Infliximab in Rheumatoid Arthritis: An Exploratory Study.

    Directory of Open Access Journals (Sweden)

    Ignacio Ortea

    Full Text Available Response to treatment of rheumatoid arthritis shows large inter-individual variability. This heterogeneity is observed with all the anti-rheumatic drugs, including the commonly used TNF inhibitors. It seems that drug-specific and target-specific factors lead individual patients to respond or not to a given drug, although this point has been challenged. The search of biomarkers distinguishing responders from non-responders has included shotgun proteomics of serum, as a previous study of response to infliximab, an anti-TNF antibody. Here, we have used the same study design and technology to search biomarkers of response to a different anti-TNF antibody, adalimumab, and we have compared the results obtained for the two anti-TNF drugs. Search of biomarkers of response to adalimumab included depletion of the most abundant serum proteins, 8-plex isobaric tag for relative and absolute quantitation (iTRAQ labeling, two-dimensional liquid chromatography fractionation and relative quantification with a hybrid Orbitrap mass spectrometer. With this approach, 264 proteins were identified in all the samples with at least 2 peptides and 95% confidence. Nine proteins showed differences between non-responders and responders (P < 0.05, representing putative biomarkers of response to adalimumab. These results were compared with the previous study of infliximab. Surprisingly, the non-responder/responder differences in the two studies were not correlated (rs = 0.07; P = 0.40. This overall independence with all the proteins showed two identifiable components. On one side, the putative biomarkers of response to either adalimumab or infliximab, which were not shared and showed an inverse correlation (rs = -0.69; P = 0.0023. On the other, eight proteins showing significant non-responder/responder differences in the analysis combining data of response to the two drugs. These results identify new putative biomarkers of response to treatment of rheumatoid arthritis and

  17. Identification and Characterization of GH Receptor and Serum GH-binding Protein in Chinese Sturgeon (Acipenser sinensis)

    Institute of Scientific and Technical Information of China (English)

    Zhi-Yong LIAO; Shang-Quan ZHU


    Chinese sturgeon, a kind of cartilage ganoid, has a history of over one billion years and it is called the living fossil of aquatic biology since it keeps some evolutionary trace. Here, we characterized the growth hormone receptor (GHR) and serum growth hormone binding protein (GHBP) of Chinese sturgeon. It was shown that GHR was expressed in various tissues, mainly in hepatic, kidney and intestine tissues. GHR on the hepatic membrane has high and specific affinity for bream GH (brGH) and Scatchard analysis of the binding data showed a single class of high affinity binding site with an association constant Ka of 3.1×109 M-1. A specific band around 94 kD was detected by SDS-PAGE in cross-linking studies of membrane receptors. After incubation of Chinese sturgeon serum with 125I-brGH, a 125I-brGH-GHBP complex was identified by Sephadex G-75, indicating that in the serum exists GHBP specially binding to brGH.

  18. Increased serum levels of C-reactive protein and matrix metalloproteinase-9 in obstructive sleep apnea syndrome

    Institute of Scientific and Technical Information of China (English)

    YE Jin; LIU Hui; LI Yuan; LIU Xian; ZHU Jie-ming


    Background Obstructive sleep apnea syndrome (OSAS), characterized by intermittent hypoxia/reoxygenation (IHR),has been identified as an independent risk factor for cardiovascular diseases (CVD). The CVD biomarkers associated with OSAS have not been thoroughly investigated.Methods Fifty-one men with OSAS recently diagnosed by polysomnography were classified into two groups according to the severity of apnea: moderate to severe OSAS group (n= 28) and mild OSAS group (n= 23). Twenty-five obese men,of comparable age and body mass index (BMI), without OSAS were chosen as control subjects. Serum metabolic variables, C-reactive protein (CRP) and matrix metalloproteinase-9 (MMP-9) were measured. Spearman correlation and regression analysis were performed.Results Serum concentrations of CRP and MMP-9 were significantly higher in 51 OSAS patients than in 25 control subjects. Levels of CRP and MMP-9 were significantly higher in patients with moderate to severe OSAS than in patients with mild OSAS or in obese control subjects. A positive correlation was found between levels of CRP and MMP-9 in OSAS patients. Regression analysis showed that after adjusting for age and BMI, apnea/hypopnea index (AHI) significantly correlated with serum concentrations of CRP and MMP-9 in patients with OSAS.Conclusions AHI, mirroring the frequency of IHR, was a predictor of enhanced circulating CVD biomarkers MMP-9 and CRP. Our data support the theory that IHR contributes to the upregulation of the inflammatory factors in OSAS patients.

  19. Serum thymidine kinase 1 and C-reactive protein as biomarkers for screening clinically healthy dogs for occult disease. (United States)

    Selting, K A; Sharp, C R; Ringold, R; Knouse, J


    Thymidine kinase (TK1) is a biomarker that correlates well with diagnosis and prognosis in certain canine cancers. Canine C-reactive protein (cCRP) is a widely accepted marker of inflammation correlated with increased risk and severity of various diseases. We evaluated serum TK1 and cCRP concentrations in apparently healthy dogs (n = 360). All dogs were followed up for a minimum of 6 months by health questionnaire. All dogs with cancer were identified using a proprietary dual-biomarker algorithm [termed Neoplasia Index (NI)]. Specificity of positive NI is 0.91 and high positive is 0.98. All-cause mortality was 20% in dogs with elevated cCRP and 3% in dogs with low cCRP. The performance of serum TK1 and cCRP as tools for screening for occult cancer is improved when evaluated together. Serum TK1 and cCRP (unified in the NI) are useful in the screening of occult canine cancer. cCRP is useful in screening for other serious diseases.

  20. Serum C-reactive protein predicts early mortality in hospitalized patients with HBV-related decompensated cirrhosis. (United States)

    Zhu, ShaoMing; Waili, Yulituzi; Qi, XiaoTing; Chen, YueMei; Lou, YuFeng; Chen, Bo


    The serum C-reactive protein (CRP) is an inflammatory marker. The aim of the present study was to elucidate whether CRP could serve as a potential surrogate marker for 30-day mortality in hospitalized patients with HBV-related decompensated cirrhosis (HBV-DeCi).This was a retrospective cohort study that included 140 patients with HBV-DeCi. All patients were followed up for 1-month. A panel of clinical and biochemical variables were analyzed for potential associations with outcomes using multiple regression models.The serum CRP was significantly higher in nonsurviving patients than in surviving patients. Multivariate analysis demonstrated that CRP levels (odds ratio: 1.047, P = 0.002) and the model for end-stage liver disease score (odds ratio: 1.370, P = 0.001) were independent predictors for mortality.Serum CRP is a simple marker that may serve as an additional predictor of 1-month mortality in hospitalized patients with HBV-DeCi.

  1. Dynamic change of serum protein S100b and its clinical significance in patients with traumatic brain injury

    Institute of Scientific and Technical Information of China (English)

    CHEN Da-qing; ZHU Lie-lie


    Objective: To analyze the dynamic change of serum protein S100b in patients with traumatic brain injury and its clinical value in assessing brain damage. Methods: According to Glasgow coma scale (GCS), 102 cases of traumatic brain injury were divided into mild brain injury group (GCS≥13, n=31, Group A), moderate brain injury group (8serum S100b, with earlier increase and longer duration, than the mild and moderate brain injury groups. The patients with higher S100b exhibited lower GCS scores and poor clinical prognosis. The increase in S100b could emerge before clinical image evidence indicated so. Conclusions: Serum S100b can be used as a sensitive index for assessment and prediction of traumatic brain injury severity and prognosis.

  2. Nanochemistry of protein-based delivery agents

    Directory of Open Access Journals (Sweden)

    Subin R.C.K. Rajendran


    Full Text Available The past decade has seen an increased interest in the conversion of food proteins into functional biomaterials, including their use for loading and delivery of physiologically active compounds such as nutraceuticals and pharmaceuticals. Proteins possess a competitive advantage over other platforms for the development of nanodelivery systems since they are biocompatible, amphipathic, and widely available. Proteins also have unique molecular structures and diverse functional groups that can be selectively modified to alter encapsulation and release properties. A number of physical and chemical methods have been used for preparing protein nanoformulations, each based on different underlying protein chemistry. This review focuses on the chemistry of the reorganization and/or modification of proteins into functional nanostructures for delivery, from the perspective of their preparation, functionality, stability and physiological behavior.

  3. Nanochemistry of protein-based delivery agents (United States)

    Rajendran, Subin; Udenigwe, Chibuike; Yada, Rickey


    The past decade has seen an increased interest in the conversion of food proteins into functional biomaterials, including their use for loading and delivery of physiologically active compounds such as nutraceuticals and pharmaceuticals. Proteins possess a competitive advantage over other platforms for the development of nanodelivery systems since they are biocompatible, amphipathic, and widely available. Proteins also have unique molecular structures and diverse functional groups that can be selectively modified to alter encapsulation and release properties. A number of physical and chemical methods have been used for preparing protein nanoformulations, each based on different underlying protein chemistry. This review focuses on the chemistry of the reorganization and/or modification of proteins into functional nanostructures for delivery, from the perspective of their preparation, functionality, stability and physiological behavior.

  4. New serum biomarkers for detection of HBV-induced liver cirrhosis using SELDT protein chip technology

    Institute of Scientific and Technical Information of China (English)

    Xiao-Dong Zhu; Wei-Hua Zhang; Cheng-Lin Li; Yang Xu; Wei-Jiang Liang; Po Tien


    AIM: To find new serum biomarkers for liver cirrhosis (LC)in chronic carriers of hepatitis B virus (HBV).METHODS: Surface enhanced laser desorption/ionization time-of-flight (SELDI-TOF) mass spectrometry was used to discover biomarkers for differentiating HBV induced LC from non-cirrhotic cohorts. A training population of 25 patients with HBV-induced LC, 20 patients with HCC, and 25 closely age-matched healthy men, was studied.RESULTS: Two biomarkers with Mr 7 772 and 3 933 were detected in sera of non-cirrhotic cohorts, but not in patients with HBV-induced LC. A sensitivity of 80% for all LC patients,a specificity of 81.8% for all non-cirrhotic cohorts and a positive predictive value of 75% for the study population were obtained.CONCLUSION: These two serum biomarkers for HBVinduced LC might be used for diagnosis and assessment of disease progression.

  5. [New topics regarding equations for GFR estimation based on serum creatinine and cystatin C]. (United States)

    Horio, Masaru


    Japanese GFR equations and CKD-EPI equations based on standardized serum creatinine and standardized cystatin C are recommended in recent Japanese CKD guides and KDIGO guidelines for CKD management, respectively. CKD-EPIcreat overestimates GFR in Japanese subjects, probably due to the difference in muscle mass between Japanese and Caucasians. Unlike CKD-EPIcreat, CKD-EPIcys performs well in Japanese subjects, indicating the advantages of using cystatin C as a GFR marker. KDIGO guidelines suggest measuring eGFRcys in adults with eGFRcreat of 45-59 ml/min/1.73 m2 who do not have markers of kidney damage if confirmation of CKD is required. Creatinine is excreted by glomerular filtration, but also secreted by the tubules. Alteration of the tubular secretion of creatinine may influence the performance of GFR equations based on serum creatinine. Multivariate analysis showed that GFR and serum albumin levels were independent parameters affecting the fractional excretion of creatinine (FE-Cr). Alteration of FE-Cr according to the serum albumin levels may be one of the reasons for the bias of GFR equations based on serum creatinine. Low GFR is a risk factor for all-cause and cardiovascular mortality in a general population. However, the relationship between eGFR and the hazard risk of events is different depending on whether cystatin C or creatinine is used to calculate eGFR. The association between eGFRcys and the hazard risk is much stronger compared with eGFRcreat. Cystatin C may be a useful alternative to creatinine for detecting a high risk of complications in a general population and subjects with CKD.

  6. Functional resting-state fMRI connectivity correlates with serum levels of the S100B protein in the acute phase of traumatic brain injury

    Directory of Open Access Journals (Sweden)

    William Hedley Thompson


    Full Text Available The S100B protein is an intra-cellular calcium-binding protein that mainly resides in astrocytes in the central nervous system. The serum level of S100B is used as biomarker for the severity of brain damage in traumatic brain injury (TBI patients. In this study we investigated the relationship between intrinsic resting-state brain connectivity, measured 1–22 days (mean 8 days after trauma, and serum levels of S100B in a patient cohort with mild-to-severe TBI in need of neuro-intensive care in the acute phase. In line with previous investigations, our results show that the peak level of S100B acquired during the acute phase of TBI was negatively correlated with behavioral measures (Glasgow Outcome Score, GOS of functional outcome assessed 6 to 12 months post injury. Using a multi-variate pattern analysis-informed seed-based correlation analysis, we show that the strength of resting-state brain connectivity in multiple resting-state networks was negatively correlated with the peak of serum levels of S100B. A negative correspondence between S100B peak levels recorded 12–36 h after trauma and intrinsic connectivity was found for brain regions located in the default mode, fronto-parietal, visual and motor resting-state networks. Our results suggest that resting-state brain connectivity measures acquired during the acute phase of TBI is concordant with results obtained from molecular biomarkers and that it may hold a capacity to predict long-term cognitive outcome in TBI patients.

  7. Serum protein changes in lead exposed mice infected with Hymenolepis nana. (United States)

    Agrawal, R; Johri, G N


    Significant decrease in serum globulin took place in Hymenolepis nana mouse model treated with lead prior to infection. Decrease of alpha, beta and gamma globulins was maximum on day 4 in comparison to that of control, gradually tapering off by day 100 when suppression was minimum with the experimental values almost reaching the same level as those of the control. Thus, suppression of globulins was neither total nor permanent.

  8. Protein-calixarene interactions: complexation of Bovine Serum Albumin by sulfonatocalix[n]arenes. (United States)

    Memmi, L; Lazar, A; Brioude, A; Ball, V; Coleman, A W


    The complexation of Bovine Serum Albumin with sulfonatocalix[n]arenes has been demonstrated by means of electrospray mass spectrometry, dynamic light scattering and atomic force microscopy; with sulfonatocalix[4]arene one strong and two weaker binding sites are detected; the effects on the structure of thin films formed by surface deposition of BSA show that the sulfonatocalix[n]arenes act to reticulate the films and produce essentially planar systems.



    Zaiter Samir; Kulikova EA; Garyuk GI


    Some kinds of interleikines of patients with chronic laryngitis disease were investigated. There is “cytokines explosion” of the patients with chronic laryngitis with persistent herpes simplex virus. Comparative investigation cytokine profile in serum blood is demonstrated: balanced reaction cytokines profile of patients with chronic laryngitis without persistent herpes simplex virus and dysbalanced reaction of patients with laryngitis (hyperergation). Increased content IL-6 and low content g...

  10. The prognostic value ofserum C-reactive protein-bound serum amyloid A inearly-stage lung cancer

    Institute of Scientific and Technical Information of China (English)

    XueYanZhang; GeZhang; YingJiang; DanLiu; ManZhiLi; QianZhong; ShanQiZeng; WanLiLiu; MuShengZeng


    Background:Elevated levels of serum C‑reactive protein (CRP) have been reported to have prognostic signiifcance in lung cancer patients. This study aimed to further identify CRP‑bound components as prognostic markers for lung cancer and validate their prognostic value. Methods:CRP‑bound components obtained from the serum samples from lung cancer patients or healthy controls were analyzed by differential proteomics analysis. CRP‑bound serum amyloid A (CRP‑SAA) was evaluated by co‑immunoprecipitation (IP). Serum samples from two independent cohorts with lung cancer (retrospective cohort, 242 patients; prospective cohort, 222 patients) and healthy controls (159 subjects) were used to evaluate the prognostic value of CRP‑SAA by enzyme‑linked immunosorbent assay. Results:CRP‑SAA was identiifed speciifcally in serum samples from lung cancer patients by proteomic analysis. CRP binding to SAA was conifrmed by co‑IP in serum samples from lung cancer patients and cell culture media. The level of CRP‑SAA was signiifcantly higher in patients than in healthy controls (0.37±0.58 vs. 0.03±0.04,P<0.001). Elevated CRP‑SAA levels were signiifcantly associated with severe clinical features of lung cancer. The elevation of CRP‑SAA was associated with lower survival rates for both the retrospective (hazard ration [HR]=2.181, 95% conifdence interval [CI]=1.641–2.897,P<0.001) and the prospective cohorts (HR=2.744, 95% CI=1.810–4.161,P<0.001). Multivariate Cox analysis showed that CRP‑SAA was an independent prognostic marker for lung cancer. Remarkably, in stages I–II patients, only CRP‑SAA, not total SAA or CRP, showed signiifcant association with overall survival in two cohorts. Moreover, univariate and multivariate Cox analyses also showed that only CRP‑SAA could be used as an inde‑pendent prognostic marker for early‑stage lung cancer patients. Conclusion:CRP‑SAA could be a better prognostic marker for lung cancer than total SAA or CRP

  11. 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. (United States)

    García-Villalobos, Héctor; Morales-Trejo, Adriana; Araiza-Piña, Benedicto A; Htoo, John K; Cervantes-Ramírez, Miguel


    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 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 dietary amino acids, affect the expression of cationic amino acid transporters in pigs fed wheat-based diets.

  12. Low-protein amino acid-supplemented diets for growing pigs: effect on expression of amino acid transporters, serum concentration, performance, and carcass composition. (United States)

    Morales, A; Buenabad, L; Castillo, G; Arce, N; Araiza, B A; Htoo, J K; Cervantes, M


    Pigs fed protein-bound AA appear to have a higher abundance of AA transporters for their absorption in the jejunum compared with the duodenum. However, there is limited data about the effect of dietary free AA, readily available in the duodenum, on the duodenal abundance of AA transporters and its impact on pig performance. Forty-eight pigs (24.3 kg initial BW) distributed in 4 treatments were used to evaluate the effect of the CP level and form (free vs. protein bound) in which AA are added to diets on the expression of AA transporters in the 3 small intestine segments, serum concentration of AA, and performance. Dietary treatments based on wheat and soybean meal (SBM) were 1) low-CP (14%) diet supplemented with L-Lys, L-Thr, DL-Met, L-Leu, L-Ile, L-Val, L-His, L-Trp, and L-Phe (LPAA); 2) as in the LPAA but with added L-Gly as a N source (LPAA+N); 3) intermediate CP content (16%) supplemented with L-Lys HCl, L-Thr, and DL-Met (MPAA); and 4) high-CP (22%) diet (HP) without free AA. At the end of the experiment, 8 pigs from LPAA and HP were sacrificed to collect intestinal mucosa and blood samples and to dissect the carcasses. There were no differences in ADG, ADFI, G:F, and weights of carcass components and some visceral organs between treatments. Weights of the large intestine and kidney were higher in HP pigs (P protein in HP wheat-SBM diets without affecting pig performance; nonessential N does not seem to be limiting in very low-protein wheat-SBM diets for growing pigs. Also, the inclusion of free AA in the diet appears to affect their serum concentration and the expression of the AA transporter b0,+ in the duodenum of pigs.

  13. Prognostic Significance of Initial Serum Albumin and 24 Hour Daily Protein Excretion before Treatment in Multiple Myeloma.

    Directory of Open Access Journals (Sweden)

    Jia-Hong Chen

    Full Text Available 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 β2-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.

  14. Effects of aerobic exercise intervention on serum cartilage oligomeric matrix protein levels and lymphocyte dna damage in obese elderly females (United States)

    Cho, Su Youn; Roh, Hee Tae


    [Purpose] The aim of the reported research was to investigate the effects of regular aerobic exercise on cartilage oligomeric matrix protein and oxidative DNA damage in obese, elderly females. [Subjects and Methods] Sixteen class I obese, elderly females, according to World Health Organization criteria, were randomly and equally assigned to a control group (n=8) or an exercise group (n=8). The exercise group participated in exercise sessions of 60 minutes per day, 3 days per week, for a period of 8 weeks. [Results] After aerobic exercise intervention, weight, body mass index, body fat, waist circumference, and DNA damage (Tail moment) were significantly decreased, compared with baseline values. In contrast, serum cartilage oligomeric matrix protein levels were not significantly different among any groups or time-points. [Conclusion] Regular aerobic exercise may be effective for reducing obesity-induced high DNA damage levels in obese females, without causing the deformation or degradation of lower extremity articular cartilage. PMID:27390441

  15. Serum Retinol-Binding Protein 4 Concentration and Its Ratio to Serum Retinol Are Associated with Obesity and Metabolic Syndrome Components in Children

    NARCIS (Netherlands)

    Aeberli, I.; Molinari, L.; Spinas, G.; Lehmann, R.; Allemand, l' D.; Zimmermann, M.B.


    Objective: The objective of the study was to measure serum RBP4, serum retinol (SR), the RBP4-to-SR molar ratio, and dietary VA intakes in normal-weight and overweight children and investigate the relationship of these variables to IR, subclinical inflammation, and the metabolic syndrome in this age

  16. Lipopolysaccharide binding protein and serum amyloid A secretion by human intestinal epithelial cells during the acute phase response. (United States)

    Vreugdenhil, A C; Dentener, M A; Snoek, A M; Greve, J W; Buurman, W A


    The acute phase proteins LPS binding protein (LBP) and serum amyloid A (SAA) are produced by the liver and are present in the circulation. Both proteins have been shown to participate in the immune response to endotoxins. The intestinal mucosa forms a large surface that is continuously exposed to these microbial products. By secretion of antimicrobial and immunomodulating agents, the intestinal epithelium contributes to the defense against bacteria and their products. The aim of this study was to explore the influence of the inflammatory mediators TNF-alpha, IL-6, and IL-1beta on the release of LBP and SAA by intestinal epithelial cells (IEC). In addition, the induction of LBP and SAA release by cell lines of intestinal epithelial cells and hepatic cells was compared. The data obtained show that in addition to liver cells, IEC also expressed LBP mRNA and released bioactive LBP and SAA upon stimulation. Regulation of LBP and SAA release by IEC and hepatocytes was typical for class 1 acute phase proteins, although differences in regulation between the cell types were observed. Endotoxin did not induce LBP and SAA release. Glucocorticoids were demonstrated to strongly enhance the cytokine-induced release of LBP and SAA by IEC, corresponding to hepatocytes. The data from this study, which imply that human IEC can produce LBP and SAA, suggest a role for these proteins in the local defense mechanism of the gut to endotoxin. Furthermore, the results demonstrate that tissues other than the liver are involved in the acute phase response.

  17. A comparison of chemical and electrophoretic methods of serum protein determinations in clinically normal domestic animals of various ages. (United States)

    Green, S A; Jenkins, S J; Clark, P A


    The biuret total protein method and a bromcresol green (BCG) albumin method were used on the Abbott ABA-100 chemistry analyzer to assay serum proteins in clinically normal cattle, sheep, ponies, pigs, and ducks. Total proteins were also read on a refractometer and mylar supported cellulose acetate electrophoresis was performed. Globulins and A/G ratios were calculated from the chemical method and the results compared with the electrophoretic method. Total protein, albumin and A/G ratios in the ponies, sheep and older cattle were in agreement between the two methods. The younger cattle and all the pigs had higher albumin levels and A/G ratios with the chemical BCG method. Ducks had slightly higher albumin values and A/G ratios with the electrophoretic method and the presence of pre-albumin was detected. Typical mylar supported cellulose acetate electrophoretic patterns are presented which show the excellent separation using these membranes. Means and range for normal animals are given and changes of proteins with age are discussed.

  18. Periodontitis increases rheumatic factor serum levels and citrullinated proteins in gingival tissues and alter cytokine balance in arthritic rats (United States)

    Corrêa, Mônica G.; Sacchetti, Silvana B.; Ribeiro, Fernanda Vieira; Pimentel, Suzana Peres; Casarin, Renato Corrêa Viana; Cirano, Fabiano Ribeiro; Casati, Marcio Z.


    This study investigated some immunological features by experimental periodontitis (EP) and rheumatoid arthritis (RA) disease interact in destructive processes in arthritic rats. Rats were assigned to the following groups: EP +RA; RA; EP; and Negative Control. RA was induced by immunizations with type-II collagen and a local immunization with Complete Freund’s adjuvant in the paw. Periodontitis was induced by ligating the right first molars. The serum level of rheumatoid factor (RF) and anti-citrullinated protein antibody (ACCPA) were measured before the induction of EP (T1) and at 28 days after (T2) by ELISA assay. ACCPA levels were also measured in the gingival tissue at T2. The specimens were processed for morphometric analysis of bone loss, and the gingival tissue surrounding the first molar was collected for the quantification of interleukin IL-1β, IL-4, IL-6, IL-17 and TNF-α using a Luminex/MAGpix assay. Paw edema was analyzed using a plethysmometer. Periodontitis increased the RF and ACCPA levels in the serum and in the gingival tissue, respectively. Besides, the level of paw swelling was increased by EP and remained in progress until the end of the experiment, when EP was associated with RA. Greater values of IL-17 were observed only when RA was present, in spite of PE. It can be concluded that periodontitis increases rheumatic factor serum levels and citrullinated proteins level in gingival tissues and alter cytokine balance in arthritic rats; at the same time, arthritis increases periodontal destruction, confirming the bidirectional interaction between diseases. PMID:28358812


    Directory of Open Access Journals (Sweden)

    M. Tariq Javed, F, Ahmad. N, Z, Rafique1 and M, Bashir


    Full Text Available Effects of higher levels of chromium alone and in combination with copper were investigated in broiler chicks divided into seven equal groups viz. A, B, C, D, E, F and G. Group G served as control receiving no treatment. Groups A, B and F received chromium chloride at the rate of 2 g/kg and nicotinic acid 150 mg/kg feed while C, D and F received chromium chloride 8 g/kg and nicotinic acid 150mg/kg. Broilers of groups A and C received copper sulfate at the rate of 200 mg/kg while groups Band D 400 mg/kg feed. Haematological parameters studied revealed non-significant difference between treatment groups and control in haemoglobin concentration and total erythrocyte counts. However, only at 4th week, lower PCV was observed in birds fed higher levels of chromium chloride alone. Increase in TLC was observed in birds fed low chromium alone or' with low levels of copper. Results of serum proteins including total protein, albumin and globulin during first three weeks showed significantly or relatively lower values in treatment groups than control. Serum globulins generally revealed non-significant difference between treatment groups and control.

  20. High Serum Adipocyte Fatty Acid Binding Protein Is Associated with Metabolic Syndrome in Patients with Type 2 Diabetes

    Directory of Open Access Journals (Sweden)

    Jer-Chuan Li


    Full Text Available Adipocyte fatty acid binding protein (A-FABP is a key mediator of obesity-related metabolic syndrome (MetS. The aim of this study was to evaluate the relationship between A-FABP concentration and MetS in type 2 diabetes mellitus (DM patients. Fasting blood samples were obtained from 165 type 2 DM volunteers. MetS and its components were defined using diagnostic criteria from the International Diabetes Federation. Among 165 DM patients, 113 patients (68.5% had MetS. Diabetic persons who had MetS had significantly higher A-FABP levels (P<0.001 than those without MetS. Female DM persons had higher A-FABP level than man (P<0.001. No statistically significant differences in A-FABP levels were found in use of statin, fibrate, or antidiabetic drugs. Multivariate forward stepwise linear regression analysis revealed that body fat mass (P<0.001, logarithmically transformed creatinine (log-creatinine; P<0.001, female DM patients (P<0.001, and logarithmically transformed high sensitive C-reactive protein (log-hs-CRP; P=0.013 were positively correlated, while albumin (P=0.004 and glomerular filtration rate (GFR; P=0.043 were negatively correlated with serum A-FABP levels in type 2 DM patients. In this study, higher serum A-FABP level was positively associated with MetS in type 2 DM patients.

  1. Efficacy of mineral and organic adsorbent in alleviating harmful effects of zearalenone on pig blood serum protein status

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    Nešić Ksenija


    Full Text Available The influence of zearalenone on blood serum protein status and the feasibility of utilizing a modified clinoptilolite and esterified glucomannan to alleviate its harmful effects was examined in two trials, 31 and 29 days long, conducted on a total of 64 pigs (32 each 60 days old, divided into four groups, each containing 8 pigs. Control groups (K received noncontaminated feed, while experimental groups received feed supplemented with 3.84 mg/kg in the first trial and 5.12 mg/kg of zearalenone in the second trial. Pigs in the first experimental groups (O-I were given feed with toxin only. Modified clinoptilolite in the amount of 0.2% and esterified glucomannan in the amount of 0.1% were introduced in contaminated feed of the second (O-II and the third experimental groups (O-III of both trials. With the use of contaminated feed, a declining trend of the A/G ratio was observed: decrease of albumin content and increase of globulin content on account of the _ globulin fraction. A decrease of the _ globulin fraction was detected at the same time. Total protein concentration was also lower. The application of adsorbents successfully alleviated harmful effects of the F-2 toxin on the affected biochemical parameters in blood serum.

  2. Targeting proteins with toxic azo dyes: a microcalorimetric characterization of the interaction of the food colorant amaranth with serum proteins. (United States)

    Basu, Anirban; Kumar, Gopinatha Suresh


    The interaction of amaranth with two homologous serum albumins from human and bovine (HSA and BSA) was studied by microcalorimetry. The binding stoichiometry for the complexation of amaranth to both BSA and HSA was around 1, and the equilibrium constants were (5.79 ± 0.07) × 10(5) and (1.76 ± 0.05) × 10(5) M(-1), respectively. The binding reaction to HSA at 298.15 K was driven by a large negative enthalpic contribution and a small but positive entropic contribution, while to BSA, it was entirely enthalpy-driven and the entropic contribution was unfavorable. Parsing of the standard molar Gibbs energy revealed that the complexation was dominated by non-polyelectrolytic forces. Temperature-dependent isothermal titration calorimetry studies revealed that the enthalpic contribution increased and the entropic contribution decreased with the rise in the temperature but the Gibbs energy change remained almost unaltered. Differential scanning calorimetry results revealed that the binding reaction stabilized the serum albumins significantly against thermal unfolding.

  3. Relationship of serum RBP, NEFA and FKN content with urinary protein excretion and glomerular filtration function damage in patients with early diabetic nephropathy

    Institute of Scientific and Technical Information of China (English)

    Zhi Yang; Ji Zhang; Li-Ping Tang


    Objective:To analyze the relationship of serum RBP, NEFA and FKN content with urinary protein excretion and glomerular filtration function damage in patients with early diabetic nephropathy.Methods: A total of 76 patients with early diabetic nephropathy were included in observation group and 80 healthy subjects were included in control group. Differences in retinol-binding protein (RBP), non-esterified fatty acid (NEFA) and chemokine Fractalkine (FKN) content in serum as well as urinary protein excretion indexes and glomerular filtration function indexes were compared between the two groups immediately after admission, and the correlation of RBP, NEFA and FKN content with the disease was further analyzed.Results:Serum RBP, NEFA and FKN content of observation group were higher than those of control group; 24 h total urine volume, 24 h urinary albumin and urinary albumin excretion rate as well as urine-specific proteins Kim-1, NGAL, L-FABP and CysC content of observation group were higher than those of control group and positively correlated with serum RBP, NEFA and FKN content; GFR level of observation group was lower than that of control group and negatively correlated with serum RBP, NEFA and FKN content while serum Cr, BUN, VF, LP-a,β2-MG and HA content were higher than those of control group and positively correlated with serum RBP, NEFA and FKN content.Conclusions:Serum RBP, NEFA and FKN content significantly increase in patients with early diabetic nephropathy and are correlated with urinary protein excretion and glomerular filtration function damage.

  4. Difficulties in diagnosing acute kidney injury post liver transplantation using serum creatinine based diagnostic criteria

    Institute of Scientific and Technical Information of China (English)

    Banwari; Agarwal; Andrew; Davenport


    Renal function in patients with advanced cirrhosis is an important prognostic factor for survival both prior to and following liver transplantation. The importance of renal function is reflected by the introduction of the model for end stage liver disease(MELD) score, which includes serum creatinine. The MELD score has been shown to predict the short term risk of death for transplant wait listed patients and is currently used by many countries to allocate liver transplants on the basis of severity of underlying illness. Changes in serum creatinine are also used to stage acute kidney injury. However prior to liver transplantation the serum creatinine typically over estimates underlying renal function, particularly when a colorimetric Jaffe based assay is used, and paradoxically then under estimates renal function post liver transplantation, particularly when immunophyllins are started early as part of transplant immunosuppression. As acute kidney injury is defined by changes in serum creatinine, this potentially leads to over estimation of the incidence and severity of acute kidney injury in the immediate post-operative period.

  5. Correlation of Serum Zinc Level with Simple Febrile Seizures: A Hospital based Prospective Case Control Study

    Directory of Open Access Journals (Sweden)

    Imran Gattoo


    Full Text Available Background: Febrile seizures are one of the most common neurological conditions of childhood. It seems that zinc deficiency is associated with increased risk of febrile seizures.Aim: To estimate the serum Zinc level in children with simple Febrile seizures and to find the correlation between serum zinc level and simple Febrile seizures.Materials and Methods: The proposed study was a hospital based prospective case control study which included infants and children aged between 6 months to 5 years, at Post Graduate Department of Pediatrics, (SMGS Hospital, GMC Jammu, northern India. A total of 200 infants and children fulfilling the inclusion criteria were included. Patients were divided into 100(cases in Group A with simple febrile seizure and 100(controls in Group B of children with acute febrile illness without seizure. All patients were subjected to detailed history and thorough clinical examination followed by relevant investigations.Results: Our study had slight male prepondance of 62% in cases and 58% in controls . Mean serum zinc level in cases was 61.53±15.87 ugm/dl and in controls it was 71.90+18.50 ugm/dl .Serum zinc level was found significantly low in cases of simple febrile seizures as compaired to controls ,with p value of

  6. Electrochemical immunoassay of cotinine in serum based on nanoparticle probe and immunochromatographic strip

    Energy Technology Data Exchange (ETDEWEB)

    Nian Hungchi [Pacific Norwest National Laboratory, Richland, WA 99352 (United States); Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, Hsinchu, 30013, Taiwan (China); Wang Jun; Wu Hong [Pacific Norwest National Laboratory, Richland, WA 99352 (United States); Lo, Jiunn-Guang [Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, Hsinchu, 30013, Taiwan (China); Chiu, Kong-Hwa [Department of Applied Science, National DongHwa University, Hualien, 970, 30013, Taiwan (China); Pounds, Joel G. [Pacific Norwest National Laboratory, Richland, WA 99352 (United States); Lin Yuehe, E-mail: [Pacific Norwest National Laboratory, Richland, WA 99352 (United States)


    Highlights: Black-Right-Pointing-Pointer Using nanoparticle probe in immunochromatographic strip. Black-Right-Pointing-Pointer Enhanced sensitivity by using nanoparticle label. Black-Right-Pointing-Pointer Rapid and sensitive detection of cotinine in serum. - Abstract: A disposable sensor for the determination of cotinine in human serum was developed based on immunochromatographic test strip and quantum dot label. In this assay, cotinine linked with quantum dot competes with cotinine in sample to bind to anti-cotinine antibody in the test strip and the quantum dots serve as signal vehicles for electrochemical readout. Some parameters governing the performance of the sensor were optimized. The sensor shows a wide linear range from 1 ng mL{sup -1} to 100 ng mL{sup -1} cotinine with a detection limit of 1.0 ng mL{sup -1}. The sensor was validated with spiked human serum samples and it was found that this method was reliable in measuring cotinine in human serum. The results demonstrate that this sensor is rapid, accurate, and less expensive and has the potential for point of care (POC) detection of cotinine and fast screening of tobacco smoke exposure.

  7. Binding of complement proteins C1q and C4bp to serum amyloid P component (SAP) in solid contra liquid phase

    DEFF Research Database (Denmark)

    Sørensen, Inge Juul; Nielsen, EH; Andersen, Ove;


    Serum amyloid P component (SAP), a member of the conserved pentraxin family of plasma proteins, binds calcium dependently to its ligands. The authors investigated SAPs interaction with the complement proteins C4b binding protein (C4bp) and C1q by ELISA, immunoelectrophoresis and electron microscopy...... affinity, did not interfere with the subsequent binding of C4bp or C1q to SAP. In contrast, collagen I and IV showed partial competition with the binding of C1q to SAP. Using fresh serum, immobilized native SAP bound C4bp whereas binding of C1q/C1 could not be demonstrated. Altogether the results indicate...

  8. How does fatty acid influence anti-thyroid drugs binding and specificity toward protein human serum albumin? A blind docking simulation study

    Indian Academy of Sciences (India)

    Bijan K Paul; Nikhil Guchhait


    This study reports an AutoDock-based blind docking simulation investigation to characterize the binding interaction of a series of anti-thyroid drugs (2-mercapto-1-methylimidazole (MMI), 2-thiouracil (TU), 6-methyl-2-thiouracil (MTU), 6--propyl-2-thiouracil (PTU) with a model plasma protein Human SerumAlbumin (HSA) in the presence and absence of fatty acid (FA). The drug-protein binding efficiency is characterized in terms of binding free energy and the association constant (Ka, which is estimated as the reciprocal of the inhibition constant, Ki) of the drugs to the transport protein. The study also unveils the substantial impact of the presence of fatty acid (FA) on the binding interaction process. It is shown that in the presence of FA the drug-protein binding efficiency is markedly enhanced (except for MTU) and the binding location is changed. Hydrogen bonding interaction appears to play a governing role in the process of FA-induced modifications of binding efficiency and location.

  9. Glial and neuronal proteins in serum predict outcome after severe traumatic brain injury.

    NARCIS (Netherlands)

    Vos, P.E.; Lamers, K.J.B.; Hendriks, J.C.M.; Haaren, M. van; Beems, T.; Zimmerman, C.; Geel, W.J.A. van; Reus, H.P.M. de; Biert, J.; Verbeek, M.M.


    OBJECTIVE: To study the ability of glial (glial fibrillary acidic protein [GFAP] and S100b) and neuronal (neuron specific enolase [NSE]) protein levels in peripheral blood to predict outcome after severe traumatic brain injury. METHODS: Eighty-five patients with severe traumatic brain injury (admiss

  10. Temperature-Triggered Protein Adsorption on Polymer-Coated Nanoparticles in Serum

    NARCIS (Netherlands)

    Koshkina, O.; Lang, T.; Thiermann, R.; Docter, D.; Stauber, R.H.; Secker, C.; Schlaad, H.; Weidner, S.; Mohr, B.; Maskos, M.; Bertin, A.


    The protein corona, which forms on the nanoparticle's surface in most biological media, determines the nanoparticle's physicochemical characteristics. The formation of the protein corona has a significant impact on the biodistribution and clearance of nanoparticles in vivo. Therefore, the ability to


    Directory of Open Access Journals (Sweden)

    Zaiter Samir


    Full Text Available Some kinds of interleikines of patients with chronic laryngitis disease were investigated. There is “cytokines explosion” of the patients with chronic laryngitis with persistent herpes simplex virus. Comparative investigation cytokine profile in serum blood is demonstrated: balanced reaction cytokines profile of patients with chronic laryngitis without persistent herpes simplex virus and dysbalanced reaction of patients with laryngitis (hyperergation. Increased content IL-6 and low content g-interferon and tumor necrosis factor (TNF-a are predisposition of chronisation inflammation processes in larynges. This situation needs sighting correction.

  12. CLP-based protein fragment assembly

    CERN Document Server

    Palu', Alessandro Dal; Fogolari, Federico; Pontelli, Enrico; 10.1017/S1471068410000372


    The paper investigates a novel approach, based on Constraint Logic Programming (CLP), to predict the 3D conformation of a protein via fragments assembly. The fragments are extracted by a preprocessor-also developed for this work- from a database of known protein structures that clusters and classifies the fragments according to similarity and frequency. The problem of assembling fragments into a complete conformation is mapped to a constraint solving problem and solved using CLP. The constraint-based model uses a medium discretization degree Ca-side chain centroid protein model that offers efficiency and a good approximation for space filling. The approach adapts existing energy models to the protein representation used and applies a large neighboring search strategy. The results shows the feasibility and efficiency of the method. The declarative nature of the solution allows to include future extensions, e.g., different size fragments for better accuracy.

  13. MALDI-TOF MS Combined With Magnetic Beads for Detecting Serum Protein Biomarkers and Establishment of Boosting Decision Tree Model for Diagnosis of Colorectal Cancer

    Directory of Open Access Journals (Sweden)

    Chibo Liu, Chunqin Pan, Jianmin Shen, Haibao Wang, Liang Yong


    Full Text Available The aim of present study is to study the serum protein fingerprint of patients with colorectal cancer (CRC and to screen protein molecules that are closely related to colorectal cancer during the onset and progression of the disease with Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS. Serum samples from 144 patients with CRC and 120 healthy volunteers were adopted in present study. Weak cation exchange (WCX magnetic beads and PBSII-C protein chips reader (Ciphergen Biosystems Ins. were used. The protein fingerprint expression of all the Serum samples and the resulted profiles between cancer and normal groups were analyzed with Biomarker Wizard system. Several proteomic peaks were detected and four potential biomarkers with different expression profiles were identified with their relative molecular weights of 2870.7Da, 3084Da, 9180.5Da, and 13748.8Da, respectively. Among the four proteins, two proteins with m/z 2870.7 and 3084 were down-regulated, and the other two with m/z 9180.5 and 13748.8 were up-regulated in serum samples from CRC patients. The present diagnostic model could distinguish CRC from healthy controls with the sensitivity of 92.85% and the specificity of 91.25%. Blind test data indicated a sensitivity of 86.95% and a specificity of 85%. The result suggested that MALDI technology could be used to screen critical proteins with differential expression in the serum of CRC patients. These differentially regulated proteins were considered as potential biomarkers for the patients with CRC in the serum and of the potential value for further investigation.

  14. Discovery of binding proteins for a protein target using protein-protein docking-based virtual screening. (United States)

    Zhang, Changsheng; Tang, Bo; Wang, Qian; Lai, Luhua


    Target structure-based virtual screening, which employs protein-small molecule docking to identify potential ligands, has been widely used in small-molecule drug discovery. In the present study, we used a protein-protein docking program to identify proteins that bind to a specific target protein. In the testing phase, an all-to-all protein-protein docking run on a large dataset was performed. The three-dimensional rigid docking program SDOCK was used to examine protein-protein docking on all protein pairs in the dataset. Both the binding affinity and features of the binding energy landscape were considered in the scoring function in order to distinguish positive binding pairs from negative binding pairs. Thus, the lowest docking score, the average Z-score, and convergency of the low-score solutions were incorporated in the analysis. The hybrid scoring function was optimized in the all-to-all docking test. The docking method and the hybrid scoring function were then used to screen for proteins that bind to tumor necrosis factor-α (TNFα), which is a well-known therapeutic target for rheumatoid arthritis and other autoimmune diseases. A protein library containing 677 proteins was used for the screen. Proteins with scores among the top 20% were further examined. Sixteen proteins from the top-ranking 67 proteins were selected for experimental study. Two of these proteins showed significant binding to TNFα in an in vitro binding study. The results of the present study demonstrate the power and potential application of protein-protein docking for the discovery of novel binding proteins for specific protein targets.

  15. Serum- and glucocorticoid-induced protein kinase 1 (SGK1) increases the cystic fibrosis transmembrane conductance regulator (CFTR) in airway epithelial cells by phosphorylating Shank2E protein. (United States)

    Koeppen, Katja; Coutermarsh, Bonita A; Madden, Dean R; Stanton, Bruce A


    The glucocorticoid dexamethasone increases cystic fibrosis transmembrane conductance regulator (CFTR) abundance in human airway epithelial cells by a mechanism that requires serum- and glucocorticoid-induced protein kinase 1 (SGK1) activity. The goal of this study was to determine whether SGK1 increases CFTR abundance by phosphorylating Shank2E, a PDZ domain protein that contains two SGK1 phosphorylation consensus sites. We found that SGK1 phosphorylates Shank2E as well as a peptide containing the first SGK1 consensus motif of Shank2E. The dexamethasone-induced increase in CFTR abundance was diminished by overexpression of a dominant-negative Shank2E in which the SGK1 phosphorylation sites had been mutated. siRNA-mediated reduction of Shank2E also reduced the dexamethasone-induced increase in CFTR abundance. Taken together, these data demonstrate that the glucocorticoid-induced increase in CFTR abundance requires phosphorylation of Shank2E at an SGK1 consensus site.

  16. Vagus nerve electrical stimulation inhibits serum levels of S100A8 protein in septic shock rats. (United States)

    Lei, Ming; Liu, Xin-Xin


    The vagus nerve and the released acetylcholine exert anti-inflammatory effects and inhibit septic shock. However, their detailed mechanisms remain to be elucidated. The present study aimed to investigate the effects of vagus nerve electrical stimulation on serum S100A8 levels in septic shock rats. A total of 36 male Sprague-Dawley rats were randomly divided into six equal groups: i) Sham group, receiving sham operation; ii) CLP group, subjected to cecal ligation and puncture (CLP) to establish a model of polymicrobial sepsis; iii) VGX group, subjected to CLP and bilateral cervical vagotomy; iv) STM group, subjected to CLP, bilateral cervical vagotomy and electrical stimulation on the left vagus nerve trunk; v) α‑bungarotoxin (BGT) group was administered α‑BGT prior to electrical stimulation; vi) Anti‑receptor for advanced glycation end products (RAGE) group, administered intraperitoneal injection of anti‑RAGE antibody prior to electrical stimulation. The right carotid artery was cannulated to monitor mean artery pressure (MAP). The serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were measured to assess the liver function. Serum S100A8 and advanced glycation end product (AGE) levels were measured using enzyme‑linked immunosorbent assays. The expression of hepatic RAGE was determined by western blotting. The present study revealed that Sprague‑Dawley rats exhibited progressive hypotension and significantly increased serum AST and ALT levels following CLP challenge compared with the sham group. The levels of S100A8 and AGEs, and the protein expression of hepatic RAGE were significantly increased following CLP compared with the sham group. Vagus nerve electrical stimulation significantly prevented the development of CLP‑induced hypotension, alleviated the hepatic damage, reduced serum S100A8 and AGEs production, and reduced the expression of hepatic RAGE. The inhibitory effect of vagus nerve electrical

  17. Fluorescent sensors based on bacterial fusion proteins (United States)

    Prats Mateu, Batirtze; Kainz, Birgit; Pum, Dietmar; Sleytr, Uwe B.; Toca-Herrera, José L.


    Fluorescence proteins are widely used as markers for biomedical and technological purposes. Therefore, the aim of this project was to create a fluorescent sensor, based in the green and cyan fluorescent protein, using bacterial S-layers proteins as scaffold for the fluorescent tag. We report the cloning, expression and purification of three S-layer fluorescent proteins: SgsE-EGFP, SgsE-ECFP and SgsE-13aa-ECFP, this last containing a 13-amino acid rigid linker. The pH dependence of the fluorescence intensity of the S-layer fusion proteins, monitored by fluorescence spectroscopy, showed that the ECFP tag was more stable than EGFP. Furthermore, the fluorescent fusion proteins were reassembled on silica particles modified with cationic and anionic polyelectrolytes. Zeta potential measurements confirmed the particle coatings and indicated their colloidal stability. Flow cytometry and fluorescence microscopy showed that the fluorescence of the fusion proteins was pH dependent and sensitive to the underlying polyelectrolyte coating. This might suggest that the fluorescent tag is not completely exposed to the bulk media as an independent moiety. Finally, it was found out that viscosity enhanced the fluorescence intensity of the three fluorescent S-layer proteins.

  18. Protein-protein interaction network-based detection of functionally similar proteins within species. (United States)

    Song, Baoxing; Wang, Fen; Guo, Yang; Sang, Qing; Liu, Min; Li, Dengyun; Fang, Wei; Zhang, Deli


    Although functionally similar proteins across species have been widely studied, functionally similar proteins within species showing low sequence similarity have not been examined in detail. Identification of these proteins is of significant importance for understanding biological functions, evolution of protein families, progression of co-evolution, and convergent evolution and others which cannot be obtained by detection of functionally similar proteins across species. Here, we explored a method of detecting functionally similar proteins within species based on graph theory. After denoting protein-protein interaction networks using graphs, we split the graphs into subgraphs using the 1-hop method. Proteins with functional similarities in a species were detected using a method of modified shortest path to compare these subgraphs and to find the eligible optimal results. Using seven protein-protein interaction networks and this method, some functionally similar proteins with low sequence similarity that cannot detected by sequence alignment were identified. By analyzing the results, we found that, sometimes, it is difficult to separate homologous from convergent evolution. Evaluation of the performance of our method by gene ontology term overlap showed that the precision of our method was excellent.

  19. A protein relational database and protein family knowledge bases to facilitate structure-based design analyses. (United States)

    Mobilio, Dominick; Walker, Gary; Brooijmans, Natasja; Nilakantan, Ramaswamy; Denny, R Aldrin; Dejoannis, Jason; Feyfant, Eric; Kowticwar, Rupesh K; Mankala, Jyoti; Palli, Satish; Punyamantula, Sairam; Tatipally, Maneesh; John, Reji K; Humblet, Christine


    The Protein Data Bank is the most comprehensive source of experimental macromolecular structures. It can, however, be difficult at times to locate relevant structures with the Protein Data Bank search interface. This is particularly true when searching for complexes containing specific interactions between protein and ligand atoms. Moreover, searching within a family of proteins can be tedious. For example, one cannot search for some conserved residue as residue numbers vary across structures. We describe herein three databases, Protein Relational Database, Kinase Knowledge Base, and Matrix Metalloproteinase Knowledge Base, containing protein structures from the Protein Data Bank. In Protein Relational Database, atom-atom distances between protein and ligand have been precalculated allowing for millisecond retrieval based on atom identity and distance constraints. Ring centroids, centroid-centroid and centroid-atom distances and angles have also been included permitting queries for pi-stacking interactions and other structural motifs involving rings. Other geometric features can be searched through the inclusion of residue pair and triplet distances. In Kinase Knowledge Base and Matrix Metalloproteinase Knowledge Base, the catalytic domains have been aligned into common residue numbering schemes. Thus, by searching across Protein Relational Database and Kinase Knowledge Base, one can easily retrieve structures wherein, for example, a ligand of interest is making contact with the gatekeeper residue.

  20. Identification of high immunoreactive proteins from Streptococcus agalactiae isolates recognized by human serum antibodies. (United States)

    Brzychczy-Wloch, Monika; Gorska, Sabina; Brzozowska, Ewa; Gamian, Andrzej; Heczko, Piotr B; Bulanda, Malgorzata


    The aim of the studies was to identify immunogenic proteins of Streptococcus agalactiae (group B streptococcus; GBS) isolates. Investigation of the immunoreactivity with human sera allowed us to determine major immunogenic proteins which might be potential candidates for the development of vaccine. For the study, we have selected 60 genetically different, well-characterized GBS clinical isolates. The proteins immunoreactivity with 24 human sera from patients with GBS infections, carriers, and control group without GBS was detected by SDS-PAGE and Western blotting. As a result, some major immunogenic proteins were identified, of which four proteins with molecular masses of about 45 to 50 kDa, which exhibited the highest immunoreactivity features, were analyzed by LC-MS/MS. The proteins were identified by comparative analysis of peptides masses using MASCOT and statistical analysis. The results showed known molecules such as enolase (47.4 kDa), aldehyde dehydrogenase (50.6 kDa), and ones not previously described such as trigger factor (47 kDa) and elongation factor Tu (44 kDa). The preliminary results indicated that some GBS proteins that elicit protective immunity hold promise not only as components in a vaccine as antigens but also as carriers or adjuvants in polysaccharide conjugate vaccines, but more studies are needed.

  1. Expression analysis of bone morphogenetic protein 4 between fat and lean birds in adipose tissue and serum. (United States)

    Cheng, B H; Leng, L; Wu, M Q; Zhang, Q; Zhang, X Y; Xu, S S; Cao, Z P; Li, Y M; Luan, P; Li, H


    The objectives of the present study were to characterize the tissue expression of chicken (Gallus gallus) bone morphogenetic protein 4 (BMP4) and compare differences in its expression in abdominal fat tissue and serum between fat and lean birds and to determine a potential relationship between the expression of BMP4 and abdominal fat tissue growth and development. The results showed that chicken BMP4 messenger RNA (mRNA) and protein were expressed in various tissues, and the expression levels of BMP4 transcript and protein were relatively higher in adipose tissues. In addition, the mRNA and protein expression levels of BMP4 in abdominal fat tissue of fat males were lower than those of lean males at 1, 2, 5, and 7 wk of age (P fat males was lower than that of lean males at 7 wk of age (P fat deposition through differences in its expression level. The results of this study will provide basic molecular information for studying the role of BMP4 in the regulation of adipogenesis in avian species.

  2. Expression and bioactivity of recombinant human serum albumin and dTMP fusion proteins in CHO cells. (United States)

    Ru, Yi; Zhi, Dejuan; Guo, Dingding; Wang, Yong; Li, Yang; Wang, Meizhu; Wei, Suzhen; Wang, Haiqing; Wang, Na; Che, Jingmin; Li, Hongyu


    The 14-amino acid (IEGPTLRQWLAARA) thrombopoietin mimetic peptide (TMP) shares no sequence homology with native thrombopoietin (TPO). When dimerized, it displays a high-binding affinity for the TPO receptor and has equipotent bioactivity with recombinant human TPO (rhTPO) in stimulating proliferation and maturation of megakaryocytes in vitro. However, TMP is limited for clinical usage because of its short half-life in vivo. In this study, fusion proteins that composed of tandem dimer of TMP (dTMP) genetically fused at the C- or N-terminus of human serum albumin (HSA) were separately expressed in Chinese hamster ovary (CHO) cells. In vitro bioactivity assays showed that purified fusion proteins promoted the proliferation of megakaryocytes in a dose-dependent manner and activated signal transducer and activator of transcription (STAT) pathway in TPO receptor-dependent manner. Following subcutaneous administration, both HSA-dTMP and dTMP-HSA significantly elevated peripheral platelet counts in normal mice in a dose-dependent manner. In addition, fusion with HSA successfully prolonged dTMP half-life in mice. However, when HSA was fused at the C-terminus of dTMP, the bioactivity of dTMP-HSA was about half of that of HSA-dTMP. In conclusion, these results suggested that HSA/dTMP fusion proteins might be potential drugs for thrombocytopenia and, when HSA was fused at the N-terminus of dTMP, the fusion protein had a higher activity.

  3. Role of pH-induced structural change in protein aggregation in foam fractionation of bovine serum albumin

    Directory of Open Access Journals (Sweden)

    Rui Li


    Full Text Available For reducing protein aggregation in foam fractionation, the role of pH-induced structural change in the interface-induced protein aggregation was analyzed using bovine serum albumin (BSA as a model protein. The results show that the decrease in pH from 7.0 to 3.0 gradually unfolded the BSA structure to increase the molecular size and the relative content of β-sheet and thus reduced the stability of BSA in the aqueous solution. At the isoelectric point (pH 4.7, BSA suffered the lowest level in protein aggregation induced by the gas–liquid interface. In the pH range from 7.0 to 4.7, most BSA aggregates were formed in the defoaming process while in the pH range from 4.7 to 3.0, the BSA aggregates were formed at the gas–liquid interface due to the unfolded BSA structure and they further aggregated to form insoluble ones in the desorption process.

  4. Multi-parametric surface plasmon resonance platform for studying liposome-serum interactions and protein corona formation. (United States)

    Kari, Otto K; Rojalin, Tatu; Salmaso, Stefano; Barattin, Michela; Jarva, Hanna; Meri, Seppo; Yliperttula, Marjo; Viitala, Tapani; Urtti, Arto


    When nanocarriers are administered into the blood circulation, a complex biomolecular layer known as the "protein corona" associates with their surface. Although the drivers of corona formation are not known, it is widely accepted that this layer mediates biological interactions of the nanocarrier with its surroundings. Label-free optical methods can be used to study protein corona formation without interfering with its dynamics. We demonstrate the proof-of-concept for a multi-parametric surface plasmon resonance (MP-SPR) technique in monitoring the formation of a protein corona on surface-immobilized liposomes subjected to flowing 100 % human serum. We observed the formation of formulation-dependent "hard" and "soft" coronas with distinct refractive indices, layer thicknesses, and surface mass densities. MP-SPR was also employed to determine the affinity (K D ) of a complement system molecule (C3b) with cationic liposomes with and without polyethylene glycol. Tendency to create a thick corona correlated with a higher affinity of opsonin C3b for the surface. The label-free platform provides a fast and robust preclinical tool for tuning nanocarrier surface architecture and composition to control protein corona formation.

  5. The possible link between the elevated serum levels of neurokinin A and anti-ribosomal P protein antibodies in children with autism

    Directory of Open Access Journals (Sweden)

    Mostafa Gehan A


    Full Text Available Abstract 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 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.

  6. A novel serum metabolomics-based diagnostic approach for colorectal cancer.

    Directory of Open Access Journals (Sweden)

    Shin Nishiumi

    Full Text Available BACKGROUND: To improve the quality of life of colorectal cancer patients, it is important to establish new screening methods for early diagnosis of colorectal cancer. METHODOLOGY/PRINCIPAL FINDINGS: We performed serum metabolome analysis using gas-chromatography/mass-spectrometry (GC/MS. First, the accuracy of our GC/MS-based serum metabolomic analytical method was evaluated by calculating the RSD% values of serum levels of various metabolites. Second, the intra-day (morning, daytime, and night and inter-day (among 3 days variances of serum metabolite levels were examined. Then, serum metabolite levels were compared between colorectal cancer patients (N = 60; N = 12 for each stage from 0 to 4 and age- and sex-matched healthy volunteers (N = 60 as a training set. The metabolites whose levels displayed significant changes were subjected to multiple logistic regression analysis using the stepwise variable selection method, and a colorectal cancer prediction model was established. The prediction model was composed of 2-hydroxybutyrate, aspartic acid, kynurenine, and cystamine, and its AUC, sensitivity, specificity, and accuracy were 0.9097, 85.0%, 85.0%, and 85.0%, respectively, according to the training set data. In contrast, the sensitivity, specificity, and accuracy of CEA were 35.0%, 96.7%, and 65.8%, respectively, and those of CA19-9 were 16.7%, 100%, and 58.3%, respectively. The validity of the prediction model was confirmed using colorectal cancer patients (N = 59 and healthy volunteers (N = 63 as a validation set. At the validation set, the sensitivity, specificity, and accuracy of the prediction model were 83.1%, 81.0%, and 82.0%, respectively, and these values were almost the same as those obtained with the training set. In addition, the model displayed high sensitivity for detecting stage 0-2 colorectal cancer (82.8%. CONCLUSIONS/SIGNIFICANCE: Our prediction model established via GC/MS-based serum metabolomic analysis

  7. Flow cytometry-based assay to evaluate human serum MUC1-Tn antibodies

    DEFF Research Database (Denmark)

    Van Elssen, Catharina H M J; Clausen, Henrik; Germeraad, Wilfred T V;


    to the underglycosylation of MUC1, cancer-specific MUC1-Tn/STn antigens, which are highly immunogenic, become exposed. We aimed at developing a system that allows detection of antibodies directed to the native form of MUC1 and the underglycosylated MUC1-Tn epitopes. To this end, we made use of the Chinese Hamster Ovary......-ldlD MUC1 system was used to detect serum MUC1 and MUC1-Tn antibodies. Using this system, we could confirm the presence of MUC1-Tn antibodies in the serum of a patient vaccinated with a truncated MUC1 peptide. This indicates that the CHO-ldlD MUC1 system represents a flow cytometry-based technique...... in vaccination studies as well as for functional assays....

  8. A direct electron transfer-based glucose/oxygen biofuel cell operating in human serum

    Energy Technology Data Exchange (ETDEWEB)

    Coman, V.; Gorton, L. [Department of Analytical Chemistry/Biochemistry, Lund University, 22100 Lund (Sweden); Ludwig, R. [Research Centre Applied Biocatalysis, 8010 Graz (Austria); Department of Food Sciences and Technology, BOKU-University of Natural Resources and Applied Life Sciences, 1190 Wien (Austria); Harreither, W.; Haltrich, D. [Department of Food Sciences and Technology, BOKU-University of Natural Resources and Applied Life Sciences, 1190 Wien (Austria); Ruzgas, T. [Biomedical Laboratory Science, Health and Society, Malmoe University, 20506 Malmoe (Sweden); Laboratory of Chemical Enzymology, A.N. Bach Institute of Biochemistry, 119071 Moscow (Russian Federation); Shleev, S.


    We report on the fabrication and characterisation of the very first direct electron transfer-based glucose/oxygen biofuel cell (BFC) operating in neutral glucose-containing buffer and human serum. Corynascus thermophilus cellobiose dehydrogenase and Myrothecium verrucaria bilirubin oxidase were used as anodic and cathodic bioelements, respectively. The following characteristics of the mediator-, separator- and membrane-less, a priori, non-toxic and simple miniature BFC, was obtained: an open-circuit voltage of 0.62 and 0.58 V, a maximum power density of ca. 3 and 4 {mu}W cm{sup -2} at 0.37 and 0.19 V of cell voltage, in phosphate buffer and human serum, respectively. (Abstract Copyright [2010], Wiley Periodicals, Inc.)

  9. Prognostic Significance of Initial Serum Albumin and 24 Hour Daily Protein Excretion before Treatment in Multiple Myeloma. (United States)

    Chen, Jia-Hong; Hsu, Shun-Neng; Huang, Tzu-Chuan; Wu, Yi-Ying; Lin, Chin; Chang, Ping-Ying; Chen, Yeu-Chin; Ho, Ching-Liang


    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 β2-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 (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.

  10. Hepatic DNA Damage and Abnormality in Serum Protein Pattern Due to Long Term Use of Tramadol in Rats

    Directory of Open Access Journals (Sweden)

    Laila Abd El kawy


    Full Text Available Tramadol is a widely used analgesic that stimulates the μ opioid receptor and inhibits serotonin and noradrenalin reuptake. In this study, we investigate the effect of chronic administration of 200 and 400mg of tramal for 15,30 ,45 days followed by withdrawal periods(w15 on protein activity in male rats as manifested by changes in electrophoretic serum protein patterns and gene expression manifested by DNA damage, measured by Comet Assay . Materials and methods:100 male Wistar rats (100-150 g were included and divided into three groups, control group (n = 20, Tramal(I group (n = 40, received the drug orally at doses of 200mg/kg/day for 15,30,45 days of the study, respectively(10 rats for each subgroup.TR45 group was followed by 15 withdrawal period(W15 ,Tramal(II group (n = 40, received the drug orally at doses of400mg/kg/day for 15,30,45 days of the study, respectively. TR45 group was followed by 15 withdrawal period (W15. Results:Results exhibited major changes in the protein pattern which included changes in the molecular weight of the control bands and the relative percentage of protein fraction as well as the total number of bands, as a result of disappearance of some original bands and appearance of other new one. Serum protein fraction revealed an increase in total number of protein fractions being in Tr200mg (14 and 16 bands, whileTR400mg revealed a decrease in total number of bands (except TR15 which exhibited an increase in bands (14. The changes were observed all over the treated groups as well as in the withdrawal groups In this study, the alkaline version of the comet assay has been used to determine the effect of tramal administration (200 and 400mg/Kg on peroxide-initiated free radical-mediated DNA damage in rat liver cells. Indeed, levels of strand breaks in rat liver cell exposed to tramal400mg/Kg were significantly higher than in cells exposed to 200mg/Kg, especially after a long administration period (TR45 days. The intensity

  11. Unique surface adsorption behaviors of serum proteins on chemically uniform and alternating surfaces (United States)

    Song, Sheng

    With increasing interests of studying proteins adsorption on the surfaces with nanoscale features in biomedical field, it is crucial to have fundamental understandings on how the proteins are adsorbed on such a surface and what factors contribute to the driving forces of adsorption. Besides, exploring more available nanoscale templates would greatly offer more possibilities one could design surface bio-detection methods with favorable protein-surface interactions. Thus, to fulfill the purpose, the work in this dissertation has been made into three major sections. First, to probe the intermediate states which possibly exist between stable and unstable phases described in mean-field theory diagram, a solvent vapor annealing method is chosen to slowly induce the copolymer polystyrene-block-polyvinylpyridine (PS-b-PVP)'s both blocks undergoing micro-phase separations from initial spherical nanodomains into terminal cylindrical nanodomains. During this process, real time atomic force microscopy (AFM) has been conducted to capture other six intermediate states with different morphologies on the polymeric film surfaces. Secondly, upon recognizing each intermediate state, the solution of immunoglobulin gamma (IgG) proteins has been deposited on the surface and been rinsed off with buffer solution before the protein-bounded surface is imaged by AFM. It has been found IgG showing a strong adsorption preference on PS over P4VP block. Among all the six intermediate states, the proteins are almost exclusively adsorbed on PS nanodomains regardless the concentration and deposition time. Thirdly, a trinodular shape protein fibrinogen (Fg) is selected for investigating how geometry and surface charge of proteins would interplay with cylindrical nanodomains on a surface developed from Polystyrene -block-Poly-(methyl methacrylate) PS-b-PMMA. Also, Fg adsorptions on chemically homogeneous surfaces are included here to have a better contrast of showing how much difference it can make

  12. Long-term effects of insulin-like growth factor (IGF)-I on serum IGF-I, IGF-binding protein-3 and acid labile subunit in Laron syndrome patients with normal growth hormone binding protein. (United States)

    Kanety, H; Silbergeld, A; Klinger, B; Karasik, A; Baxter, R C; Laron, Z


    A minority of patients with Laron syndrome have normal serum GH binding protein (GHBP), indicating that the defect is elsewhere than in the extracellular domain of the GH receptor. We have evaluated the effect of long-term IGF-I treatment on serum IGF-binding protein (IGFBP)-3 and the acid-labile subunit (ALS) in three sibling with Laron syndrome caused by a GH post-receptor defect and with normal GHBP. The children (a boy aged 3 years, a girl aged 4 years and a boy aged 10 years) were treated by daily s.c. injection of IGF-I in a dose of 150 micrograms/kg. IGFBP-3 was measured by RIA and Western ligand blotting, ALS by RIA. Based values of IGFBP-3 and ALS were low. During IGF-I treatment, the IGFBP-3 concentrations in the girl gradually increased, whereas in the boys there was a 60% decrease during the first week, followed by gradual increase towards baseline. The ALS concentrations followed a similar pattern. We conclude that IGF-I treatment induces and initial suppression and then an increase in the IGFBP-3 and ALS concentrations, confirming data from animal experiments that IGFBP-3 synthesis is not solely under GH control. The differences in responsiveness between the female and male siblings may reflect genetic differences, or lower circulating concentrations of IGF-I in the boys compared with the girl.

  13. Serum antibody responses to Wolbachia surface protein in patients with human lymphatic filariasis. (United States)

    Shiny, Chandanapurath; Krushna, Nagampalli S A; Archana, Bairavasundaram; Farzana, Begum; Narayanan, Rangarajan B


    Wolbachia surface protein (WSP), which is the most abundantly expressed protein of Wolbachia from the human filarial parasite Brugia malayi, was chosen for the present study. B-cell epitope prediction of the WSP protein sequence indicates a high antigenicity, surface probability and hydrophilicity by DNA STAR software analysis. ProPred analysis suggests the presence of HLA class II binding regions in the WSP protein that contribute to T-cell responses and isotype reactivity. In order to validate these findings, the gene coding for endosymbiont WSP was PCR-amplified from the genomic DNA of the human filarial parasite Brugia malayi and cloned in T-7 expression vector pRSET-A. Western blot and ELISA at the total IgG level with recombiant WSP indicated a significantly elevated reactivity in CP compared to MF, EN and NEN individuals. Isotype ELISA also suggested an elevated reactivity in CP patients at the IgG1 level. In contrast, WSP-specific IgG4 levels were found to be elevated in MF patients compared to CP and EN. Besides this, WSP-specific IgE levels indicated an elevated reactivity in CP and MF patients compared to normals. Observations from ELISA supported the in silico predictions that indicate the presence of B- and T-cell epitopes. Hence, a combinatorial approach of in silico predictions and wet-lab studies provides interesting insights into the role of Wolbachia proteins in filarial pathogenesis.

  14. Assessing protein-protein interactions based on the semantic similarity of interacting proteins. (United States)

    Cui, Guangyu; Kim, Byungmin; Alguwaizani, Saud; Han, Kyungsook


    The Gene Ontology (GO) has been used in estimating the semantic similarity of proteins since it has the largest and reliable vocabulary of gene products and characteristics. We developed a new method which can assess Protein-Protein Interactions (PPI) using the branching factor and information content of the common ancestor of interacting proteins in the GO hierarchy. We performed a comparative evaluation of the measure with other GO-based similarity measures and evaluation results showed that our method outperformed others in most GO domains.

  15. Microchip-based ELISA strategy for the detection of low-level disease biomarker in serum

    Energy Technology Data Exchange (ETDEWEB)

    Liu Yun; Wang Huixiang; Huang Jingyu; Yang Jie; Liu Baohong [Department of Chemistry, Institute of Biomedical Sciences, Fudan University, Shanghai 200433 (China); Yang Pengyuan, E-mail: [Department of Chemistry, Institute of Biomedical Sciences, Fudan University, Shanghai 200433 (China)


    A simple and sensitive method has been proposed to determine a trace level of {alpha}-fetoprotein (AFP), hepatocellular carcinoma biomarker, using poly(methyl methacrylate) (PMMA) microfluidic chips coupled with electrochemical detection system. The PMMA microchannels have been modified with poly(ethyleneimine) (PEI) containing abundant NH{sub 2} groups to covalently immobilize AFP monoclonal antibody. Afterward, the antigen AFP and horseradish peroxidase (HRP)-conjugated AFP antibody can sequentially bind through antigen-antibody specific interaction. Atomic force microscopy (AFM) and confocal fluorescence microscope (CFFM) were utilized to characterize the surface topography and protein immobilization after modification. Coupled with three-electrode electrochemical detection system, the immunochip can perform the detection limit of AFP down to 1 pg mL{sup -1}, and achieve a detectable linear concentration range of 1-500 pg mL{sup -1} by differential pulse voltammetry (DPV). The on-chip immunoassay platform can not only provide rapid and sensitive detection for target proteins but also be resistant to non-specific adsorption of proteins, which contributes to the detection of low-level protein in real sample. Finally, AFP existing in healthy human serum was detected to demonstrate the utility of the immunochip. The result shows that the proposed approach is feasible and has the potential application in clinical analysis and diagnosis.

  16. LC-MS/MS analysis of Δ9-tetrahydrocannabinolic acid A in serum after protein precipitation using an in-house synthesized deuterated internal standard. (United States)

    Wohlfarth, Ariane; Roth, Nadine; Auwärter, Volker


    An assay based on liquid chromatography/tandem mass spectrometry is presented for the fast, precise and sensitive quantitation of Δ9-tetrahydrocannabinolic acid A (THCA) in serum. THCA is the biogenetic precursor of Δ9-tetrahydrocannabinol in cannabis and has aroused interest in the pharmacological and forensic field especially as a potential marker for recent cannabis use. After addition of deuterated THCA, synthesized from D(3)-THC as starting material, and protein precipitation, the analytes were separated using gradient elution on a Luna C18 column (150 × 2.0 mm × 5 µm) with 0.1% formic acid and acetonitrile/0.1% formic acid. Data acquisition was performed on a triple quadrupole linear ion trap mass spectrometer in multiple reaction monitoring mode with negative electrospray ionization. After optimization, the following sample preparation procedure was used: 200 μL serum was spiked with internal standard solution and methanol and then precipitated 'in fractions' with 500 μL ice-cold acetonitrile. After storage and centrifugation, the supernatant was evaporated and the residue redissolved in mobile phase. The assay was fully validated according to international guidelines including, for the first time, the assessment of matrix effects and stability experiments. Limit of detection was 0.1 ng/mL, and limit of quantification was 1.0 ng/mL. The method was found to be selective and proved to be linear over a range of 1.0 to 100 ng/mL using a 1/x weighted calibration model with regression coefficients >0.9996. Accuracy and precision data were within the required limits (RSD ≤ 8.6%, bias: 2.4 to 11.4%), extractive yield was greater than 84%. The analytes were stable in serum samples after three freeze/thaw cycles and storage at -20 °C for one month.

  17. Proteinograma sérico de bovinos da raça Curraleir Serum protein concentration in Curraleiro bovine breed

    Directory of Open Access Journals (Sweden)

    R.S. Juliano


    Full Text Available Estudou-se o perfil eletroforético das proteínas séricas de bovinos sadios da raça Curraleiro por meio da técnica de eletroforese em gel de acrilamida contendo dodecil sulfato de sódio. Utilizaram-se amostras de soro sanguíneo de 228 bovinos da raça Curraleiro, 51 machos e 177 fêmeas, com idades entre sete meses e 12 anos, pertencentes a dois rebanhos localizados nos Estados de Goiás e Tocantins. Foram quantificadas proteína total e concentração plasmática de fibrinogênio. Verificaram-se variações nas concentrações das diferentes frações proteicas. Foram detectadas 26 proteínas e identificadas 10 delas. A ceruloplasmina esteve ausente em 78,1% dos indivíduos, e a α-antitripsina não foi detectada em nenhum animal. Proteína total, globulina, IgA, IgG e fibrinogênio aumentaram com a idade e houve correlação positiva entre os níveis séricos de haptoglobina e α1-glicoproteína ácida.The eletrophoretic serum protein profile in healthy Curraleiro bovine breed was studied by dodecyl sulphate-polyacrylamide gel electrophoresis. A total of 228 serum samples from Curraleiro cattle, being 51 males and 177 females were analyzed. They were from seven month to 12-year-old and were raised in two farms of Goiás and Tocantins states. Total protein and plasma fibrinogen quantification were performed. It was possible to verify variation in proteins fractions concentration. Twenty-six proteins were detected and ten of them were identified. Ceruloplasmin was absente in 78,10% of animals and α-antitrypsin was not detected. The total protein, globulin, IgA, IgG, and fibrinogen increased with age and there was a positive correlaction between haptoglobin and α1-acid glycoprotein.

  18. Systematic mutagenesis of genes encoding predicted autotransported proteins of Burkholderia pseudomallei identifies factors mediating virulence in mice, net intracellular replication and a novel protein conferring serum resistance. (United States)

    Lazar Adler, Natalie R; Stevens, Mark P; Dean, Rachel E; Saint, Richard J; Pankhania, Depesh; Prior, Joann L; Atkins, Timothy P; Kessler, Bianca; Nithichanon, Arnone; Lertmemongkolchai, Ganjana; Galyov, Edouard E


    Burkholderia pseudomallei is the causative agent of the severe tropical disease melioidosis, which commonly presents as sepsis. The B. pseudomallei K96243 genome encodes eleven predicted autotransporters, a diverse family of secreted and outer membrane proteins often associated with virulence. In a systematic study of these autotransporters, we constructed insertion mutants in each gene predicted to encode an autotransporter and assessed them for three pathogenesis-associated phenotypes: virulence in the BALB/c intra-peritoneal mouse melioidosis model, net intracellular replication in J774.2 murine macrophage-like cells and survival in 45% (v/v) normal human serum. From the complete repertoire of eleven autotransporter mutants, we identified eight mutants which exhibited an increase in median lethal dose of 1 to 2-log10 compared to the isogenic parent strain (bcaA, boaA, boaB, bpaA, bpaC, bpaE, bpaF and bimA). Four mutants, all demonstrating attenuation for virulence, exhibited reduced net intracellular replication in J774.2 macrophage-like cells (bimA, boaB, bpaC and bpaE). A single mutant (bpaC) was identified that exhibited significantly reduced serum survival compared to wild-type. The bpaC mutant, which demonstrated attenuation for virulence and net intracellular replication, was sensitive to complement-mediated killing via the classical and/or lectin pathway. Serum resistance was rescued by in trans complementation. Subsequently, we expressed recombinant proteins of the passenger domain of four predicted autotransporters representing each of the phenotypic groups identified: those attenuated for virulence (BcaA), those attenuated for virulence and net intracellular replication (BpaE), the BpaC mutant with defects in virulence, net intracellular replication and serum resistance and those displaying wild-type phenotypes (BatA). Only BcaA and BpaE elicited a strong IFN-γ response in a restimulation assay using whole blood from seropositive donors and were

  19. Systematic mutagenesis of genes encoding predicted autotransported proteins of Burkholderia pseudomallei identifies factors mediating virulence in mice, net intracellular replication and a novel protein conferring serum resistance.

    Directory of Open Access Journals (Sweden)

    Natalie R Lazar Adler

    Full Text Available Burkholderia pseudomallei is the causative agent of the severe tropical disease melioidosis, which commonly presents as sepsis. The B. pseudomallei K96243 genome encodes eleven predicted autotransporters, a diverse family of secreted and outer membrane proteins often associated with virulence. In a systematic study of these autotransporters, we constructed insertion mutants in each gene predicted to encode an autotransporter and assessed them for three pathogenesis-associated phenotypes: virulence in the BALB/c intra-peritoneal mouse melioidosis model, net intracellular replication in J774.2 murine macrophage-like cells and survival in 45% (v/v normal human serum. From the complete repertoire of eleven autotransporter mutants, we identified eight mutants which exhibited an increase in median lethal dose of 1 to 2-log10 compared to the isogenic parent strain (bcaA, boaA, boaB, bpaA, bpaC, bpaE, bpaF and bimA. Four mutants, all demonstrating attenuation for virulence, exhibited reduced net intracellular replication in J774.2 macrophage-like cells (bimA, boaB, bpaC and bpaE. A single mutant (bpaC was identified that exhibited significantly reduced serum survival compared to wild-type. The bpaC mutant, which demonstrated attenuation for virulence and net intracellular replication, was sensitive to complement-mediated killing via the classical and/or lectin pathway. Serum resistance was rescued by in trans complementation. Subsequently, we expressed recombinant proteins of the passenger domain of four predicted autotransporters representing each of the phenotypic groups identified: those attenuated for virulence (BcaA, those attenuated for virulence and net intracellular replication (BpaE, the BpaC mutant with defects in virulence, net intracellular replication and serum resistance and those displaying wild-type phenotypes (BatA. Only BcaA and BpaE elicited a strong IFN-γ response in a restimulation assay using whole blood from seropositive donors

  20. Effect of soluble calcium and lactose on limiting flux and serum protein removal during skim milk microfiltration. (United States)

    Adams, Michael C; Hurt, Emily E; Barbano, David M


    The tendency of calcium to promote microfiltration (MF) membrane fouling is well documented, but the role of lactose has not been studied. Milk protein concentrate that is 85% protein on a dry basis (MPC85) contains less calcium and lactose than skim milk. Our objectives were to determine the effects of skim milk soluble calcium and lactose concentrations on the limiting fluxes (LF) and serum protein (SP) removal factors of 0.1-µm ceramic graded permeability membranes. The MF was fed with 3 different milks: skim milk, liquid MPC85 that had been standardized to the protein content of skim milk with reverse osmosis water (MPC), and liquid MPC85 that had been standardized to the protein and lactose contents of skim milk with reverse osmosis water and lactose monohydrate (MPC+L). Retentate and permeate were continuously recycled to the feed tank. The LF for each feed was determined by increasing flux once per hour from 55 kg·m(-2)·h(-1) until flux did not increase with increasing transmembrane pressure. Temperature, pressure drop across the membrane length, and protein concentration in the retentate recirculation loop were maintained at 50°C, 220 kPa, and 8.77 ± 0.2%, respectively. Experiments were replicated 3 times and the Proc GLM procedure of SAS was used for statistical analysis. An increase in LF between skim milk (91 kg·m(-2)·h(-1)) and MPC+L (124 kg·m(-2)·h(-1)) was associated with a reduction in soluble calcium. The LF of MPC+L was lower than the LF of MPC (137 kg·m(-2)·h(-1)) due to the higher viscosity contributed by lactose. Permeates produced from the MPC and MPC+L contained more protein than the skim milk permeate due to the transfer of caseins from the micelles into the reduced-calcium sera of the MPC and MPC+L. A SP removal factor was calculated by dividing true protein in the permeate by SP in the permeate portion of the feed to describe the ease of SP passage through the membrane. No differences in SP removal factors were detected among the

  1. Alexithymia, Suicide Ideation, C-Reactive Protein, and Serum Lipid Levels Among Outpatients with Generalized Anxiety Disorder. (United States)

    De Berardis, Domenico; Serroni, Nicola; Campanella, Daniela; Marini, Stefano; Rapini, Gabriella; Valchera, Alessandro; Iasevoli, Felice; Mazza, Monica; Fornaro, Michele; Perna, Giampaolo; Di Iorio, Giuseppe; Martinotti, Giovanni; Di Giannantonio, Massimo


    The aim of the present study was to investigate the relationships between alexithymia, suicide ideation, C-Reactive Protein (CRP), and serum lipid levels in adult outpatients with a DSM-IV diagnosis of Generalized Anxiety Disorder (GAD). Seventy consecutive patients with GAD were recruited and evaluated. Measures were the Hamilton Anxiety Scale, the Toronto Alexithymia Scale (TAS-20), the Scale of Suicide Ideation (SSI), and the Montgomery Åsberg Depression Rating Scale (MADRS). All patients were assessed for: CRP, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), total cholesterol (TC), triglyceridaemia (TG), and very-low-density lipoprotein cholesterol (VLDL-C). TC/HDL-C and LDL-C/HDL-C ratios were also evaluated. Alexithymic patients showed higher scores on almost all rating scales and altered serum CRP and lipid levels vs. non-alexithymics. In the hierarchical regression model, the presence of higher MADRS scores together with higher scores at the Difficulty in Identifying Feelings dimension of TAS-20 were associated with higher rates of suicide ideation. Although alexithymic subjects with GAD may show a CRP and cholesterol dysregulation, this latter seems independent on increased suicide ideation, rather to Difficulty in Identifying Feelings, and subthreshold depressive symptoms. Study limitations and future research implications are discussed.

  2. Bovine serum albumin detection and quantitation based on capacitance measurements of liquid crystals (United States)

    Lin, Chi-Hao; Lee, Mon-Juan; Lee, Wei


    Liquid crystal (LC)-based biosensing is generally limited by the lack of accurate quantitative strategies. This study exploits the unique electric capacitance properties of LCs to establish quantitative assay methods for bovine serum albumin (BSA) biomolecules. By measuring the voltage-dependent electric capacitance of LCs under an alternating-current field with increasing amplitude, positive correlations were derived between the BSA concentration and the electric capacitance parameters of LCs. This study demonstrates that quantitative analysis can be achieved in LC-based biosensing through electric capacitance measurements extensively employed in LCD research and development.

  3. Characterization of granulations of calcium and apatite in serum as pleomorphic mineralo-protein complexes and as precursors of putative nanobacteria.

    Directory of Open Access Journals (Sweden)

    John D Young

    Full Text Available Calcium and apatite granulations are demonstrated here to form in both human and fetal bovine serum in response to the simple addition of either calcium or phosphate, or a combination of both. These granulations are shown to represent precipitating complexes of protein and hydroxyapatite (HAP that display marked pleomorphism, appearing as round, laminated particles, spindles, and films. These same complexes can be found in normal untreated serum, albeit at much lower amounts, and appear to result from the progressive binding of serum proteins with apatite until reaching saturation, upon which the mineralo-protein complexes precipitate. Chemically and morphologically, these complexes are virtually identical to the so-called nanobacteria (NB implicated in numerous diseases and considered unusual for their small size, pleomorphism, and the presence of HAP. Like NB, serum granulations can seed particles upon transfer to serum-free medium, and their main protein constituents include albumin, complement components 3 and 4A, fetuin-A, and apolipoproteins A1 and B100, as well as other calcium and apatite binding proteins found in the serum. However, these serum mineralo-protein complexes are formed from the direct chemical binding of inorganic and organic phases, bypassing the need for any biological processes, including the long cultivation in cell culture conditions deemed necessary for the demonstration of NB. Thus, these serum granulations may result from physiologically inherent processes that become amplified with calcium phosphate loading or when subjected to culturing in medium. They may be viewed as simple mineralo-protein complexes formed from the deployment of calcification-inhibitory pathways used by the body to cope with excess calcium phosphate so as to prevent unwarranted calcification. Rather than representing novel pathophysiological mechanisms or exotic lifeforms, these results indicate that the entities described earlier as NB most

  4. Plant Protein Intake Is Associated with Fibroblast Growth Factor 23 and Serum Bicarbonate in Patients with CKD: The Chronic Renal Insufficiency Cohort Study (United States)

    Scialla, Julia J.; Appel, Lawrence J; Wolf, Myles; Yang, Wei; Zhang, Xiaoming; Sozio, Stephen M.; Miller, Edgar R.; Bazzano, Lydia A.; Cuevas, Magdalena; Glenn, Melanie J.; Lustigova, Eva; Kallem, Radhakrishna R.; Porter, Anna C.; Townsend, Raymond R.; Weir, Matthew R.; Anderson, Cheryl A.M.


    Background Protein from plant, as opposed to animal, sources may be preferred in chronic kidney disease (CKD), due to lower bioavailability of phosphate and lower nonvolatile acid load. Study Design Observational cross-sectional study. Setting & Participants 2938 participants with chronic kidney disease and information on dietary intake at the baseline visit in the Chronic Renal Insufficiency Cohort Study. Predictors Percentage of total protein from plant sources (% plant protein) was determined by scoring individual food items from the National Cancer Institute Diet History Questionnaire (DHQ). Outcomes Metabolic parameters, including serum phosphate, bicarbonate (HCO3), potassium, and albumin, plasma fibroblast growth factor 23 (FGF23), and parathyroid hormone (PTH), and hemoglobin. Measurements We modeled the association between % plant protein and metabolic parameters using linear regression. Models were adjusted for age, sex, race, diabetes, body mass index, eGFR, income, smoking, total energy intake, total protein intake, 24 hour urinary sodium, use of angiotensin converting enzyme inhibitors/angiotensin receptor blockers and use of diuretics. Results Higher % plant protein was associated with lower FGF23 (p=0.05) and higher HCO3 (p=0.01), but not with serum phosphate or PTH (p=0.9 and 0.5, respectively). Higher % plant protein was not associated with higher serum potassium (p=0.2), lower serum albumin (p=0.2) or lower hemoglobin (p=0.3). The associations of % plant protein with FGF23 and HCO3 did not differ by diabetes status, sex, race, CKD stage (2/3 vs. 4/5) or total protein intake (≤ 0.8 g/kg/d vs. >0.8 g/kg/d) (p-interaction > 0.10 for each). Limitations Cross-sectional study; Determination of % plant protein using the DHQ has not been validated. Conclusions Consumption of a higher percentage of protein from plant sources may lower FGF23 and raise HCO3 in patients with CKD. PMID:22480598

  5. Widely predicting specific protein functions based on protein-protein interaction data and gene expression profile

    Institute of Scientific and Technical Information of China (English)

    GAO Lei; LI Xia; GUO Zheng; ZHU MingZhu; LI YanHui; RAO ShaoQi


    GESTs (gene expression similarity and taxonomy similarity), a gene functional prediction approach previously proposed by us, is based on gene expression similarity and concept similarity of functional classes defined in Gene Ontology (GO). In this paper, we extend this method to protein-protein interaction data by introducing several methods to filter the neighbors in protein interaction networks for a protein of unknown function(s). Unlike other conventional methods, the proposed approach automatically selects the most appropriate functional classes as specific as possible during the learning process, and calls on genes annotated to nearby classes to support the predictions to some small-sized specific classes in GO. Based on the yeast protein-protein interaction information from MIPS and a dataset of gene expression profiles, we assess the performances of our approach for predicting protein functions to "biology process" by three measures particularly designed for functional classes organized in GO. Results show that our method is powerful for widely predicting gene functions with very specific functional terms. Based on the GO database published in December 2004, we predict some proteins whose functions were unknown at that time, and some of the predictions have been confirmed by the new SGD annotation data published in April, 2006.

  6. Widely predicting specific protein functions based on protein-protein interaction data and gene expression profile

    Institute of Scientific and Technical Information of China (English)


    GESTs (gene expression similarity and taxonomy similarity), a gene functional prediction approach previously proposed by us, is based on gene expression similarity and concept similarity of functional classes defined in Gene Ontology (GO). In this paper, we extend this method to protein-protein interac-tion data by introducing several methods to filter the neighbors in protein interaction networks for a protein of unknown function(s). Unlike other conventional methods, the proposed approach automati-cally selects the most appropriate functional classes as specific as possible during the learning proc-ess, and calls on genes annotated to nearby classes to support the predictions to some small-sized specific classes in GO. Based on the yeast protein-protein interaction information from MIPS and a dataset of gene expression profiles, we assess the performances of our approach for predicting protein functions to “biology process” by three measures particularly designed for functional classes organ-ized in GO. Results show that our method is powerful for widely predicting gene functions with very specific functional terms. Based on the GO database published in December 2004, we predict some proteins whose functions were unknown at that time, and some of the predictions have been confirmed by the new SGD annotation data published in April, 2006.

  7. Adsorption of bovine serum albumin on CoCrMo surface: effect of temperature and protein concentration. (United States)

    Valero Vidal, C; Olmo Juan, A; Igual Muñoz, A


    The adsorption of bovine serum albumin (BSA) onto CoCrMo surface has been studied as a function of concentration of BSA and temperature by electrochemical techniques. The electrochemical impedance spectroscopy (EIS) technique was used to investigate the interfacial behaviour of BSA at open circuit potential (OCP). The charge transfer resistance was very sensitive to the amount of adsorbed protein, indicating that the adsorption process was accompanied by the transfer of charge and influenced the mechanism and kinetics of the corrosion reaction. At all the temperatures studied, adsorption of BSA onto the CoCrMo surface was successfully described with a Langmuir adsorption isotherm. EIS study was also carried out for determine the surface charge density, resulting from protein adsorption, and it was shown to be directly proportional to the amount of adsorbed protein (surface concentration). Thermodynamic data of adsorption was obtained for analyzing the adsorption of BSA onto CoCrMo surface. Gibbs free energy of adsorption, DeltaG(ADS) values, for BSA in the investigated temperature range (-51kJmol(-1)) showed that the molecules have a strong affinity for the CoCrMo surface. Enthalpy (DeltaH(ADS)) and entropy (DeltaS(ADS)) of adsorption suggested that the adsorption process of BSA onto the CoCrMo surface is an endothermic process and the molecule suffers structural changes when adsorbing on the metallic surface.

  8. Analyses of cardiac blood cells and serum proteins with regard to cause of death in forensic autopsy cases. (United States)

    Quan, Li; Ishikawa, Takaki; Michiue, Tomomi; Li, Dong-Ri; Zhao, Dong; Yoshida, Chiemi; Chen, Jian-Hua; Komatsu, Ayumi; Azuma, Yoko; Sakoda, Shigeki; Zhu, Bao-Li; Maeda, Hitoshi


    To investigate hematological and serum protein profiles of cadaveric heart blood with regard to the cause of death, serial forensic autopsy cases (n=308, >18 years of age, within 48 h postmortem) were examined. Red blood cells (Rbc), hemoglobin (Hb), platelets (Plt), white blood cells (Wbc), total protein (TP) and albumin (Alb) were examined in bilateral cardiac blood. Blood cell counts, collected after turning the bodies at autopsy, approximated to the clinical values. Postmortem changes were not significant for these markers. In non-head blunt injury cases, Rbc counts, Hb, TP and Alb levels in bilateral cardiac blood were lower in subacute deaths (survival time, 1-12 h) than in acute deaths (survival time blood were significantly higher for non-head injury than for head injury in subacute deaths. In fire fatality cases, Plt count was markedly higher with an automated hematology analyzer than by using a blood smear test, suggesting Rbc fragmentation caused by deep burns, while increases in Wbc count and decreases in Alb levels were seen for subacute deaths. For asphyxiation, Rbc count, Hb, TP and Alb levels in bilateral cardiac blood were higher than other groups, and TP and Alb levels in the right cardiac blood were higher for hanging than for strangulation. These findings suggest that analyses of blood cells and proteins are useful for investigating the cause of death.

  9. Comparable autoantibody serum levels against amyloid- and inflammation-associated proteins in Parkinson's disease patients and controls.

    Directory of Open Access Journals (Sweden)

    Walter Maetzler

    Full Text Available Naturally occurring autoantibodies (NAbs against a number of potentially disease-associated cellular proteins, including Amyloid-beta1-42 (Abeta1-42, Alpha-synuclein (Asyn, myelin basic protein (MBP, myelin oligodendrocyte glycoprotein (MOG, and S100 calcium binding protein B (S100B have been suggested to be associated with neurodegenerative disorders, in particular Alzheimer's (AD and Parkinson's disease (PD. Whereas the (reduced occurrence of specific NAbs in AD is widely accepted, previous literature examining the relation of these NAb titres between PD patients and controls, as well as comparing these levels with demographic and clinical parameters in PD patients have produced inconsistent findings. We therefore aimed, in a cross-sectional approach, to determine serum titres of the above NAbs in a cohort of 93 PD patients (31 of them demented and 194 controls. Levels were correlated with demographic and clinical variables, cerebrospinal fluid Abeta1-42, total tau and phospho-tau levels, as well as with single nucleotide polymorphisms (SNPs of genes which either have been reported to influence the immune system, the amyloid cascade or the occurrence of PD (ApoE, GSK3B, HLA-DRA, HSPA5, SNCA, and STK39. The investigated NAb titres were neither significantly associated with the occurrence of PD, nor with demographic and clinical parameters, neurodegenerative markers or genetic variables. These results argue against a major potential of blood-borne parameters of the adaptive immune system to serve as trait or state markers in PD.

  10. Increased serum and musculotendinous fibrogenic proteins following persistent low-grade inflammation in a rat model of long-term upper extremity overuse.

    Directory of Open Access Journals (Sweden)

    Helen G L Gao

    Full Text Available We examined the relationship between grip strength declines and muscle-tendon responses induced by long-term performance of a high-repetition, low-force (HRLF reaching task in rats. We hypothesized that grip strength declines would correlate with inflammation, fibrosis and degradation in flexor digitorum muscles and tendons. Grip strength declined after training, and further in weeks 18 and 24, in reach limbs of HRLF rats. Flexor digitorum tissues of reach limbs showed low-grade increases in inflammatory cytokines: IL-1β after training and in week 18, IL-1α in week 18, TNF-α and IL-6 after training and in week 24, and IL-10 in week 24, with greater increases in tendons than muscles. Similar cytokine increases were detected in serum with HRLF: IL-1α and IL-10 in week 18, and TNF-α and IL-6 in week 24. Grip strength correlated inversely with IL-6 in muscles, tendons and serum, and TNF-α in muscles and serum. Four fibrogenic proteins, TGFB1, CTGF, PDGFab and PDGFbb, and hydroxyproline, a marker of collagen synthesis, increased in serum in HRLF weeks 18 or 24, concomitant with epitendon thickening, increased muscle and tendon TGFB1 and CTGF. A collagenolytic gelatinase, MMP2, increased by week 18 in serum, tendons and muscles of HRLF rats. Grip strength correlated inversely with TGFB1 in muscles, tendons and serum; with CTGF-immunoreactive fibroblasts in tendons; and with MMP2 in tendons and serum. Thus, motor declines correlated with low-grade systemic and musculotendinous inflammation throughout task performance, and increased fibrogenic and degradative proteins with prolonged task performance. Serum TNF-α, IL-6, TGFB1, CTGF and MMP2 may serve as serum biomarkers of work-related musculoskeletal disorders, although further studies in humans are needed.

  11. Controlled Modulation of Serum Protein Binding and Biodistribution of Asymmetric Cyanine Dyes by Variation of the Number of Sulfonate Groups

    Directory of Open Access Journals (Sweden)

    Franziska M. Hamann


    Full Text Available To assess the suitability of asymmetric cyanine dyes for in vivo fluoro-optical molecular imaging, a comprehensive study on the influence of the number of negatively charged sulfonate groups governing the hydrophilicity of the DY-67x family of asymmetric cyanines was performed. Special attention was devoted to the plasma protein binding capacity and related pharmacokinetic properties. Four members of the DY-67x cyanine family composed of the same main chromophore, but substituted with a sequentially increasing number of sulfonate groups (n = 1−4; DY-675, DY-676, DY-677, DY-678, respectively, were incubated with plasma proteins dissolved in phosphate-buffered saline. Protein binding was assessed by absorption spectroscopy, gel electrophoresis, ultrafiltration, and dialysis. Distribution of dye in organs was studied by intraveneous injection of 62 nmol dye/kg body weight into mice (n = 12; up to 180 minutes postinjection using whole-body near-infrared fluorescence imaging. Spectroscopic studies, gel electrophoresis, and dialysis demonstrated reduced protein binding with increasing number of sulfonate groups. The bovine serum albumin binding constant of the most hydrophobic dye, DY-675, is 18 times higher than that of the most hydrophilic fluorophore, DY-678. In vivo biodistribution analysis underlined a considerable influence of dye hydrophilicity on biodistribution and excretion pathways, with the more hydrophobic dyes, DY-675 and DY-676, accumulating in the liver, followed by strong fluorescence signals in bile and gut owing to accumulation in feces and comparatively hydrophilic DY-678-COOH accumulating in the bladder. Our results demonstrate the possibility of selectively controlling dye-protein interactions and, thus, biodistribution and excretion pathways via proper choice of the fluorophore's substitution pattern. This underlines the importance of structure-property relationships for fluorescent labels. Moreover, our data could provide the

  12. The diagnostic and prognostic value of serum bone Gla protein (osteocalcin) in patients with recurrent breast cancer

    DEFF Research Database (Denmark)

    Kamby, C; Egsmose, C; Söletormos, G


    bone survey. The sites of recurrence were bone (61%), bone marrow (46%), soft tissue (52%), lung (13%), pleura (11%), liver (4%), and brain (2%). Radiology and bone biopsy served as key diagnoses as to the presence or absence of bone metastases. The diagnostic efficiency of B-scan and S-AP was greater...... than that of S-BGP, and the result of BGP measurement was associated with neither extent nor number of bone metastases. However, the BGP values were significantly lower in patients who had visceral metastases, and the median duration of survival after recurrence was 13 months for patients with low S......Serum bone Gla protein (S-BGP), a marker of bone metabolism, was measured in 60 patients included in a staging programme for recurrent breast cancer. Other diagnostic procedures comprised S-alkaline phosphatase (S-AP), bone scan (B-scan), bilateral iliac crest bone marrow biopsies, and radiological...

  13. Differential expression of serum glycodelin and insulin-like growth factor binding protein 1 in early pregnancy. (United States)

    Douglas, Nataki C; Thornton, Melvin H; Nurudeen, Sahadat K; Bucur, Maria; Lobo, Rogerio A; Sauer, Mark V


    This prospective study evaluated whether serum glycodelin and insulin-like growth factor binding protein 1 (IGFBP-1) predict the likelihood of embryo implantation in recipients undergoing donor egg in vitro fertilization. We measured glycodelin and IGFBP-1 at 6 points from lining check to lutenizing hormone (LH) + 31. β-Human chorionic gonadotropin levels were first measured at LH + 17. The recipients were divided into those without embryo implantation (group 1, n = 6) and those with successful implantation (group 2, n = 30). Although this is a negative study in that neither glycodelin nor IGFBP-1 alone reflected endometrial (EM) receptivity, the glycodelin/IGFBP-1 ratio on the day of blastocyst transfer was higher in recipients who achieved pregnancy (P = .05). At LH + 17, glycodelin was higher (P = .04), and IGFBP-1 was lower (P = .004) in recipients who achieved pregnancy when compared to those who did not. These observations are likely due to EM changes induced by successful embryo implantation.

  14. Formulation of antiacne serum based on lime peel essential oil and in vitro antibacterial activity test against Propionibacterium acnes (United States)

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


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

  15. Ag/Au纳米粒子的没食子酸还原制备及其共振瑞利散射光谱测定人血清总蛋白%Gallic Acid Based Synthesis of Silver/Gold Nanoparticles and Resonance Rayleigh Scattering Detection of Human Serum Proteins

    Institute of Scientific and Technical Information of China (English)

    王文星; 黄玉萍; 徐淑坤


    以没食子酸为还原剂和稳定剂,用种子生长法制备出粒径均匀、单分散性和稳定性好、近球形的Ag/Au 核壳纳米粒子.高分辨透射电镜(HRTEM)与 X-射线能量色散光谱仪(EDX)测试表明,在Ag/Au摩尔比为1:1.6时,Au已完全包裹在Ag纳米粒子表面时,平均粒径为25 nm.以此摩尔比制备的Ag/Au核壳纳米粒子为探针用共振瑞利散射光谱测定人血清总蛋白,在NaAc-HAc缓冲液(pH 4.4)及0.05 mol/L NaCl介质中,Ag/Au核壳纳米粒子与HSA形成稳定的复合物;Ag/Au-HSA纳米复合物的相对散射强度ΔI390 nm与HSA浓度在0.0011~0.35 mg/L范围内呈线性关系,其回归方程为ΔI390 nm=-0.54+494.82c(r=0.9994),检出限为0.36 μg/L.本方法有较好的选择性,可用于人血清蛋白分析,结果与考马斯亮蓝G-250法一致,回收率在98.2%~102.3%之间,相对标准偏差为2.3%.%Nearly spherical Ag/Au core/shell nanoparticles with good monodispersity and stability were synthesized in aqueous solution by seed growth method using gallic acid as reductant and stabilizer. High resolution transmission electron microscopic (HRTEM) image and energy dispersive X-ray spectrometry (EDX) showed complete coating of gold on the surface of Ag nanoparticles and the presence of Au and Ag in individual nanoparticle with the average size of about 25 nm. The Ag/Au core/shell nanoparticles with Au/Ag molar ratio of 1:1.6 were used as probes to determine human serum proteins by resonance Rayleigh scattering (RRS) spectrometry. The results indicated that in pH 4.4 NaAc-HAc buffer solutions and in the presence of NsC1 of 0.05 mol/L, HSA was combined with Ag/Au core/shell nanoparticles to form stable complex. The enhanced resonance scattering intensity at 390 nm (△I390nm) was linear to HSA concentration in the range of 0. 0011-0.35 mg/L, with the regress equation of △I390nm = -0. 54+494. 82C(r= 0. 994) and the detection limit of 0.36 μg/L.Co-existing metal ions and amino acids

  16. Facile internalization of paclitaxel on titania nanoparticles in human lung carcinoma cells after adsorption of serum proteins

    Energy Technology Data Exchange (ETDEWEB)

    Seo, Ji Hye; Dembereldorj, Uuriintuya; Park, Jin; Kim, Mira; Kim, Semi; Joo, Sang-Woo, E-mail: [Soongsil University, Department of Chemistry (Korea, Republic of)


    The enhanced damage of human carcinoma cells was achieved using paclitaxel (PTX)-assembled TiO{sub 2} nanoparticles (NPs). The crystalline structures and light absorption properties of the TiO{sub 2} NPs were examined by X-ray diffraction patterns and ultraviolet-visible absorption spectroscopic tools. The fabrication of PTX and serum proteins on TiO{sub 2} NPs via self-assembly was checked by diffuse reflectance infrared Fourier transform spectroscopy and dynamic light scattering measurements. The PTX-coated TiO{sub 2} NPs appeared to be well adsorbed by serum proteins. The PTX-coated TiO{sub 2} NPs were found to be well internalized in cancer cells as indicated by transmission electron microscopy. The intracellular aggregation of PTX-coated TiO{sub 2} NPs was assumed to occur after endocytosis leading to an entrapment of NPs in an endosomal or a lysosomal structure. Human lung carcinoma A549 cancer cells were chosen to examine their viability after the cellular uptake of the PTX-coated TiO{sub 2} NPs. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell viability assay test indicated that the cell viability should be affected by the concentration of TiO{sub 2} NPs. After applying PTX-coated TiO{sub 2} NPs (50 ppm ({mu}g/mL)) incubating for 24 h, we observed a 30 % further decrease of cell viability for PTX alone in the low concentration range of 0.1-10 nM.Graphical Abstract.

  17. Serum Heat Shock Protein 70 Concentration in Relation to Polycystic Ovary Syndrome in a Non-Obese Chinese Population.

    Directory of Open Access Journals (Sweden)

    Hui Gao

    Full Text Available Polycystic ovary syndrome (PCOS represents the most common cause of anovulatory infertility and affects 6-15% of women of reproductive age. However, the underlying etiology is still poorly understood. In this study, we attempted to examine the association between circulating heat shock protein 70 (Hsp70 concentrations and PCOS in a non-obese Chinese population.Human peripheral blood from 52 patients with PCOS and 57 healthy controls, matched for age and BMI, were analyzed. Women with PCOS were found to have significantly higher fasting insulin (FI levels, as well as Insulin resistance index (HOMA-IR (P < 0.05. Identically, markers of oxidative stress (malondialdehyde (MDA, 8-Hydroxy-desoxyguanosine (8-OHdG, Nitric oxide (NO and inflammation (tumor necrosis factor-alpha (TNF-α, C-reactive protein (CRP were markedly increased when compared to controls (P < 0.05. Elevated serum Hsp70 was positively correlated with IR, oxidative stress and inflammation in PCOS, even after adjustment for age, BMI and gynecologic inflammation (GI. The receiver-operating characteristic curve (ROC analysis yielded notably different discriminative value for PCOS, with or without an addition of Hsp70 (areas under the curves were 0.884 (95% CI 0.822-0.946 vs. 0.822 (95% CI 0.744-0.900; P for difference = 0.015.Increased serum Hsp70 levels are associated with the combination of IR, oxidative stress and low-grade chronic inflammation in PCOS individuals, which provides supportive evidence that Hsp70 plays a key role in the pathogenesis of PCOS. More consequent studies were warranted to confirm the clinical utility of circulating Hsp70, especially in diagnosis and prognosis of PCOS and its long-term health cost.

  18. Effect of alprostadil combined with Shenkang injection on urine protein, renal function and serum inflammatory in patients with chronic nephritis

    Institute of Scientific and Technical Information of China (English)

    Chen Wang; Zhi-Feng Gu; Shuo Wang; Liang-Lan Shen; Fen Zhang


    Objective:To study the effect of alprostadil combined with Shenkang injection on urine protein, renal function and serum inflammatory in patients with chronic nephritis.Methods:A total of 96 patients with chronic nephritis in our hospital from May 2013 to May 2016 were enrolled in this study. The subjects were divided into control group (n=48) and treatment group (n=48) randomly. Patients in control group were treated with Shenkang injection, the treatment group were treated with alprostadil combined with Shenkang injection. The two groups were treated for 12 days. The levels of 24 h Upro, Uβ2-MG, SCr, BUN, UAER, hs-CRP, TNF-α, IL-6, IL-8 and IL-18 of the two groups before and after treatment were compared. Results:There were no significantly differences of the levels of 24 h Upro, Uβ2-MG, SCr, BUN, UAER, hs-CRP, TNF-α, IL-6, IL-8 and IL-18 of the two groups before treatment (P>0.05). The levels of 24 h Upro, Uβ2-MG, SCr, BUN and UAER of the two groups after treatment were significantly lower than before treatment (P<0.05), and that of experiment were significantly lower than control group (P<0.05). The levels of hs-CRP, TNF-α, IL-6, IL-8 and IL-18 of the two groups after treatment were significantly lower than before treatment (P<0.05), and that of experiment were significantly lower than control group (P<0.05).Conclusions: Alprostadil combined with Shenkang injection can significantly reduce urine protein and serum inflammatory, protect renal function of patients with chronic nephritis, and it is worthy clinical application.

  19. Effects of serum containing natural cerebrolysin on glucose-regulated protein 78 and CCAAT enhancer-binding protein homologous protein expression in neuronal PC12 cells following tunicamycin-induced endoplasmic reticulum stress

    Institute of Scientific and Technical Information of China (English)

    Zhengzhi Wu; Ming Li; Andrew C.J. HuangO; Xiuqing Jia; Yinghong Li; Manyin Chen


    BACKGROUND: Glucose-regulated protein 78 (GRP78), a marker of endoplasmic reticulum stress, can prolong cell survival. Alternatively, CCAAT enhancer-binding protein homologous protein (CHOP), a transcription factor specific for endopiasmic reticulum stress, can cause cell cycle arrest and cell apoptosis.OBJECTIVE: To study the protective effects of serum containing natural cerebrolysin on endoplasmic reticulum stress in tunicamycin-induced neuronal PC12 cells, and analyze the influence on GRP78 and CHOP expressions.DESIGN, TIME AND SETTING: A parallel controlled study was performed at the Institute of Integrated Western and Traditional Chinese Medicine, Shenzhen Hospital, Southern Medical University, between March 2006 and August 2008.MATERIALS: Adult Sprague-Dawley rats were perfused with natural Cerebrelysin aqueous extract (0.185 g/kg/d) to produce serum containing natural Cerebrolysin. Physiological saline was used to produce blank serum. PC12 cell line was provided by Shanghai Institute of Cell Biology, Chinese Academy of Science. Tunicamycin was provided by Sigma (St. Louis, USA), and natural Cerebrolysin, containing ginseng, rhizoma gastrodiae, and gingko leaf (1:2:2), by Shengzhen Institute of Integrated Western and Traditional Chinese Medicine.METHODS: PC12 cells were treated with DMEM culture media containing 10% blank serum (normal control group), tunicamycin (1μg/mL; model group), and 5%, 10%, and 15% serum containing natural cerebrolysin and tunicamycin (1μg/mL; low-, moderate-, and high-dose serum containing natural cerebrelysin groups), for 2 hours.MAIN OUTCOME MEASURES: PC12 cells were treated with tunicamycin for 48 hours after which apoptosis was measured using the TUNEL method to calculate apoptotic index. GRP78 expression was detected using immunocytochemistry. After 24 hours of treatment with tunicamycin, GRP78 and CHOP mRNA expressions were measured using RT-PCR.RESULTS: The apoptotic index and CHOP mRNA expression were in the model group

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

    NARCIS (Netherlands)

    Westaby, S; Saatvedt, K; White, S; Katsumata, T; van Oeveren, W; Bhatnagar, NK; Brown, S; Halligan, PW


    Objectives: Over the past decade, the glial protein S-100 beta has been used to detect cerebral injury in a number of clinical settings including cardiac surgery. Previous investigations suggest that S-100 beta is capable of identifying patients with cerebral dysfunction after cardiopulmonary bypass

  1. Introducing Proteomics in the Undergraduate Curriculum: A Simple 2D Gel Electrophoresis Exercise with Serum Proteins (United States)

    Kim, Thomas D.; Craig, Paul A.


    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…

  2. Relative contributions of measles virus hemagglutinin- and fusion protein- specific serum antibodies to virus neutralization.

    NARCIS (Netherlands)

    R.L. de Swart (Rik); S. Yüksel (Selma); A.D.M.E. Osterhaus (Albert)


    textabstractThe relative contribution of measles virus hemagglutinin (H)- or fusion protein (F)-specific antibodies to virus neutralization (VN) has not been demonstrated. We have depleted these specific antibodies from sera collected from young adults, who had been vaccinated during childhood, by p

  3. Serum surfactant protein D is correlated to development of dementia and augmented mortality

    DEFF Research Database (Denmark)

    Nybo, Mads; Andersen, Kjeld; Sørensen, Grith Lykke;


    Development of dementia, including Alzheimer's disease (AD), is associated with lipid dysregulation and inflammation. As the host defense lectin surfactant protein D (SP-D) has multiple effects in lipid homeostasis and inflammation, the correlation between SP-D concentrations and development of d...

  4. Micro-ELISA for the quantitation of human urinary and serum retinol-binding protein

    DEFF Research Database (Denmark)

    Jensen, T; Deckert, M; Dawnay, A;


    ) and dilution of urine was linear. The within-assay coefficient of variation ranged from 1.2-3.1% and the day-to-day coefficient of variation from 9.2-10.5% depending on concentration. The correlation with urinary retinol-binding protein determined by radioimmunoassay was good (n = 90, r = 0.95). In vitro...

  5. MALDI-MS-Based Profiling of Serum Proteome: Detection of Changes Related to Progression of Cancer and Response to Anticancer Treatment

    Directory of Open Access Journals (Sweden)

    Monika Pietrowska


    Full Text Available Mass spectrometry-based analyses of the low-molecular-weight fraction of serum proteome allow identifying proteome profiles (signatures that are potentially useful in detection and classification of cancer. Several published studies have shown that multipeptide signatures selected in numerical tests have potential values for diagnostics of different types of cancer. However due to apparent problems with standardization of methodological details, both experimental and computational, none of the proposed peptide signatures analyzed directly by MALDI/SELDI-ToF spectrometry has been approved for routine diagnostics. Noteworthy, several components of proposed cancer signatures, especially those characteristic for advanced cancer, were identified as fragments of blood proteins involved in the acute phase and inflammatory response. This indicated that among cancer biomarker candidates to be possibly identified by serum proteome profiling were rather those reflecting overall influence of a disease (and the therapy upon the human organism, than products of cancer-specific genes. Current paper focuses on changes in serum proteome that are related to response of patient’s organism to progressing malignancy and toxicity of anticancer treatment. In addition, several methodological issues that affect robustness and interlaboratory reproducibility of MS-based serum proteome profiling are discussed.

  6. NMR based serum metabolomics reveals a distinctive signature in patients with Lupus Nephritis (United States)

    Guleria, Anupam; Pratap, Avadhesh; Dubey, Durgesh; Rawat, Atul; Chaurasia, Smriti; Sukesh, Edavalath; Phatak, Sanat; Ajmani, Sajal; Kumar, Umesh; Khetrapal, Chunni Lal; Bacon, Paul; Misra, Ramnath; Kumar, Dinesh


    Management of patient with Lupus Nephritis (LN) continues to remain a challenge for the treating physicians because of considerable morbidity and even mortality. The search of biomarkers in serum and urine is a focus of researchers to unravel new targets for therapy. In the present study, the utility of NMR-based serum metabolomics has been evaluated for the first time in discriminating LN patients from non-nephritis lupus patients (SLE) and further to get new insights into the underlying disease processes for better clinical management. Metabolic profiling of sera obtained from 22 SLE patients, 40 LN patients and 30 healthy controls (HC) were performed using high resolution 1D 1H-CPMG and diffusion edited NMR spectra to identify the potential molecular biomarkers. Using multivariate analysis, we could distinguish SLE and LN patients from HC and LN from SLE patients. Compared to SLE patients, the LN patients had increased serum levels of lipid metabolites (including LDL/VLDL lipoproteins), creatinine and decreased levels of acetate. Our results revealed that metabolic markers especially lipids and acetate derived from NMR spectroscopy has high sensitivity and specificity to distinguish LN among SLE patients and has the potential to be a useful adjunctive tool in diagnosis and clinical management of LN. PMID:27739464

  7. Electrochemical Immunoassay of Cotinine in Serum Based on Nanoparticle Probe and Immunochromatographic Strip

    Energy Technology Data Exchange (ETDEWEB)

    Nian, Hung-Chi; Wang, Jun; Wu, Hong J.; Lo, Jiunn-Guang; Chiu, Kong-Hwa; Pounds, Joel G.; Lin, Yuehe


    A disposable sensor for the determination of cotinine in human serum was developed based on immunochromatographic test strip and quantum dot label. In this assay, cotinine modified on quantum dot competes with cotinine in sample to bind to anti-cotinine antibody in the test strip and the quantum dots serve as signal vehicles for electrochemical readout. Some parameters governing the performance of the sensor were optimized. The sensor shows a wide linear range from 1 ng mL-1 to 100 ng mL-1 cotinine with a detection limit of 1.0 ng mL-1. The relative standard deviation (R.S.D.) of the sensor was less than 2% for cotinine. The sensor was validated with spiked human serum samples and it was found that this method was reliable in measuring cotinine in human serum with average recovery of 100.99%. The results demonstrate that this sensor is a rapid, clinically accurate, and less expensive and has the potential for point of care (POC) detection of cotinine and fast screening of tobacco smoke exposure.

  8. The relation between serum vitamin D levels and blood pressure: a population-based study.

    Directory of Open Access Journals (Sweden)

    Fatemeh Ardeshir Larijani


    Full Text Available Vitamin D deficiency has been proposed as an associating factor with increased blood pressure. We studied the relationship between serum vitamin D and blood pressure in a large representative sample of Iranian population. In this cross-sectional study, based on the data of 2508 adults (aged between 20 and 70 years from the Iran Multicenter Osteoporosis Study (IMOS, the association between serum vitamin D and blood pressure was investigated. There was a significant difference between mean (±SD vitamin D levels of the individuals with stage I hypertension and that of the three other groups (Normal: 32.9 (±27.5; Prehypertension: 34.4 (±27.2; Stage-I: 38.7 (±29.2; Stage-II: 34.7 (±24.0 ng/ml; P<0.05. In multivariate regression models, the weak positive association of vitamin D and systolic blood pressure values disappeared after age and Body Mass Index (BMI adjustment. We found a statistically positive but weak association between vitamin D serum concentration and systolic blood pressure. Considering the difference noted between our results and previous studies, further research is needed to assess the potential effect of ethnicity and genetic factors on these findings.

  9. The relation between serum vitamin D levels and blood pressure: a population-based study. (United States)

    Ardeshir Larijani, Fatemeh; Kalantar Motamedi, Seyed Mohammad; Keshtkar, Abbas Ali; Khashayar, Patricia; Koleini, Zahra; Rahim, Fakher; Larijani, Bagher


    Vitamin D deficiency has been proposed as an associating factor with increased blood pressure. We studied the relationship between serum vitamin D and blood pressure in a large representative sample of Iranian population. In this cross-sectional study, based on the data of 2508 adults (aged between 20 and 70 years) from the Iran Multicenter Osteoporosis Study (IMOS), the association between serum vitamin D and blood pressure was investigated. There was a significan